The chicken β-defensin-1(Gallinacin-1) cDNA fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from the bone marrow cells of Yuehuang chicken. The RT-PCR products were inserted into the pGEM-T cloning vector, and then was sequenced. The result showed that the obtained 198 bp DNA fragment is identical to the Gal-1 cDNA sequence registered in GenBank. Compared with the other β-defensin registered in GenBank, Gal-1 and THP-1 had the highest homology of nucleotide and putative amino acid sequence, were 85.4% and 72.7%, respectively. Then the fragment of mature Gallinacin-1 gene was inserted into the pPICZα-C expression vector and pPICZα-C-gal-1 was acquired, then the recombinant plasmid was electro-transformed into pichia pastoris -X-33 strain. The positive transformed strains were cultured and induced by addition of 0.5% methanol every 24 h. Expression products with molecular weight approximately 4.5 ku were displayed on the gel during Tricine-SDS-PAGE. The expression products had antimicrobial activity against E. coli, salmonella and ataphylococcosis aureas. The retention time of recombinant Gal-1 peptides in RP-HPLC was identical to that of synthetic Gal1 peptides, which further showed the Gal-1 gene was expressed in the pichia pastoris.
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Bayesian-MCMC mothed was performed to detect QTL for binary traits under different family structures in animal outbred population in our study. During linkage analysis, three samplers of MCMC, including Gibbs sampling, Metropolis algorithm and reversible jump MCMC，were implemented to generate the joint posterior distribution of all unknowns so that the QTL parameters were obtained by Bayesian statistical inferring under the Identity-by-Descent-Based variance component random model. The results showed that Bayesian-MCMC approach could work well and robust under different family structures, especially for estimating the number of QTL which is one of the most important QTL parameters.

To find out the impact of intrauterine growth retardation on the development of small intestinal enzyme in newborn piglets, 14 normal piglets and 14 piglets with IUGR were selected from 14 litters using natural select. They were randomly assigned to three groups: Newborn group (D0), Group naturally suckled for 3 days (D3) and Group naturally suckled for 7 days (D7), and were slaughtered on the end of the experiment. At birth, both of total activity and specific activity of lactase in mucous membrane of small intestine in normal piglets were significantly higher than those with IUGR. Both of total activity and specific activity of aminopeptidase in piglets with IUGR were lower than those in normal ones, and the difference of total activity between them is greatly significant. The total activity of maltase in IUGR piglets was significantly lower than that in normal piglets, but the difference of specific activity of maltase between them was not significant. The difference of alkaline phosphatase activity was also not significant. After naturally suckled with their dams for 3 days, the differences of mucosal maltase and alkaline phosphatase activities between normal piglets and piglets with IUGR were all not significant, but the difference of total activity of lactase and AP was significant. At 7 days age, there were no significances in mucosal lactase, maltase, aminopeptidase and alkaline phosphatase activities between them. All experiment results suggested that IUGR blocked the maturation of lactase and aminopeptidase in small intestine, and resulted in the premature of maltase and alkaline phosphatase. At the same time, it also suggested that the development of intestinal function in intrauterine growth retarded piglet partly realized catchup growth during postnatal 1 week, and the catch-up growth is step by step with increased age.

Twelve male adult local sheep with permanent rumen cannula were used to study the effect of four levels of Sarsa-saponine (0，100，200，300 mg/kg)on rumen fermentation parameters and blood biochemical traits in a completely randomized block design. The pH，VFA and NH₃ concentration of rumen liquid were sampled at 0，2，4，6 and 8 h after feeding. The result showed that pH were not distinctly influenced at the same time points in each group（P>0.05）. The variety current of pH have delayed by concentration of Sarsa-saponine supplement increased ;the current of acetic concentration was decreased in each group，but the propionic acids concentration was increased. 300 mg/kg group compared with control ,the acetic concentration decreased by 15.1% and 19.8% , the propionic acids concentration increased by 22.7% and 22.9%，after feeding 4 h and 6 h（P<0.05）.The groups showed no difference of the volatile fatty acids concentration（P>0.05）;The inhibitory rate of ammonia concentration of 100，200，300 mg/kg group were 17.17%、29.84% and 27.12%，and the 200 mg/kg group were distinctly difference compared with control（P<0.05）.The result showed that the blood biochemical traits of four groups were in normal range and were not distinctly varied（P>0.05）after feeding 28 d.

