Genome-wide association study (GWAS) is an effective method to detect complex diseases and traits genetics by the genetic markers in the range of the genome，based on association study.To find new SNPs，QTLs and candidate genes，researchers employ GWAS to study main traits in pigs.It is important to research on molecular genetics of pig breeding and study on related functional genes and QTNs.This review will provide reference for research progress of GWAS about important traits in pig.Furthermore，the paper introduces the research progress of QTN briefly.

Heterogeneous nuclear ribonucleoproteins (hnRNPs) is a family of highly conserved RNA-binding proteins，which widely exist in most kinds of tissues and cells.HnRNPs primarily participate in pre-mRNA slicing，mRNA transport，mRNA stability and post-transcriptional regulation in physiological condition.For the past few years，studies have found that hnRNPs could affect viral replication through multiple mechanisms，including binding with viral nuclei acid or viral protein，and regulating apoptosis of host cells.This paper reviews the progress in the study of the relation between heterogeneous nuclear ribonucleoprotein and virus replication.

To explore the pathogenesis of the rooster asthenospermia，the expression profiles of 6 candidate genes (COX7B，PTGDS，hPGDS，SPAG6，WNT2 and CCNF) selected from the previous digital gene expression profiling (DGE) and bioinformatics analysis，were analyzed.A total of 300 Beijing-You roosters of 42 weeks were detected for 4 weeks for semen quality including semen volume，sperm mobility，etc.Fifteen asthenospermia and 15 normal roosters were selected accordingly.Real-time quantitative PCR were performed to determine the expression of the 6 genes in the testicular of those roosters.The results indicated that COX7B and PTGDS were significantly high-expressed in the asthenospermia group (P<0.01)，while the WNT2，hPGDS and CCNF were significantly low-expressed in the asthenospermia group (P<0.05).However，the expression of SPAG6 gene was not significantly different in the 2 groups.These results were in accordance with the previous DGE analysis.The expression patterns of the 6 genes identified from DGE for asthenospermia in chickens were validated in the present study.They can be important candidate genes for further functional study and revealing the mechanism of chicken asthenospermia.

The aims of this study were to identify the main regulatory regions of the chicken Stra8 gene promoter，to predict the binding sites of the transcription elements，and to investigate the regulatory effects of Am80 and TSA on the chicken Stra8 gene promoter activity.A series of promoter missing mutants were directly subcloned into pGL3-Basic vector，and the recombinant plasmids were transfected into DF-1 and GC-1 cells.By detecting luciferase activity and predicting binding sites of regulatory elements of promoter regulatory region，the optimal promoter fragment was selected to construct recombinant plasmid Stra8-EGFP.Am80 and TSA were applied to screen the best induced concentration，Am80 and TSA with optimal dose singly and in combination were used to induce transfected cells，and the green fluorescence expression levels in induced groups were detected.The chicken Stra8 gene promoter fragment -1 055-+54 bp had stronger activity，and had the binding sites of RARα，RXRα and RARβ；Am80 and TSA singly and in combination were applied to induce the transfected cells，and the luciferase activity was highest in the cells induced by Am80 and TSA in combination.After the cells were transfected by Stra8-EGFP，the expression of green fluorescence were the highest in cells induced by Am80（10^-5 mol•L^-1） and TSA（10^-6 mol•L^-1） in combination.The activity and binding sites of regulatory elements of chicken Stra8 gene promoter were successfully analyzed，and combined actions of Am80 and TSA significantly could promote the activity of Stra8 gene promoter.

This study was conducted to study the effect of S.Enteritidis infection on early immune performance between two chicken breeds(the Beijing-You chicken(BJY)，a Chinese indigenous breed，and the White Leghorn(WL)，an imported laying type breed).40 BJY chickens and 40 WL chickens on 1-day-old were reared in isolators during the same period and were challenged by intramuscular injection with 8.7×10⁸ cfu•mL^-1 per chick(0.5 mL) of S.Enteritidis suspension，the concentrations of cytokines，antibodies and the S.Enteritidis colonization level in the blood and the expression of TLR4 and methyltransferase genes were determined in each group at 12，24，72 and 144 h after post-infection (12 hpi，24 hpi，72 hpi，144 hpi).The results showed that：（1）After challenged by S.Enteritidis，the two breeds showed significant differences in the serum antibody levels of IgY，IL-6，TNF-α，the antibody levels in BJY were higher(P<0.05)；（2） The SE colonization level in the blood of BJY was significantly lower than that of WL at 144 hpi (P<0.05)，while there were no differences at 12 hpi，24 hpi，72 hpi between two breeds(P>0.05)；（3） At 12 hpi the relative quantity of TLR4 in caecum in WL chickens was higher than that in BJY chickens，while at 24 hpi BJY chickens was significantly higher than that in WL chickens(P<0.05).There were significant differences for methyltransferase genes (DNMT1，DNMT3A and DNMT3B) expression in caecum between the two breeds(P<0.05).The results demonstrated that the two breeds differed markedly in immune traits after S.Enteritidis infection.BJY chicken was superior than WL chickens in considering of immune index.

