The authenticity of the so-called “chicken Leptin gene” was questioned since it was first reported in 1998.Actually it is not sure whether the Leptin gene exists in poultry genome until now.Although Leptin receptor gene had been widely confirmed in poultry，its physiological functions were still ambiguous due to the lack of endogenous Leptin.With the coming of the post genome age，it’s possible to look into the Leptin gene matter of avian species in wide scale and great depth.In 2014，3 laboratories independently reported the cloning and functional verification of Leptin gene in some wild birds.The findings greatly stimulated research interests in avian Leptin and Leptin receptor.We reviewed the latest advances in structure characteristics，origin and evolution，tissue distribution and “physiological functions” of Leptin and Leptin receptor gene in avian.

The aim of this study was to analyze the expression of the sheep BTG2/3 genes in the mid to late embryonic skeletal muscle and to investigate the relationship between MSTN and BTG2/3.Longissimus dorsi of 85 dpc(day postconception)，100 dpc，120 dpc and 135 dpc in Texel and Ujimqin sheep were collected as tested subjects.Real-time PCR was used to detect the expression of MSTN and BTG2/3 in the longissimus dorsi.Semimembranosus，longissimus dorsi，semitendinosus and quadriceps femoris of 100 dpc in the Texel and Ujimqin sheep were selected to detect the expression of BTG2/3.Lentiviral vectors pFU-GW-myostatin were constructed to infect sheep myoblasts at the proliferation and differentiation stage stably and the expression of BTG2/3 were detected at different stages.The results showed that a downward expression trend was emerged of MSTN in Texel fetuses，whereas the expression level in Ujimqin increased after decline at 100 dpc.The expression of BTG2 and BTG3 in Texel had downward trend after rising on 100 dpc，but BTG2 represented decrease-increase-decrease trend and BTG3 represented opposite trend to BTG2 in Ujimqin.Expression levels in different muscle on 100 dpc in the Texel and Ujimqin showed that，BTG2 gene expression was significantly higher(P<0.01) in Texel longissimus dorsi and semitendinosus than that in Ujimqin，and BTG3 gene expression was significantly lower(P<0.01) in Texel semimembranosus and longissimus dorsi than that in Ujimqin.Transfection and interference efficiency of lentiviral vector were very high.After transfected myoblast at proliferation and differentiation stage，respectively，expression levels of BTG2 and BTG3 also were decreased significantly(P<0.01) at both stages.The results indicate that BTG2 and BTG3 play an important role in sheep skeletal muscle growth from the mid to late embryonic stage and may be involved in myostatin regulatory pathway.Our study provide an insight into the molecular regulation mechanism of embryonic development in sheep.

This paper aimed to study the possible role of endogenous jaagsiekte sheep retrovirus（enJSRV） envelope protein in sheep placenta formation process.Chorionic trophoblast cells from Mongolian sheep were cultured in vitro.RT-PCR technique was used to amplify enJSRV-env gene as well as the full-length gene of its receptor Hyal2.The genes were then directionally cloned into the eukaryotic expression vector pEGFP-C1.The electric transfection conditions of the chorionic trophoblast cells were optimized and the electrical transfer efficiency was measured.The constructed eukaryotic expression plasmids were transfected into chorionic trophoblast cells by electroporation，and transient expressions of enJSRV envelope protein and its receptor Hyal2 were observed under a fluorescence microscope.The activity of cell fusion in chorionic trophoblast cells under high expression and RNA interference silencing of enJSRV-env gene was also observed.The results showed that eukaryotic expression plasmids were successfully constructed and named as pEGFP-C1/enJSRV-env and pEGFP-C1/Hyal2，respectively.The best transfection conditions were pulse voltage 150 V，pulse time 5.0 ms，electric shock number for 2 times with 50 ms interval.The number of multinucleated cells were significantly increased in the trophoblast cells which co-transfected pEGFP-C1/Hyal2 with the transfection of pEGFP-C1/enJSRV-env plasmid and pEGFP-C1/enJSRV-env.We observed 2.3 and 1.8 multinucleated cells in average per field.The enJSRV-env had a certain role in promoting the cell fusion of chorionic trophoblast cells.This study provides an experimental basis for further exploring the structure and function of enJSRV envelope protein and the fusion mechanism of chorionic trophoblast cells.

