Adipose tissue，an important factor that affects the porcine carcass quality，is closely related with the pork quality.Understanding the regulation mechanisms of adipose tissue development and fat metabolism are important to the animal husbandry production and disease treatment.miRNAs(microRNAs) are a class of endogenous，non-coding，small RNA molecules，which are typically 18-24 nucleotides(nt) in length.miRNAs play an important role in the regulation of gene expression at the post-transcriptional level.A large number of evidences demonstrated that miRNAs have influence on diverse biological processes，such as preadipocyte differentiation，adipogenesis，fatty acid metabolism and cholesterol biosynthesis.However，only 326 porcine miRNAs has been currently reported and rare studies could be read about the porcine miRNAs that regulate the fat development.Herein，it is reviewed that the studies on the discovery and identification of fat-associated miRNAs in pigs in this paper，and the progress on the research of porcine miRNAs that regulate fat development are also summarized.

Ubiquitylation is a covalent post-translational modification that regulates protein stability and is involved in many biological functions.Many poxviruses have evolved multiple mechanisms to perturb the cell ubiquitin system and manipulate it to their own benefit.Such as the poxvirus family encode ubiquitin ligases with intrinsic activity，including the membrane-associated RING-CH(MARCH) domain，p28/Really Interesting New Gene(RING) finger，ankyrin-repeat/F-box and Broad-complex，Tramtrack and Bric-a-Brac(BTB)/Kelch subgroups of the E3 Ub ligase superfamily.Here we describe and discuss the proteins with E3 ubiquitin ligase function encoded by poxvirus.

 In order to further explore the biological function of pig LTβR gene，the coding sequence(CDS) of LTβR gene was amplified from the cDNA of porcine lymphoid tissue and the protein structure and function，GO function，regulatory pathways of porcine LTβR were further analyzed by bioinformatics tools.Real-time PCR was used to detect the expression level of porcine LTβR gene in different tissues of Yorkshire.Meanwhile LTβR gene was cloned into the eukaryotic expression vector pEGFP-C1，then transfected into HEK293 cell and small intestinal epithelial cell IPEC-J2，respectively.The expression of recombinant plasmid was detected in both cells through microscopic observation.The results showed that the pig LTβR cDNA full length was 1 209 bp encoding 402 amino acids.LTβR was a fat-soluble，hydrophilic and unstable protein，containing a transmembrane structure between 205 and 227 amino acids，meanwhile no signal peptide and subcellular localization indicated that LTβR protein was non-secretory.LTβR protein had 2 glycosylation sites，18 potential phosphorylation sites including 10 conservative binding sites of specific protein kinase such as PKC，CKI，CDC2，GSK3，CDK5，INSR，etc.Pig LTβR protein had 2 conservative domains named TNFR superfamily，which were located in the region between 32 and 125 amino acid，between 128 and 192 amino acid，respectively.Besides the mutation C422T>A141V occurred in the conservative domain.KEGG and GO analysis showed LTβR gene participated in 12 GO function classifications，5 regulatory pathways such as NF-kappa B signaling pathway，intestinal immune network for IgA production，etc.Real-time PCR analysis showed the expression level of LTβR gene in Yorkshire lung tissue was very significantly higher than other tissues(P<0.01).Moreover，LTβR gene was also highly expressed in both pig immune organs such as kidney，spleen and intestinal tissue such as duodenum，jejunum.Transfection experiments and microscopic observation showed the pEGFP-C1-LTβR recombinant plasmid was expressed in both HEK293 and IPEC-J2 cells.This study will provides materials and basis for studying the function of LTβR gene and LTβR-mediated signaling pathways，therefore it is necessary to further verify and analyze the regulatory mechanism and important role of LTβR gene and signaling pathways at cellular level，meanwhile we should systematically analyze the mutation C422T as a potential genetic marker for pig disease-resistant breeding.

In order to provide data support for genetic parameters of piglet uniformity in breeding，to understand the impacts of non-genetic factors on raising pigs，and to provide references for production management of pig farms，we have evaluated the genetic parameters of piglet uniformity and statistically analyzed the fixed effects (years，mating seasons and parity) affecting the uniformity of piglets in this study.Statistical analysis of 2 766 purebred Large White sows’ farrowing records were conducted and the heritabilities and genetic correlations among important reproductive traits in groups were estimated using REML (restricted maximum likelihood) method.Moreover，least squares analysis for the influence factors of piglet uniformity were set up using fixed-effect model.The heritability of litter uniformity，total number of newborns，number born alive，number born dead and total litter weight at birth of piglets was 0.096，0.107，0.101，0.068 and 0.171，respectively.The genetic correlation coefficient between piglet uniformity and total number of newborns and between piglet uniformity and number born dead were 0.205 and 0.157，respectively.Fixed-effect analysis revealed that the seasons had significant impacts (P=0.023 6) on the uniformity of piglets.Parity and number of newborns had significant impacts (P=0.001) on the uniformity of piglets.The uniformity of piglets is heritable and its heritability is similar to the heritability of number of newborns.The uniformity of piglets can be improved by a reasonable breeding program.As uniformity of piglets has a positive genetic correlation with the total number of newborns and death rate，breeders should not pursue the high yield blindly.Uniformity of piglets，piglet death rate and sow annual productivity should also be considered.