The genetic diversity and genetic structure of Jinnan cattle，Jiaxian Red cattle，Luxi cattle and Qinchuan cattle were detected by microsatellite technology.Allele frequencies and distribution of 16 microsatellite markers were detected in 4 cattle populations.Number of effective alleles of the other 15 loci was from 2 to 8 except HEL9 as a monomorphic loci in all individuals，and the average number of effective alleles was 3.067 6.BM1818 was low genetic polymorphisms (PIC=0.083 0，PIC<0.25)，and the others were high genetic polymorphisms (PIC＞0.5)，with the highest PIC value at HUAT24 locus (PIC=0.727 5).80 alleles were identified in 4 populations (63 alleles in Jinnan cattle，65 in Jiaxian Red cattle，65 in Luxi cattle and 68 in Qinchuan cattle，respectively).The allele B at IDVGA46 was detected only in Jinnan cattle，and the allele B at TGLA44 was only lack in Jinnan cattle.Mean observed heterozygosity，mean expected heterozygosity and mean heterozygosity were 0.385 2，0.640 3 and 0.578 0，respectively in 4 populations.Jinnan cattle was independently clustered in the NJ tree and the genetic distance (D_A) with Luxi cattle，Jiaxian Red cattle and Qinchuan cattle were 0.809 3，0.759 8 and 0.807 1.There were special genetic characterics and sufficient genetic diversity in Jinnan cattle，and it was a relatively closed population in its evolutionary process.

To elucidate the roles of KiSS-1 and RFRP genes in goat，their expression profilings in tissues and the expression levels in hypothalamus-pituitary-ovary (and uterus) were determined between Jining Grey goat (year-round estrus) and Liaoning Cashmere goat (seasonal estrus) by using RT-PCR and qRT-PCR methods，respectively.The results showed that：(i) KiSS-1 mRNA was detected in goat hypothalamus，pituitary，pineal，kidney，ovary，subcutaneous fat and thyroid，while RFRP mRNA was detected in goat hypothalamus，cerebral cortex，cerebellum，ovary and pituitary，both of them had different tissue expression profilings between two goat breeds；(ii) KiSS-1 mRNA expression level was higher in hypothalamus for Jining Grey than that in Liaoning Cashmere goats (P<0.05)，while the expression had no significant difference in ovary and pituitary between 2 breeds (P>0.05)；(iii) The expression level of RFRP mRNA was higher in hypothalamus of Jining Grey than that in Liaoning Cashmere goat (P<0.05)，while its expression in ovary and pituitary was opposite between 2 breeds (P<0.05).The present study suggests that KiSS-1 gene may involve in regulating seasonal breeding in goats，but the effect (or the acting manner) of RFRP gene in goat seasonal breeding need be further researched.The results lay the foundation for elucidating the function of goat KiSS-1 and RFRP genes and provide reference for revealing the genetic mechanism of reproductive seasonality in goat.

This experiment was conducted to study the effect of follicle stimulating hormone(FSH)，luteinizing hormone(LH) on apoptosis and steroids hormone secretion of yak’s granulosa cells cultured in vitro，reveal the mechanism of gonadotropin regulating follicle development.In vitro culture system was established and the growth characteristics，apoptotic morphology were observed using cell culture and micro examination technology.The effect of FSH，LH on apoptosis and estradiol (E₂)，progesterone (P) secreting of yak’s granulosa cells cultured in vitro was analyzed using flow cytometry (FCM) and radioimmunoassay (RIA).The results showed that：Cultured yak’s granulosa cells demonstrated typical epithelioid growth characteristic and had “S” shape growth curve.Occasionally，the apoptotic morphological characteristics such as nuclear pyknotic and margination，endoplasmic reticulum loosen can be found in the cell culture dishes.With the increase of FSH concentration from 0.050 to 5.000 IU•mL^-1，the apoptotic rate decreased and the E₂，P level be on the rise.When FSH reached 5.000 IU•mL^-1，the apoptotic rate down to 7.3% which was very significantly from control group (P<0.01).The apoptosis of granulosa cells were inhibited and the E₂，P secreting were prompted when LH was added in a small quantity (0.050-0.500 IU•mL^-1)，on the other hand，high concentration of LH(5.000 IU•mL^-1) prompted the apoptosis of granulosa cells and inconspicuously effected the E₂，P secreting.These results indicated that FSH and LH played an important role in the regulation of follicle development and ovulation through adjust the apoptosis and steroids hormone secretion of granulosa cells in yak.

