In order to amplify cDNA of Mongolia sheep ghrelin, the differential primers were designed according to the GenBank sequence. Total RNA was extracted from the abomasums of Mongolia sheep and the cDNA encoding Mongolia sheep ghrelin was obtained by the reverse transcription PCR(RT-PCR).The purified RT-PCR product was cloned into pTZ19 vector.The results of restriction endonuclease pattern analysis of recombinant plasmid and DNA sequencing demonstrated that the Mongolia sheep ghrelin cDNA was part of ghrelin, because blasted sequencing result with published sheep ghrelin in NCBI web, only one base was different in 205 bases.The difference didn’t influence polypeptide sequence after translation. The cDNA contained a ORF of 168 bases which encoded a 56 amino acid prepropeptide.The research to Mongolia sheep ghrelin will help us to open out its physiological and pathological function, and it will make farreaching sense in preventing and curing some ruminant diseases.

Total RNA was extracted from the tougue epithelia of a reindeer and the cDNA encoding reindeer β-defensin (reBD-1) was amplified by the reverse transcription PCR(RT-PCR).The purified RT-PCR product was cloned in pBlueselect T vector. The results of restriction endonuclease pattern analysis of recombinant plasmid and DNA sequencing demonstrated that the reBD-1 is belong to the family of β-defensin because the cDNA contain a open reading frame (ORF) of 192 bases which encoded a 64 amino acid prepro-peptide and the prepro-peptide contained the β-defensin consensus sequence of six invariantly spaced cysteine residues. The reBD-1 is the first β-defensin found in reindeers. This finding of the reindeer β-defensin facilitate a better understanding of mucosal defense mechanisms in reindeers.

One pair of primer for goat MSTN gene intron 2 was designed according to goat MSTN gene sequence (Accession No. DQ167575) and PCR-RFLP was carried out in a total number of 102 individuals including Boer goat and its backcrossed offspring to Tangshan dairy goat. The result of sequencing showed that there were two substitution mutations in the fragments. Statistical analysis indicated that AA genotype had significant higher weaning weight than BB and AB genotype (P<0.05).CC genotype had significant higher birth weight than CD genotype (P<0.05). It could be inferred that the MSTN gene maybe a major gene or linked to the major gene affecting the body weight. The polymorphic site could be used to marker assisted selection program for body weight.

123 geese were clustered using between-groups linkage based on 12 common factors of 40 goose carcass and feather-down traits and divided into 6 groups. By analysing difference among groups, productivity of Wanxi White goose×Sichuan White goose were estimated. The results showed that the difference among groups was significant while all geese were divided into 6 groups. The factor scorings of meat power, feather-down power, and eviscerated percentage were higher in group 1 and group 2, which indicated that geese in group 1 and group 2 had a superiority in meat-and feather-yield capability, and had lower fat, better meat hold-water capability, and lower down hydrophilicity. The factor scorings of fatty acid quality, fat power, leg muscle percentage, meat water loss, digestion, heart function, down quality, and down hydrophilicity were higher in group 5 and group 6, which indicated that geese in group 5 and group 6 had a preponderance in goose meat and down quality, and had leg muscle upgrowth, fatness, and lower hold-water capability. Group 3 and group 4 had higher factor scoring in freather-type, and other factor scorings were middlebrow.

Retinoic acid receptor-gamma（RARG）gene was studied as a candidate gene for the prolificacy of Small Tailed Han sheep. The single nucleotide polymorphism of RARG gene in both high fecundity breeds (Small Tailed Han and Hu sheep) and low fecundity breeds (Texel，Dorset and Suffolk sheep) was analyzed by PCR-SSCP. There was PCR-SSCP polymorphism for primer 1 in five sheep breeds. AA genotype was only detected in Hu sheep, AB and BB genotypes were detected in five sheep breeds. The ewes with genotype BB had 0.41 (P>0.05) lambs more than those with genotype AB in Small Tailed Han sheep. There was PCR-SSCP polymorphism for primer 2 in five sheep breeds. CC and CD genotypes were detected in five sheep breeds. The ewes with genotype CC had 0.55 (P<0.05) lambs more than those with genotype CD in Small Tailed Han sheep.

