Feeding is a reflective activity in animals，and regulation of feeding behavior is associated with brain-gut axis.After feeding，brain-gut peptides were released from gastrointestinal after receiving signal stimuli of nutrients，vagal afferent neurons received signal of brain-gut peptides and transferred them to central nervous system，which integrated these signals to effector organ and promoted or inhibited feeding.Brain-gut peptides，important gastrointestinal hormones involved in food intake，are playing a critical role in the regulation of feeding behavior of brain-gut axis.The advances on food intake regulation of brain-gut axis are reviewed in this article.

This study aimed to detect polymorphisms of porcine IRS4 gene and its expression pattern in 17 tissues，and to identify gene markers associated with the QTL for fat deposition traits.Comparative sequence analysis of the IRS4 revealed 3 SNPs，including g.96C>G in the promoter region，a synonymous substitution g.1829T>C and a missense substitution g.1794T>C (p.Phe432Leu).Through RT-PCR assay，we found that IRS4 gene was abundantly expressed in hypothalamus and pituitary，but weakly expressed in other tissues.Furthermore，we genotyped the 3 SNPs in White Duroc×Erhualian F₂ intercross，Sutai pigs，Erhualian pigs and DLY［Duroc×(Landrace×Yorkshire)］three-way crossbred pigs and Chinese local wild boar by PCR-RFLP，then calculated allele frequencies and estimated the associations between those SNPs and backfat thicknesses (BFTs) at shoulder，chest，waist and hip，abdominal fat weight (AFW)，intramuscular fat content (IMF) and loin eye area (LEA) in these populations.The SNP g.96C>G was almost fixed in the F₂ intercross，so it was first excluded as a causative mutation (QTN) underlying this QTL.For both SNPs g.1794T>C and g.1829T>C，all western breeds (White Duroc and DLY) only had TT genotype，while，in the Erhualian population，the frequencies of CC genotypes at g.1794T>C and g.1829T>C were 65% and 100%，respectively.The results of association analysis showed that g.1794T>C was highly associated with all investigated traits in the F₂ intercross (P<0.01)，but associated only with IMF in Sutai pigs（P<0.05）and had no significant effect on Erhualian’s phenotypes；g.1829T>C was significantly associated with several fatness traits (e.g.BFT and LEA) in the F₂ intercross and Sutai pigs.Moreover，bioinformatics analysis demonstrated that the potential influence of the missense variant g.1794T>C on IRS4 protein function was limited.The results suggest that the SNP g.1794T>C is unlikely to be QTN，and the SNP g.1829T>C associated with fat deposition traits across populations could be used as marker for selection of these traits.

In this study，we exposed pig intestinal epithelial cells (IPEC-J2) to 0.1 and 1 μg•mL^-1 LPS for 2，4 and 6 h，respectively.Then，we estimated the relative mRNA expression of TLR4 and TLR4 signaling pathway-related genes (CD14，MyD88， TNF-α，IL-1β，IFN-α) using qPCR，which preliminary revealed the mechanism of the molecules related to inflammatory reactions in pig intestinal epithelial cells induced by enterotoxigenic Escherichia coli.Both concentrations of LPS upregulated the expression of TLR4 and its signaling pathway-related genes，and the expression level of all genes sharply increased from 4 to 6 h.The fold change of mRNA expression induced by 1.0 μg•mL^-1 LPS was significantly higher than that induced by 0.1 μg•mL^-1 LPS.The former stimulated the intestinal tract to produce stronger immune responses and more rapid development of inflammatory reactions.Above results suggested that LPS was released in the pig intestinal tract with E.coli infection，and upregulated the LPS receptor TLR4，leading to activation of the TLR4 signaling pathway.Given the dependency on myeloid differentiation factor 88 (MyD88) during signaling，stable upregulation of MyD88 and the cytokine cascade results in the release of large amounts of proinflammatory cytokines，causing inflammatory reactions，diarrhea and edema disease.