 The research was designed to study the activity of goat ovarian maintenance gene FOXL2 promoter and explore the gene’s regulation mechanism.FOXL2 promoter sequence was retrieved from the NCBI database，bioinformatics software was adopted to predict its core promoter and transcription factors.PCR technology was used to clone FOXL2 gene promoter sequence and construct a series of deletion vectors，293T and A375 cells were transiently transfected，dual luciferase gene detector was used to measure the relative luciferase activity value.The results indicated that there were 2 typical CpG islands in the gene promoter region，which were located at （-920/+51(972 bp)） and （+125/+555(430 bp)） regions；the result of Kpn I and Hind Ⅲ dual enzyme digestion test suggested that the recombinant plasmid was constructed correctly；FOXL2 gene promoter fragments with different lengths were inserted into the cells.When the promoter 5′was truncated，luciferase transcriptional activity firstly increased and then decreased.The result indicate that （-934/+324） region has transcriptional activity，（-32/+324） region contain the basic elements of transcription；（-934/-456） region negatively regulates FOXL2 gene during the transcription process，（-456/-192） region is a positive regulatory region.

The study aimed to screen the selection signatures on X chromosome of Debao pony.The Illumina Equine 65K SNP BeadChip was used to perform a X-chromosomal scan on Debao pony，Yili horses and Mongolian horses and obtain 2 339 SNPs with high quality.The result showed that，based on these SNPs，we detected 2 regions(4.0-39.9 and 87.1-123.5 Mb) under strong selection on the X chromosome of Debao pony by 2 different methods，F_ST and XP-EHH.The 2 detected regions contained 64 outliers overlapping genes including BCOR，PHEX，PNPLA4 and GPC3，which are related to the body size and the bone development.The results indicate that genes associated with growth on X chromosome are subject to strong artificial selection pressure during the breeding of the smallest Chinese pony.A few of them are not reported before and could be important candidate genes in future study.

The study is focused on screening the proteins interacting with cocaine-and amphetamine-regulated transcript(CART) during follicle development in bovine.Four 10-month-old healthy cows，treated for estrus synchronization，were monitored by B-ultrasonic machine to find the ovarian follicles with the diameter of 8-12 mm，which were collected followed by cows being slaughtered.The granulosa cells(GCs) were isolated from those ovarian follicles and then total protein was extracted.CART and the binding protein were deposited by primary antibodies(Rabbit anti CART) using Protein G-Agarose magnetic beads，followed by salt and miscellaneous protein being removed by washing repeatedly.Protein complexes was analyzed by LC-ESI-Q-TOF mass spectrometry(MS) and alignment followed by being eluted and digested by trypsin.Results showed that 111 proteins were identified，in which 81 proteins were functional in conformity with the interaction network modules by Cytoscape V3.1.0 analysis.For the function enrichment analysis，those 81 proteins were classified into 34 groups in 3 categories：molecular function(11.73%)，biological processes(46.93%)，cell component(41.34%).The correlation coefficient of the interaction between A2M，VIM and CART was maximum.It was also found CART was negatively regulated by NALP1，SERPINH1 and PDIA6，which suggested that they might participate in regulating follicle development in bovine.

The aim of this study was to clone Nanog gene promoter of chicken and construct the expression vector containing dual luciferase report gene to analyze the promoter activity，find out its core regulatory region，explore the effect of 5-Azadc(5-Aza-2′-deoxycytidine) and TSA(Trichostatin A)on its promoter activity and ESC pluripotent maintaining in vitro condition，so as to elucidate preliminarily the regulating mechanisms of Nanog gene and provide the theory basis for the future research.The different length fragments Nanog gene promoter was amplified by PCR technology and cloned into pGL3-basic vector or pEGFP-N1 directly to construct a recombinant vector.After the recombinant vector was transfected into DF-1 cells for 24 h and the protein was collected，the transcriptional activity of Nanog gene was measured by dual luciferase assay system to search for the basic transcriptional regulatory region；The recombinant vector was transfected into DF-1 cells，and methylation inhibitor 5-Azadc or histone deacetylase inhibitor TSA was added to detect its effect on the promoter activity.The third generation ESC was selected and grown into the medium supplemented with the optimum concentration of 5-Azadc and TSA，the cells were observed every 2 d to analyze the pluripotent maintaining of ESC in vitro culture condition，and indirect immune fluorescence of SSEA-1 was detected on the tenth days induction.The results showed that 3 dual luciferase report gene expression vector was constructed successfully.The strongest dual luciferase activity was shown by pGL3/1967；the transcription activity of Nanog gene was enhanced when 5-Azadc and TSA was added single or together.ESC was cultured in the medium with 5-Azadc and TSA in vitro，a great number of cells differentiated after 6 d of induction and cell colony was not maintained in control group，while the cell colony in 5-Azadc and TSA induction group was significant higher than the control group；On the tenth days induction，the cells in control group were fully differentiated，there were still a large number of cell clone in the 5-Azadc + TSA group，the number of colony were significantly more than the other 3 groups；the results of indirect immunofluorescence showed there was no cell colony in the control group，but more colonies were observed in the 5-Azadc and TSA induction group.It further showed that 5-Azadc and TSA could effectively maintain ESC pluripotence of chicken in vitro culture condition by increasing the expression of Nanog gene.

