The traditional Chinese medicine network pharmacology(TCMNP) may supply new methods to the research of Chinese materia medica and traditional Chinese veterinary medicine(TCVM).In order to introduce network-based relating pharmacology into the research of Chinese materia medica and TCVM，an overview related to the TCMNP was developed in four different aspects，including the current situation，the methods，the application and the prospection of the TCMNP.

The objective of this study was to analyze the association of SNPs between Myf5 gene and chicken growth and reproductive traits.The SNPs of Myf5 gene in Jinghai Yellow chicken were detected by PCR-SSCP method and then the haplotype analysis was done.As a result，8 SNPs were identified in exons of the Myf5 gene.Nine haplotypes were detected in the group of 379 Jinghai Yellow chicken.For growth traits，the least square analysis showed that H1H5 had significant effects on 8 and 12 week-age-weight (P<0.05)，H2H6 had significant effects on 12 and 14 week-age-weight (P<0.05).For reproductive traits，H1H5 had higher body weight in first egg than H1H4，H2H4 and H1H3（P＜0.05 or P＜0.01），of which，the difference between H1H5 and H1H3 was significant(P<0.01).H1H3 had a poor performance in average egg weight of 300 days.On the other hand，H1H3 had an advantage in egg number of 300 days，and higher than that of H1H4(P<0.01).The results show that SNPs of Myf5 have certain effects on growth and reproductive traits in the Jinghai Yellow chicken，and which can be used in marker-assisted selection to accelerate the chicken breeding progress.

This study was designed to interfere Piwi gene in chicken primordial germ cells (PGCs) and explore its interaction with CVH，Dazl，CDH, Sox2, Nanog and PouV genes.PGCs was isolated from 19 stage chicken embryo，and after identification of PGCs，we analyzed expression of CVH，Dazl，CDH，Nanog，Sox2 and PouV under the condition of Piwi interference in PGCs by using real time quantitative PCR.The results showed that expression of CVH，Dazl and Nanog genes increased significantly(P<0.05)，while the expression of PouV and Sox2 genes decreased significantly followed with Piwi interference in PGCs (P<0.05).Collectively，we constructed a Piwi gene interference model in vitro and found,and which influenced expression of CVH，Dazl，CDH, Sox2，Nanog and PouV，which provided new basis for studying Piwi gene function in reproductive biology.

To develop a visible，accurate and sensitive method for zinc-finger nucleases (ZFN) screening，a screening system based on GFP (green fluorescence protein) was constructed in eukaryotic cells.A general screening vector pEGFP-ATG was constructed by removing the initiation codon of the GFP gene to the upstream of multiple cloning sites (MCS) in pEGFP-N1 vector.The target sequence of ZFN was then inserted into the upstream of the GFP sequence，the initiation codon and GFP gene were in the different ORF.The screening vector pEGFP-728 was constructed.Hela cells were transfected with pEGFP-728 vector and selected by G418 to produce the stable Hela-728 cell lines，which were thereafter transfected by the ZFNs.The fluorescence in transfected cells within 48 hours was observed to determine the efficiency of the ZFNs.Those cells with fluorescence were cloned in 96-well plate.Their genomic DNA was extracted，and the target sequence amplified by PCR was sequenced.Sequencing analysis showed that the general vector pEGFP-ATG and the screening vector pEGFP-728 were constructed correctly.The green fluorescence was not observed in Hela cells transfected with pEGFP-728，indicating that the frameshift took place in the GFP gene after the target sequence was inserted.However，the green fluorescence was observed in Hela-728 transfected with ZFNs in 48 hours，suggesting that the ZFNs were effective.Six fluorescent cell clones were selected；PCR and sequencing analysis showed that deletion mutations appeared in the target sequences of four clones.A ZFN screening system was constructed successfully in eukaryotic cells.It was more accurate，visible and sensitive than other screening methods previously reported，and more suitable for screening ZFNs in animal and plant cells.Furthermore，this system can also be applied in TALEN and Cas9 selection.