The aim of this study was to investigate the effect of Tibet chicken as male and female parents on hypoxia development of hybrid eggs.Eggs from Tibet × Tibet (TT)，Chahua × Chahua (CC)，Tibet × Chahua (TC) and Chahua × Tibet (CT) chickens were incubated under hypoxic condition.Then the hatchability of eggs were examined.The results showed that the hatchability of TC (36.41%) and CT (39.57%) groups were significantly higher than that of CC group (16.97%) under 13% oxygen concentration (equivalently 3 980 m altitude).Eggs from Tibet hens (including TT and CT groups) had a higher egg weight and a lower water loss during incubation than eggs from Chahua hens (including CC and TC groups).The results indicated that the egg weight and the water loss during incubation were mainly determined by female parent.The intercrossing eggs between Tibet chicken and Chahua chicken had higher hatchability and had stronger heterosis in traits of hypoxic hatchability.There was no significant difference in hypoxic hatchability of eggs between TC and CT groups.The embryo development of eggs from Tibet chicken as female parent was better at early stage and embryo development of eggs from Tibet chicken as male parent was better at last stage under hypoxic incubation.

This research evaluated the availability of 5 “housekeeping genes”as reference genes for gene expression detection in different early developmental stages of muscle tissues in chicken，which would provide scientific evidences for selecting reference genes in the molecules detection related to muscle development of chicken.Real-Time quantitative PCR (RT-qPCR) and Western blot were used to test the mRNA and protein levels of β-actin (ACTB)，β-tubulin (TUBB)，TATA-binding-protein (TBP)，Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Heat shock 70 kDa protein (HSP70) in breast muscle of chicken at embryonic day 12 and 17，day 1 and day 14 post-hatch，respectively.The results showed that，only mRNA and protein levels of HSP70 didn’t significantly change in the early developmental stages of breast muscle in chicken，while that of other genes significantly altered (P<0.05 or P<0.01).HSP70 was more suitably used as reference gene for gene or protein expression detection in the early developmental stages of muscle tissues in chicken.This study provides methodological basis for functional genes and proteins studies of muscle development in chicken.It also could play a referential role in other species.

This study focused on the cloning of ovine UCP2 gene and its seasonal expression differences in adipose tissues.The results would reveal the effects of seasonal regulation on the UCP2 expression.The full-length cDNA was cloned by RACE method.Real-time PCR and immunohistochemistry techniques were used to study the expressions of UCP2 mRNA and its coding protein in 6 adipose tissues of South African Mutton Merino and Shanxi Mutton Sheep Dam Line during two season periods.The results showed that the ovine UCP2 cDNA was 1 641 bp in length with a 930 bp open reading frame.It contained 8 exons which encoded 309 amino acids.The mRNA expressed in all 6 adipose tissues detected in this study.Higher expression in deep than in superficial depots (P<0.05) was found as suggested by the highest expression in perirenal and lowest in subcutaneous fat.UCP2 expressed in similar trend at mRNA and protein levels，and distributed on the membrane of adipocytes.Both mRNA and protein expressions were higher in winter-spring period than that in summer-autumn period (P<0.05).These results indicate that UCP2 differentially expression in adipose tissues is related to seasonal changes and functions in maintaining body temperature and regulating energy balance.

The study was conducted to investigate the expression characteristics and bioinformatics analysis of beta-defensin family genes in the testes and epididymal caput of bucks.The data were from the transcriptomes of three testes of 7-day-old kids，testes and epididymal caput of three adult bucks of known fertility analyzed by Illumina HiSeq^TM 2000 high-throughput RNA sequencing.All beta-defensin family genes were sorted out according to annotation of unigenes and contigs for analysis of different expression.Protein profile of beta-defensins was analyzed by bioinformatics methods.The results showed that a total of 7，12 and 33 β-defensins were discovered in testes of 7-day-old kids，testes and epididymal caput of adult bucks respectively，while gDB123，gDB119 and gDB124 were the highest expression in the testes of 7-day-old kids，testes and epididymal caput of adult bucks respectively.gDB33 was specifically expressed in testes of adult bucks.Twenty-one β-defensins were specifically expressed in epididymal caput of adult bucks.The molecular weight of β-defensins was from 6.89 to 13.85 ku.The bioinformatics analysis showed that goat β-defensins protein were small molecular，polar，hydrophobicity and basic polypeptide with positive charge.A high degrees of conservation in spacing pattern was 6 cysteines in peptides sequence of β-defensins，which consensus pattern was -C-X_2-45-C-X_3-6-C-X_3-13-C-X_4-7-CC-.However，the pattern of 24 β-defensins was -C-X_5，6-C-X_3，4-C-X₉-C-X_5，6-CC-.A total of 16 β-defensins were secreted glycoprotein.Except for gDB126，the rest of 24 β-defensins had the phosphorylation sites.The goat β-defensins family protein were also the antimicrobial cationic peptides.The rich specific-epididymis β-defensins were expressed in epididymal caput，which could coat sperm plasma membrane with glycocalyx in sperm maturation and play a vital role in making sperm motion.And β-defensins maybe were cell signal in some pathways.