This study was aimed to identify the availability of early embryo blood vessels microinjection of lentiviral vector as a new，effective way in making transgenic birds，here，we combined some of the transgenic technology such as primordial germ cells(PGCs)，microinjection and lentiviral vector together according to the characteristic of bird development.The number of PGCs in the bloodstream was maximal when the chicken embryoes developed to Hamburger-Hamilton stage 13-15(HH13-15)，1 μL lentiviral vector，containing an enhanced-green fluorescent protein(eGFP)，was microinjected into the blood vessels of quail embryos at stage HH13-15 with a titer of 1×10⁹ TU•mL^-1.A total of 80 embryos were injected and 48 quails(60%) were successfully hatched.In newly hatched quails，eGFP expression was shown as the presence of green fluorescence in the beak，feather，eye，brain，blood vessels，heart，liver，spleen，lung，kidney，glandular stomach，mesenterium，small intestine，large intestine，oviduct，muscle and claws，especially in gonads.In 5 out of 21 mature G0 male quails，the semen was eGFP-positive(5/21，23.8%)，as detected by polymerase chain reaction(PCR).Southern blot and genetic analyses revealed that in the 46 G1 offspring produced by G0 quail，6 were transgenic(6/46，13.0%) according to the PCR and Southern blot results.In conclusion，transgenic germ line chimeras was successfully generated by injection of lentiviral vector into embryonic blood vessel at stage HH 13-15.It indicated that infection of PGCs with lentiviral vector via direct injection into blood vessels has the potential to provide a more convenient and efficient way to produce transgenic birds.

The effect of IFN-γ on melanocytes proliferation and melanogenesis was studied.Different concentrations of IFN-γ (0.0，0.5，2.0，8.0，16.0，32.0，64.0 ng•mL^-1) treated separately melanocytes of Alpaca skin at 24，48，72 h.The activity of melanocytes was detected using MTT method，the melanin content was tested with ultraviolet spectrophotometric method and qRT-PCR was used to detect the changes of MC1R，TYR，TYRP1，MITF，DCT and NOS2 on mRNA level.The results showed that the melanocytes activity was inhibited after 24 h，melanocytes activity recovered and the melanogenesis was enhanced after 72 h.IFN-γ promoted the increase of melanin content and 32.0 ng•mL^-1 IFN-γ increased significantly.The relative mRNA expression increased significantly on TYRP1，MITF，DCT，NOS2 (P<0.05)，but there were no significant differences on MC1R，TYR (P>0.05).IFN-γ promoted the growth of melanocyte dendrites，which may contribute to melanin transport；IFN-γ induced melanogenesis，which may be related to NOS2 and MITF high expression in melanoma cells and the melanin may be produced through NO/cGMP/PKG-mediated signal pathway.

To monitor the body temperature of cows is important significance to estrus identification，pregnancy diagnosis and other physical activity，as well as to the diseases forecasting and monitoring.Since there is no expeditious means to measure the surface temperature，we used body temperature monitoring device developed by ourselves，to monitor the surface temperature of 5 Simmental cows in continuous 3 days，in the meanwhile we collected the rectal temperature data.For the first time，we revealed the diurnal variation rule of cow body surface temperature in 1 day through statistic analysis.Through the linear and nonlinear regression studies，we obtained the optimal correction formula：Z^-1=0.026 7+0.000 256lnX/X-2.047 4×10^-8×Y³(Z representing rectal temperature，X representing time point and Y representing surface temperature)，The average difference between the surface temperature after correction and the rectal temperature was 0.001 22 ℃，The biggest difference was not more than 0.3 ℃，which indicated that the surface temperature obtained from our self-development automatic detected devices could reflect the real body temperature of cows.It has great significance to the further research on the relationships between surface temperature and the cow physiology，and surface temperature and diseases.