In order to explore the biological functions of TGF-β1 in coat color formation in alpaca,the experiment was conducted to clone alpaca TGF-β1 gene，investigate the expression of TGF-β1 in different tissues/organs，and examine the relationship between the TGF-β1 expression and alpaca coat color.The TGF-β1 cDNA was cloned from alpaca by 3′ and 5′ RACE.The sequence analysis was performed by DNAman，PROSITE and megAlign.The expression levels of TGF-β1 gene in different organs/tissues were analyzed by qRT-PCR.The full length of TGF-β1 cDNA in alpaca was 1 364 bp (GenBank Accession No.KF887499) with 1 173 bp ORF coding 390 amino acids.Sequence analysis showed alpaca TGF-β1 has typical structures of TGF-β1 family，showing high similarity with responsed proteins from other mammals.Phylogenetic analysis showed that alpaca TGF-β1 gene is much closed to wild boar(Sus scrofa).qRT-PCR showed that the relative gene expression level of TGF-β1 in brown alpaca was 3.5 times than that in white alpaca.TGF-β1 gene was expressed widely in tested tissues in goat while the expression levels were different in those tissues and highest in lymphoid tissue.Cloning and sequence characterization of alpaca TGF-β1 gene plus the expression analysis in different tisssues/organs indicate that TGF-β1 may play important role in alpaca coat color formation，provide the related DNA elements and experimental basic for further investigating on the molecular mechanism of TGF-β1 functions in animal coat color formation and regulation.

The present study utilized three staining assays (Annexin-V，mitochondrial membrane potential (JC-1)，and TUNEL) for flow cytometric analysis of apoptosis in sex-sorted sperm from four different bulls (A，B，C and D).Correlations between sperm quality and IVF efficiency were then assessed to determine which assay provided the best prediction of IVF efficiency.The results showed that: (1) the percentage of viable sperm (AN^-•PI^-) in X-sorted semen from bull A (89.30%±3.04%) was significantly higher than that from other bulls ((74.39%±3.24%)-(81.17%±4.25%); P<0.05)，while the percentage of DNA fragmentation sperm from bull A (12.24%±1.25%) was significantly lower than that from other bulls ((19.09%±1.27%)-(27.56%±3.17%); P<0.05); (2) the percentage of cells reaching the cleavage and blastocyst stages after in vitro fertilization (IVF) in sex-sorted sperm from bull A（ （76.82%±3.00%），（20.45%±1.44%）） were significantly greater than those from the other bulls ((22.00%±6.85%)-(31.88%±6.69%)，(3.33%±2.67%)-(4.00%±3.26%); P<0.05); (3) a significant positive correlation was observed between viable sperm and the percentage of cells at the cleavage or blastocyst stages (P<0.05) and a negative correlation was found between early apoptotic sperm and the percentage of cells at the cleavage or blastocyst stages (P<0.05).In conclusion，these results indicated that the Annexin-V assay was the most reliable technique for the prediction of the IVF success of sex-sorted bovine sperm.

Expression pattern of TR alpha in different developmental stages of porcine follicules，in vitro matured oocytes and early embryos were systemically investigated in the present study.The THs concentration of the different diameter sizes of pig follicular was determined by chemiluminescence immunoassay (CLIA) technique.And the localization of TR alpha in pig ovary were determined by immunohistochemical techniques.The expression pattern of TR alpha in pig oocytes and pre-implantation embryos were determined with immunofluorescence and QRT-PCR.The results showed that T3 concentration in pig follicles was rised at first and then declined following the increase of follicle diameter.TR alpha-immunoreactive substance was mainly distributed over granular cells，theca cells and oocytes．The quantity of the TR alpha-immunoreactive and TR alpha-immunoreactive cells were increased following the development of follicles．The results of QRT-PCR showed that TR alpha was expressed through out the process of in vitro matured oocytes，and the relative expression level in 1-cell，2-cell and 4-cell stages were significant higher than that of 8-cell morula and blastula stage.Immunohistochemical analysis results showed that TRa clearly expressed following the increase of follicles，and reached peak at 24-36 h.TR alpha protein expressed in different stages of early embryos，and its expression level was upregulated at 8-cell，morula and blastocyst stage.THs existed in the pig follicle fluid，TR alpha expressed in different stages of COCs and early embryos，THs may be affected pig folliculogenesis and early embryonic development by banding with the TR alpha.