The purpose of this study is to investigate the effects of LDL from pigeon egg yolk on expression of HSP70 and apoptosis related genes in cold-shocked and frozen-thawed boar spermatozoa.This study divided into 5 groups：fresh sperm group，3% LDL（-80 and -196 ℃） treatment groups，9%LDL（-80 and -196 ℃）treatment groups.The addition of LDL with different concentrations has effect on biological features of the spermatozoa before and after cold-shock and frozen-thawed treatments ，which was analysed by CASA，and changes in expression of HSP70 and apoptosis-related genes were analysed and checked by SDS-PAGE，Western blotting and PCR.The results showed that：HSP70 expression was higher than that of fresh sperm after freezing，there was no significant difference between HSP70 expression of frozen-thawed stored at -196 ℃ with addition of 9% pigeon egg and fresh spermatozoa in treatment group（P＞0.05）；in the experiment of anti-apoptosis gene Bcl-x1，Bcl-21，gene expression in fresh sperm treatment group was higher than other treatment groups（P＞0.05）；in the expression of pro-apoptotic Bak gene，the expression of fresh sperm was the lowest，there was no significant difference between Bak gene of the 9% LDL(-196 ℃) treatment group and fresh spermatozoa in treatment group（P＞0.05）.Addition of LDL from pigeon egg yolk could reduce the damage caused by freezing and thawing，maintain biological features and improve the survival of the pig spermatozoa.

The purpose of this study was to establish a method for melanocytes culture and identification system in vitro，and to provide an ideal biological material for investigating the melanocytes functions.Peritoneum of Silky Fowl was collected and digested by dispaseⅡ and trypsin-EDTA mixed digestive solution under different digestive time.The pure melanocytes were obtained after continuous passages.Through the observation of cell morphology and cell growth curve showed that the primary melanocytes grew well under the specific medium.The cell specificities were further confirmed by the morphology，ultrastructure observation under electron transmission microscope，genes expression associated with melanin synthesis，and cell characteristic by DOPA stain and immunocytochemistry.The successfully cultured melanocytes sustained the normal biological characteristics on passage 10 demonstrating that the culture of primary melanocytes was accomplished under this in vitro system.All together，the in vitro culture system of melanocytes from Silky Fowl was successfully established in our study.

The aim of the present study was to determine whether STX4 is associated with coat color formation in mice. The expression and localization of STX4 in mice skins and melanocytes were investigated. Three different coat color skins, white, grey and black, were collected from the back of 6 white Kunming mice, the abdomen of 6 C57BL/6 mice and the back of 6 C57BL/6 mice, respectively. Melanocytes were cultured in vitro. The expression of STX4 in these skin and cell samples were analyzed by standard RT-PCR, quantitative real-time PCR, immunohistochemical staining and Western blot. The 897 bp sequence of STX4 CDS region was successfully amplified from 3 different coat color mice skins and melanocytes cultured in vitro by RT-PCR. Real-time PCR analysis revealed that STX4 was expressed in mice skins of all coat colors. STX4 showed the highest expression in black skin, which was 3.44 times higher than white skin. The expression of STX4 in grey skin was 1.92 times higher than white skin. Immunohistochemical analysis showed that STX4 was expressed in whole hair follicles in white and black skins including keratinocytes, as well as in melanocytes cultured in vitro. Western blot results showed positive STX4 bands in white, grey and black skin samples and melanocytes cultured in vitro. These results were consistent with the real-time PCR results. In conclusion, STX4 is expressed in mice skins, hair follicles, keratinocytes, and melanocytes, and the expression of STX4 increases as the coat color deepens. Therefore, it is speculated that STX4 is positively correlated to coat color formation in mice.