The aim of this study was to investigate the adaption mechanism of plateau animals to extreme environment of the Qinghai-Tibet plateau，and to provide some basic data for exploring the adaption mechanism of plateau animals.The coding region sequence of yak AQP9 was cloned by molecular cloning technique and analyzed by bioinformatics in this study.Some characteristics of the AQP9 gene and encoded protein sequences were predicted and analyzed in the following aspects as the general physical and chemical properties，hydrophobicity，transmembrane structure and secondary structure.The results showed that the CDS of yak AQP9 contains a complete ORF (885 bp) encoding 295 amino acids.The putative molecular weight and theory isoelectric point of AQP9 gene coding protein in yak were 31.89 ku and 6.21，respectively.The protein encoded by yak AQP9 contains 6 transmembrane domains，and it was a hydrophobic protein.The secondary structures are mainly composed of α-helix，extended strand，β-turn and random coil.Deduced amino acid sequences of AQP9 gene between yak and cattle，sheep and other species were high homology，and the phylogenetic distance consistent with their genetic relationship.This study will provide certain basic data for studying hypoxia adaptation of yak.

This study was designed to investigate the pluripotency-related markers of bovine ES cells so as to provide a basis for optimizing culture conditions of bovine ES cells and promoting its research progress.The inner cell mass (ICM) were separated from bovine IVF blastocysts，seeded on mouse embryonic fibroblasts cultured in 2i/LIF medium and then passaged mechanically.The pluripotent markers of the ICM-derived colonies were detected at passage 1 and passage 10 by immunostaining.Additionally，the ICM and trophectoderm (TE) isolated from bovine blastocysts by magnetic activated cell sorting were used for qRT-PCR to validate the differences in gene expression between ICM and TE.We have derived ICM colonies from bovine IVF blastocysts and cultured them to passage 10，which displayed typical stem cell morphology and expressed specific markers such as OCT4，SOX2，NANOG，SSEA1，SSEA4 and TRA-1-60.Moreover，the results of qRT-PCR showed that OCT4，SOX2 and NANOG were differentially expressed between ICM and TE，SOX2 showed the most significant difference.In conclusion，2i/LIF culture medium is helpful for the culture of the ICM-derived colonies from bovine IVF blastocysts，and SOX2 is more likely to be a candidate pluripotency-related marker gene of bovine ICM-derived colonies.

In order to investigate whether the Bactrian camel’s special metabolic pathways and unique detoxic capabilities were caused by particularities of CYP gene family，the Bactrian camel’s whole genome sequencing data were systemically analyzed and annotated，and then CYP gene family was searched from the whole protein database，and compared with CYP gene families of cattle，horse，chicken and human，respectively.The results showed that the total of 63 CYP gene copies were found in Bactrian camel’s whole genome，and were classified into 17 families and 38 subfamilies.Among them，9 multi gene families were found，and CYP2，CYP3 and CYP4 had 27，6 and 7 subfamilies，accounting for 43%，10% and 11% in camel CYP，respectively.Compared with cattle，chicken，horse and human，the distribution of CYP gene subfamilies in camel was different，with more CYP2J and CYP3A copies in Bactrian camel，which was probably associated with Bactrian camel’s some specific biological characteristics and metabolic pathways.Therefore，the certain special biological characteristics and metabolic pathways of Bactrian camels are related to their high copy number of CYP2J and CYP3A genes，and the present analytical results will lay a good foundation for further revealing of Bactrian camel’s unique properties of salt tolerance and detoxification.