Leydig cells from 2～3 week old piglets were collected to study the mechanism of HCG regulating testosterone synthesis. Some precursors including cholesterol(5 mg/mL), pregnenolone(500 ng/mL), dehydroepiandrosterone (500ng/mL), androstenedione (500ng/mL)and 22 R-Hydroxycholesterol(3 μg/mL) was added into these cultivating bottles and cocultivated with Leydig cells for 48 hours. Then, certain concentrations of HCG were added to each bottle and go on cultivating up to 2hours.The concentrations of testosterone and cAMP were measured by radioimmunoassay, but the level of steroidogenic acute regulatory(StAR) protein and mRNA were measured by immmunoblotting and RT-PCR respectively. The results were as follows: (1) HCG increased not only the production of testosterone, but also the concentration of cAMP of cells;(2) Although the conversion from cholesterol to testosterone could significantly be promoted, the conversion from other precursors to testosterone were not affected significantly .(3) The levels of both protein and mRNA of StAR were increased by HCG in 2hours.The results showed that in the acute period, HCG could increase the concentration of testosterone by promoting StAR expression.

The total DNAs were extracted from peripheral whole-blood of three Tibetan sheep and three goats respectively. The PrP^c genes were amplified by PCR using two pairs of primers, and then were cloned into pMD 18-T Vector. The sequencing showed that the genes were 771 bp in length. All the entire PrP coding sequences had the complete ORFs contained within a single exon and were basically conserved with the published gene sequences. The sequence of the sheep and goat PrP genes cloned in this experiment contained five copies of a short, G-C-rich element, which encoded the octapeptide Pro-His-Gly-Gly-Gly-Trp-Gly-Gln or the nonapeptide Pro-Gln/His-Gly-Gly-Gly-Gly-Trp-Gly-Gln. The comparison of these genes each other revealed that the nucleotide and putative amino acid sequence identities ranged from 99.0% to 100.0 % and from 98.1% to 100.0 %, respectively. Of the total 17 base substitutions in the genes, six substitutions were synonymous mutation, and eleven produced amino acid mutation. Except for S240P of SY200301,SY200302 and SY200303 as well as M112T of MY200301, all other amino acid mutations including MY200301’s G129S, MY200302’s T191R, MY200303’s G127S, SY200302 and SY200303’s H143R and R211G, respectively were lied in residues 125～231 of spherical structural domain on the C-termination. All the six PrP genes were PrPARQ alleles at codons 136,154 and 171
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In this study, the chicken type II interferon gene(ChIFNγ) and infectious bronchitis virus (IBV) S1 gene were inserted into fowlpox virus transferring vector, for constructing pSY-ChIFNγS1 which contained these two genes. Then by lipofectin, the pSY-ChIFNγS1 was transfected into the chicken embryo fibroblasts (CEF) which was pre-infected with wide type fowlpox virus. After eight times selections of blue plagues on the CEF overlaid using X-gal as substrate, we obtained the purified rFPV-ChIFNγS1 which could express the S1 and ChIFNγ protein at the same time. Special antibody to IBV S1 protein can be identified by ELISA about one week after immunization of 28 days SPF chickens with rFPV-ChIFNγS1.The percentages of CD4⁺ , CD8⁺ and γδTCR positive T lymphocyte in peripheral blood of immune group are remarkably higher than that of Mock. Four weeks after immunization, the two groups were challenged with violent IBV LX4 strain, and only 1/16 chicken appeared light respiratory symptom in rFPV-ChIFNγS1 immunized group, but 16/16 chickens appeared in Mock, and 2/16 chickens died. The result of animal trail showed, rFPV-ChIFNγS1 can protect immunized chickens well.