This study aimed to ascertain the differences of hypoxia adaptation between Yunnan plateau Wujin pig and Yuedawu pig.Blood samples of 30 Wujin pigs and 30 Yuedawu pigs were collected（Half male and female）on 60 days.The blood HGB，HCT，RBC，MCH，MCHC and oxygen saturation were determined.The gene expressions of HIF-1α ，VEGF and EPO were determined by real-time PCR in heart，liver，lung，kidney，jejunum，ileum，duodenum，skin，muscle，pancreas，testes and ovaries of Yuedawu and Wujin pigs（half male and female）.The results showed that HGB and RBC of Wujin pigs was significant higher than that of Yuedawu pigs(P<0.01)，and hematocrit (HCT) and MCH was lower than Yuedawu pigs.Oxygen saturation of Wujin pigs was higher than that of Yuedawu pigs，without significant differences(P>0.05).HIF-1α，VEGF， EPO mRNA were detected in various tissues of Yuedawu and Wujin pigs.The gene expressions of HIF-1α，VEGF and EPO in most tissues of Yuedawu pigs were higher than that in Wujin pigs，and main expression tissues for HIF-1α and VEGF were liver，lungs and pancreas in 2 pig breeds，whereas EPO was mainly expressed in kidney and liver，the trends of HIF-1α，VEGF and EPO genes expression in 2 pig breeds is consistent.In 2 pig breeds，the hypoxia adaptation gene expressions of sows were significantly higher than that of boar.The HGB，RBC and oxygen saturation of Wujin pigs was significantly higher than that of Yuedawu pigs.Physiological characterization parameters of hypoxia adaptation of Wujin pig were significantly higher than Yuedawu pig.Hypoxia adaptation gene expression in most tissues of Yuedawu pig were higher than that in Wujin pig.But the mechanisms of hypoxia adaptation need to be researched in the further.

This experiment was conducted to investigate the effects of low crude protein diet supplemented rumen-protected methionine (RPM) and lysine (RPL) on growth performance，nutrient digesitibility and serum biochemical indices in sika deer during the wintering season.Sixteen healthy 8-month-old sika deer were randomly divided into 4 groups with 4 replicates per group and 1 sika deer per replicate.Sika deers were fed 4 diets：Control group (GroupⅠ) fed 16.52% crude protein (CP) basal diet，tests group fed 13.70% CP diet with 0.30% RPL and 0 (Group Ⅱ)，0.08% (Group Ⅲ)，0.16%(Group Ⅳ) RPM.The results showed that average daily gain (ADG) of control group and group Ⅳ were higher than group Ⅱ，and average daily feed intake (ADFI) of group Ⅲ was lower than control group and group Ⅳ，and feed to gain ratio (F/G) of group Ⅱ was higher than other groups(P＜0.05)；digestibility of phosphorus (P) of control group was lower than other groups (P＜0.01) and group Ⅱ was lower than group Ⅲ (P＜0.05)，CP and ether extract(EE) digestibility of group Ⅱ were lower than group Ⅳ；serum urea nitrogen content of control group was higher than group Ⅲ (P＜0.05) and group Ⅳ (P＜0.01)，glucose and uric acid content of control group were higher than other groups (P＜0.01)；other index had no significant influence (P＞0.05).It is concluded that reducing diet CP level from 16.52% to 13.70% reduced nitrogen and phosphorus emission，slowed fawn growth，however，0.16% RPM and 0.30% RPL supplementation would enhance fawn growth，promoted nutrient digestion and metabolism balance.

The objectives of this study were to express the native structure of fusion protein (F) and hemagglutinin glycoprotein (H) of canine distemper virus (CDV).The F and H gene of lesser panda attenuated strain were amplified by PCR and cloned into pFastBac^TM1 vector.The recombinant plasmids were sequenced，and then were transformed into competent DH10Bac^TM E.coli cells to construct shuttle plasmids，rBacmid F and rBacmid H，by homologous recombination.The recombinant plasmids were then transfected into Sf9 cells to construct recombinant baculoviruses and the expression products of rF and rH were identified with IFA and Western blot.The expression of rF and rH in insect cells infected with recombinant baculoviruses were identified by IFA with dog anti-CDV hyperimmune serum and were detected by Western blot with mouse polyclonal antibody against F and H major epitopes of CDV.The molecular weight of expressed F and H protein were identified as 63 and 68 kD，which were consistent with the expected value.The results show that both envelope glycoproteins are successfully expressed in baculovirus-insect cell expression system and have a good immunoreactivity.Our study laid the foundation for the development of the CDV virus-like particle vaccine.