The objective of this study was to investigate the variant types of goat Agouti gene spliceosome in goat skin with different coat color.The sequences and mRNA of goat Agouti gene were obtained and analyzed from goat skin with different coat color by PCR and 5′RACE methods，respectively.The 27 793 and 27 858 bp of partial Agouti gene sequences were obtained from white and black goat skin，respectively.Four fragments，88，180，81 and 79 bp，in which the 180，81 and 79 bp were first detected in goat，were matched with the sequences of sheep mRNA and corresponded with the length of exon 1 spliceosome obtained by 5′RACE.The 5 types（It，It′，IA，2C and IE） of spliceosome were detected in goat skin with white coat color，and only 2 types（It and It′） were found in goat skin with black coat color.There was length variation within and among spliceosomes.The result indicate that the IA，2C and IE might be related with the formation of goat coat color.

The purpose of this experiment was to study the expression of miR-133 and its target gene SRF in skeletal muscle tissue and cells in goat，and the effects of miR-133 on target gene（SRF）.The qRT-PCR was used to detect different expression of miR-133 and its target gene in different tissues of newborn，7 months and 18 months old Anhui white goat and Boer goat（heart，liver，spleen，lung，kidney，small intestine，fat，longissimus dorsi） and different goat skeletal muscle satellite cells（F6，F9，F12）；qRT-PCR and the Western blot was used to detect the mRNA of target gene and protein expression level.The results of qRT-PCR showed that the expression level of miR-133 in myocardium and longissimus dorsi muscle was higher than that in other tissues，the expression level of the target gene SRF was different from miR-133 in goat；The expression level of miR-133 in skeletal muscle satellite cells increased with the cell generations；That regulation of miR-133 on SRF protein expression occurred at the post transcriptional level by qRT-PCR and Western blot.The results showed that miR-133 was expected to become a new marker of goat skeletal muscle growth and development；miR-133 was involved in the regulation of proliferation and differentiation of goat by inhibiting the expression of SRF protein in the post transcriptional level，thereby affecting the goat skeletal muscle development.

The prediction and analysis of bta-miR-320a target genes by bioinformatics were performed to explore its effect on the fat deposition in bovine.In this study，bioinformatics tools such as Promoter Scan，TargetScan，DAVID，Cytoscape and databases for example miRBase，Ensemble，NCBI，miRWalk，et al were used to perform the prediction of the transcription factor binding site and the analysis of its target genes，involved Gene ontology and KEGG pathway.As a result，miR-320(a) was very conservative among various species.There were many different transcription factor binding sites in the bta-miR-320a promoter area.Eighty-four target genes were obtained in total which involved in multiple biological processes such as negative regulation of cell differentiation，regulation of cell cycle and negative regulation of growth and participated in p53，cell cycle and MAPK signal pathways.We assumed that bta-miR-320a is regulated by different transcription factors，and it can also regulate the adipocyte differentiation and fat deposition in bovine by post transcriptional inhibiting of its target genes involved in p53，cell cycle and MAPK signal pathways.

This study was performed to screen genes associated with follicular development from ODF1（The largest follicle at onset of deviation）and ODF2(The second largest follicle at onset of deviation)in transcript levels of bovine.The largest and second largest follicle were collected from onset of deviation then separated granulosa cells from which extracted total RNA.High-throughput deep sequencing was carried out by sequencing platform of Illumina.According to the results of assessment analysis in significant enrichment analysis of Gene Ontology function and differential expression in gene screening.Further，screening differential expression gene by ODF1/ODF2 or ODF2/ODF1>2 as the candidate genes.By real-time fluorescent quantitative PCR(qRT-PCR) was performed to detect the gene expression pattern between DF and SF granulose cells of the first follicular wave during estrous cycle in bovine.Eight candidate genes related to follicular development were screened and 7 genes（BEX2，UBN1，SIK1，SPARCL1，LOC784256，LOC789231，LOC785462）from ODF1/ODF2 and one （SAFB2）from ODF2/ODF1，BEX2，UBN1，LOC784256，LOC789231 expressed in DF was extremely significantly higher than that in SF(P<0.01).But the expression levels of SAFB2 was extremely significantly higher in SF than those in DF(P<0.01) and SIK1，SPARCL1 were significantly greater in SF compared with DF(P<0.05).This indifferences trends was consistent between qRT-PCR test results of BEX2，UBN1，LOC784256，LOC789231，SAFB2 and high-throughput sequencing results of RPKM in ODF1 and ODF2.The qRT-PCR test results of SIK1，SPARCL1 and LOC785462 was opposite with the high-throughput sequencing of the gene RPKM in ODF1 and ODF2.