This experiment aims to detect the effect of down-regulation of GDF9 on reproduction and growth in GDF9 shRNA transgenic mice through statistical analysis of the litter size and weight gain of transgenic mice and detection of biomolecular and blood biochemical indexes such as GDF9 shRNA，GDF9 and serum FSH and GH level after reproduction.The levels of GDF9 shRNA and GDF9 mRNA expression were detected in hypothalamus，pituitary and ovary via RT-PCR，and serum FSH and GH level were examined using ELISA.The results showed that 16 GDF9 shRNA transgenic mice obtained were positive，and GDF9 shRNA in transgenic mice was widely expressed in various tissues.Litter sizes between transgenic and wild mice were significantly different (P<0.05)；Body weight of transgenic male mice was higher than that of control mice with significant difference (P<0.05) in the seventh week，and body weight of female mice did not show significant difference.Compared with the negative mouse，GDF9 gene expression in transgenic mice in hypothalamus and pituitary were decreased by 35.4% and 39.8% (P<0.05)，respectively.In transgenic female mice，GDF9 expression in ovaries was not significantly decreased (P>0.05)，but its serum FSH level was 55% higher than the control (P<0.01).The present study showed down-regulation expression of GDF9 gene can improve the ability of animal reproduction.It may provide a favorable material and molecular target for producing transgenic large livestock with high reproduction.

The objectives of this study were to evaluate the effects of RNAi on MC1R expression and melanin synthesis in the melanocytes of alpaca.Three pairs of specific siRNA for MC1R gene were designed for RNA interference in the melanocytes of alpaca.The expression of MC1R siRNA was detected by using qRT-PCR technology.The location of MC1R was detected by immunohistochemistry.The amount of melanin synthesis in melanocytes was detected by the absorbance value.The results showed that the relative expression of MC1R mRNA in the siRNA2 and siRNA3 groups was significantly lower (P<0.01) than that in the blank control group，but there were no significant differences at the relative expression of MC1R mRNA in the siRNA1 and negative control group (P>0.05)；the production of melanin synthesis of siRNA2 and siRNA3 groups were significantly lower than that in the blank control group (P<0.01).The study was successfully transfected MC1R siRNA into alpaca melanocytes and the effective MC1R siRNA was screened from 3 groups of specific siRNAs.The inhibition of MC1R is consistent with the amount of melanin synthesis.MC1R of melanocytes played a regulatory role in the synthesis of melanin，and the mechanism need further research.

This experiment was conducted to examine the effects of light color on the production and carcass performance and sexual characteristics of Yellow-feathered broilers，aimed to provide theoretical and practical guidance for poultry production.A total of 720 one-day-old Beijing-You chicks were reared under white light，green light (λ = 525 nm)，blue light (λ = 460 nm) and red light (λ = 630 nm) from light-emitting diode lamps as light sources.Each treatment included 6 replicate groups with 30 chicks per group.Feed and water were provided for ad libitum consumption.All birds were exposed to 20 lx of light intensity for the first week，and then 5 lx till the end.Feed consumption was recorded daily.At the end of 4，8 and 13 weeks of age，body weight was recorded.At the end of 13 weeks of age，the chosen birds were sacrificed and carcass data were collected.The present study found that in the brooding and growth period，the birds in red light group grew slowly，and reached statistical significanc groups(P<0.05）at the end of 8 weeks of age，but then the red light group grew rapidly and caught up with the other groups (P>0.05) at the end of 13 weeks age.Compared with the white light group，the blue and red color light improved the survival rate.Breast muscle weight and percentage of red light group were lower than that of green light group(P＜0.05)，while thigh muscle weight and percentage had no significant differences(P>0.05).Fat deposition capability of monochromatic light groups was higher than that of white light group.The red light increased the weight of testis.The result indicate that，in the brooding period，the intelligent use of blue light improves survival rate，and then the red light enhances the chickens’ body growth and gonadal development in the fattening period.