The aim of this study was to reveal the effect of LPS on expression level of TLR4，and downstream inflammatory cytokines，IL-1β and IL-6 in hypothalamus，oviduct and ovary.36 healthy New Zealand female rabbits，4 animals were non-treatment as 0.0 h，16 animals were injected with LPS according to the dosage of 0.5 mg•kg^-1 body weight as LPS group，and other 16 ones were injected with saline (LPS carrier) as control group.The hypothalamus，ovary and oviduct were collected at 0.0，1.5 ，3.0，6.0 and 12.0 h after injection.The expression level of TLR4，IL-1β and IL-6 were analyzed by real-time PCR and the concentration of IL-1β and IL-6 were examined by ELISA.The results showed：TLR4 was expressed in all experimental tissue samples，but LPS induced the significant up-regulation of TLR4 mRNA only in the ovary (P<0.05).And LPS significantly induced the expression level of IL-1β and IL-6 in the hypothalamus，ovary and oviduct (P<0.05)，but the immunosuppressant of LPS had obviously different in three organs.Namely，immunoresponse in the hypothalamus response to LPS was more sensitive and the duration of immunoresponse was longer than in the ovary and oviduct.According to the results we speculate ：LPS immune stress can increase TLR4，IL-1β，IL-6 expression in the hypothalamus and also increase expression of IL-1β，IL-6 in the ovaries，and oviduct.

In order to find out the possible causes of the cryptorchidism in cloned pigs which produced by our research group.The RT-PCR techniques and bisulfite salt sequencing analysis(BSP) was used to analysis the expression and promoter methylation status of the promoter CpG sites of WT1 and FGF9 genes in the testis of cloned boars.The results showed that，the expression of both genes in cloned pig C1 and C2 were higher than those of control group N，expression of WT1 and FGF9 genes in cloned pig C1 was much higher (3.49，9.83 vs 1.00).Bisulfite sequencing PCR results showed that there were no significant change of WT1 gene methylation in cloned pig C1 and C2，and the methylation level of the both CpG inslands in FGF9 gene of cloned pig C1 was much higher than those in N（94.54% vs 18.18%，71.11% vs 26.67%）.The cloned pig C2 was not significant.Results indicated that the abnormal expression of WT1 gene may be one of the reasons for cryptorchidism of the cloned pigs，but the WT1 gene methylation had no relation on this abnormal expression.The change of DNA methylation reprogramming of FGF9 gene occurred in the testis of cloned pigs，which effected normal expression levels of the FGF9 gene，suggested that it may be an important factor for the occurrence of cryptorchidism in the cloned pigs.

In order to investigate the effects of Ghrelin on the development of early ovine embryos.Early ovine embryos were prepared by in vitro fertilization.The fertilized embryos were cultured in four groups of culture medium:①The group of adding gradiently 300，600，1 000，4 000，10 000 ng•mL^-1 Ghrelin;②The group of adding gradiently 1，10，200，400 ng•mL^-1 Ghrelin receptor antagonist;③The group of containing 400 ng•mL^-1 Ghrelin receptor antagonist and adding gradiently 300，1 000，4 000 ng•mL^-1 Ghrelin;④The control group.Embryo development was observed on the 2nd，3rd，4th，5th day，and the cleavage rates of ovine embryos were calculated at 2-cell，4-cell,8-cell and 16-cell stages.The results showed that: Compared with control group，ovine embryo cleavage rates had no significant change in the group adding Ghrelin (P>0.05)；Embryo cleavage rate had no obvious change at 2-cell，16-cell stages (P>0.05)，but decreased significantly at 4-cell，8-cell stages，and extremely significant difference at 200，400 ng•mL^-1 in the group adding Ghrelin receptor antagonist (P<0.01)；Embryo cleavage rates was recovered slowly on 4-cell，8-cell stages by gradiently adding Ghrelin but still significant difference in the group containing Ghrelin receptor antagonist(P<0.05).These results indicate: Ghrelin promote ovine embryo development at 4-cell and 8-cell stages.