 To evaluate the quality of bovine blastocysts efficiently and accurately，a triple immunofluorescence staining method was established in bovine blastocyst.The trophectoderm(TE) cells and apoptotic cells were identified with labeling of CDX2 protein and caspase-3 protein，respectively.All of nuclei in blastocyst cells were stained with Hochest 33342.Triple staining was designed as 3 schemes.PI was used to stain nuclei of TE cells and apoptotic cells were detected by TUNEL method in double staining.The results showed that the triple immunofluorescence staining，with 3 schemes，could distinguish the TE and apoptotic cells accurately.Additionally，co-incubation of 2 first antibodys/second antibodys was the most optimal method to triple staining in terms of accuracy，efficiency and stain duration.Therefore，in same blastocyst，identification the TE and apoptotic cells by labeling CDX2 and caspase-3 protein respectively was available to evaluate the quality of bovine blastocysts.Also，there were great advantages in triple staining compared with double staining.

To explore the reason for the slow growth of Guizhou pony，sequences of exon 10 in GHR gene were cloned from the genome of Guizhou pony by polymerase chain reaction（PCR） and gene cloning methods.Distribution of 5 single nucleotide polymorphism（SNP） sites was determined by allele-specific PCR（AS-PCR） and single-strand conformation polymorphism（SSCP） methods.Taking Ili horse as control，the relationship between the genotypes of GHR and body sizes of Guizhou pony and Ili horse was calculated based on population genetics and bioinformatics algorithms.The results showed that a total of 10 haplotypes of exon 10 of GHR gene were sequenced from Guizhou pony，in which contained 10 SNP sites.Among them，5 sites，1028，1267，1471，1697 and 1732，changed the amino acid residues of GHR.Frequencies of 3 sites，1028，1267 and 1471，were tested by AS-PCR method.Compared with Ili horse，allele A at site 1028 was dominant and related with the lower rump height and higher cannon circumference in Guizhou pony.The percentage of allele E at 1471 site in Guizhou pony was higher than that in Ili horse and related to higher chest girth ratio of pony.Heterozygous genotypes of 1697 and 1732 sites were detected from the shortest male pony with 106 cm in height，6 years old.The V343A change resulted from 1028 site located in the ubiquitin-dependent endocytosis（UbE） motif and might affect the GHR internalization and degradation.Changes at S491G，Y566C and R578G sites resulted from SNP sites of 1471，1697 and 1732，might directly or indirectly impact the phosphorylation of GHR and influence the activation of signal transduction pathways.It suggested that the polymorphisms in exon 10 of GHR gene might have an effect on the growth and development of Guizhou pony.

The objective of this study was to analyze the reasons of different functions of NF-κB1/2 and to identify the evolutionary status of its domains.The sequence information of NF-κB1/2 was analyzed by following bioinformatic methods，including multiple sequence alignment phylogenetic tree construction and selective pressure prediction.The results showed that there was no intersection among vertebrate NF-κB1/2 during evolution，and RHD and IPT were highly conserved；ANK had diverged among both orthologous and paralogous structures；DD showed a high conservation with orthologous structures but a large difference with paralogous structures.Our results suggested that NF-κB1/2 were highly conservative among vertebrates，and the sequence variation of ANK might be an important factor causing the different functions of NF-κB1/2 in vertebrate，and DD might be one of the reasons for the divergence of NF-κB1/2 genes.