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of H7 and N9 subtype avian influenza viruses (AIV).According to the conservative sequences of hemagglutinin (HA) and neuraminidase (NA) genes of H7 and N9 subtype AIV in GenBank，two sets of primers were designed，then the reaction conditions were optimized.The specificity and sensitivity of the RT-LAMP assay were evaluated.RT-LAMP amplification was completed in 1 h by incubating all of the reagents in a single tube by real-time turbidimeter at 63 ℃.The specific test demonstrated that the RT-LAMP assay was specific for detecting H7 and N9 subtypes of AIV，and no cross reaction with other subtypes of AIV and avian respiratory pathogens.The detection limit of the RT-LAMP assay was 10 copies using vitro transcription RNA of target genes.The newly developed RT-LAMP assay is specific，sensitive，simple，rapid and can identify H7，N9 and H7N9 subtype AIV visually，and it will be a very useful screening assay for prevention and control these diseases.

 This study was undertaken to reveal the possible resistance mechanism of B19 and B21 haplotype SPF chickens against MDV.One day-old B19 and B21 haplotype chickens were infected with MDV Md5 strain by intramuscular as 800 PFU per feather.The transcription levels of targets genes including BNK，BLec1，IFN-γ，IL-4，IL-10 and IL-18 were measured at different time points after MDV treatment.Results showed that gene transcription levels of BNK，BLec1 and the four cytokines in B19 and B21 chickens decreased compared to the control group on the 4th and 7th day post infection (dpi).On the 10th dpi，BNK，BLec1， IFN-γ，IL-4 and IL-18 gene transcriptions highly increased in B21 chicken in different levels，while decreased in B19 chickens.On the 13th dpi，gene transcriptions of BNK，BLec1，IL-18 and IL-10 in MDV treated B21 chickens also rose in different levels compared to the control group，while decreased in MDV treated B19 chickens.In conclusion，the present study demonstrate that B21 haplotype SPF chickens express high levels of BNK，BLec1 and cytokines than B19 ones and the resistant difference against MD between MD tolerant and sensitive chickens is related to the gene transcriptions of BNK， BLec1 and cytokines.

In this study，the pathogenicity of a very virulent Marek’s disease virus (MDV) SD2012-1 strain was analyzed.sixty SPF chickens were randomly divided into 3 groups：non-immunized group，HVT immunized group and CVI988 immunized group.Chickens were vaccinated with different types of MD vaccine at 1-day-old，and were challenged with SD2012-1 strain at 10-day-old.The challenged chickens were housed in separate isolators，and were sampled for clinic，histopathology and PCR detection of MDV.Clinical and histopathologic lesions in the infected chickens showed：slight histopathologic lesions were developed at 2 weeks post challenge，characteristic histopathologic lesions with scattered clumps of tumor cells were developed at the 6th weeks post challenge，and grossly obvious tumor lesions with large number of tumor cell aggregates were developed at the 9th weeks post challenge；SD2012-1 could break through the protection of CVI988 vaccine，causing MD with up to 30% rate in immunized chickens.PCR tracking of MDV in the infected chickens showed：MDV could be detected in non-immunized group and HVT group at 5 days post challenge；the positive rates of MDV were 100% in non-immunized and HVT group，30% in CVI988 group，at 10 days post challenge；the positive rates in the three groups were all 100% at 20 days post challenge；samples，including liver，kidney，muscular stomach，mesentery，duodenum，heart and bursa of Fabricius collected from clinical chickens，were all MDV positive by PCR，feather pulp samples collected from the infected living chickens were all MDV positive by PCR.When infected with MDV SD2012-1，chickens immunized with HVT hardly had protective effect on this virus，and the long-term existence of SD2012-1 in chickens immunized with CVI988 vaccine could break through its immune protection，which proposed a severe challenge for Marek’s disease prevention and control.