The safety of recombinant fowlpox virus (rFPV-B-) expressing glycoprotein B (gB) gene of infectious larngotracheitis virus (ILTV) and F gene of Newcastle disease Virus(NDV) were evaluated. 100 30-day- old SPF chickens were divided into five groups, and inoculated with 50~5×10⁴ PFU/0.1 mL rFPV-B- F, respectively. 30 days after inoculation, chickens were divided and challenged with ILTV of Wanggang strain and NDV of F48E9 strain, respectively. Antibody titer to FPV was all positive in a dose dependent manner. And in the all range of all the inoculation, it can protect chickens against the challenge of the virulent ILTV and NDV, 7/10 chickens can be protected from clinical symptom in all the inoculation group, 8/10 of chickens inoculated survived from lethal challenge of NDV F48E9 in 500~5000PFU inoculation group, all these results lay the foundation for the further evaluation the efficiency of rFPV- gB- F.

BALB/c mice were immunized with pcDNA-PRRSV-ORF5 DNA vaccine by different routes of gene gun bombardment and intramuscular injection, with PBS and pcDNA3.1(+) as controls. Fluorescence activated cell sorter（FACS）was used to detect the number of〖JP2〗 CD4+ and CD8+ T lymphocytes. T lymphocyte proliferation test(MTT) and indirect ELISA methods were employed respectively to detect the proliferation of peripheral T lymphocyte and antiPRRSV antibody. Peripheral T lymphocytes responses to ConA of the experiment group and the control group were significantly different (P<0.01). In contrast with the control group, the number of CD4+ T lymphocyte 7 days after vaccination and CD8+ T lymphocyte 28 days after vaccination of the experiment group were significantly higher. The titers of specific IgG of the experiment group were higher than that of the control group. The results demonstrated that pcDNA-PRRSV-ORF5 DNA vaccine could induce good humoral and cellular immune response in mice. The titers of specific IgG of the group vaccinated by gene gun bombardment were higher than that by intramuscular injection.

Based on the NcSRS2 gene sequence of Neospora caninum, a pair of primers containing Kozak sequence, PstⅠand XbaⅠenzyme digestion sites were designed .Using the plasmid P43 which contains NcSRS2 gene as template,the ORF region of NcSRS2 gene was amplified by polymerase chain reaction (PCR), then the PCR product was digested with PstⅠand XbaⅠ,and the eukaryotic expression vector pcDNA3.1(+)was also done ,then the gene was cloned into the vector.Finally a recombinant plasmid named pcNcSRS2 was obtained, and was identified by PCR, restriction enzyme analysis and sequencing

The rat adipocytes were treated with 5,10,15 ng/mL tumor necrosis factor α (TNF-α) for 0，12， 24， 36 and 48 hours respectively. The apoptotic rates were determined by means of fluorescent staining and flow cytometry. Ultrastructure of adipocytes was observed by transmission electronic microscopy (TEM) and the DNA fragments were analyzed through agarose gel electrophoresis. The results showed that TNF-α induced the apoptosis of adipocytes and the apoptotic indexes were dependent on the concentration and time. Thus the optimum concentration and time for TNF-α inducing apoptosis was determined at the level of 10 ng/mL and 36 hours. Under the electronic microscope,the characteristics of apoptosis,such as the chromatin condensed and marginated ,and apoptotic bodies were observed in adipocytes. DNA ladders appeared in 24 hours after treatment,whereas,the ladders did not appeared in 0 and 12 hours treatment groups.