This study was conducted to investigate the response of ingestive behaviour of sheep to the restriction of time at pasture combined with indoor feeding.Thirty castrated male Ujumuqin lambs were randomly assigned to 5 equal groups：（i） no grazing（0H；control），（ii） 2 h grazing（2H），（iii） 4 h grazing（4H），（iv） 8 h grazing（8H），and（v） 12 h grazing（12H）.Behavioural activities of two lambs from each treatment were monitored daily by 2 observers during the first 10 days of July，August and September（investigating periods），respectively.Observations of each group were conducted for a 2 d period and daily observation time was 15 h from 06：00 to 21：00 throughout the whole investigating period.The results showed as follow：（1） Restrictions at pasture significantly affected animal behavioural pattern.With the time at grazing reduction，the proportion time spent grazing increased（P<0.001），while the time spent resting and walking，and walking distance decreased significantly（P<0.001）.2H and 4H treatments had no time to rest during grazing.（2） Chewing rate was significantly affected by treatments（P=0.003）.0H treatment present the lowest chewing rate and 4H，8H and 12H treatments present the highest，however，lambs in the 2H treatment had a lower chewing rate than the 4H，8H and 12H treatments.Biting rate tended to be affected by treatment（P=0.067），and lambs allocated to the 2H and 4H treatments had higher biting rates（+3.6 and +1.8 bites•min^-1） compared to the 12H treatment（51 bites•min^-1）.（3） Behavioural activities were significantly affected by the season.The time spent on grazing in July was more than（P<0.001） that in August and September.A significant（P<0.001） reduction in walking distance was observed in 8H and 12H treatments from July to September，but no difference was found between 2H and 4H treatments.Lambs decreased（P<0.001） their biting rates and increased（P<0.001） their chewing rates as the season progressed.Animals restricted access of 4 h per day at pasture has the strong ability to adapt their behavioural activity through increasing the proportion of grazing time，decreasing the proportion of resting time，and accelerating the chewing rate to improve grazing efficiency in compensation for the reduced access time to pasture.

The objective of this study was to measure the effects of dietary energy levels of pre-partum Holstein dairy cows on dry matter intake，milk yield，body weight and blood biochemical indicators at postpartum stage.21 healthy pre-partum Holstein cows in 2-4 parities with similar date of delivery and body weights were randomly divided into 3 groups（n=7）.The cows were fed with diets of different energy levels but similar crude protein content.The NE_L and crude protein levels in diets for different groups were，6.12 MJ•kg^-1 and 13.03% for group A，5.80 MJ•kg^-1 and 13.01% for group B，5.47 MJ•kg^-1 and 13.03% for group C，respectively.All the cows were fed with the same ration of ab libitum after delivery.The experiment lasted 49 d including 28 d pre-partum and 21 d postpartum.The results showed that：（1） In the beginning 1-7 d of lactation，DMI did not show obvious differences（P>0.05） among 3 groups.Compared with group A，DMI of group C was increased by 9.93%（P<0.01） in the following 8-14 d and increased by 8.87%（P<0.01） during 15-21 d.（2）The milk yield had the extremely significant difference among groups（P<0.01） at 7 d postpartum（groups C > B > A）.Compared with group A，milk yield of group C was increased by 4.71%（P<0.01） and 8.02%（P<0.05） at 14 d and 21 d postpartum.（3）The body weights among the 3 groups were not significantly different（P>0.05） during 1-14 d.At the final experimental period，the body weights were also significantly different（P<0.01） among 3 groups at this stage（groups C > B >A）.（4） The triglyceride（TG） was not vary obviously different（P>0.05） among groups.The highest Glucose（Glu） was observed in group A in the pre-natal stage，but group C at the postpartum stage.In comparison with group A，the nonesterified fatty acids（NEFA） levels of group B and C were reduced by 3.80%（P<0.01） and 6.30%（P<0.01） at 7 d after parturition，3.94%（P<0.01） and 6.01%（P<0.01） at 14 d，respectively.Decreasing the energy level of pre-partum diet not only improve the DMI，milk yield and blood glucose concentration，but also significantly reduce cows’ weight loss and NEFA concentrations in the blood at postpartum.It all suggests that low energy level of pre-partum diet could alleviate negative energy balance（NEB） of perinatal cows.