The study was conducted to clarify the mechanism of low dietary cation-anion difference (DCAD) level preventing hypocalcemia in transition cows by investigating the influence of DCAD levels on blood Ca concentration and calcium binding protein-9kDa (CaBP-D9k) expression level in gastrointestinal tract of transition goats.Twenty-seven Guizhou Black goats in transition period were randomly allocated to 3 treatments with 9 goats each treatment and were fed 3 diets with varying DCAD levels：high DCAD (HD，+325)，control (CON，+179) and low DCAD (LD，-110) groups，respectively.Goats were blooded on d 10 before lambing，d 0 for lambing and d 3 after lambing，respectively，and were slaughtered to detect the plasma parameters and CaBP-D9k expression level of gastrointestinal tract.Compared with high DCAD，feed intake was unaffected by low DCAD treatment (P>0.05)，urine pH of goats fed LD was reduced with decreasing DCAD level (P<0.05).Compared to CON and HD,reducing DCAD level induced higher plasma Cl^- and Ca²⁺ concentration of goats during the experiment (P<0.05).Greater CaBP-D9k mRNA expression level was observed in whole gastrointestinal tract of LD-fed goats except for ileum，cecum and rectum compared with goats in HD group (P<0.05).Moreover，feeding LD diet relieved the decline of CaBP-D9k mRNA expression level in abomasum，jejunum and colon after lambing (P<0.05).In conclusion，reducing DCAD could up-regulate CaBP-D9k expression level in stomach，proximal intestine and colon，and kept Ca²⁺ concentration for transition goats.The results indicated that Ca²⁺ absorption in the above gastrointestinal tracts was increased and would be the possible mechanism of low DCAD enhancing blood Ca homeostasis in dairy cows during periparturient period.

This experiment was conducted to investigate the nitrogen apparent digestibility of different feeding levels and net protein requirement for maintenance of Dorper × Thin-tailed Han crossbred ewes in different physiological periods.According to the weight,fifteen ewes with synchronous estrus and artificial insemination were assigned into three groups，one group fed ad libitum(100%),and the other two groups fed eighty percents(80%) or sixty percents(60%) of ad libitum,and another nine non-pregnant ewes were assigned into blank control group.Digestibility trials was performed in the non-pregnant condition and 40th,100th,130th day of pregnancy and 20th,50th,80th day of lactation,to determine the metabolic parameters of nitrogen and net protein requirement for maintenance.The apparent digestibilities of nitrogen in the non-pregnancy and 40th,100th,130th day of pregnancy and 20th,50th,80th day of lactation were 45.21%-51.34%；59.71%-64.14%,69.98%-71.91%,67.34%-68.80%；73.72%-82.71%,72.79%-80.51%,73.57%-76.47%，respectively，which only in the non-pregnancy,mid- and late-lactation had significant differences (P＜0.01).The regression equations between nitrogen intake (N intake，g•kg^-1 W^0.75•d^-1) and nitrogen retention (RN，g•kg^-1 W^0.75•d^-1) in the non-pregnancy and 40th,100th,130th day of pregnancy and 20th,50th,80th day of lactation were RN=0.341 5NI-0.269 7；RN=0.489 5NI-0.303 7,RN=0.384 5NI-0.323 4,RN=0.457 4NI-0.496 2；RN=0.252 1NI-0.338 0,RN=0.282 1NI-0.313 6,RN=0.286 3NI-0.298 2(P＜0.01)，respectively.The net nitrogen requirements for maintenance of ewes in the non-pregnancy and 40th,100th,130th day of pregnancy and 20th,50th,80th day of lactation were 269.7,303.7,323.4,496.2,338.0,313.6 and 298.2 mg•kg^-1 W^0.75•d^-1 ，respectively.

The objective of this paper was to investigate the effects of three protein feeds (soybean meal，rapeseed meal and cottonseed meal) on quantities of ruminal proteobacteria (R.amylophilus，B.fibrisolvens,P.ruminicoal and S.bovis) adhered to solid fractions in Sichuan Xuanhan Yellow cattle.Four rumen-cannulated Xuanhan Yellow cattle with similar healthy and weight were each fed four diets with different protein feeds over a period in a 4×4 Latin Square design，which were basal diet(CN)，replaced 15% concentration of basal diet with soybean meal (SBM)，rapeseed meal (RSM) and cottonseed meal (CSM)，respectively.The PCR technique was used to analyze the change of the population of ruminal proteobacteria adhensive to solid fractions.The results showed that: 1)the quantities of R.amylophilus，B.fibrisolvens，P.ruminicoal and S.Bovis in group SBM reached the highest point at 9，9，5 and 5 h after morning feeding，the group RSM at 3，7，7 and 1 h，the group CSM at 9，9，7 and 1 h.2)the highest mean value was found in group SBM while the lowest in the group CSM on R.amylophilus(P<0.05) quantity; the highest mean value was recorded in group CSM while the lowest in SBM on the quantities of B.fibrisolvens，P.ruminicoal and S.Bovis (P<0.05) quantities.In conclusion，different protein feeds significantly affected the quantities of ruminal proteobacteria，and cottonseed meal had a better promoting effect on ruminal proteobacteria adhensive to solid fractions.