The development of skin and hair follicle traits have been studied on the Inner Mongolian Arbas goats in the fetal period. Skin samples were obtained from 10 different body parts of fetuses at the age of 55 days to birth. The results showed that the complete structure of epidermis layer formed at the age of 55-65 days and epidermis was the thickest at the age of 85-95 days; Dermis thickness increased rapidly at the age of 95-125 days. The primary hair follicle was seen first at the age of 55 days while secondary hair follicle at the age of 65 days. Moreover, an important fact that secondary hair follicle is a branch of primary hair follicle has been found first on cashmere goat. The sebaceous gland and sweat gland were seen at the age of 85 and 95 days, respectively. Wool and cashmere began to grow at the age of 95 days. Wool came out of the skin surface at the age of 95 days while cashmere at the age of 115 days. The primary and secondary follicle grew very fast at the age of 95-115 days and 105-125 days, respectively. The density of primary follicle showed highest at the age of 75-85 days and secondary follicle at the age of 105-125 days. The value of S/P was the biggest at the age of 115 days to birth. The hair follicle group pattern formed at the age of the 105 days, trio group is dominant.

The effects of 5-amino-4-imidazolecarboxamide riboside（AICAR）and the antidiabetic drug metformin on AMP-activated protein kinase (AMPK) activity and its downstream effector, acetyl-CoA carboxylase (ACC),β-hydroxy-β-methylglutary-CoA reductase（HMGR）activities in laying hens’ primary cultured hepatocytes were studied. Four treatments were employed in this experiment AICAR（0.5mmol/L）, metformin（0.5mmol/L）, and metformin（1 mmol/L）were added respectively into the standard culture medium(control). At 100 min after incubation, AICAR or metformin activated AMPK significantly ( P＜0.05), inactivated ACC and HMGR which are the major enzymes involved in fatty acid and cholesterol biosyntheses. AMPK regulated ACC and HMGR activities negatively. AICAR or metformin decreased hepatic lipid and cholesterol contents by activating AMPK. AMPK negatively regulated hepatic lipid and cholesterol contents. These findings suggest that the regulation of hepatic lipid and the cholesterol metabolism of primary hepatocytes in laying hens can be mediated by AMPK. 

The objective of this study was to determine the effects of chelated chromium (chromium-picolinate or chromium-lysine ) supplementation on lactation performance and some blood components of Holstein cows during the lactating period. Seventy five multiparous lactating Holstein cows (80 days post-partum) used in this study were divided equally into three groups (25 cows/treatment) and were randomly allocated to one of three treatment diets. Diet A, basal diet, with chromium-picolinate added at manufacturer’s recommended level (0.6 g /head per day). Diet B, basal diet, with chromium-lysine(0.076 g /head per day). Diet C, was basal diet without chromium (Cr) added and served as a control. The result of this study showed the supplement of chromium to the diet of cattle contributed to the increase of milk yield and may contribute to the improvement of the immune status of stressed dairy cows. In comparison with the diet C, milk production of diet A and diet B were both increased（P<0.01）,insulin concentration in blood of diet A led to depressed（P<0.01）and glucose concentrations in blood of diet A and diet B were higher than that of treatment diet C（P<0.01）.The mean Cortisol concentration in blood of diet A and diet B decreased by 7.21 nmol/L, 4.92 nmol/L, respectively, and the mean IgG concentration in blood of diet A and diet B increased by 2.1 g/L and 3.1 g/L , respectively, compared with diet C.

This study was carried out to determine the effect of infusion of Fructooligosaccharides(FOS) on rumen fermentation of growing sheep by nutrients infusion technique.Six Inner Mongolian wethers with permanent ruminal cannulas were randomly alloted to two groups which were infused FOS at 1.00% and 2.00% of dry matter intake respectively. The results indicated that there were significant decreases（P<0.05） in pH value and NH₃-N concentration in the rumen by the infusion of FOS at both 1.00% and 2.00%,at the same time MCP production and VFA concentration in the rumen were significantly enhanced（P<0.05）. There was a higer decrease （P<0.05）of NH₃-N concentration in the rumen of the sheep infused 2.00% FOS than those infused 1.00% FOS .However,no significant difference （P>0.05） was observed in pH value , VFA concentration and MCP production between the two group sheep infused different level FOS .