In this study，the effects of weaning stress on blood serum index and body growth at different ages foals were investigated to get the best weaned time.A total of 15 Thoroughbred mare-foal pairs were divided into 3 groups（each group，n=5） according to the weaning age.Foals and mares were separated by using the abrupt-weaned method，the changes of 10 blood serum biochemical indices and body weight of foals were measured on pre-weaning day，weaning day and post-weaning day.The results showed as follows：The concentrations of GLU in all foals on weaning day more significantly increased than those on pre-weaning day and post-weaning day（P<0.05）.On weaning day，the concentration of GLU for 8-month foals was the highest and that of 6-month foals was the least（P<0.05）.The daily weight gain of 6-month foals was the highest（0.78±0.05 kg） and that of 8-month foals was the least（0.13±0.52 kg） from weaning day to post-weaning day.Results of the present study indicated that 6-month foals had less weaning stress and rapid weight gain compared with 7-month and 8-month foals by using the abrupt-weaned method.

In order to assess the possibility of developing one-plasmid rescue systems of rabies virus(RABV)，this study was undertaken to generate a single-infectious cDNA clone by inserting Picornaviral IRES elements.Five full-length cDNA clones were constructed by inserting encephalomyocarditis virus(EMCV) IRES sequences，PV IRES sequences or TaV 2A-like sequences to upstream of the respective ORFs.BSR T7/5 cells in 6-well plates were directly transfected with 10 μg of either full-length clones or co-transfected with pTiT-N，pTiT-P or pTiT-L using CaPO₄ as the control.Two recombinant RABVs were successfully rescued from these constructs，yet the rescue of SAD-NeNePeL was significantly more efficient than the rescue of SAD-NeNtPeL.Passaged SAD-NeNtPeL was genetic stable whereas passaged SAD-NeNePeL was recombinant in the RNA level.Both viruses were strongly attenuated，only growing to titers reduced about 2-log steps in comparison to the wild-type virus，SAD L16 strain.The rescue of RABVs from these single infectious cDNA clones provide a powerful tool to investigate the molecular biology，neurotropism，and pathogenicity of RABV.

The study was aimed to prepare monoclonal antibody against BEV-2 and develop ELISA for the detection of antigen of bovine enterovirus type 2 (BEV-2) based on the monoclonal antibody.The purified BEV-2 VP1⁺ recombinant protein was used to immunize BALB/c mice，of which splenocytes were fused with SP2/0 cells by PEG4000．The selection of positive hybridomas was conducted.Two hybridomas (designated as 1G1 and 5G6) secreting monoclonal antibodies(mAb) against BEV-2 VP1⁺ protein were selected.Based on the monoclonal antibodies against BEV-2，a sandwich ELISA was established to detect BEV-2 through a series of optimization.The sensitivity of the sandwich ELISA was 100 TCID₅₀•0.1 mL^-1，and this method had no cross-reaction with BVDV，IBRV and BLV.The results showed that the sandwich ELISA was specific，sensitive and reproducible.This is the first domestic report of establishment of sandwich ELISA for the detection of BEV-2 based on the preparation of the monoclonal antibodies，and it provided a basis for development of kit for rapid diagnosing BEV-2.

