Exosome exist in various kinds of body fluid，including milk，serum，urea，etc.It was reported that proteins，mRNAs，microRNAs，lipids，etc in exosome are transferred into receptor cells to play some physiological roles.In recent years，although there are a number of researchs on miRNA in animals，relatively a few studies illustrated miRNAs in milk.In this review，by collecting and collating some reports published，we confirmed the presence of miRNAs in the milk and they enriched in exosome，then compared the similarities and differences of miRNAs with high expression in different animal milk.We also concentrated on miRNAs highly expressed in exosome，and then drew some general conclusions.Moreover，we analyzed the other miRNAs binding forms in milk，then discussed the conservation of miRNAs in different animal milk and possible physiological function of miRNAs.

Long noncoding RNA (lncRNA) is one kind of RNA which is longer than 200 nt and does not show the potential of coding protein.It has been known to play vital roles in eukaryotic gene regulation.Transcriptional capacities of sex chromosomes and some imprinted genes is regulated in cis by lncRNAs which vary among species.The researches of lncRNAs regulation in reproduction mainly focused on screening lncRNAs candidates，identifying their target genes and the biological functions，clarifying the molecular mechanisms of those lncRNAs regulation.The present review concerned with the role of lncRNAs in processes vital to reprodution，such as gametogenesis，placentation and pregnancy，sex hormone responses，early embryonic development，sex determination and gonad development stages during reproduction process.These results can give scientists new insights into the regulation mechanism of animal reproduction.

Mycotoxins are secondary metabolites of some fungi that distributed widely in nature，especially in feedstuffs and animal products，with great harm to animals and humans.This review introduces several mycotoxins that commonly presents in feedstuffs and animal products and their risks to animals and humans，summarizes the research progress on the rapid detection technology of mycotoxins，analyzes the main influence factors of antibody preparation against mycotoxins，and prospects the further development of the rapid detection technology of mycotoxins as well.

Extracellular traps (ETs)，composed of DNA and granular proteins，act as a sort of new innate immune phagocytosis-independent extracellular defense mechanism which was discovered in recent years.As the third killing mechanisms of innate immune cells after phagocytosis and autophagocytosis，ETs can ensnare varied kinds of invading microbes inducing bacteria，virus，fungi，and protozoa to prevent local diffusion and kill them in vitro，and also in vivo.In this review，we will briefly describe the recent progress of the concept and molecular mechanism of ETs，and its role in host defense.This review has substantial significance on the new understanding of defense strategy in innate immune cells.

In order to study the role of RB1 gene on chicken preadipocyte differentiation and proliferation.CRISPR/Cas9 system was established and conducted to knockout the RB1 gene in the chicken preadipocyte.The proliferation and differentiation of chicken preadipocytes and the expression of related marker genes were detected.Results showed that the CRISPR/Cas9 system could effectively mediate RB1 knockout and cause truncated translation in chicken at the rate of 10%.Oil Red O and CCK-8 were used to evaluate the ability of differentiation and proliferation in chicken preadipocyte respectively.Knockout of RB1 in chicken preadipocyte resulted in enhanced proliferation capability and decreased differentiation capability in chicken preadipocyte.RB1 knockout decreased the transcriptional expression of adipocyte differentiation associated genes such as G0S2，FAS and A-FABP.Especially，the expression of G0S2 was significantly decreased at all stages of preadipocyte differentiation.Further， RB1 knockout increased the expression of cell proliferation associated genes such as CyclinD1，E2F1，Ki67 and PCNA, which was corresponded to the enhanced proliferation capability in the RB1 knockout cells.In summary，RB1 may be a key regulator on differentiation and proliferation of chicken preadipocyte.