This study was undertaken to investigate the prevalence and molecular variation of infectious bronchitis virus in the northern area of China.Thirteen infectious bronchitis viruses (IBV) strains obtained from commercial chickens in northern China between 2012 and 2013 were isolated and identified by the sequencing of the entire S1 gene.The variants were distinguished by phylogenetic analysis and alignment in comparison with the vaccination-challenge tests that were performed using heterogeneous strains.The results showed that both the S1 and N gene of the prevailing IBV shared higher homologies.Furthermore，S1 gene shared nucleotide (amino acid) homology of 94.0%-100% (93.1%-100%)，and N gene had nucleotide (amino acid) 98.4%-99.1% homology (98.1%-99.8%)，receptively.Compared with the classical vaccine strain，there were at least three hyper-variable regions were found in S1 gene，and point mutations，deletions and insertions of IBV strains were also occurred at the genomic level，which were concentrated in the 53-150，and 250-290.Four amino acids were found missing in GM/13 at 62-65.Further analysis showed that the IBV epidemic strains shared highest homology （94.9%-97.2%） with A2 strains and the nucleotide (nt) mutation frequency averaged 2.4 nt×10^-3/year in view of S1 gene；while，the N gene was the LX4 strains of the homologous highest (93.0%-93.5%)，the average annual variation in frequency 5.1 nt×10^-3/year.Those findings suggest that the recent IBV epidemic strains may be recombinant of A2 and LX4 type strains，and N gene mutation frequency is greater than of the S1 gene.

In this study，non-coding RNA rli87 gene deletion strain of LM was constructed and identified，and growth characteristics of deletion strain was studied.The rli87 gene deletion mutant was constructed by fusion PCR method，and then pKSV7-Δrli87 shuttle vector was constructed，which was transformed into competent cells LM-SB5.Homologous recombination was conducted using temperature (42 ℃) and chloramphenicol (10 μg•mL^-1) resistance to achieve the rli87 gene deletion strain，and growth curves of wild strain and deletion strain were assayed at 37 ℃.The PCR and sequencing results confirmed that LM-Δrli87 was successfully obtained.PCR identification results showed that the deletion strain has good genetic stability through 25 generations of continuous passage.Comparison with the wild strain，there was no significant difference (P>0.05) in growth between deletion mutant and wild strain at 37 ℃.The results suggest that rli87 gene did not play significant role in regulatory growth under the conditions of 37 ℃.

The infectious status of echinococcosis in wild rodents was investigated at different areas of Alashankou Port.The wild rodent samples were collected from rural，suburb and urban area of Ala-shankou Port during 2013-2014.Then，pathological sections of liver were prepared and stained with H&E；the genomic DNA of livers and cysts were extracted.Cytochrome C oxidase 1 (Cox1)，NADH dehydrogenase subunit 1 (ND1) in Echinococcus mitochondria were amplified from the DNA and sequenced.Results were as follows：eight samples with cyst were found in the process of autopsy，and the positive rate was 2.08%.Ten positive samples were detected positive by PCR，and the positive rate was 2.60%.The analysis of Blast showed that 8 samples of our sequences were clustered with that of Echinococcus sp. (Accession No.JQ 690286.1)，and their identities were 85%.The remaining 2 samples of our sequences were clustered with that of Echinococcus multilocularis (Accession NO：AB 720065.1)，and their identities were 100%.The results of statitical analysis conveyed us the infection rates of echinococcosis at rural and suburb area were 2.74% and 2.38%，respectively，while no positive sample was detected at urban area.The results presented here demonstrate that the wild rodents at rural area of Ala-shankou Port showed the highest infection rate，and its susceptible host was the Rhombomys opimus.