The objective of this study was to investigate the role of cellular autophagy in bovine viral diarrhea virus (BVDV) infection.We used Oregon C24V strain to determine whether BVDV infection induced autophagy by using western blot，indirect immunofluorescent assay and transmission electron microscopy.To analyze the effect of autophagy on the replication of BVDV in MDBK cell，wortmannin and small interfering RNAs targeting Beclin-1 were used to inhibit autophagy.The results showed that BVDV infection results in LC3-Ⅰ/Ⅱ conversion，p62 degradation，an increased accumulation of punctate GFP-LC3-expressing cells，and a higher number of autophagosome-like double-membrane vesicles in the cytoplasm of host cells.Inhibition of autophagy or interfering expression of Beclin-1 led to a significant reduction in BVDV titers and protein expression.These results demonstrate that BVDV infection induces autophagy which in turn enhances BVDV replication.

The aim of this study was to express FnBPA-A（A domain of fibrinogen-binding region A）and FnBPA-BCD (BCD domain of fibrinogen-binding region A different domains of FnBPA) of Staphylococcus aureus，and compare the adhesion inhibition activities of their antiserum for prevention of bovine mastitis caused by S.aureus.FnBPA-A and FnBPA-BCD encoding genes were cloned and inserted into pET-28a vector.Then the recombinant proteins were expressed and purified to immune Newzeland rabbits and mice.The adhesin binding ability，adhesion inhibition activity of their serum antibody and challenge protection ability were tested.The results showed that the antibody level，the binding ability to Fg and Fn and adhesion inhibition activity of FnBPA-A and FnBPA-BCD co-immunization group were better than that of single immunization groups.The mice antibody level of FnBPA-A immunization group and co-immunization group were better than that of FnBPA-BCD immunization group.In addition，FnBPA-A and co-immunization group showed better adhesion inhibition activity to Fg，FnBPA-BCD and co-immunization group showed better adhesion inhibition activity to Fn.The immunological protection result showed that each group had good protective effect and FnBPA-A immunization group was the best one.The results of this study may provide the basis for design of new vaccines of bovine mastitis caused by S.aureus.

According to the bidirectional development of the protoscolex (PSC) of Echinococcus granulosus，the study was conducted to screen the specifical genes of E.granulosus PSC development between a larval cyst and adult worm.The subtractive PCR products of the SSH library of E.granulosus PSC-adult (AW) and E.granulosus PSC-cyst (CW) were connected to the pGEM-T easy vector，and were cloned and sequenced.Totally，280 and 200 clones were obtained from PSC-CW and PSC-AM libraries，respectively，with the length of fragments 200 to 1000 bp.The clones were sequenced.Sequence alignment analysis showed that these sequences belong to 16 and 10 gene clusters representing 16 and 10 unique genes from PSC-CM and PSC-AW libraries respectively.As the genes are specifically or differentially expressed during the development of PSC to either adult worm or cyst，they are important for the parasite development and differentiation，thus can be candidates for drug and vaccine development.

The swine major histocompatibility complex class Ⅰ or swine leukocyte antigen class Ⅰ (SLA-Ⅰ)，made of the heavy chain (α chain) and light chain (β chain)，is involved in the process of endogenous antigen presenting.In the present study，we first amplified the genes of the extracellular region in SLA-Ⅰα chain and β chain from porcine alveolar macrophage (PAM) using RT-PCR.The two genes were then cloned into the prokaryotic expression vectors and induced to express in the form of inclusion bodies.After being washed，refolded and purified，the recombinant proteins were confirmed by western blot and mass spectrometer.Then，the rabbit antiserum against the recombinant proteins was prepared with titers of 1：256 000 by an indirect ELISA.Finally，the antiserum was used in Western blot and indirect immunofluorescence assay，showing that the antiserum can specifically react with SLA-Ⅰ α chain and β chain expressed in both PAM and PAM cell line 3D4/21.This study not only provides useful reagents for analyzing the function of SLA-Ⅰ，but also develops the basis for further study on the mechanisms associated with immune responses induced by viral infections.