The objective of the present study was to obtain recombinant chicken Visfatin protein with biological activity.The total RNA was extracted from livers of AA broilers.The Visfatin cDNA was first amplified by qRT-PCR and cloned into pGEM-T vector.The cDNA was then cut out from pGEM-T vector by digestion with endonuclease NcoⅠ and XhoⅠand inserted into corresponding cloning sites of the plasmid pET-30a.The positive recombinant plasmid designated as pET-30a-Visfatin was transformed into E.coli BL21 (DE3) competent cells.The cells containing pET-30a-Visfatin were identified.The conditions of IPTG induction were optimized by examining the expression level of recombinant protein using SDS-PAGE analysis.The recombinant protein was purified by Nickel ion affinity chromatography.The identification of the purified recombinant protein was checked by Western blot and the bioactivity was examined by looking into differentiation status of 3T3-L1 cells.A fragment which was the same size to prediction was observed when the recombinant plasmid pET-30a-Visfatin was digested with endonuclease NcoⅠ and XhoⅠ.It indicated that the recombinant expression vector was constructed successfully.The SDS-PAGE analysis revealed that the optimum conditions of induction were 0.4 mmol•L^-1 IPTG，pH 8.0，at 30 ℃ for 10-12 h.The molecular weight of the recombinant protein was about 60 ku，which was the same to prediction.The Western blot analysis indicated that the recombinant protein could be recognized by anti-His antibody.Additionally，the Oil Red O stain assay showed that there were more lipid droplets formation in 3T3-L1 cells induced by Visfatin than that in the controls.Meanwhile，the results of qRT-PCR revealed that the expression levels of genes involved in cell differentiation such as PPARγ， aP2，FAS and C/EBPα increased significantly (P<0.05) when 3T3-L1 cells induced by Visfatin.This study successfully established a standardized expression program for recombinant chicken Visfatin protein production，and obtained the purified recombinant protein with biological activity，which laid a foundation for further research in the field of poultry.

The aim of this study was to accurately evaluate comprehensive carcass and meat quality characteristics of Duroc×(Landrace×Yorkshire) (DLY) three-way crossed pigs and (Pietrain×Duroc)×(Landrace×Yorkshire) (PDLY) four-way crossed pigs，and to estimate the effects of breed and sex on these traits.In this study，a total of 304 DLY pigs and 213 PDLY pigs with the average age of approximately 180 days were slaughtered at the same abattoir，and then phenotype for different carcass and meat quality traits were determined，including backfat thickness，loin eye area，lean meat percentage，pH，electrical conductivity，drip loss，etc.The data showed that the lean meat percentage of DLY and PDLY populations were higher than 55% and the slaughter rate within the 2 populations were above 84%.The coefficients of variation in lean meat percentage and slaughter rate were less than 6.2%，while the coefficients of variation in electrical conductivity (24 h) and drip loss in both 2 populations were higher than 45%，implying that the drip loss could be improved to some extent by within-bred selection.Comparison between 2 populations，the average of loin eye area and loin eye depth of PDLY population were better than that of DLY population (P<0.01)，while DLY had higher slaughter rate than that of PDLY population (P<0.001).Compared to sows，castrated boars had higher live weight (P<0.01)，while sows had higher lean meat percentage (P<0.001) and lower backfat thickness (P<0.001).In addition，the lean meat percentage of the 2 populations were negatively correlated with live weight and backfat thickness (|r|>0.45，P<0.001).However，loin eye area was positively correlated with loin eye depth (r>0.70，P<0.001).pH24 h was significantly negatively correlated with drip loss and electrical conductivity in the 2 populations （|r|>0.22，P<0.05）.The results of this study indicate that PDLY pigs have more excellent production performance than DLY pigs，while DLY pigs have much better pork quality characteristics than PDLY pigs.Sows have better meat quality than castrated boars in both breeds.In addition，improving pH would reduce drip loss，thus increasing the sensory and freshness of pork.The results of this study contributes to the further understanding of carcass and meat quality characteristics of DLY and PDLY populations，and provides an important reference for future genetic improvement of commercial pigs.

The aim of this study was to construct an insulin-induced gene-1 (INSIG1) recombinant adenovirus overexpression vector of Xinong Saanen dairy goat，and determine its effect on lipid synthesis in goat mammary epithelial cells (GMEC)，to lay the foundation for its functional study in regulating lipid synthesis in goat mammary gland.The primers were designed according to the INSIG1 sequence in GenBank (accession number：JQ665439)，then its coding sequences (CDS) was cloned by PCR.The gene fragments were inserted into shuttle vector pAdTrack-CMV to obtain pAdTrack-CMV-INSIG1 vector.After being linearized by PmeⅠ，pAdTrack-CMV-INSIG1 vector was transformed into Escherichia coli BJ5183 competent cells to obtain the recombinant adenovirus overexpression vector pAdEasy-INSIG1 by homologous recombination.Next，the pAdEasy-INSIG1 vector，linearized by Pac Ⅰ，was transfected into 293A cells for viral packaging and amplification.LaSRT was used for titer assay.Finally，after infecting the goat mammary gland epithelial cells(GMEC) with Ad-INSIG1 recombinant adenovirus，qRT-PCR was used to measure the mRNA expression of INSIG1 and genes related to lipid synthesis，and the GPO-Trinder enzyme reaction was used to measure the cellular triacylglycerol (TAG) content.The result showed that INSIG1 overexpression recombinant adenovirus vector was successfully constructed，and the recombinant adenovirus Ad-INSIG1 with a high titer of 2×108 U•mL^-1 was obtained.Compared with Ad-GFP infected group，the mRNA expression of INSIG1 increased by about 500-fold after GMEC incubated with Ad-INSIG1 for 48 h.No obvious changes were observed on the mRNA expression of sterol regulatory element binding protein 1 (SREBP1) and SREBP cleavage-activating protein (SCAP)，however，there was a significant decrease in the expression genes related to fatty acid de novo synthesis and desaturasion：acetyl-CoA carboxylas α (ACCα)，fatty acid synthase (FASN) and stearoyl-CoA desaturase 1 (SCD1) (P<0.05).Among the 3 key genes involved in TAG synthesis，the transcript abundance of glycerol-3-phosphate acyltransferase (GPAM) and diacylgycerol acyltransferase 2 (DGAT2) were significantly decreased (P<0.05)，and 1-acyl-sn-glycerol-3-phosphate acyltransferase 6 (AGPAT6) had no obvious change.Meanwhile，the mRNA expression of adipose triacylglycerol lipase (ATGL)，an enzyme catalyzes the initial step of TAG hydrolysis，also decreased significantly (P<0.05).No significant difference was found in the cellular TAG content between 2 groups.In conclusion，INSIG1 can inhibit the expression of genes related to lipid synthesis in GMEC，and may have a regulatory function on lipid synthesis in goat mammary gland.