This experiment was conducted to explore the pathological changes of the peste des petits ruminants (PPR) of sheep.On the basis of clinical diagnosis，then the six doubtful sheep of the PPR were definite diagnosed by RT-PCR detection technology.The pathological changes were observed by macroscopy and light microscope and special staining technique.The results showed that the specific N gene fragment of 350 bp of peste des petits ruminants virus (PPRV) were amplified and the PPRV was definite diagnosed as the pathogen of the six sick sheep.The anatomopathological changes showed there were a lot of yellow mucopurulent nasal discharge in nasal cavities of the sheep and there were also many secretions around the eyes.The ulcers of oral cavity mucous membrane were frequently observed.The sheep had pneumonia of different degree and there were numerous yellowish-white lesions from pinpoint-shaped to millet-shaped on the surface of the livers and spleen.The lymphoglandulae of the sheep were obviously swelling and showed serous lymphadenitis and hemorrhagic lymphadenitis.Hemorrhagic stripes，plaques and ulcers were observed on intestinal mucosa and especially on small intestinal mucosa.The histopathological changes indicated that a great number of multiple nuclear giant cells or syncytial cells were observed in lung，bronchus，liver，spleen and lymphoglandulae.The multiple nuclear giant cells or syncytial cells showed decentralized and concentrated distribution in the tissues and organs and there were eosinophil viral inclusions in their cytoplasm.Lung lesions of bronchointerstitial pneumonia were seen obviously in the sheep.There were numerous macrophages，lymphocytes in alveoli and interstitial tissue of the lungs.The parenchymal cells of the liver，kidney and spleen showed widely degeneration and necrosis.A great number of macrophages，plasma cells and lymphocytes were seen in the interstitial tissue of the liver，kidney and spleen.Those results confirmed the proliferation of multiple nuclear giant cells or syncytial cells and bronchointerstitial pneumonia were importantly histopathological changes of the peste des petits ruminants of sheep.

The aim of this study was to study the drug resistance of chicken isolated Escherichia coli and role of traditional chinese medicines in the drug resistance elimination.Twenty kinds of E.coli isolated from chicken around Shanxi province were tested to determine the resistant rate to 22 kinds of antibiotic by Kirby-Bauer way，the plasmid analysis and transformation test were conducted.Based on the resistance test results to antibiotic，we also evaluated the elimination effects of several traditional Chinese medicine (TCM) extracts on the isolated chicken E.coli.The results showed that the percent of E.coli resistant to above 5 kinds of antibiotics reached 90%.The isolated E.coli exhibited the strongest resistant rate to Penicillin G (90%)，followed by Enrofloxacin (85%)，while exhibited the highest sensitivity to Colimycin and Furadantin.According to the plasmid analysis results，12 kinds of plasmid patterns in total were found.Plasmids collected from the near farms with each other were likely to have the similar patterns，while different types among diverse areas.The transformation test results showed that one plasmid could encode single or several resistance genes，and different plasmids could have same resistance gene as well.TCM extracts，especially Scutellaria baicalensis，had significant elimination function to the drug resistant E.coli.Majority of the TCM treated plasmids maintain the same patterns，while two plasmids change their profiles after treating with coptis chinensis and shuanghuanglian，losing two DNA bind respectively.After all，TCM extract treatment recovered the susceptibility of chicken E.coli to drugs，which support the permitted use of TCM on the chicken E.coli disease control.

To study the efficacy of combined application of cefpodoxime proxetil and Origanum oil in nanoscale level，compound nanoemulsion of cefpodoxime proxetil was innovatively prepared.Furthermore，antibacterial activity tests in vitro and acute toxicity tests were conducted.Preliminary prescription was sieved on the base of traditional method of preparation of nanoemulsion.Then the multi-index orthogonal experiment was proceeded with L₉（3⁴） orthogonal test table，regarding bacteriostatic circle diameter and nanoemulsion stability constant Ke as examining index.Three elements，ratio of the mass fraction of cefpodoxime proxetil and Origanum oil，pH of aqueous phase and emulsifying temperature are selected as factors.Antibacterial activity tests in vitro and acute toxicity tests were done through trace broth dilution method and maximal tolerance dose test.Then histopathological examination was measured.The optimal prescription is composed of 0.33% of cefpodoxime proxetil，2.67% of Origanum oil，1.13% of 1，2-propylene glycol，22.51% of EL-40，73.36% of distilled water for enteropathogenic E.coli and Salmonella and pH value of nanoemulsion is 6.17.Compared with the active pharmaceutical ingredients，both of the minimum inhibitory concentrations and minimum bactericidal concentrations of test drug were significantly different（P<0.05）；the maximum tolerated dose of the oral acute toxicity of mice is higher than 1 800 mg•kg^-1，showing low toxicity.Histopathological examinations show no toxicity of kidney and small intestine，but can do pathological damage to the liver.When in the presence of Origanum oil，enteropathogenic E.coli，Salmonella，Proteus mirabilis，Pasteurella，Shigella dysenteriae，Staphylococcus，Streptococcus agalactiae are more sensitive to CFP-OR-NE than cefpodoxime proxetil.