The aim of this study was to study the role of rfaE gene of Haemophilus parasuis lipooligosaccharide (LOS) which induced pro-inflammatory cytokine (interleukin (IL)-1α，IL-1β，IL-6，IL-8 and tumor necrosis factor (TNF)-α) mRNA transcription in porcine alveolar macrophages (PAMs).The LOS of H.parasuis SC096 strain，rfaE mutant and its complementation was extracted.The PAMs were stimulated with LOS (5 μg and 10 μg) from H.parasuis SC096，ΔrfaE mutant (ΔrfaE) and its complementation (cΔrfaE) and LPS from Escherichia coli for 2，4，6，12 and 24 h.The RNA was extracted and reversed into cDNA.The mRNA transcription of IL-1α，IL-1β，IL-6，IL-8 and TNF-α were then detected by Real-time PCR.The result showed that the mRNA transcription of IL-1α in PAMs induced by 5 μg LOS from H.parasuis for 2，4，6，12 and 24 h were up-regulated，and significantly higher than the mRNA transcription of IL-1α in PAMs induced by 5 μg LOS from ΔrfaE (P<0.05)， the mRNA transcription of IL-1β，IL-6，IL-8 in PAMs induced by 5 μg LOS from H.parasuis for 12 and 24 h were up-regulated，and remarkably higher than the mRNA transcription of IL-1α in PAMs induced by 5 μg LOS from ΔrfaE (P<0.05)，the mRNA transcription of TNF-α in PAMs induced by 5 μg LOS from H.parasuis for 24 h was up-regulated，and notably higher than the mRNA transcription of IL-1α in PAMs induced by 5 μg LOS from ΔrfaE (P<0.05)；the 10 μg LOS from H.parasuis up-regulated mRNA transcription of IL-1α，IL-1β，IL-6，IL-8 and TNF-α in PAMs，and were significantly higher than that of 10 μg LOS from ΔrfaE mutant (P<0.05).At the same time，the cytokine mRNA transcription levels in PAMs induced by LOS from cΔrfaE were restored to the level in PAMs induced by LOS from H.parasuis.The date firstly confirmed that the rfaE gene were involved in the pro-inflammatory cytokine mRNA transcription in PAMs induced by H.parasuis LOS，suggesting that the rfaE gene play an important role in the pathogenesis of H.parasuis LOS.

Milk fever (MF) is a seriously nutrition metabolism disease which is divide into clinical and subclinical hypocalcemia (SH).The level of Ca in blood combining the clinical signal were always used to diagnose MF.But in the total development progress of this disease the pathogenesis was not clearly.In this study we screened the biomarkers of MF and SH，and want to reveal the possibly pathogenesis.Twenty-seven dairy cows were selected as the experiment animal，and there were 3 groups MF，SH and C according to the blood calcium concentration and the clinical signal of these dairy cows.Every 3 samples were mixed into a hybrid sample in each group，and 9 hybrid samples were crossed into the 2D-DIGE.Eighty of 110 spots，which were found via 2D-DIGE，were selected into mass spectrum.Sixty-six spots were found，and were identified into 16 proteins.A2M，HP，PON1 were selected in verification test by using Western blotting，and the results support the results of DIGE experiment.In this study 16 proteins were found among three groups，and 3 proteins of them were verified first time.Collectively，the results indicate that A2M，HP，PON1 are 3 novel biomarkers for MF，and they may contribute to explore the possible mechanism of MF.

In order to investigate the immune-enhancing activity of ophiopogon polysaccharide liposome (OPL)，two experiments were carried out.In vitro，the effects of OPL on splenocyte proliferation of chicken were determined by MTT method，and ophiopogon polysaccharide (OP) was used as control.In vivo，four hundred 11-day-old chickens were randomly assigned into 8 groups，and the chickens were injected with cyclophosphamide to build the immunosuppression model.At 14-day-old，the chickens were vaccinated with Newcastle disease vaccine.At the same time of the first vaccination，the chickens were injected respectively with OPL at different dosages.At various time points after the first vaccination，the peripheral lymphocyte proliferation，serum antibody titer and the contents of cytokines were measured.The results showed that OPL could significantly promote splenocyte proliferation singly or synergistically with PHA and LPS at 7.813-125 μg•mL^-1 in vitro，and the efficacy was superior to OP (P<0.05)；In vivo，OPL at high and medium doses could significantly promote lymphocyte proliferation，enhance antibody titer and improve the contents of IL-2 and IFN-γ compared with OP at most time points (P<0.05).These results indicated that OPL could significantly improve the immune-enhancing activity of OP，OPL would be expected to exploit into a new-type preparation of OP.