This experiment was conducted to estimate the variance component and genetic parameters including heritability and genetic correlation for economic important traits of 2 939 Simmental beef cattle that were born between 2008 and 2013.These parameters were estimated using derivative-free restricted maximum likelihood method.We found that the estimation of heritability for birth weight，weaning weight，marketing weight，carcass weight，dressing percentage and meat percentage were 0.48，0.44，0.43，0.38，0.31 and 0.39，respectively.Furthermore，the genetic correlation between birth weight and weaning weight，marketing weight and carcass weight，dressing percentage and meat percentage were 0.57，0.94 and 0.89，respectively.The heritabilities of these development traits and carcass traits in Chinese Simmental beef cattle population belong to medium and high heritability，and these traits had high genetic correlation values.Our study conducted the system analysis on genetic parameters of important economic traits in Chinese Simmental beef cattle.And the results lay the foundation for further breeding scheme and genetic evaluation.

 The aim of the study is to investigate the impact of the variations of GPR54 promoter on the expression levels of GPR54 gene in Holstein (HT)，Wandong cattle (WD) and Jianghuai buffalo (JH)，therefore further unveil the association between the polymorphisms of GPR54 gene promoter and the sexual precocity.Total RNAs in leukocytes of HT，WD and JH were extracted and the polymorphism of the promoter region were detected by PCR and sequencing.Furthermore，the efficiency of the promoters were determined via RT-PCR and luciferase expression from the fusion reporter gene.Various SNPs were detected in the promoter regions of GPR54 genes in HT，WD and JH with significant difference.Moreover，it was shown that the promoter efficiency of HT was 4.44 times and 1.99 times higher compared to JH buffalo and WD respectively.Meanwhile the promoter efficiency of WD was 2.24 times higher than JH.Notably，the RT-PCR results showed that GPR54 gene expression levels in youngsters were dramatically higher than that of the adults especially in HT than in WD and JH.JH showed the weakest expression level of GPR54 while the cows exhibited the strongest GPR54 expression level compared to ox (P<0.01).These results indicated that the polymorphisms of GPR54 promoter and the bovine sexual precocity were evidently correlated.

For elaborate the general physiological molecular features of the estrus ovary of yaks and particularity of yak reproduction.RNA-seq technology was applied to analyze transcriptome data comparatively between the yak and plain cattle estrus ovaries in this study.Compared with cattle，genome-wide divergent expression patterns during the ovary transcriptome data revealed that 1 307 genes were significantly and differentially expressed，in which 661 genes were upregulated and 646 genes were downregulated.Functional analysis showed that the differentially expressed genes were involved in various GO categories and KEGG pathways.GO annotations indicated that the genes were closely related to cell adhesion，hormonal biological processes，whereas the calcium ion binding，cation transmembrane transport molecular events were significantly active.KEGG pathway analysis showed that the complement and coagulation cascade pathway was the most enriched，followed by the cytochrome P450 related pathways.Moreover，several novel pathways，such as circadian rhythm，were significantly enriched despite having no evident associations with the reproductive function.This study is the first to compare the ovary transcriptome data between yak and cattle and to identify and analyze the related differential genes.Our findings provide a theory support for further investigation of the molecular mechanism of yak ovary and offer new insights to understand comprehensively the specificity of yak reproduction.