Corn stalks from Zea mays L, were served as experimental material to study the effects of N-fertilizer (urea 0,150,300,450 kg/ha) on the nutritional value by using chemical analysis, in vitro two-stage digestion, in vitro gas production and sheep feeding trial. The results showed that the corn stalks had a linear (P<0.001) increase in the contents of CP, true protein, amino acids, nonessential amino acids, essential amino acids and Ca as quantity of urea fertilized increased, and total saccharide content did not change with urea under 300 kg/ha and had a negative linear correlation with urea above 300kg/ha. N-fertilizer had little or no effect on EE, ADF content of corn stalks. The in vitro digestibilities of DM, CP, NDF and ADF of corn stalks increased (P<0.01) in a linear (P<0.001) with the application of N-fertilizer, but when urea dose was upon 300 kg/ha, the function was not enhanced. Daily gain, feed conversion and economic return increased after the lambs (Fat-tailed Han-yang) were fed the silage of the corn stalks fertilized with urea. When corn plants were fertilized with urea 300 kg/ha, the result of feeding was the best. In conclusion, the contents of chemical components of corn stalks were alternated, and moreover, nutritional value of corn stalks was increased after corn plants had been applied with N-fertilizer.

In this article, the IgG was purified from the positive antiserum to FMDV-NSP 3ABC. The IgG was used as target molecule to screen the phage display 12mer random peptide library for 4 rounds by the binding-eluting-amplification method. After 4 rounds of screening, 20 clones were picked out randomly and amplified.Their antigenicities were tested by ELISA. Among 20 phage clones, 8 phage clones had specific reactions with the purified IgG.The ssDNA of the positive clones were abstracted for DNA sequencing and the sequences of peptides displayed on the positive clones were analyzed by ExPASy method.The results showed that the peptides displayed on the 7 phage clones were highly homologous. 22 dubitable porcine sera were tested by 8 positive clones which were used as target molecules respectively, and 5 clones showed high correlation with traditional Kit. These results were successfully provided a foundation for research of the structural epitope of the FMDV-NSP 3ABC and to establish a new diagnostic method for the natural infection of the FMDV.

The purpose of this study was to construct PRRS-CSF-PCV2 diagnostic DNA microarray. Target genes were chose from PRRSV, CSFV and PCV2 based on their sequences, and corresponding probes were prepared. Then the DNA fragments were spotted onto slide to form DNA microarray. The results showed that the diagnostic DNA microarray can detect PRRS, CSF and PCV2 simultaneously , and can distinguish genotype of PRRSV. The positive standard was formulated as SNR≥1.5 or Intensity≥1 000 based on quantification patterns of total signal intensity and signal Intensity median, respectively. It was high specificity, high sensitivity and reutilization. 20 clinical samples were detected by PRRS-CSF-PCV2 diagnostic DNA microarray. 15%(3/20) , 5% (1/20), 0(0/20) samples were positive for CSFV, PCV, PRRSV, respectively. Co-infection ratios were 15%(3/20) for PRRSV and CSFV, 35%(7/20) for PRRSV and PCV2, 15%(3/20) for PRRSV,CSFV and PCV2.

The objective of the present study was to construct a cDNA library for male adult of Ascaris suum with SMART(switching mechanism at 5′end of RNA transcript) technique using Creator^TM SMART^TM cDNA library construction kit. The total RNA was extracted from A. suum male adult using TriPure isolation reagent and mRNA was purified using Poly(A)Purist^TM kit. Single-strand cDNA was synthesized using PowerScript^TM reverse transcriptase, and then doublestrand cDNA was synthesized and amplified by longdistance PCR (LD-PCR). The PCR products were digested by proteinase K and purified. After digestion with SfiⅠ and size fractionation using CHROMA SPIN-400TM columns, SMART cDNA was ligated to the Sfi I-digested, dephosphorylated pDNR-LIB vector. The ligation mixture was transformed into E.coli DH5α by electroporation. The constructed cDNA library contained 7.26×10⁵ independent clones. The recombination rate was 96.7%. The average cDNA insert size was 1 kb. After the library was amplified, its capacity was 6.359×10⁹ cfu/mL. A cDNA library for A. suum male adult was successfully constructed using SMART technology, which provides foundation for the screening and isolation of male-specific genes from A. suum.