 This study was designed to explore the impact of Oyster Polysaccharides on immune stress piglets of NF-κB signaling pathway-related genes A total of thirty Duroc × Landrace × Yorkshire castrated piglets of 28±1 d were randomly allocated into five groups，namely Blank control (Ⅰ)，Immunological stress control (Ⅱ) and Oyster polysaccharide Low (Ⅲ)，Medium (Ⅳ) and High (Ⅴ) dose group，with six replicates per group according to the principle of similar weight.Piglets were fed basal diet (Control) or 0.5%，0.8%，1.2% OPS (OPS Low，Medium and High dose group) for 30 days.The piglets were injected i.p with a dose of Escherichia coli LPS (100 μg•kg^-1 BW) except the Blank control which were injected with the same dose of normal saline.Three hours later，the liver，spleen，adrenal gland，lymph nodes and thymus were collected for detecting the relative transcription level of TLR-4，p50 and p300.Results were as follows：(1) Compared with the GroupⅠ，p300 relative transcript levels of GroupⅡ were significantly decreased in Lymph Node，while the other indicators were significantly increased in various organs (P<0.01).The relative transcript levels of TLR-4 in lymph nodes，p50 in adrenal gland and p300 in liver，adrenal gland and lymph nodes were significantly decreased (P<0.01) while the relative transcript levels of p50 in adrenal gland and p300 in liver，adrenal gland and lymph node were significantly increased (P<0.01) in Group Ⅲ than GroupⅠ.The relative transcript levels of TLR-4 in lymph nodes and spleen，p300 in liver，adrenal gland and lymph nodes were significantly decreased (P<0.05 or P<0.01) while the relative transcript levels of TLR-4 in liver，adrenal gland and thymus and p50 in liver，thymus were significantly increased (P<0.05 or P<0.01) in Group Ⅳ than GroupⅠ.The relative transcript levels of TLR-4 in spleen，p300 in liver，lymph nodes，thymus and adrenal gland were significantly decreased (P<0.05 or P<0.01) while the relative transcript levels of TLR-4 in liver，adrenal gland and lymph nodes and p50 in liver，thymus and adrenal gland were significantly increased in Group Ⅴ than Group Ⅰ(P<0.05 or P<0.01).(2)Compared with the Group Ⅱ，the relative transcription of TLR-4 levels were significantly decreased in these organs of Group Ⅲ，Ⅳ and Ⅴ (P<0.05 or P<0.01)other than in lymph node of Group Ⅴ (P>0.05).The relative transcription of p50 levels were significantly decreased in adrenal glands，lymph glands and spleen of Group Ⅲ，Ⅳ，and Ⅴ(P<0.01)；and were significantly decreased in liver and significantly increased in thymus of Group Ⅲ(P<0.01)；and that were significantly increased in thymus of Group Ⅴ (P<0.01).The relative transcription of p300 mRNA levels were significantly decreased in liver，adrenal glands，thymus and spleen of Group Ⅲ，Ⅳ，and Ⅴ(P<0.01)；and that were significantly increased in lymph nodes of Group Ⅳ (P<0.01).Oyster polysaccharides can relieve stress piglets transcription of NF-κB signaling pathway-related genes.

The experiment was conducted to study the effects of se-enriched Ligustrum lucidium Fruit on proliferation，apoptosis and antioxidant activity of mammary epithelial cells cultured under heat circumstance in vitro.The heat stress model temperature was set as 42 °C and the temperature 37 °C was set as the control.In the experiment，heat-stressed mammary epithelial cells were cultured with different concentration of water extract，protein and oleanolic acid extract of Se-LLF.The growth and apoptosis rate of cultured mammary epithelial cells were detected by MTT and flow cytometry，respectively，and the content of SOD in the supernatant fluid of cultured mammary epithelial cells was detected by xanthine oxidase.The results showed that the proliferation of mammary epithelial cells cultured under high temperature was significantly promoted in each experimental concentration of water extract，protein and oleanolic acid extract of Se-LLF (P<0.05).The apoptosis rate decreased significantly (P<0.05)，the activity of SOD (P<0.05) was increased significantly，and the results were better than those of the ordinary Ligustrum lucidum group.In conclusion，water extract，protein and oleanolic acid extract of Se-LLF can not only improve the proliferation and apoptosis rate of heat-stressed mammary epithelial cells，but also improve their antioxidant capacity.