The aim of the present study was to assess the changes of miRNA expression levels and serum biochemical indexes of cattle in heat stress.Twenty Red Angus bulls with similar body condition and consistent feeding and management selected from Chongqing Improved Beef Cattle Farm were used in the experiment.Blood samples were collected from each bull in heat stress and non heat-stress to analyze miRNA expression levels and serum biochemical indexes.The results showed that the serum concentrations of Cl-(P<0.05) and malondialdehyde (MDA) (P<0.01) and the activities of lactate dehydrogenase (LDH) (P<0.05) were significantly increased in heat stress in comparison with control.Whereas，the activities of total antioxidation (T-AOC) (P<0.05)，glutathione peroxidase (GSH-Px) (P<0.05) and superoxide dismutase (SOD) (P<0.01) as well as the contents of IgA (P<0.05)，IgG (P<0.05) and interleukin-2 (IL-2) (P<0.01) were remarkably decreased.Meanwhile，the expression levels of miR-181a (P<0.05) and miR-486 (P<0.05) were significantly downregulated and positively correlated with the contents of IgA (P<0.05)，IL-2 (P<0.05) and IgG (P<0.01)，respectively，but that of miR-1246 were significantly upregulated (P<0.01) and positively correlated with serum contents of MDA (P<0.05).In conclusion，the current results suggested that miR-1246，miR-181a，miR-486 might be involved in regulating the immune response and antioxidation process of cattle in heat stress，and could be as a molecular indicator of anti-heat stress.

The objective of this study was to find novel imprinted long noncoding RNAs (lncRNAs) in cattle Dlk1-Dio3 domain.Transcripts mapping to this region between Meg8 and Meg9 were under a systematic search and 2 novel lncRNAs were identified，which were named as Linc24062 and Linc24063 according to annotated bibliography of genecode.The expression patterns of Linc24062 and Linc24063 in adult cattle tissues were characterized using reverse transcription polymerase chain reaction (RT-PCR).The results indicated that Linc24062 and Linc24063 were expressed in all 8 tested tissues，including heart，liver，spleen，lung，kidney，subcutaneous fat，skeletal muscle and brain.The imprinting status of Linc24062 and Linc24063 was assessed based on a polymorphism-based sequencing approach.The results showed that Linc24062 and Linc24063 exhibited monoallelic expression in all examined cattle tissues by comparing sequence chromatograms between genomic DNA and cDNA levels at single nucleotide polymorphism (SNP) site，suggesting that Linc24062 and Linc24063 were imprinted in cattle.

The aim of the present study was to screen differential expression proteins associated with inflammatory model of bovine mammary epithelial cells (BMECs) induced by lipopolysaccharide (LPS) using isobaric tags for relative and absolute quantification (iTRAQ) combined with 2-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS).Normal and LPS-treated bovine mammary epithelial cells were analyzed using iTRAQ combined with 2D-LC-MS/MS.Bioinformatics analyses of differentially expressed proteins were performed by means of Gene Ontology and PPI (Protein protein interaction）analysis.According to iTRAQ results，a total of 2 487 proteins were identified.Compared with normal BMECs，163 proteins were significantly up-regulated while 178 proteins were significantly down-regulated in response to LPS treatment.The bioinformatics analyses highlighted the effects of LPS treatment on milk proteins and fat synthesis such as αs1-casein，κ-casein and fatty acid synthase.Down-regulation of αs1-casein (CSN1S1) in BMECs was confirmed by Western blotting；α_s1-casein，κ-casein (CSN3) and fatty acid synthase (FASN) in culture medium were confirmed by ELISA.The results showed that LPS treatment significantly decreased the expression of αs1-casein in BMECs (P<0.05)，and also decreased the concentration of α_s1-caseins，κ-casein and fatty acid synthase in culture medium (P<0.05).In conclusion，LPS treatment could affect the synthesis of protein and fat in BMECs.

 In order to provide the foundation for breeding large transgenic animals in futher，in this study，the genetic traits of mammary gland-specific PRL transgenic mice produced by pronuclear microinjection，were analyzed from the genetic integrity，the copy number and the inserting site of exogenous gene.The transgenic mice deleted partially exogenous gene were screened by genomic DNA PCR method，and were verified by Southern blot to detect the genetic stability of exogenous gene.The copy number of exogenous gene in transgenic mice was detected by absolute real-time fluorescence quantitative PCR method.The inserting site of exogenous gene in transgenic mice was analyzed by Genome walking method.The results showed that the losing rate of exogenous gene fragment was 1.91%.The green fluorescence signal was observed under the long-wave UV lamp in the transgenic mice integrated higher copies of exogenous gene，while not observed in the transgenic mice integrated 2 copies of exogenous genes.The exogenous gene expressed normally in the 2 025-2 026 bp site of NW_001073945.1 gene and the 10 760 020-10 760 021 bp site of NW_001030574.1 gene.The result indicated that the exogenous gene could be inherited stablely in the transgenic mice produced by pronuclear microinjection，the expression level was positively correlated with the copy number of exogenous gene in certain range，and it was beneficial for the exogenous gene expression while there was no other genes nearby the inserting site.