Porcine reproductive and respiratory syndrome（PRRS） is an important viral infectious disease in swine industry worldwide.The causative agent is porcine reproductive and respiratory syndrome virus（PRRSV），which can inhibit host innate and adaptive immune response，cause immunosuppression，and lead to persistent infection.So it is difficult to control and eradicate PRRS.As a critical part of innate immune，NLRP3 inflammasome plays an extremely important role in host anti-viral immunity.Previous studies have shown that PRRSV activated the NLRP3 inflammasome.However，whether there are components of PRRSV which suppress NLRP3 inflammasome remains unknown.Therefore，in the present study，we first reconstructed NLRP3 inflammasome through co-transfecting expression plasmids encoding NLRP3，ASC，procaspase-1，and pro-IL-1β in HEK293T cells which are deficient in endogenous inflammasomes.Then，HEK293T cells and porcine alveolar macrophages（PAMs） were transfected with expression plasmid encoding nsp1α of PRRSV.The results indicated that nsp1α can apparently block NLRP3 inflammasome activation.The further mutation experiments demonstrated that deletion or mutation of Zinc-Finger（ZF） domain in N-terminal of nsp1α fails to activate the NLRP3 inflammasome.Our study first demonstrated that nsp1α had the ability to suppress the NLRP3 inflammasome-mediated IL-1β secretion and ZF domain was essential for nsp1α to inhibit the IL-1β induction.Our study reveals a new mechanism that PRRSV antagonize host innate immune responses and may provide some insights into the research on molecular targets of anti-PRRSV drugs and prevention of PRRS.

To investigate the regulative effect of expression level of three proteins gene (AP2α2，Elf4 and ISCU) derived from porcine kidney cell line (PK-15) on porcine circovirus type 2 (PCV2) propagation efficiency,the genes of three proteins were amplified by RT-PCR and then three eukaryotic expression vectors were constructed.The RNAi fragments of these proteins were designed and cloned into pGPU6-GFP vector to construct shRNA vectors respectively.The efficiency of interference was determined by real-time PCR and three shRNA expression plasmids with higher efficiency of interference were determined.The PK-15 cells transfected with eukaryotic expression vectors or shRNA were screened by antibiotics of G418.The propagation efficiency of strains PCV2 LG in the cell lines was investigated using a capture enzyme-linked immunosorbent assay (ELISA) and an immunoperoxidase monolayer assay (IPMA).The results showed that Elf4 and ISCU overexpression increased PCV2 replication.In contrast，overexpression of AP2α2 exhibited no significant influence on PCV2 replication.In contrast，AP2α2，ISCU and Elf4 down-regulation by shRNA resulted in decreased PCV2 propagation，which demonstrated that these three proteins play a regulative role in PCV2 replication.

To study the integration site of endogenous avian leukosis virus（ALV） elements in the genome of Hy-Line Variety Brown，a non-joined PCR amplicon（named HL-E1） was obtained using a B family DNA polymerase from hypothermophilic archaea Thermococcus sp.4557（TXW-PolB）.HL-E1 contains three essential genes including gag，pol，and env and the 3’-LTRs.And it lacks 93，1 960 and 805 base pairs in the gag，pol and env genes，respectively.A 4 663 bp functional mutant of ALV integrated in the Hy-Line Variety Brown genome.The conserved env（highly similar with endogenous avian leucosis virus） and LTR genes have identities of 96.8% and 97.4% to ev1 site.This indicates a newly identified endogenous ALV sequence integrated in an unknown site in the chicken genome.

The lgtF encodes a glucosyltransferase responsible for adding a glucose to get heptose I in the synthesis of Lipooligosaccharide （LOS）.The lgtF gene is an important virulence-associated gene in some Gram-negative bacteria.However，the function of lgtF gene in Haemophilus parasuis was still unknown.We constructed an ΔlgtF mutant （ΔlgtF） of H.parasuis SC096 using a natural transformation system.The LOS of H.parasuis SC096 strain，ΔlgtF mutant was extracted by the hot phenol-water method.The LOS was addressed by SDS-PAGE and sliver stain.The growth curve，Serum resistance activity，adhesion and invasion ability in porcine kidney epithelial cells （PK-15） and porcine umbilical vein endothelial cells （PUVEC） of wild type SC096 and ΔlgtF mutant were measured.Compared to the wild-type SC096 strain，the ΔlgtF mutant exhibited obvious truncation of LOS structure and slight growth defect.Furthermore，the mutant had a greater sensitivity to the bactericidal action of porcine and rabbit serum （P<0.05），and displayed a significantly reduced ability to adhere to and invade PK-15 and PUVEC （P<0.05）.The above findings suggested that the lgtF gene is a virulence-associated gene of the H.parasuis SC096 strain.