To identify a feasible insertion site in the foot-and-mouth disease virus (FMDV) and obtain a tagged recombinant virus，in this study，we have used reverse genetics to introduce a histidine (His)tag into the ninth site in the downstream of RGD motif in FMDV and obtain His tagged full-length-cDNA clone， named prAsia1-9His.We transfected the prAsia1-9His into BHK-21 cells and typical cytopathic effect (CPE) were found in the transfacted BHK-21 cells.Then，RT-PCR，indirect immunofluorescence assay and sequence analysis were performed，the results showed that the recombinant virus named as rAsia1-9His was successfully rescued and the His tag was stably maintained.Finally，we compared rAsia1-9His with the parental virus (named rAsia1) for pathogenicity to BHK-21 cells and suckling mouse，the results showed that they had similarity in the biological characteristics.The successful rescue of tagged FMDV lays a foundation for further study of molecular pathogenesis and marker vaccine of FMDV.

The current study was undertaken to study the infection status of EMCV from different pig breed.Modified “cell inoculation and RT-PCR” technology was used，and the 5 EMCV strains were isolated from local pig，domestic boar and improved breed pig.The full length genome of the EMCV strains were acquired and sequenced，and their molecular characteristic were analyzed.The results showed that the full-genome sequence of 5 EMCV isolates were 7724-7735 bp in length，and shared 99.3%-99.8% nucleotide identity each other，79.9%-99.9% identity with reference EMCV strains from different animal sources，and 99.4% with Chinese strains from pig and mice.Among all gene fragments，VP1 and 2A are easy to mutation，VP2 and 3D are most conserved.The phylogenetic tree based on the full length genome，ORF and VP1 gene sequences showed that the EMCV were divided into G1，G2 and G3 groups.The isolates from pig belong to G1 and G2 groups，the ones from mice distribute in G1 and G3 group，5 isolates strains in this study belong to G1 group with other Chinese reference strains.The results identified that the EMCV infection could cause severe clinical symptoms in local pig and domestic boar，which remind us of taking fully consideration of the zoonosis ecology in the course of Wild animal breeding activities.Mice may play an important medium role in incross infection of EMCV at the different pig breed.A big regional difference exists in EMCV and the transmission is limited in a range of area，moreover，some mutation may occur in EMCV infection to adapt new hosts.

The aim of this study was to study the expression and immunogenicity of H protein of peste des petits ruminants virus (PPRV).The H gene of PPRV Nigeria75/1 strain was amplified by RT-PCR，and cloned into the donor plasmid pFB-LIC-Bse.After being sequenced，the recombinant plasmid pFB-LIC-Bse-PPRV-H was transformed into competent cells of E.coli DH10Bac to get recombinant shuttle plasmid.The recombinant shuttle plasmid was then transfected into sf21 cells to get recombinant baculovirus.Western blot，IFA and immunity tests in mice were performed to identify the immunoreactivity and immunogenicity of the expression products.The recombinant H protein with molecular mass of approximately 68 kDa was detected with His tag monoclonal antibody and PPRV positive serum.The exprsssion products possessed a favorable immunogenicity and fall immunized mice could produce anti-PPRV neutralizing antibody.The cell immune responses were examined by lymphocyte proliferative assay using MTS method.The results showed that the rBacmid-H was able to express H proteins in Vero cells.Furthermore，specific antibodies were induced in the second week after primary vaccination.In conclusion，the recombinant H protein had good immunogenicity and immunoreactivity，which laid a foundation of developing a new vaccine against PPR.