The objective of this study was to investigate the effects of protein with different sources in milk replacer on nutrient digestion and rumen fermentations of suckling calves.Fifty Chinese Holstein bull calves with similar body weight and day age were randomly assigned to 5 groups (1 control group and 4 treatment groups) with 10 calves for each group.The protein source of control group (MP) was from milk protein.Calves in 4 treatment groups were fed with milk-replacer whose protein contained 70% vegetable protein and 30% milk protein of crude protein.Vegetable proteins were respectively soybean protein concentrate (SP)，hydrolyzed wheat protein (WP)，peanut protein concentrate (PP) and rice protein isolate (RP).Digestibility trials were respectively performed on 4-5 weeks and 8-9 weeks after birth.Blood serum were respectively extracted on 3，5，7 and 9 weeks after birth.The results showed as follows：At the age of 28 days，fecal energy (FE)，urinary energy (UE)，digestible energy (DE) and metabolic energy (ME) of groups of vegetable protein were 2.01-2.60，4.37-5.53，12.49-13.09，6.98-8.73 MJ•d^-1，and the digestibility and retention of nitrogen (N) were 68.94%-82.65%，33.79%-41.67%，respectively.Thereinto，FE of WP，PP，RP groups were significant higher than MP group (P＜0.05).DE and ME/GE (gross energy) of WP，PP groups were significant lower than MP group (P＜0.05).DE/GE of WP，PP，RP groups were significant lower than MP group (P＜0.05).ME and ME/DE of PP group were significant lower than MP group (P＜0.05).Nitrogen digestibility (ND) and P retention (PR) of all vegetable protein groups were significant lower than MP group(P＜0.05).Nitrogen retention (NR)，Ca retention (CaR)，CaR/CaI (Ca retention/Ca intake)，PD /PI (P digestibility/P intake)，PR/PI，PR/PD of WP，PP groups were significant lower than MP group (P＜0.05).CaR/CaD (Ca retention/Ca digestibility) of WP group was significant lower than MP group (P＜0.05).At the age of 63 days，FE，UE，DE，ME of vegetable protein groups were 4.75-5.48，10.00-11.02，23.93-24.89，13.20-14.56 MJ•d^-1，and ND，NR were between 71.86%-78.58%，40.70%-44.01%，respectively.Thereinto，FE of WP，PP groups were significant higher than MP group (P＜0.05).DE/GE of WP，PP groups were significant lower than MP group (P＜0.05).ND of WP，PP，RP groups were significant lower than MP group (P＜0.05).CaR，CaR/CaI of SP，WP，PP groups were significant lower than MP group (P＜0.05).CaD/CaI of SP，WP groups were significant lower than MP group (P＜0.05).PR of PP group were significant lower than MP group (P＜0.05).PR/PI，PR/PD of SP，RP groups were significant higher than MP group (P＜0.05).For serum immune indexes，IgG concentration of WP，PP groups were significant lower than MP group (P＜0.05)，and IgM concentration of SP，WP，PP groups were significant lower than MP group at 21 days (P＜0.05).IgA concentration of WP，PP groups were significant higher than MP group at 35 days (P＜0.05).IgG concentration of WP，PP groups were significant lower than MP group (P＜0.05)，and IgM concentration of PP group was significant higher than MP group at 49 days (P＜0.05).IgA concentration of PP group was significant lower than MP group at 63 days (P＜0.05).IL-2 concentration had no significant difference among all groups during the whole trial period.In conclusion，protein with different sources in milk replacer have effects on metabolism of energy，nitrogen，Ca，P and immune function.Vegetable protein in milk replacer could significantly reduce metabolic level of energy，nitrogen，Ca and P than milk protein，and soybean and rice protein have smaller influence.From the level of IgG，IgA，IgM and IL-2 in blood serum，stress of calves fed milk，soybean，rice protein were weaker than wheat，peanut protein.With the age of calves rising，the effects of protein sources were continually reduced on metabolism of energy，nitrogen，Ca，P and immune function，and the ability of using vegetable proteins also were continually enhanced.

