In order to investigate the effect of E3 ubiquitin ligase Nrdp1 and SOCS-1 genes on apoptosis during Brucella infection macrophages.The cell models of interference and over expression of Nrdp1 and SOCS-1 genes (pLL3.7-N1,pLL3.7-S1 and pLEX-Nrdp1,pLEX-SOCS-1) were constructed.Normal RAW264.7 cell,pLL3.7-N1,pLEX-Nrdp1,pLL3.7-S1 and pLEX-SOCS-1 group cells were infected with Brucella melitensis 16M (referred to 16M),the expression of Bax,Bcl-2,TNF-α genes were detected by qRT-PCR and apoptosis rate was detected by flow cytometry.pLL3.7-N1,pLEX-Nrdp1,pLL3.7-S1,pLEX-SOCS-1 cell model of the works best of interference and overexpression Nrdp1,SOCS-1 were successfully constructed and screened.After 16M infecting each group cells,compared with the control group,the expression of Bcl-2 and Bax mRNA in pLL3.7-N1,pLEX-Nrdp1,pLL3.7-S1 and pLEX-SOCS-1 groups were significantly different in different time periods (P < 0.05).The expression of TNF-α mRNA in pLL3.7-S1 group was significantly lower (P < 0.05),while the pLEX-SOCS-1 group was significantly higher (P < 0.05).Apoptosis rates in pLEX-Nrdp1,pLEX-SOCS-1 groups were significantly higher (P < 0.05),while pLL3.7-S1 group was lower (P < 0.05).The results showed that the Nrdp1 and SOCS-1 genes were closely related to apoptosis with 16M induction,the research laid the foundation for the study of 16M intracellular parasitism mechanisms.

The aim of this study was to screen effective shRNA fragments targeting pig zonula occludens-1 (ZO-1) gene,which would lay foundation to further explore its function in pig oocytes in vitro maturation (IVM) by RNAi technology.Pig ZO-1 gene was cloned and its eukaryotic expression vector was constructed,its effective shRNA fragments were screened.The results showed that the CDS length of cloned pig ZO-1 gene was 3 036 bp.The results of sequence multialigned showed that the sequence of pig ZO-1 gene shared 88%,88%,87% and 85% homology with Taurus,Ovis aries, Equns caballus and Homo sapiens,respectively.Clear RFP red fluorescent was observed in transfected cells with the pDsRed-N1-ZO-1 eukaryotic expression plasmid transfected into HEK293T cells by X-tremeGENE HP DNA transfection reagent.Two shRNA fragments targeting pig ZO-1 gene were designed and synthesized,weaker RFP red fluorescent was observed in co-transfected cells with the target and interfered shRNA plasmid groups.The Real-time PCR results showed that the two designed shRNAs could effectively inhibit the expression of pig ZO-1 mRNA,shRNA201 fragment had significantly higher inhibition effect than that of shRNA1276 fragment (77.8% and 67.0%,P < 0.05).

The study was conducted to explore the potential different characters between Blue-shelled chicken and White leghorn.Global genome microRNA was combined the identified microRNA with complementary lab-predicted microRNA.Then the two breed chicken's SNP data got by GGRS were mapped to the microRNA and focused on SNP that deliberately located in mature-microRNA.Bioinformatics method was adopted for target prediction on microRNA which had SNPs.By further gene enrichment analysis,the study found these genes enriched in 22 GO terms,10 KEGG pathways,and 3 IPA important networks.And they enriched in traits which associated with growth,such as mTOR signaling pathways,Wnt signaling pathways,growth hormone receptor networks and insulin-like growth factor Ⅰ receptor networks.And they also enriched in some laying traits,such as oocyte meiotic signaling pathways and progesterone mature oocytes signaling pathways.The methods and the results might provide references for further studies.