In order to investigate the expression of p-p38 in central nervous system in rats anesthetized with the Combined Anaesthetic for Miniature Pigs (XFM)，30 rats were divided randomly into two groups：control group and XFM group.XFM group consisted of four subgroups：M1 subgroup (rats received XFM intraperitoneally and then waiting the disappearance of right reflection)；M2 subgroup (rats received XFM intraperitoneally and then waiting for 1 h after the disappearance of right reflection)；M3 subgroup (rats received XFM intraperitoneally and then waiting the recovery of right reflection) and M4 subgroup (rats received XFM intraperitoneally and then waiting for 1 h after the recovery of right reflection).The brain tissues were taken when reaching each time point.With the method of RT-PCR，expression of p38 mRNA were detected，and the expression of p-p38 in central nervous system was detected with Western blot.The results showed that，compared with the control group，the transcription of p38 mRNA on cerebellum and thalamus in experiment groups were significant lower than that in control group (P<0.05)，and p38 mRNA increased at group M3 and group M4，but significantly lower than control group (P<0.05).In group M1 and group M2，the expression of p-p38 in cerebellum and thalamus were significantly lower than that in the control group (P<0.05)，but the expression of p-p38 in cerebellum，hippocampus，brainstem increased significantly after administration of XFM (P<0.05) in group M1，group M2，and group M3，compared with the control group，and in group M4，the expression of p-p38 decreased，were no difference with the control group (P>0.05).These results indicated that the decrease of the expression of p-p38 in cerebellum and thalamus were induced by XFM，and these changes of p38 mRNA and p-p38 maybe one of the anaesthetic mechanisms of XFM.

This experiment was conducted to screen the transcription factor binding sites of MyoDI promoter in Guizhou Guanling cattle．According to the sequence of bovine MyoDI gene published in GenBank，the specific primer was designed to amplify the promoter region of MyoDI gene in Guanling cattle．The PCR products was used to construct the recombinant cloning vector pUCM-T-MyoDI-pro．Positive plasmids were authenticated by sequencing.Then screening experiment and bioinformatics analysis were applied to screen the transcription factor binding sites of MyoDI promoter in Guanling cattle．The results showed that the transcription factor binding sites of MyoDI promoter in Guizhou Guanling cattle had SATB1，Xbp，MEF2，Pax-3，Pbx1，PPAR，TFⅡD，COUP-TF，Gfi-1，HNF-1，HOX4C，NRF2(ARE)，MEF1，RXR，SMUC，Snail，MyoD and VDR．Combined with online software analysis and literature，we ultimately screened out the MyoD，TFIID，Pax3，MEF1，VDR and MEF2 in MyoDI promoter.These transcription factors play an important role in regulation of promoter activity．The result indicate that the Guanling cattle MyoDI promoter contain MyoD，TFIID，Pax3，MEF1，VDR and MEF2 transcription factor binding sites.

An epidemic disease characterized by neurological symptoms，diarrhea and acute death phenomenon in different ages of mink occurred in many fur farms of China since 2014.One pseudorabies virus (PRV) was identified by cell culture，plaque purification，morphology observation and animal inoculation experiment，named Mink 1.The isolate was able to cause typical CPE in MDCK cells，and cause itch，death in experimentally infected rabbit.Virus particles were round，with envelope，diameter of about 150 nm under the electron microscope.The gE of the Mink 1 was amplified by PCR and the results showed that PRV were found in detected animal.Sequence alignments indicated that the gE genes from the mink shared 99.4% identity with swine PRV isolated in 2012，located in a relatively independent branch.PRVs isolated had the same sequence features with two consecutive bases insertions of GAC and ACG at sites of 142-144 and 1 487-1 490 in gE compared with the classical strains.The above experimental results confirmed that the case is caused by PRV infection.