The objective of this study was to evaluate the effects of fibrolytic enzymes，isobutyrate or their mixture on digestive enzyme activity of small intestine and growth axis hormone receptor gene expression of liver in dairy calves.Thirty-six Holstein male calves of 30 days of age with similar weight were randomly divided into 4 groups.Calves in control group were fed basal ration.Calves in treatments were supplemented fibrolytic enzymes (FE，1.83 g per calf per day)，isobutyrate (IB，98.5% of isobutyrate，6 g per calf per day) and their mixtures (IBFE，FE + IB)，respectively.At 45 and 90 days of age,3 calves from each group were randomly selected and slaughtered.Samples of small intestine contents and liver were collected in the morning before feeding on 45 and 90 days of age for determining intestinal digestive enzyme activity，GHR，IGF-IR and IR mRNA expression in liver.The results showed that on 45 days of age，amylase activities of proximal jejunum digesta，distal jejunum digesta and ileum digesta in IBFE group were significantly higher than FE and control groups(P＜0.05).Trypsin activity of duodenum digesta，proximal jejunum digesta，distal jejunum digesta and ileum digesta in IBFE group were significantly higher than other groups(P＜0.05).Lipase activities of proximal jejunum digesta and distal jejunum digesta in IBFE and IB groups were significantly higher than control group(P＜0.05).The mRNA expression of GHR and IGF-IR in liver in IBFE and IB groups were significantly higher than that of FE and control groups(P＜0.05).The mRNA expression of IR in liver in IBFE group was significantly higher than other groups (P＜0.05).On 90 days of age，amylase activities of duodenum digesta and ileum digesta in IBFE group were significantly higher than that of FE and control groups(P＜0.05).Trypsin activities of proximal jejunum digesta，distal jejunum digesta and ileum digesta in IBFE group were significantly higher than other groups(P＜0.05).Lipase activities of duodenum digesta，proximal jejunum digesta，distal jejunum digesta and ileum digesta in IBFE group were significantly higher than that of FE and control groups(P＜0.05).The mRNA expression of GHR in liver in IBFE，IB and FE groups were significantly higher than that of control group(P＜0.05).The mRNA expression of IR in liver in IBFE group was significantly higher than other groups(P＜0.05).The mRNA expression of IGF-ⅠR of liver in IBFE and IB groups were significantly higher than that of FE and control groups(P＜0.05).Intestinal digestive enzyme activity，mRNA expression of GHR，IGF-IR and IR in liver were improved by the supplementation of the mixture of fibrolytic enzymes and isobutyrate in calves.

The contribution of the gene lpxL to cell membrane structures and bionomics of avian pathogenic Escherichia coli (APEC) was studied to reveal the pathogenic mechanism of APEC to avian.An isogenic lpxL mutant was constructed from APEC O2 strain E058,and then compared its structure，biological characteristics，and pathogenicity with E058.The results showed that compared with E058，the lpxL mutant had a smoother surface，slower growth rate and reduction of endotoxin.Nitric oxide production reduction and cytokine gene transcription downregulation also were observed in HD11 treated with mutant-derived LPS relative to that in cells treated with E058-derived LPS at different times.The histopathological lesions in visceral organs of birds challenged with the wild-type strain were more severe than in birds infected with the mutant.These results indicated that the lpxL gene is important for the pathogenicity and biological activity of APEC strain E058.

Mycoplasma hyorhinis (M.hyorhinis) is a swine pathogen that can cause multiple chronic inflammation including polyserositis，pneumonia，arthritis and otitis media.It is also a potential pathogen for human beings，associated with various human cancers.In previous study，vlp family has been revealed to play a role in mediating cytoadhesion of M.hyorhinis.Herein，we furtherly studied the cytoadhesion function of vlpA，one of the vlp family member，particularly how the repeat time of repetitive unit impact the function.According to the sequence published on GenBank，the vlpA gene was amplified from bacterial genome.Furthermore，vlpA genes containing 0，3，6，9 and 12 times of repetitive unit in region Ⅲ were artificially synthesized.The genes were inserted into pET-32a(+) and expressed in Escherichia coli.At the same time，a peptide containing 2 times of repetitive unit in region Ⅲ was artificially synthesized.All the recombinant proteins，as well as the peptide were detected for their adherence to PK-15 cell by using the indirect immunofluorescence assay and microtiter plate adherence assay.All the recombinant proteins have been successfully induced for expression and purified by affinity chromatography.Adherence of vlpA to PK-15 cell was observed by the indirect immunofluorescence assay.vlpA0 could bind PK-15 cell in the microtiter plate adherence assay，but not the peptide containing 2 times of repetitive unit in region Ⅲ.Furthermore，we detected the adherence of recombinant proteins vlpA3，vlpA6，vlpA9，vlpA12 containing 3，6，9 and 12 times of repetitive unit in region Ⅲ by using the microtiter plate adherence assay，the results showed that all of the four recombinant proteins could adhere to the cell，but the adherence was lower than the recombinant protein vlpA0.Additionally，as the repeat times increased，the adherence significantly decreased.The results of this study suggest that vlpA is one of the adhesion factors of M.hyorhinis.The region Ⅱof VlpA contains cytoadhesion site.The adhesion ability of the whole molecule significantly decreased as the times of repetitive unit in region Ⅲ increased.The results of this study laid the foundation for further study on the pathogenic mechanism of M.hyorhinis.