This experiment was conducted to investigate the effects of different levels of lactoferrin(LF) on growth performance，intestinal protection and the body resistance to disease of Diansa weaning piglets.Two hundred and forty healthy Diansa weaning piglets，(28±2) days of age with(6.56±0.75) kg of average body weight，were allocated randomly to 5 groups，48 for each group，and each had 6 replicates with 8 piglets per replicate.They were fed with basal diet(control group)，basal diet + 125 mg•kg^-1 LF，basal diet + 250 mg•kg^-1 LF，basal diet + 500 mg•kg^-1 LF and antibiotics positive group(50 mg•kg^-1 terramycin)，respectively.The trial period lasted 42 d.At the end，24 piglets，with 4 piglets in each replicate per each group，were selected，and the test piglets were collected blood from precava，then serum were separated.The IgG and NO content，LDH，NOS and LSZ activity and TIBC of serum were detected.Six piglets，1 piglet in each replicate per each group，were slaughtered.The heart，liver and spleen were collected to detect Hepcidin and β defense 1(pBD-1)，β defense 2(pBD-2) and β defense 3(pBD-3) genes mRNA expression.The results showed that：1) The final weight，ADG and ADFI of 250 mg•kg^-1 LF group were significantly higher than those of control group and terramycin group（P<0.05），feed conversion and diarrhea rate were significantly lower than those of control group and terramycin group（P<0.05）.2) IgG levels，NO content of 250 mg•kg^-1 LF group were significantly higher than those of control group（P<0.05），TIBC of 250 mg•kg^-1 LF group were extremely significantly higher than those of control group and terramycin group（P<0.01）.V/C of duodenum，villus height and V/C of jejunum，villus height and V/C of ileum in 250 mg•kg^-1 LF group were extremely significantly higher than those in control group and terramycin group（P<0.01），crypt depth of ileum in 250 mg•kg^-1 LF group were extremely significantly lower than those in control group（P<0.01）.Results showed that diarrhea rate，IgG，NO，LSZ，crypt depth of duodenum，V/C of duodenum，villus height of jejunum，crypt depth of jejunum，V/C of jejunum increased with a quadratic curve by the LF adding levels(P<0.05)；3) Hepcidin mRNA expression in heart and liver of piglets in 250 mg•kg^-1 LF group were extremely significantly higher than those of control group and terramycin group（P<0.01）.Hepcidin mRNA expression in liver in 125 mg•kg^-1 LF group were significantly higher than those of control group and terramycin group（P<0.05）.The Hepcidin mRNA expression in liver was extremely significantly higher than that in the heart and spleen(P<0.01).The pBD-1 mRNA expression in the heart and spleen in 250 mg•kg^-1 LF group were extremely significantly higher than those of the other groups(P<0.01).The pBD-2 mRNA expression in liver was significantly or extremely significant higher than those in heart and the spleen(P<0.05，P<0.01).When the adding level was 250 mg•kg^-1，the pBD-3 mRNA expression in heart and spleen were extremely significantly higher than the other groups(P<0.01).Results suggest that adding LF in the diet can obviously improve the growth performance，serum immunological parameters and small intestinal morphology of Diansa weaning piglets，and enhance piglets disease resistance through inducing genes expression of Hepcidin，pBD-1，pBD-2，pBD-3 in heart，liver and spleen of Diansa weaning piglets.In this experimental conditions，based on quadratic curve results of growth performance，serum immunological and small intestinal morphology parameters，the adequate amount of LF in Diansa weaning piglets diet was 250-300 mg•kg^-1.

The objective of this study was to evaluate the effects of 2-methylbutyrate supplementation on development of rumen by determining rumen fermentation，enzyme activities and cellulolytic bacteria in pre- and post-weaning dairy calves.Thirty-two Holstein male calves with 15-day-old and body weight(46.45±0.37) kg were selected and allocated randomly to 4 groups.Calves in control were fed on milk during pre-weaning and on calf-concentrate and alfalfa hay during post-weaning.Calves in treatments were supplemented 2-methylbutyrate at levels of 3，6 and 9 g•d^-1，respectively.All calves were weaned at 45-day-old.At 30- and 90-day-old，4 calves in each group were selected at random and slaughtered at morning before feeding，and samples of ruminal liquid and rumen mucosa were collected and determined.The results showed that total VFA concentrations，ratio of acetate to propionate，activities of filter paper enzymes，carboxymethyl cellulose，cellobiose enzymes，xylanase，α-amylase，β-amylase and protease，populations of R.albus，R.flavefaciens，B.fibrisolvens and F.succinogenes in calves at 90-day-old were increased compared with calves at 30-day-old(P<0.05)，while ruminal pH and ammonia N concentration were decreased(P<0.05).When calves were supplemented 2-methylbutyrate at 6 and 9 g•d^-1，ruminal pH were decreased compared with control，ammonia N concentration were decreased compared with control and calves supplemented 2-methylbutyrate at 3 g•d^-1(P<0.05).When calves were supplemented 2-methylbutyrate at 9 g•d^-1，ratio of acetate to propionate was enhanced，activities of filter paper enzymes，carboxymethyl cellulose，cellobiose enzymes，xylanase，α-amylase and β-amylase were increased，populations of R.albus，R.flavefaciens，B.fibrisolvens and F.succinogenes were improved compared with control and calves supplemented 2-methylbutyrate at 3 g•d^-1，but activities of protease were not affected.The results indicated that the growth of cellulolytic bacteria and amylolytic bacteria were stimulated by supplemental 2-methylbutyrate，and then activities of cellulolytic and amylolytic enzymes were enhanced，which improved the rumen fermentation.The optimum daily dose of 2-methylbutyrate was at 6 g•d^-1.