This experiment was conducted to study the invasion and location in host cells of different virulent Mycoplasma hyopneumoniae.The ability of wild strain XLW-1 to invade PK15 cells at different infection time interval was investigated firstly by using indirect immunofluorescence technique and laser scanning confocal microscope.Then，the invasion to swine tracheal epithelial cells (STEC)，PK15，SJPL and 3D4/21 of different virulent M.hyopneumoniae strains was observed.The results showed that 1) M.hyopneumoniae wild strains XLW-1 can intrude into PK15 cells up to 10 h post infection and no internalization was observed before 8 h; 2) The amount of intracellular M.hyopneumoniae XLW-1 was increased as infection time; 3) M.hyopneumoniae wild strainsXLW-1，168 attenuated strains and vaccine strains all could invade STEC，PK15，SJPL and 3D4/21 cells; 4) The ingested wild strainsXLW-1，168 attenuated strains and vaccine strains were randomly distributed in the cytoplasm of STEC，and mainly distributed in the cytoplasm of long and narrow two polars and cytoplasm near cytomembrane in PK15; The internalized wild strainsXLW-1 and 168 vaccine strains principally concentrated in the cytoplasm close to medial surface of SJPL cells membrane，but 168 attenuated strains randomly interspersed in the cytoplasm of SJPL cells; The incursive wild strainsXLW-1 and 168 attenuated strains principally distributed in the cytoplasm close to inboard surface of 3D4/21 cytomembrane，and 168 vaccine strains randomly spreaded in the cytoplasm of 3D4/21 cells.These results demonstated that M.hyopneumoniae of different virulence all can invade host cells STEC，PK15，SJPL and 3D4/21.

Two H6N6 subtype avian influenza viruses (AIV)，A/Duck/Guangdong/C45/2010(H6N6) and A/Duck/Guangdong/C120/2011(H6N6)，were isolated from the live-poultry markets of Guangdong during 2010 to 2011.The sequence analysis of the hemagglutinin (HA) gene showed that the isolates were categorized as low pathogenic AIV，with the typical HA cleavage site of P-Q-I-E-T-R-G.The HA gene of the two isolates was closely related to that of A/Duck/Guangxi/GXd-5/2010(H6N1).The NA genes of the two isolates were closely related to that of A/Duck/Fujian/3242/2007(H6N6) and A/Duck/Fujian/6159/2007(H6N6)，respectively.The phylogenic analysis indicated that the eight genes of the two isolates belonged to the Eurasian avian lineage.The HA and NP genes of them belonged to ST2853-like lineage and HN573-like lineage，respectively.However，the other genes of them belonged to ST339-like lineage.Those indicated that the two isolates were recombinant viruses resulted from different genotype viruses.Therefore，we should focus on studying the pathogenic evolution and genetic variation of H6N6 subtype AIV to prevent and control the AIV.

The aim of this study was to clone and express the chicken β-defensin Gal-9 gene，and detect the antibacterial activity of the expressed product.The Gal-9 gene was amplified from chicken’s esophagus by RT-PCR，with primers based on the chicken Gal-9 (NM_001001611.2) sequence in GeneBank，then it was cloned into pET-32a(+) vector using the restriction sites of EcoRⅠ/XhoⅠ.The recombinant plasmid was transformed into E.coli BL21 (DE3) and induced at 37 ℃.The length of Gal-9 cDNA was 204 bp，and encoding 42 amino acid mature peptide.The product was confirmed to be about 25 kD in size by SDS-PAGE.It was mainly expressed in the form of inclusion body.The recombinant chicken Gal-9 protein showed bacteriostatic action on Staphylococcus aureus (ATCC 25923)，Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 25922)，and yeast (GS115).The minimum inhibitory concentration was 62.50，31.75，125.00，31.75 mg•L^－1.The results presented here demonstrate that the Gal-9 mature peptide gene，chicken β-defensin，was successfully expressed.The expressed protein showed broad-spectrum antimicrobial activity，which indicated the recombinant chicken Gal-9 protein could be potentially used for prevention and treatment.