This experiment was conducted to establish a simple，sensitive and rapid detection method for the biosecurity control of the porcine circovirus 2 (PCV2) diseases.The Cap gene of PCV2 was used as the signature sequence for cross amplification primers design and the constructed Cap gene plasmid DNA was used as the standard template.Amplification temperature，time and constituents of the reaction，including concentration of primers，MgSO4，dNTP，Betaine and Bst DNA polymerase were systematically optimized.Used the nucleic acid test strip detect the amplified products，compared the sensitivity，specificity and consistency of CPA，PCR and ELISA by 127 standard clinical samples.The detection results showed that the cross amplification could be realized at 58 ℃ within 50 min，could specifically detect PCV2 with a sensitivity that was about 10 times higher than conventional PCR method.The detection limit of recombinant plasmid was 7.6 copies•μL^-1 and the assay gave no amplification from PCV1，PRV，CSFV，PRRSV，PEDV，TGEV and RV.The final optimal CPA condition established for rapid detection of PCV2 was as follows：0.5 μmol•L^-1 each of F3 and B3，1.0 μmol•L^-1 of CPR，0.7 μmol•L^-1 of DF5F，0.9 μmol•L^-1 of DF5B，0.075 mol•L^-1 Betaine，3 mmol•L^-1 MgSO₄， 0.4 mmol•L^-1 dNTPs，9.6 U Bst DNA polymerase，2 μL 10×ThermoPol Reaction Buffer，4 μL templates and sterilized double-distilled water in 20 μL volumes.The sensitivity，specificity and consistency were 78.08% to 94.52%，87.03% to 92.59% and 84.25% to 91.34% among the three methods，respectively.Receiver operating characteristic curve showed that CPA was the best detection method. PCV2 cross priming amplification method does not require expensive equipment and skilled technicians，the visual detection could provide an intuitive and objective decision for people，and could also effectively avoid the aerosol pollution，which could be used in the on-site diagnosis for effective prevention and control of porcine circovirus 2 diseases.

 To find transcriptome difference of female and male Trichuris suis，and to enrich pig whipworm transcriptome data，the transcriptomes were analyzed by Illumina Hiseq 2000 high-throughput RNA sequencing.All of the assembled Unigenes were annotated using BLAST search against NR，GO，COG and KEGG databases.The differentially expressed genes were enriched in gene ontology and KEGG pathway.The results showed that 21 026 and 28 886 unigenes were got from female and male T.suis transcriptomes which were assembled from 59 657 854 and 48 414 184 clean reads respectively.The annotated Unigenes of female T.suis to NR，GO，COG and KEGG databases were 11 700，2 287，1 650 and 9 690 respectively，while the data of male T.suis were 12 902，2 414，1 791 and 10 377.There were 4 320 differentially expressed genes found in female and male T.suis，and more than 84.90% of the information were derived from pig whipworm in NR database.Take female T.suis for reference，3 496 genes were up-regulated and 824 genes were down-regulated in male T.suis.There were 906 differentially expressed genes enriched in KEGG pathways，and 7 of them were significantly enriched.Many important genes associated with reproduction including Major sperm protein，Histone H2A/H2B and Vitellogenin，and genes related to pathogenisis，immunity and inflammatory regulation such as protease，Serpin and Cystatin were found in transcriptome of T.suis，in addition to these，there were many genes of unknown functions.This study revealed the number of differentially expressed genes in female and male T.suis，and obtained the function，classification and metabolic pathways of the differentially expressed genes，which enriched T.suis transcriptome information，and provide a solid basis for the study on T.suis-host interactions and developing anti-whipworm drugs.