The purpose of this study was to construct the recombinant adenovirus expressing two serotypes of vesicular stomatitis virus(VSV) G proteins with human adenovirus expression vector.The gene VSV-IN-G-NJ-G was amplified by fusion-PCR and cloned into the adenovirus shuttle vector pacAd-CMV K-NpA.The recombinant shuttle plasmid and adenovirus backbone plasmid linearized by digestion with PacⅠ，and then were transfacted into AAV-293 cells to obtain recombinant adenovirus rAd-VSV-IN-G-NJ-G.The recombinant adenovirus was stable in AAV-293 cells after serial passage to 20 generations.By indirect immunofluorescence and western blot detection，G proteins were expressed in recombinant adenovirus.The mice were inoculated with recombinant adenovirus for evaluating the humoral immune responses by indirect ELISA and neutralization experiment.The results showed that the recombinant adenovirus could induce VSV specific antibodies and the neutralizing antibody level of inoculated group was 1：32.The result of cellular immune showed that the recombinant adenovirus can cause a strong lymphocyte proliferation of the inoculated mice.These results suggested that the recombinant adenovirus can cause a certain degree of humoral and cellular immune responses in inoculated mice.The research laid foundations for further studies of VSV gene-engineering vaccines.

 Porcine Epidemic Diarrhea Virus(PEDV)，Transmissible Gastroenteritis Virus(TGEV) and Porcine Rotavirus(PoRV) are three common pathogenies causing piglets diarrhea.A novel cDNA microarray was necessary to be developed to simultaneous detection of three diarrheal viruses successfully.To make this possible，primers and probes were designed to amplification of target genes according to the conserved sequences of PEDV-S，PEDV-M，TGEV-S，TGEV-N，PoRV-VP7 and PoRV-NSP4.Mutiplex PCR was established for labeling target genes after designing the microarray matrix and finally evaluated the specificity，sensitivity and repeatability of microarray.The positive judgement standard of microarray was determined：SNR(Signal to Noise Ratio) ≥2.0 or median signal ≥1 500.The result showed that PRRSV，CSFV，JEV and PRV could not be detected by six probes and as few as 20 pg•μL^-1 of target plasminds were detected successfully.One microarray could be tested at least 7 times.This assay provided a novel and high throughput detecting technology to identification and detection of three diarrheal viruses.

The aim of this study was to delete asd gene of Salmonella gallinarum (SG) attenuated vaccine strain 97A and construct bivalent live vaccine based on host-vector balanced lethal system to express F protein of Newcastle disease virus(NDV).Two DNA fragments flanking asd gene as allele were amplificated by PCR and asd gene was replaced by chloramphenicol resistance(Cm^R) gene to construct the suicide plasmid pGMB151Δasd：：Cm^R.After conjugation，the recombiant plasmid was transformed into SG97A from E.coli χ7213，SG97AΔasd：：Cm^R was reversely screened out under the selectable pressure of chloramphenicol，DAP-dependent strain was named SG97AΔasd.Vector-host balanced lethal system was construct by pYA3334 containing asd gene being transferred into SG97AΔasd and to expressed F protein of NDV.SG97AΔasd was successfully constructed by allelic exchanges，its growth was DAP-dependent.Vector-host balanced lethal system was constructed by pYA3334 being transferred to SG97AΔasd and could express F protein of NDV and also induce specific antibodies in infected chickens.Vector-host balanced lethal system was constructed successfully based on attenuated Salmonella SG97AΔasd in this study and could express F gene of NDV as foreign gene to induce special antibodies in animal model，it laid a foundation for bivalent live vaccine based on attenuated Salmonella.