In order to verify the development of T & B lymphocytes and subsets of the T cell in chickens within 1 month, SPF chickens were killed in ,5,7,9,11,14,17,22,27,32 days respectively. Lymphocytes were separated from blood, spleen, thymus and bursa of chickens. Then the content of Bu-1a⁺B cell, CD3⁺, CD4⁺, CD8⁺and TCRγδ⁺CD3⁺ T cells and apoptosis of blood lymphocyte were checked by using flow cytometry with double-color-staining method in different days, respectively. The results showed: T & B lymphocytes were in the early stage of differentiating, maturing and were tending a increasing way except thymus within 1 month; but found no regularity in changes of T cell subsets. The speed of developing and maturing of B lymphocyte in bursa and T lymphocyte in spleen is the most rapidly in 3-9 days. Apoptosis of blood lymphocyte were decreasing especially 9th day later. They demonstrated T & B lymphocytes were in differentiating and developing in thymus and bursa continually, and then entering into spleen and blood, made matured lymphocyte content increasing within 1 month. But Bu-1a⁺B cell and CD3⁺T，CD4⁺T，CD8⁺T were in the lowest level in blood at 7th day, expressed lowest humeral and cellular immunological level at 7th day, then went to another lower at 17th day. TCRγδ⁺CD3⁺T lymphocyte changing in a stable pattern.

75 SD rats were used and randomly divided into 5 groups of 15 rats，including 8 male and 7 female each. The selenium contents in feed of group I, II and III were 0.1, 0.1 and 0.3 mg/kg from sodium selenite respectively. 0.3 mg/kg selenium from Seenriched malt and Se-enriched yeast were supplemented to the diet of group IV and V respectively. After 32 days, rats of group II, III，IV and V were treated with the program, which named short-term animal hepatocarcinoma model, by aflatoxin B1（AFB1）. Changes of feed intake, body weight, the haemal-indicators, such as alkaline phosphates(ALP), alphafetoprotein(AFP), nitric monoxide(NO), γ-glutamyltranspeptidase(γ-GTase), superoxide dismutase (SOD) and malonadehyde (MDA) in serum, the antioxidative indices, such as glutathione peroxidase (GSH-Px), SOD and MDA in blood and tissue, the activity of hydroxyproline(HyP) in liver were measured. The data showed that effect of selenium from Se-enriched malt and yeast were better than sodium selenite in the aspects of body weight gaining, raising GSH-Px and SOD activity, clearance of kidney MDA and decreasing the activity of HyP and γ-GTase. The selenium from Se-enriched malt was better than that of yeast by raising rapidly GSH-Px in blood and decreasing the activity of HyP and γ-GTase. In conclusion, the effect of selenium from Se-enriched malt and yeast on the hepatocarcinogenesis induced by AFB was better than that of sodium selenite in stimulating the antioxidative function and stimulating the rat growth.

Radioimmunoassay was used to detect endothelin-1(ET-1), atrial natriuretic polypeptide (ANP) in plasma and ET-1 in heart, liver, lung and kidney of normal group, light ascites group and heavy ascites group of 35-days old commercial broilers. The AHI, mRVP and mPAP were measured as well.At the same time, nitrate reductase was used to detect the level of nitric oxide (NO) in serum and AHI was measured as well. It was found that, compared with the normal group, the AHI, mRVP and mPAP of AS groups were significantly higher, and the heavy ascites group was significantly higher than that of the light ascites group, the trend of increment of them was observed following the degree of ascites. Other indexes such as the level of ET-1 in plasma and tissues, ANP in plasma, NO in serum have the same tendency. The level of ET-1 of tissues was higher than that of plasma and the level of ET-1 of lung and heart was significantly higher than that of other organs. Besides, there was a significant positive correlation among plasma ANP, plasma ET-1, mPAP, heart ET-1 and AHI, but no significant correlation was found between serum NO and other indexes among the three groups. It is concluded that ET-1, ANP and NO took part in the formation and development of AS in broiler and played an important role in it.