We have investigated the effect of Sijunzi decoction on expression of cholecystokinin(CCK) and vasoactive intestinal peptide(VIP) mRNA in duck duodenum and jejunum of experimental spleen deficiency syndrome models(SDS). The models of SDS were established by injecting Reserpine and the expressions of CCK and VIP mRNA were determined by Semiquantitative reverse transcription polymerase chain reaction (RT-PCR). No significant differences in the expression levels of CCK, VIP mRNA were found in duodenum and jejunum between Sijunzi decoction preventive group and control group. The expression levels of CCK, VIP mRNA in duodenum and jejunum of SDS group were significantly lower than that of preventive group (P<0.01 or P<0.05); The expression levels of CCK, VIP mRNA of spontaneous recovery group were significantly lower than that of control group (P<0.01). In duodenum and jejunum, the expression levels of CCK, VIP mRNA in treated group were significantly higher than that of spontaneous recovery group (P<0.01 or P<0.05). These results suggest that Sijunzi decoction may exert its beneficial therapeutic action on SDS, at least in part, by upregulated CCK and VIP mRNA levels in duodenum and jejunum.

To study the immunoenhancement of genetic adjuvant GM-CSF on somatostatin (SS) DNA vaccine, a total of 40 mice at the age of 22 days were divided into 4 groups and immunized with salt solution (group 1), pcS/2SS (group 2), pGM-CSF/SS (group 3) and pGM-CSF+pcS/2SS(group 4) delivered by attenuated Salmonella at a dose of 10⁹ CFU per mouse, respectively. A booster was given 2 weeks later with same dosage and method. The results are as follows: Lymphocyte proliferation response of group 3 was higher than the others, indicating that GM-CSF can enhance lymphocyte proliferation responses; the GH level in plasma of group 3 and IGF-Ⅰ level in plasma of group 4 are higher than that of other groups, and levels of GH and IGF-Ⅰbegin to rise at the 2nd and 4th week post primary immunization, respectively. We conclude that genetic adjuvant GM-CSF can enhance immuno-function of somatostatin DNA vaccine. Immune response of fusion-expression plasmid of GM-CSF and SS is superior to that of co-immunization of pGM-CSF+pcS/2SS.

Distribution of orexin Blike immunoreactive (OXB-IR) fibers in the central nervous system(CNS) of the rabbit was studied by immunohistochemistry. The results showed that OXB-IR fibers were observed in the CNS widely. In the telencephalon, OXB-IR fibers were present in the cerebral cortex, amygdala nucleus and septal area. In the diencephalon, OXB-IR fibers were observed in the thalamus, hypothalamus, epithalamus and hypophysis with the majority in the hypothalamus and the midline nuclear group of the thalamus. In the mesencephalon, OXB-IR fibers were located in the central gray substance, superior and inferior colliculi, interpeduncular nucleus and raphe nuclei. In the pons, OXB-IR fibers were seen in the locus coeruleus, tegmental nucleus and raphe nuclei. In the medulla oblongata, OXB-IR fibers were found in the area postrema, solitary tract nucleus and dorsal motor nucleus of vagus nerve. OXB-IR fibers were also present in the cerebellum and spinal cord. Distribution of OXB-IR fibers in rabbit CNS was similar to that of OXA-IR fibers. Our results provided morphological evidence for understanding of the roles of orexins in the rabbit