 Recombinant parvovirus-like particles (PPV-VLPs) are particulate exogenous antigens that induce strong CTL response.In order to decipher the mechanisms responsible for CTL activation by such exogenous antigen and the probable endocytic pathways in PPV-VLP capture，we analyzed the influence of different antibodies on dendritic cells (DCs) endocytic uptaking recombinant parvovirus-like particles.The results analyzed by FACStar cytometer and confocal microscopy showed that the PPV-VLPs capture efficiency of spleen DCs pretreated with anti-CD64 and anti-DEC205 antibodies was not affected.However，anti-caveolin-1 antibodies treated DCs was severely reduced.DCs pretreated with anti-actin antibodies were barely to uptake PPV-VLPs.These results showed that the uptake of PPV-VLPs seems to be related to macropinocytosis and caveolae-mediated endocytosis in an actin-dependent pathway，not clathrin-coated pits or phagocytosis.

The paper aims to study the correlation between Streptococcus thermophilus and intestinal flora and brain-gut axis function during diarrhea irritable，and further reveals the regulation mechanism of nisin on monoamine neurotransmitters.By injecting three serotypes of Escherichia coli mixed bacterium suspension liquid (1∶1，1.0×109 CFU• mL－1),the diarrhea model of mice was prepared，and nisin and Mongolian medicine were used for treatment.Tissus samples were collected at 0 h，72 h post modeling and ELISA kits were used to detect the levels of 5-HT，DA and NE，as well as the caecum contents plate were diluted 10 fold into eight gradient coating on the responging medium to calculate bacterial colony number by using plat coating method.The results showed that：(1)In brain tissue,5-HT level at 72 h post modeling in the nisin，traditional Mongolian medicine（TMM） compound，TMM ingredients，and antibiotic groups were increased to 108.60%，124.04%，117.47%，115.57% of 0 h， respectively; and the duodenal 5-HT in the nisin， TMM compound and TMM ingredients groups at 72 h post modeling were increased to 289.18%，200.30%，271.59% of 0 h,respectively，nisin group was significantly higher than other groups (P<0.05).After modeling，in addition to the jejunum，compared 72 h’s with 0 h’s，nisin and TMM compound have signicantly improved 5-HT level （P<0.05） in mice serum，brain，duodenum，ileum and colon.(2) After 72 h，nisin，TMM compound，TMM ingredients and antibiotic can improve the dopamine in the intestine and brain of normal mice,and that in the ileum increased the most，and the difference was significant (P<0.05).(3)In brain tissue at 0 h and 72 h, NE concentration increased to 115.62% and 106.17%，respectively under the react of TMM compound and nisin,and which also have significant difference comparing with blank control，and the negative control group (P<0.05)；in the duodenum and ileum，the EN concentration of nisin group at by 72 h were increased by 170.97%，121.74%，especially that in the duodenum was higher than other groups and the difference was significant (P<0.05).(4) Nisin，TMM effective ingredients and TMM compound can significantly increase the number of intestinal probiotic Lactobacillus and Bifidobacterium in E.coli infected mice，and significantly reduce the number of harmful bacteria Enterobacter and Enterococcus (P<0.05).In addition，nisin and TMM compound were significantly increased bacterial infections in mice blood，brain and various intestinal segments in 5-HT，NE content，especially in the duodenum，nisin and traditional Mongolian medicine can break the balance of intestinal 5-HT，and accompanied by increase of 5-HT in the intestine and blood,the brain 5-HT content also increased.

To study the effect of vanadium on the chick cerebral cortex γ-aminobutyric acid (GABA) expression，280 1-day-old Avian broilers were chosen and randomly divided into four groups，each group divided into seven sample groups，each sample group have 10 broilers，and fed control diet and diet added vanadium (low dose group：15 mg•kg^－1；dose group：30 mg•kg^－1；high dose group：45 mg•kg^－1) for 6 weeks using the immunohistochemical method to detect the expression of chick cerebral cortex GABA Variety.The results showed that the expression level of the cerebral cortex GABA 42 d chicks，15 mg•kg^－1of vanadium-added group was the highest (P<0.01 or P<0.05)，followed by 30 mg•kg－1 of vanadium added group (P<0.01 or P<0.05)，45 mg•kg^－1 of vanadium-added group than the control group (P>0.05)；45 mg•kg－1 of vanadium-added group，when at 28 d the cerebral cortex expression levels were significantly (P <0.01) higher than the 14 and 42 d；when at the 14 cortex horizontal cells and astrocytes of GABA expression were higher than 42 d (P>0.05).In summary，it showed that the dietary vanadium and the expression of chick’s cortex GABA exist in a dose and time-effect relationship，suggesting that the regulation of expression may be related to up-regulation of GABA.