The characteristics of deposition of chicken IMP and IMF were investigated in this experiment. Arbor Acres (AA) and Beijing Fatty Chickens (n=200, respectively) were raised under the same condition and were slaughtered at 28 d, 56 d and 90 d to assay the content of IMP and IMF in breast and leg muscle. The results showed that the IMP content of Beijing Fatty Chicken was significantly higher than that of AA (P<0.01), and also significant difference existed between them in IMF (P<0.01). The IMP content in breast muscle was higher than that in leg muscle (P<0.01), while the IMF was inverse (P<0.01). IMP content of female was a little bit higher than that of male (P>0.05). IMF content varied with growth rate, and showed no association with sex. IMF content increased with age, while IMP did not.

A so called “new duck disease” with the character of necrosis of spleen and pancreas broke out in flocks of muscovy ducks at the end of 1999 in Zhejiang province. Viruses with different conformation and size, one of which was similar to reovirus were found under electronic microscope in allantoic fluid for virus isolation. Three pairs of primers were designed and synthesized based on the published S1 sequences of muscovy duck reovirus (mDRV). Specific RT-PCR products were obtained from both diseased tissues and allantoic fluid for virus isolation. Cloning and sequencing of the fragment and BLAST analysis result further verified the existence and participation of mDRV in the disease. A 1 260 bp S1 full-length sequence of the virus (ZJ99) was further amplified and sequenced. Phylogenic analysis result based on S1 sequences showed that the ZJ99 was in the same group with mDRVs of the French strain and the Fujian strain, while it had some distance to the group of the chicken reoviruses and was far more distant to that of mammalian. On the other hand, the nucleotide sequence of S1 of the ZJ99 strain had a homology of 98% with that of Fujian strain (300 bp partial sequence), while only had 90% homology to that of French strain. It was speculated that mDRVs isolated from different area of China, which had some differences with French strains, were the same strain but spreading in different area.

In order to provide the morphological evidence for investigating the physiological function of medulla oblongata of African ostrich, the study was carried out via paraffin continual sections and haematoxylin-eosine (HE) staining. There are medullary reticular nucleus, reticular gigantocellular nucleus, lateral reticular nucleus and raphe nuclei.The raphe nuclei can be divided into three subnuclei: nucleus raphe obscurus, nucleus raphe pallidus and nucleus raphe magnus. The reticular formation nuclei contain heterogeneous population of neurons, which vary in size and shape. Cells with abundant processes are in the shape of ellipse, fusiform, polygon and triangle. Small cells are little. Cell nucleus are in the middle or at the side of the cell appeared rotundity or ellipse. Nucleolus is clear. The results indicate that the reticular nucleus cytoarchitecture of medullary reticular formation of African ostrich is not much different from other birds, except relatively less nuclei, larger cell and more abundant processes; there are some differences in raphe nuclei.

Brain, spinal cord, lung， heart and liver of mouse infected by VSV was studied using immunohistochemistry method and HE staining. The results showed that VSV can infect mouse, in earlier period of infection, lesions located mainly in lung, in anaphase of infection, lesions located mainly in brain and spinal cord.
Positive cells were found in lung, brain and spinal cord. The positive rate in brain was higher.Results showed that duplication of VSV occured mainly in lung and brain,and cause corresponding pathogenicity.

Twelve goats were divided into two study groups.GroupⅠwas treated as control and groupⅡwere fed sodium fluoride(NaF) orally according to their bodyweight. After 12 months of fluoride dosing, goats of groupⅡ showed clinical signs of fluorosis .Erthrocyte were collected for analyses. The results showed that erthrocyte anti-oxidant function,ion pump function(ATPase),the contents of sialic acid(SA) and erythrocyte immune function were decreased due to excessive fluoride ingestion. A great number of erthrocyte deformed with polyhedral angel shape and star shape by the observation of scaning electronic microscope in groupⅡ. The results provide the valuable clue of the anemia mechanism in chronic fluorosis.