This experiment aimed to study the relationship between calcium signal transduction and expression of c-fos,c-myc in broiler’s Pulmonary Arterial Smooth Muscle Cell (PASMC) induced by hypoxia. The effect of calcium signal transduction on the oncogene c-fos and c-myc expression in PASMC induced by hypoxia was observed by immunohistochemistry, in situ hybridization and image analysis methods. The results showed that the antagonism of calcium channel such as verapamil (V),nifedipine(N),Cyclosporin A(C) can restrain such an expression (Expression level of c-fos mRNA was 198.1±32.8 in hypoxia group, 161.6±24.1 in hypoxia+V group, 173.7±35.8 in hypoxia+N group, 159.0±32.8 in hypoxia+C group, P<0.01; Expression level of c-myc mRNA was 187.1±22.4 in hypoxia group, 150.5±22.6 in hypoxia+V group, 155.1±25.4 in hypoxia+N group, 148.9±29.7 in hypoxia+C group, P<0.01) and the antagonism can restrain c-fos and c-myc protein expression (Expression level of c-fos protein was 225.9±37.0 in hypoxia group,162.3±33.6 in hypoxia+V group,157.4±28.9 in hypoxia+N group, 151.6±37.3 in hypoxia+C group,P<0.01;Expression level of c-myc protein was 228.8±33.4 in hypoxia group, 170.2±26.7 in hypoxia+V group, 170.7±28.5 in hypoxia+N group, 179.1±29.2 in hypoxia+C group,P<0.01)PASMC proliferation were the important cause of ascites syndrome（AS）.Verapamil and nifedipine can significantly restrain the c-fos and c-myc expression of PASMC, indicating that calcium signal transduction was important to the development of AS. Cyclosporin A can significantly restrain c-fos and c-myc expression of PASMC indicating that the Ca2+/CaM-CaN path is important to c-fos and c-myc expression. From above results the conclusion can be obtained that calcium signal transduction play an important role in the development of AS.

Beijing Fatty, Recessive White Plymouth Rock (RWPR) and a cross between Beijing Fatty(♂) and RWPR (♀) were fed to 14 weeks to investigate the development character of growth of muscle fiber by paraffin slice protocol. The results showed that the diameter of muscle fiber increased with age, while the muscle fiber density decreased. Moreover, the relationship of diameter and density was significantly negative(P＜0.000 1). The growth rate of muscle fiber varied among different breeds. The diameter of thigh muscle fibre in Beijing Fatty, cross and RWPR at 14 weeks was 42.89 μm, 49.29 μm and 56.93 μm respectively; Contrarily, the density at 14 weeks was 581.80 piece/mm², 447.53 piece/mm² and 349.87 piece/mm² respectively; From 4 weeks to 14 weeks , the diameters of muscle fibre between Beijing Fatty and RWPR were significantly different(P＜0.01). To conclude, the diameter of thigh muscle fibre was greater than that of breast muscle, while the density was smaller; The diameter of thigh and breast muscle fibre in RWPR at 6 weeks was 27.96 μm and 24.61 μm, and their density was 1 557.50 piece /mm² and 1 809.30 piece/mm^2. The heterosis rate of the diameter and density of muscle fibre did not vary steadily along with weeks. As a whole, there was hardly any heterosis in these traits in the offspring from the cross of Beijing Fatty (♂) and RWPR (♀), however, they tended to follow that of the maternal line.

Mitochondrial cytochrome b sequence were used to study the molecular evolution of Anser cygnoides（15 breed of Chinese goose）, Anser anser（2 breed of domestic European goose） and Anser albifrons（the lesser whitefronted goose, GenBank accession number AF363031）.The results revealed that insertion/deletion or frameshift mutation were not found in Cyt-b gene sequences（1 143 bp）, the probability of transition （Ts） was higher than that of tranversion （Tv） , there was the highest number of transition in the third codon. Frequences of synonymous codon usage were relatively biased. There was a significant difference between dS and dN（Z=4.713，P<0.01）, and the low ratio（ω= 0.028 4 <0.3） of dN/dS impled that there were selective constraints against the nonsynonymous sites in cyt b gene.The distribution of nonsynonymous codon substitution pattern related to Grantharm’distance indicated that the purified selection at three codon positions was different intensity. The 2nd codon positions suffered the greatest intensity of purified selection(Granthem distance is 204).