Ninety chickens were divided into E.necatrix one time infection group, E.necatrix twice infection group,and control group.We researched dynamic change of HSP-70 expression in intestinal tissue of chickens infected with E.necatrix applying immunohistochemical method.The results showed that the HSP-70 expression in intestinal tissue of twice infection chickens were lower than the control chickens by different degrees at the 5th to 9th days after primer infection,the HSP-70 expression of duodenum,jejunum and caecum were significant or very significant lower than control chickens (P<0.05 or P<0.01),later rising rapidly,higher than control chickens at the 14th day and reached the peak；the HSP-70 expression in duodenum and jejunum of chickens showing a trend of significantly lower than control chickens at first,then gradually rising and higher than control chickens after the second challenge infection.But,the HSP-70 expression in intestinal tissue of E.necatrix one time infection chickens was higher than control chickens at the 6th,10th,15th day.These indicate that the primer infection have a certain degree of inhibition of HSP-70 expression in intestinal tissue of chickens,and the increase of HSP-70 expression is closely related with the degree of damage on the body with E.necatrix infection.This study provides important reference for the further study of pathogenic mechanism of coccidian.

 As an intestinal-specific trophic factor，porcine glucagon-like peptide-2 (pGLP-2) could stimulate intestinal mucosal growth and reduce the severity of injury specificity.In order to provide a potential therapeutic agent for intestinal dysfunction and damage，the protective effects of pGLP-2 for colonic injury were measured in dextran sulfate sodium (DSS)-induced colitis in mice.Eighteen BALB/c mice were randomly assigned to three treatments for 10 days，mice of DSS group were fed 3% (w/v) DSS through their drinking water for 10 days from first day of experiment，mice of DSS+pGLP-2 group were extraperitoneally injected with pGLP-2 daily from day 3 to day 9 and drink 3% DSS.The water group received water normally.Results were as follows：DSS-induced colitis clearly decreased body weight and relative expression of zonula occludens-1 in the colon，which was significantly reduced by treatments with 3% DSS-pGLP-2.Administration of pGLP-2 to DSS-mice preserved normal colonic mucosal architecture and significantly decreased myeloperoxidase activity，the relative expression of interleukin-6，interleukin-10 and interferon-γ which induced by DSS.These results showed that pGLP-2 could reduce the severity of colonic injury in murine colitis.The potencies of this product highlight its potential as therapeutic agent for intestinal diseases in piglet.

Solid dispersions of albendazole were prepared by melt technique using polyethylene glycol 6000 (PEG6000) and poloxamer 188 (P188) in combination (1∶2).When the ratio of albendazole (ABZ) and combined carrier was 1∶5，the dissolution characteristics of solid dispersion were evaluated through measuring the phase solubility studies and dissolution studies.The solid dispersion was investigated using X-ray diffraction analysis，Fourier-transform infrared spectroscopy and scanning electron microscopy.The results showed that PEG6000 and P188 were recognised as a better combined carrier when preparing solid dispersion of albendazole，and can significantly improve the in vitro dissolution rate to 90.48%.No chemical interaction was found between ABZ and combined carrier，and solid dispersion pattern is amorphous shape.These results indicated that as a combined carrier，the proper ratio of elected PEG6000 to P188 was 1 to 2，and preparation by melt technique for solid dispersion can significantly improve the dissolution of ABZ，and the preparation is simple.

Tetraogallus himalayensis is one of precious species of wild birds，which habitat mostly in the northwest plateau region at the altitude above 2 000 meters in China.In this study，the type and distribution of aerobic bacteria in the gastrointestinal tract of Teraogallus himalayensis was investigated by bacterial identification system combined with 16S rRNA sequencing analysis.Results showed that 37 strains of aerobic bacteria，categorized in 3 types，namely，Enterobacteriaceae，Staphylococcus and Enterococcus, were isolated and purified from the gastrointestinal tract of Teraogallus himalayensis by the method of differential medium culture.Following the identification，there were 20 strains of Enterobacteriaceae，7 strains of Staphylococcus and 10 strains of Enterococcus.The distribution of 3 types of aerobic bacteria in the gastrointestinal tract was counted for 21.62% in the crop，18.92% in the proventriculus，16.22% in the duodenum，18.92% in the ileum and 24.32% in the cecum，respectively.There were lack of Staphylococcus in the ileum and cecum.The characteristics of the types and distribution of the aerobic bacteria in the gastrointestinal tract of Tetraogallus himalayensis could be related to its specific eating property and habitat in plateau areas.The paper provides basic data for the gastrointestinal micro ecology study of Tetraogallus himalayensis and its domestication and breeding.