To investigate the changes of SUMO-1 (small ubiquitin-related modifier) gene in RAW264.7 cells infected by Brucella,a lentiviral vector for the over expression of mouse SUMO-1 was constructed.RAW264.7 cells was infected by Brucella 16M or M5-90,then quantitative Real-time PCR and Western blotting were used to detect SUMO-1 in RAW264.7 cells.SUMO-1 gene fragment was inserted into the lentiviral vector pLEX-MCS in the use of recombinant,resulting into a recombinant lentivirus pLEX-SUMO-1.Recombinant lentiviral vector was transfected into 293T cells after sequencing correctly,RAW264.7 was infected by supernatant.Quantitative Real-time PCR and Western blotting were used to quantify the transcription of SUMO-1 gene.The results showed that RAW264.7 cells was infected by Brucella 16M or M5-90,SUMO-1 showed a reduced trend in 12 h,after 12 h,SUMO-1 began to rise and became higher than normal levels,and the difference was extremely significant (P < 0.01).The sequencing results showed that mouse SUMO-1 gene lentiviral vector was constructed successfully.The mRNA expression of SUMO-1 gene increased.The results indicated that the expression of SUMO-1 changed after Brucella infection.Over-expression vector targeting SUMO-1 gene was constructed successfully,an experimental basis on the SUMO-1 functional study was provided.

In this study,hypoxia inducible factor-1α(HIF-1α) gene was cloned from yak spleen by RT-PCR,and it was divided into two sections for amplification.The sequence and protein structure were analyzed using related bioinformatics tools.The length of the coding region was 2 472 bp,which encoded 823 amino acids.Its molecule mass was 92.13 ku,and PI was 5.09.It displayed as a hydrophilic protein,whose secondary structure was mainly composed of random coil and α-helix.It had 69 phosphorylation sites and 4 N-glycosylation sites.Phylogenetic tree displayed that Maiwa yak,Bos grunniens,Bos mutus and Bos taurus gathered into one cluster.Maiwa yak HIF-1α gene was successfully cloned in this study,it provided a viable reference for further study of genetic characteristics of HIF-1α.

In order to investigate the effect of AIR on inflammatory reaction infected by Brucellamelitensis (16M), the AIR domain of Tecpr1 gene of murine macrophages RAW264.7 were knocked down (I-A), overexpressed (O-A) and reversed (OA-IA). Using the chlorine fluorescein (DCFH-DA) as a probe, we detected the variation of ROS production and mitochondria distribution by confocal laser scanning microscopy. We observed the expression changes of NLRP3, ASC and Caspase-1 by qRT-PCR and the expression changes of IL-18,IL-1β and Caspase-1 in host cells by ELISA. The results showed that 16M could stimulate RAW264.7 cells to produce ROS by time-dependent pathway, and I-A group and O-A group showed more abnormal accumulation of mitochondrial. The results of qRT-PCR and ELISA suggested that it had effect on the expression levels of NLRP3, ASC,Caspase-1 and IL-18, IL-1β and Caspase-1 in cells of different groups. Those results indicated that with AIR gene deletion, the release amount of ROS changed, mitochondrial clustered abnormally, and AIR was closely related to the activation of inflammasomes and induction of inflammatory reactions.

In order to establish an indirect ELISA to detect antibody of porcine epidemic diarrhea virus (PEDV).The experiment using the recombinant and purified truncated N protein as antigen expressed in E.coli BL21(DE3),the indirect ELISA was named rnPED-ELISA.The recombinant truncated N protein antigen showed no cross-reaction with the positive sera of other 7 kinds of swine diseases,CV%of intro-batch duplicativity test and inter-batch duplicativity test were less 13%;Sensitivity and specificity of rnPED-ELISA relative to SN were 93.33% and 90.00%,respectively;rnPED-ELISA compared with TSZ PEDV antibody diagnosis Kit,91.67% concordance was obtained.200 serum samples were detected by this method,the total masculine ratio was 69.5%.Therefore,this rnPED-ELISA based on recombinant truncated N protein antigen had good sensitivity and specificity,could afforded a simple and rapidmeans for assessment of vaccination in the field and investigation of PED epidemiology.

To establish a rapid assay for Listeria monocytogenes(LM) detection,a Real-time PCR method was developed targeting iap gene of LM.The results showed that the test for 15 bacteria strains,only LM was positive,indicated that the method had high specificity.In addition,the sensitivity of Real-time PCR was 6.5 CFU/mL.Stability and reproducibility of the test showed that the coefficient of variation for the same sample repeat the Ct values were less than 2%.Furthermore,a total of 3 positive samples for LM were detected from 139 clinical samples by the method,which was in accordance with the testing result by GB 478930-2010 standard detection protocol.Therefore,the Real-time PCR method provides a novel rapid,sensitive and good repeatability detection method for LM infection.