This study aimed to confirm the pathogens from purulent lymph goats infected naturally by Arcanobacterium pyogenes. The pathogens were isolated and cultured，identified by pathological method and 16S rRNA analysis.The result of bacterium analysis showed that the pathogen was Arcanobacterium pyogenes，the credibility was 88% identified by VITEK2 Compact automated microbial analysis system and homology was 99% by 16S rRNA identification.The strain was highly sensitive to β-lactams antibiotics and fluoroquinolones，but resistant to sulfa drugs.Pathological sections showed the sick goat appeared myocardial fibrosis，lung and renal tubular lumen with serous-fibrinous degeneration，inflammatory cell infiltration induced hepatitis，spleen was serous-purulent inflammation and front lymph nodes near shoulders appeared steatosis.Thus we can summary that the pathogen of the goat lymphadenitis was Arcanobacterium pyogenes.Histopathological changes were the main organs suppurative inflammation.

To express SAG2 gene of Toxoplasma gondii by prokaryotic expression system and to identify its antigenicity，truncated SAG2 without the highly hydrophobic signal peptide and C-terminus were amplified by PCR，and recombinant prokaryotic expression plasmid (pGEX-4T-3-SAG2t) with SAG2t protein gene was constructed.Then the recombinant plasmids were transferred into E.coli DH5α，and the bacteria were induced by IPTG.The expression and immune-reactivity of SAG2t protein were detected by SDS-PAGE and Western blotting respectively.The yield of purified rSAG2t was more than 4 mg per liter of culture medium.Western blot showed that rSAG2t can be strongly recognized by mice serum infected with T.gondii ME49 strain and mice serum immunized by rSAG2t .The result showed that the soluble expression of SAG2t protein with favorable complete antigenic activity was implemented by prokaryotic expression system.

To study the population genetic diversity and genetic structure of Heterakis gallinarum population in Sichuan，the complete sequence of mitochondrial 12S gene of 58 H.gallinarum isolates from 7 different geographical regions in Sichuan were amplified and analyzed the genetic diversity.The results showed that the complete lengths of mitochondrial 12S gene sequenced from 58 H.gallinarum isolates in this study were all 699 bp.These sequences contained 38 variable sites and were classified into 34 haplotypes(HS1-HS34).The values of haplotypes diversity(Hd) and nucleotide diversity(π) were 0.822 and 0.003 95，respectively.Genetic diversity with Fst and Nm furtherly showed that there were no significant genetic differentiation and frequent gene flow among the seven populations(Fst=0.007 87，Nm=31.52).An analysis of molecular variance(AMOVA) estimated that 99.21% of the genetic variation was partitioned within the population，and only 0.79% occurred among populations.NJ phylogenetic tree and haplotype network were both revealed that 34 haplotypes from 7 different geographical regions dispersed different geographical populations，and haplotypes did not cluster according to their geographical location.The results indicated that the H.gallinarum population in Sichuan had a low level of genetic diversity and no significant genetic differentiation，it have not formed significant geographical genetic structure.

 To determine the role of Th cell epitopes，which were screened from PCV2 ORF1，and ORF3，in improving the PCV2 ORF2 Cap P1 immunogenicity when they were added to the sequence，and the feasibility of synthetic expression of PCV2 ORF2 Cap P1 as a vaccine antigen，bioinformatics and molecular biology methods were employed to screen the peptide sequences.A pan-host of T helper cell epitope peptides and three Th cell epitopes peptide sequences of PCV2-specific were identified.These 4 Th cells peptides sequences were combined with a B-cell epitope peptide sequence screened from PCV2 ORF2 Cap1 gene，and then codon was optimized and inserted the restriction sites and stop codons.After analyzing the codon adaptation index and the frequency distribution，predicted it can achieve efficient expression，further chemical synthesis or peptides were carried out.The synthetic polypeptide antigen P1 was connected to the pET-30a expression vector，and transformed into BL21(DE3) pLysS competent cells，the expression in the engineered bacteria BL21(DE3) pLysS-pET-30a-P1 was constructed.The P1 antigen was able to be expressed after IPTG inducting.The expression level was identified by SDS-PAGE，and its biological activity was identified by Western Blot，followed by mice and pigs immunization with P1.Antibody levels in mice and the peripheral blood lymphocyte proliferation in pigs were tested.The results showed that the codon adaptation index of PCV2 P1 sequence was 0.89，it can achieve to high levels of expression，and the peptide fragment is about 28.29 kDa.MTT results showed that P1 peptide antigen had good immunogenicity.In immunized pigs，the expression of IFN-γ and IL-4 can increase obviously，and were significantly different to the control group(P<0.05).These results indicated that P1 could induce a strong humoral and celluar immune response，and it will provide a theoretical basis on PCV2 P1 peptide antigen as a vaccine antigen for further studies.