The objective of this study was to screen the main immunogenic antigen of Toxoplasma gondii excreted-secreted antigen（ESA）.2-DE combined with Western blotting were used to analysis the components of T.gondii ESA.Eighteen positive protein points were detected in this study.Among them，6 protein spots were selected to identify by matrix-assisted laser desorption/lionization time of flight mass spectrometry（MALDI-TOF/TOF），belonging to microneme protein 1，microneme protein 4，cyst matrix protein and 14-3-3 protein.This study provides a new reference for looking for new candidate molecules for prevention and diagnosis of toxoplasmosis.

The purpose of this study was to prepare tosufloxacin nanoemulsion and to analyse its physical and chemical properties and acute toxicity.The oil phase，surfactant and solubilizer were selected according to the drug loading and stability of the nanoemulsion，the optimum formula was designed using pseudo-ternary phase diagrm，prepared tosufloxacin nanoemulsion.The configuration and particle size of tosufloxacin nanoemulsion were measured by TEM and LPSA.The drug content of tosufloxacin nanoemulsion was determined through High Performance Liquid Chromatography（HPLC）.The stability and shelf life were tested by stress testing，accelerated testing and long-term testing.Its acute toxicity was evaluated by gavage testing in mice.The results showed that the optimum formula of tosufloxacin nanoemulsion was composed of tosufloxacin（1.36%），origanum oil（3.24%），lactic acid（2.60%），RH-40（25.98%），and distilled water（66.82%）.The droplet shape of this nanoemulsion was spherical，with the average diameter of 16.16 nm，and its stability was good，the shelf life was 24 months.As the concentrations of tosufloxacin ranged from 3 to 100 μg•mL^-1，the concentrations and peak areas showed good linear relationship；the average recovery rate and relative standard deviation of tosufloxacin nanoemulsion were 99.33%±1.24% and 1.25%，respectively；the precision of this nanoemulsion was well.The LD₅₀ of tosufloxacin nanoemulsion was greater than 10 000 mg•kg^-1，the MTD of this nanoemulsion was 39.72 g•kg^-1.The tosufloxacin nanoemulsion was prepared successfully，and it will provide a theoretical basis for the further development of nanoscale antimicrobial agents.

This experiment was conducted to study the pharmacokinetics parameters of marbofloxacin after single intramuscular administration in infancy yellow cattle and the Post-antibiotic effects（PAE） of marbofloxacin against Pasteurella multocida in serum.Marbofloxacin was administered to 5 healthy infancy yellow cattle at a dose of 2 mg•kg^-1 of body weight，the concentrations of marbofloxacin in serum were determined by high-performance liquid chromatography and marbofloxacin concentration-time data in serum were analyzed using non-compartmental analysis of WinNolin6.2 programme.Furthermore，PAE of marbofloxacin against Pasteurella multocida were tested in serum collected at predetermined time point.The main pharmacokinetic parameters intramuscularly were as follows：C_max=2.04 μg•mL^-1，T_max=0.83 h，AUC=15.03 h•μg•mL^-1，AUMC =138.67 h•h•μg•mL^-1，MRT =9.17 h，t_1/2β=9.00 h，V_d=1.80 L•kg^-1，CL/F= 0.14 L•（h•kg）^-1.The PAE of marbofloxacin against P.multocida at the concentration levels of 2MIC，4MIC，8MIC in serum were 1.90，2.65，3.50 h respectively.It was noted that marbofloxacin was relatively rapidly absorbed，widely distributed and slowly eliminated after intramuscular administration.The PAE of marbofloxacin against P.multocida in serum was obviously prolonged and dependent on concentration.