The objective of this study was to investigate the changes of the contents of nitric oxide (NO) and endothelin-1 (ET-1)，and the related items in lung of broilers with ascites syndrome (AS) and the prevention and treatment of the compound recipe of the traditional Chinese medicine on AS.A total of 316 14-day-old Ross broilers were randomly divided into normal group (N，bred with a standard diet at room temperature)，control group (C，bred with high-energy diet derived from standard diet added with 3% lard and 4% fish meal and 0.12% Na⁺ in drinking water at 9-11 ℃ to induce AS)，compound recipe of the traditional Chinese medicine groups (T1，T2，T3，bred in the same way as group C except a dose of 2，1，0.5 mL•kg^-1 bw of the traditional Chinese medicine everyday added in drinking water，respectively)，and L-arginine group (L，bred in the same way as group C except 1% L-arginine added in diet).The results showed that the broilers in group C presented the typical clinical symptoms of AS at 35 and 45 days of age.Compared with group N，the incidence of AS，ascites heart index (AHI)，the contents of NO and ET-1，iNOS activity and the relative expression amount of ETAR mRNA in lung of broilers at 35 and 45 days of age in group C were increased significantly (P<0.01 or P<0.05)，and cNOS activity and the ratio of NO to ET-1 were decreased significantly (P<0.01) in group C.The clinical symptoms and items mentioned above were all improved in groups of L，T1 and T2 compared with group C (P<0.01 or P<0.05).The study showed that the increases of NO and ET-1 content，ETAR mRNA relative expression amount and iNOS activity and the decrease of cNOS activity in lung of broiler induced the unbalance of NO and ET-1 which participated in the pathogenesis of AS.The studied compound recipe of the traditional Chinese medicine with the role of tonifying qi-blood and activating qi to excrete water can prevent and treat AS of broiler effectively.

In order to study the action mechanism of Allium Mongolicum Regel Flavonoids(AMF) on immunological enhancement activity，the effects of AMF on lymphocyte proliferation and mRNA expression of IL-2，IL-4 and IFN-γ were determined by MTT method and real-time fluorescent quantitation PCR assay.The peripheral blood lymphocyte of sheep was used in this study.The results showed that AMF could significantly enhance lymphocyte proliferation.AMF could significantly up-regulate mRNA expression of IFN-γ，down-regulate mRNA expression of IL-4，and have no significantly effect on IL-2 mRNA expression.The result indicate that AMF has immunoregulatory activity and could enhance lymphocyte proliferation.

The aim of this study was to construct and characterize of a naive Camelus bactrianus variable domain of heavy chain of heavy-chain antibody（VHH）yeast two-hybrid library.The blood，bone marrow，spleen and lymph node were collected from a healthy non-immunized 6 month-old female Camelus Bactrianus. And peripheral lymphocytes were isolated from blood samples using Ficoll Paque PLUS reagent.Then total RNA was extracted from peripheral lymphocytes，bone marrow，spleen and Lymph nodes.Reverse-transcription was performed to generate the first-stand cDNA.VHH fragments were amplified through two rounds PCR using two pairs of specific primers.Naive Camelus Bactrianus VHH yeast two-hybrid library was successfully constructed according to Clontech Mate & Plate library construction system user manual.Briefly，5 μg VHHs fragment containing homologous arms in 20 μL were co-transformed with 6 μL linearized pGADT7-Rec plasmid into component Y187 yeast cells，and transformation products were plated on SD/-Leu plates.After 3-5 days incubation at 30 °C，the library was harvested and the library quality were evaluated by titer，recombination efficiency，diversity and complexity.The results showed that there were 2.07×10⁷ independent clones in the naive VHH yeast two-hybrid library and the library titer was 7.6×10⁸ cfu•mL^-1.PCR results showed that 43 independent clones inserted VHHs fragment in 47 randomly picked out clones，and the recombination efficiency was 91.5%.Nineteen independent clones were random picked out to sequence and sequence results indicate they all share unique sequences.In a conclusion，a naive Camelus Bactrianus variable domain of heavy chain of heavy-chain antibody yeast two-hybrid library was successfully constructed with large capacity，good diversity and complexity，which laid the foundation for preparation VHHs against interested antigens.