The aim of this study was to observe the histologic characteristics of adult yaks cryptorchidism and analyze the influence of plateau environment on the cryptorchidism reproductive microenvironment.Masson’s and Gomori’s staining，histochemistry and transmission electron methods was used to characterize the microstructure and ultrastructure of cryptorchidism，normal testis and unilateral normal testis in yak，and IPP(Image-Pro Plus) statistics method was used to quantitative statistics.The volume and weight of yak cryptorchidism were lesser than the normal testis and companied with depression of the lumen，the basement membrane was thickened，as well as the diameter of seminiferous tubule was significantly reduced(P<0.01).The spermatogenic cells desquamated and immature Sertoli cells were scattered in the tubule.Obviously，there were mitochondria degeneration and many lipofuscin granules with different sizes in the Sertoli cells.Besides，the area ratio of interstitial to lumen was significantly increased(P<0.01)，the numbers of Leydig cells were decreased，and the mitochondria in it was swelling.There were partly calcification was observed in parenchyma and not only the quantity of interstitial vascular was reduced but the vessel wall was thicken and shrunken.The seminiferous epithelium of unilateral normal testis were 3-4 layers with matured Sertoli cells，the primary spermatocytes and spermatozoa were rarely seen，and there was no difference with the normal testis in interstitial/lumen area ratios(P>0.05).The number of Leydig cells increased and the endoplasmic reticulum in it were represented as an loose and vesiculated network and the number of mitochondria were decreased.VEGF and its receptor(VEGFR2) immunoreactivities were abundantly distributed in the gonads of cryptorchidism，normal testis and unilateral normal testis，mainly associated with Sertoli cells，Leydig cells and spermatogenic cells，and weakly present throughout the vascular endothelial cells.Immunostaining analysis appeared that the relative expression of VEGF in cryptorchidism was significantly decreased than in normal testis and unilateral normal testis(P<0.05)，by contrast，the expression of VEGFR2 in cryptorchidism was stronger than the other two groups.But the expression of VEGF and VEGFR2 had no significant difference between the normal and unilateral normal testis(P>0.05).Taken together，in plateau environment，the cryptorchidism vascular of Yak was suffocated，the seminiferous function was seriously affected by the different degree of fibrosis and calcification in parenchyma and the dysplasia of the Sertoli cell；but in unilateral normal testis，companying the number of Leydig cells increased，the number of mitochondria decreased and also growth degree was decreased compared with the normal tissue.Our results suggest that the VEGF and VEGFR2 may serve as regulators to participate in the bilateral testicular spermatogenic suppression effect.

The aim of the present study was to detect the effects of boric acid on the morphology of African ostrich chick lung.Twenty-four healthy ostrich chicks were randomly divided into 6 groups：Group A(control group)，B，C，D，E，F，and supplemented with boric acid at the concentration of 0，40，80，160，320，and 640 mg•L^-1，respectively.The morphology changes of lung were observed by using light microscope and the HE staining technology，the collagen fiber changes were detected by MASSON staining，and TUNEL methods were used to label the apoptosis cells in ostrich lung.The results were as followings，compared with the control group，when the boric acid dose less than 80 mg•L^-1(group A，B，C)，the lung structure was well developed，cell morphology was intact，nucleus was stained darkly and the lung organizational structure was clearly，especially at 80 mg•L^-1.Adversely，when boric acid doses higher than 160 mg•L^-1(group D，E，F)，the high dose boric acid damaged the lung tissue structure，heterophilic granulocytes were increased，the interstitial was thickening with HE staining，especially at 640 mg•L^-1.MASSON staining showed that compared with the control group，collagen deposition was increased in high dose boric acid group(more then 320 mg•L^-1 boric acid).Compared with control group，the IOD of apoptosis cells was significantly decreased in the 40 mg•L^-1 boric acid group，whereas，the IOD of apoptosis cells was significantly evaluated in the 320 and 640 mg•L^-1 boric acid groups.Add 40 mg•L-1 boric acid can inhibit apoptosis in lung，more than 160 mg•L^-1 boric acid can promote inflammation，more than 320 mg•L^-1 boric acid can increase apoptosis and cause pulmonary fibrosis.

The aim of the present study was to study the anti-inflammatory effects and possible underlying mechanisms of Danqiao liquid in LPS-stimulated RAW264.7 cells.The cytotoxicity of Danqiao liquid was detected by MTT method.The NO kit assay was adopted to detect the effect of Danqiao liquid on NO release from LPS-induced RAW264.7 cells.ELISA was used to evaluate the production of TNF-α，IL-6 and PGE₂ in LPS-induced RAW264.7 cells.Real-time PCR was used to detect the transcription of COX-2，iNOS，TNF-α and IL-6 in LPS-induced RAW264.7 cells.The protein expression of nuclear NF-κB p65 was detected by Western blot.The result showed that the safe medication range of Danqiao liquid was less than 700 μg•mL^-1.Compared with the LPS model group，Danqiao liquid(100，300，600 μg•mL^-1) could reduce the secretion of NO，PGE₂，TNF-α，and IL-6 in cells induced by LPS，and significantly inhibit the mRNA transcription of iNOS，COX-2，TNF-α，IL-6 and the protein expression of NF-κB p65.In conclusion，this study preliminarily proves the protective effect of Danqiao liquid on LPS-induced RAW264.7 macrophages.Its action mechanism may be related to inhibit NF-κB signal pathway and the genes expressions and secretion of inflammatory mediators and cytokines.