In order to evaluate clinical efficacy of Chinese medicine on endometritis in cows，reports of randomized controlled trials（RCT） were collected from databases of CNKI，Wan Fang，CBM，Pub Med，Web of Science and Science Direct.According to the Cochrane system instruction，Meta-analysis was performed by using RevMan 5.3 software.A total of 24 RCTs，2 782 cases（including 429 cows treated with Chinese medicine，381 cows treated with terramycin，penicillin，streptomycin combined with penicillin and ciprofloxacin） were involved.Meta-analysis results showed that：the total effective rate of Chinese medicine group was better than that of control group［OR=2.40，95%CI（1.74，3.32），P<0.01］，but equal to the streptomycin combined with penicillin.The cure rate of Chinese medicine group was better than that of control group［OR=2.40，95%CI（1.74，3.32），P<0.01］，but equal to the terramycin.The pregnant rate of Chinese medicine group was better than that of control group.Publication bias analysis showed that the included literature had some certain publication bias.Based on the existed evidence，the efficacy of Chinese medicine on endometritis in Cows is better than that of antibiotics，and the Chinese medicine could improve the pregnant rate of dairy cows.

The present study was designed to examine the pattern and cellular localization of 11β-hydroxysteroid dehydrogenase（11β-HSD） enzymes and glucocorticoids receptor（GR） gene expression in the murine cervix during peripartal period，and to gain insight into the role of glucocorticoid（GC） and GR，11β-HSD-1 and 2 in the cervical ripening of peripartal mice.Mice in five groups were from 48 h before birth（-48 h） and-24 h，at term labor（0 h），12 h after birth（+12 h） and +48 h.The dynamic change of the serum corticosterone level was determined by ELISA and immunolocalization of GR，11βHSD-1 and 2 which influence glucocorticoid’s activity in cervix was assayed by immunohistochemistry.The results suggested that the levels of serum corticosterone were significantly increased before the onset of labor，reached the maximum during delivery（P＜0.01）.In addition，the 11βHSD-1，11βHSD-2 and GR protein were localized to the nuclei of cervical luminal epithelia，stromal cells，vascular and cervical smooth muscles.The quotient between mean optical density of 11βHSD-1 and 2 is highest at parturition.Furthermore，the immunoreactivity of GR also reached the peak at parturition（P＜0.01）.These results revealed that 11βHSD-1-regenerated glucocorticoids may act via its receptor to play an important role in the final cervical ripening.

To reveal the genetic basis and molecular mechanism of liver weight and color in chicken，5 traits，including liver weight，percentage of liver weight to carcass weight and liver color（lightness L*，redness a* and yellowness b*） were measured from 300 93-day-old F2 generation chickens from Chinese Academy of Agricultural Sciences（CAAS） chicken F2 resource population.Genome-wide linkage analysis for the liver weight and color traits were preformed on the basis of the CAAS chicken F2 resource population and genetic linkage map.The results of the present study showed that 11 quantitative trait loci（QTLs） for 5 traits were found，which explained 3.39%-6.43% of the phenotypic variation.A total of 3 QTLs on 3 chicken（Gallus gallus） chromosomes，at 22 cM on chicken chromosome 5（GGA5），56 cM on GGA12 and 1 cM on GGA15，for liver weight and the percentage of liver weight to carcass weight were identified.A total of 8 QTLs for liver color were found on 6 chicken chromosomes.Two interesting QTL regions for liver color were found，one region spanned 14 cM（16-30 cM） on GGA15 containing 2 QTLs for the chicken liver L* and b*，and the other region spanned 6 cM（81-87 cM） on GGA1 containing 2 QTLs for the chicken liver a* and b*.The present study provides the basis for further fine mapping and molecular marker-assisted selection for liver weight and color traits in chicken.

To investigate the function of endothelin 3（Edn3） in sheep skins and the regulation of hair color formation，the differential expression of Edn3 in skins of black versus white sheep was examined.Quantitative real-time PCR（qRT-PCR），Western blotting and immunohistochemistry methods were performed.The results showed that the quantity expression of Edn3 mRNA in black sheep was 2.89 times than in white sheep（P<0.01）；the average protein expression of Edn3 in black sheep skin was higher than in white sheep skin on the optical density（P<0.01）.The results of immunohistochemistry revealed that Edn3 was localized in the hair dermal papilla，lower of hair bulb and outer root sheath in hair follicles of sheep with white hair color，but was localized in lower of hair bulb and outer root sheath in hair follicles of sheep with black hair color.The differential expression and localization of Edn3 in skin of white versus black sheep suggests a potential role in hair color regulation.