This experiment was conducted to investigate the effects of lighting schedules on anti-stress ability，immune function and tibia of Yellow-feathered broilers with a completely randomized design.A total of 360 0-day-old male Lingnan Yellow-feathered broilers were randomly allocated to 4 treatments with each treatment having 6 replicates of 15 birds.The lighting schedules were continuous(23L：1D)，16L：8D，12L：3D：2L：3D：2L：2D and simulated natural lighting.The experiment lasted for 63 d.The total antioxidant capacity(P=0.08)，Newcastle disease antibody titer(P=0.07) and Bursa of fabricius index(P=0.06) tended to be higher in simulated natural lighting group than those in other groups on 63-day-old.Creatine kinase activity tended to be lower(P=0.09) in 3 short lighting groups than that in continuous lighting group on 42-day-old.Thymus index(P=0.06) on 21-day-old and ANAE⁺ on 21-day-old(P=0.09) and 42-day-old(P=0.06) in short intermit group tended to be higher than those in other groups.Lighting schedules didn’t affect other parameters(P＞0.05).These results show that natural or short intermit lighting condition are benefit to anti-stress ability and immune function of Yellow-feathered broilers and benefit to the health of broilers.

In order to determine the slaughter performance，physicochemical indicators of meat product and processing characteristics in donkey of different breeds，we selected Guanzhong donkey，Dezhou donkey，Guangling donkey，Qingyang donkey，Biyang donkey，Jiangyue donkey，Yunnan donkey on 7-month-old and tested their slaughter performance，moisture content，protein content，intermuscular fat content，shear force，rate of cooked meat of longissimus and rump.The results showed that Dezhou donkey on 7-month-old in live weight，carcass weight and net meat weight were significantly higher than other breeds（P＜0.05），as well as higher net meat percentage than Guanzhong donkey，Qingyang donkey and Yunnan donkey（P＜0.05）.Jiangyue donkey had higher slaughter rate than Guanzhong donkey，Qingyang donkey，Biyang donkey and Yunnan donkey（P＜0.05）.Guangling donkey and Guanzhong donkey had more than 2% intermuscular fat content，intermuscular fat content of longissimus were higher than Biyang donkey and Yunnan donkey（P＜0.05），while intermuscular fat contents of rump were higher than other breeds（P＜0.05）.The shear force of longissimus of Guangling donkey，Guanzhong donkey and Dezhou donkey were lower than Biyang donkey and Yunnan donkey（P＜0.05）.Dezhou donkey was significantly different（P＜0.05） from Guangling donkey，Guanzhong donkey and Qingyang donkey，showing a higher rate of cooked meat of longissimus，and was significantly different（P＜0.05） from Guangling donkey，Guanzhong donkey，Biyang donkey，Qingyang donkey and Jiangyue donkey，showing a higher rate of cooked meat of rump.Thus，Dezhou donkey have the excellent potential of meat production.Jiangyue donkey，Guangling donkey and Guanzhong donkey also have potential for meat development.Guangling donkey and Guanzhong donkey have good ability to store fat，which have the potential to become high-grade raw meat.

To explore the expression of endothelin 3 in different coat color of sheep skin and the effect of EDN3 on the sheep coat color，the experiment analyzed the mRNA and protein expression level of EDN3 in sheep skin of different coat colors in different coat color ship by real-time quantitative PCR，ELISA and immunohistochemistry.1）qRT-PCR showed that the relative expressive of EDN3 mRNA in white sheep skin was 5.540 6±0.030 7，while in black was 1.005 8±0.062 0.The gene expression quantity of EDN3 in white and black skin tissue of the same individuality was 4.341 9±0.087 7 and 1.150 1±0.005 3.2）ELISA results showed that the expression of EDN3 protein in white and black were 128.424 7±2.223 4 and 25.114 4±3.248 3.The protein expression of EDN3 in white and black skin of the same individuality were 93.945 3±7.562 2 and 28.606 0±9.295 9.3）The EDN3 was located in the matrix，outer root sheath and dermal papilla of hair follicular by immunohischemistry.Experiments showed that the EDN3 expressed normally in black and white sheep skin，and the expression quantity had significant differences.The research indicated that EDN3 was indispensable to maintain the presence of the melanocyte，and didn’t affect the formation of friesian sheep.