6 months old Yili foals were used as the experimental animal fed with different levels concentrate supplement to study the effects of fed with different levels concentrate supplement on digestion and metabolism,blood biochemical indexes and body weight gain by total feces and urine collection method,provide evidence for 6 months old Yili foals feeding.According to similar weight,20 weaned 6 months old Yili foals (all males) whose average weight was (145.28±1.69)kg were randomly divided into 4 groups named groups Ⅰ,Ⅱ,Ⅲ and Ⅳ,each group of 5 weaned Yili foals(all males).All Yili foals were fed with alfalfa 4 kg/d,basised on this,groups Ⅰ,Ⅱ,Ⅲ and Ⅳ were fed with 0.4,0.6,0.8,1.0 kg/d concentrate supplement.20 d digestion and metabolism experiment,comprised 16 d adaptation and 4 d collection.The results showed that as the amount of concentrate supplement feeding increased,nutrition intake,amount of nutrient digestion of DM,OM,CP,Ca,DE,ME,retention rate of N,Ca increased.There were no significant effects on the content of TP,GLB,ALB,and BUN in plasma of 6 months old Yili foals(P > 0.05),but the content of TP,GLB in plasma showed an increased trend,the concentration of BUN in plasma showed a decreased trend.The ADG of 6 months old Yili foals was gradually increased as the amount of concentrate supplement feeding increased.Therefore,increasing amount of concentrate supplement feeding of 6 months old Yili foals could increase the nutrient intake,digestion rate and retention rate,also could strengthen the body's immune ability,improved nitrogen deposition and the average daily gain.

Twelve lactating Holstein dairy cows were selected via a replicated 4 × 4 latin square design to determine the effect of feeding different levels of monensin on the production performance and nutrient digestion of lactating dairy cows.The test was divided into 4 period and each period was 28 d (7 d for preliminary feeding and 21 d for formal feeding).The cows in control group were fed with the basal diet (TMR,group A),and the diets of the three experimental groups were added with different levels of monensin (300 mg/d,group B;400 mg/d,group C;500 mg/d,group D) on the basis of the TMR diet,respectively.The results showed that the diets containing monensin increased the milk yield and lactose percentage,and reduced milk fat percentage.Compared with control group,the milk yield of group C and D were significantly increased by 4.08% and 4.57% (P < 0.05) respectively,the lactose of group D was significantly increased by 1.99% (P < 0.05),and milk fat of group D was significantly reduced by 7.45% (P < 0.05).However,adding monensin had no effect on 4% fat corrected milk (FCM),solids-non-fat (SNF) and milk protein (P > 0.05).In addition,adding monensin also significantly increased body weight gain (BWG),digestibility of crude protein (CP),neutral detergent fiber (NDF) and acid detergent fiber (ADF) in diet.The BWG of group D was significantly increased by 34.62% (P < 0.05),and the digestibility of CP in three experimental groups were respectively increased by 3.73%,4.35%,5.35% (P < 0.01),and the digestibility of NDF and ADF in group D was respectively increased by 7.24% (P < 0.05)and 9.08% (P < 0.05) compared to the control group.Moreover,the dry matter intake (DMI) and the digestibility of ether extract (EE),calcium (Ca) and phosphorus (P) were not significantly changed by monension (P > 0.05).The results suggested that supplementing 400 to 500 mg/d monensin with diet per cow was optimal for production performance,nutrient digestion and economic benefit of lactating dairy cows.

The experiment was carried out to investigate the effects of previously fermented juice (PFJ) on the nutrient composition and quality of stylo silage.The Stylosanthes guianensis Reyan No.2 were selected and there were six groups:Control group (to be silaged directly) and groups Ⅰ,Ⅱ,Ⅲ,Ⅳ and Ⅴ which were added with 1%,2%,3%,4% and 5% PFJ,respectively.All groups were stored for 30 days,and then the chemical composition and fermentation quality of the stylo silage were analyzed.The results showed that compared to control group,CP contents of groups Ⅰ,Ⅱ,Ⅲ and Ⅳ were significantly increased (P < 0.05) and NDF,ADF contents of groups Ⅱ,Ⅲ Ⅳ and Ⅴ were decreased in different degree.The RFV of groups Ⅱ,Ⅲ,Ⅳ and Ⅴ were significantly increased (P < 0.05).Compared with control group,the pH of the PFJ treatment groups were significantly decreased (P < 0.05),meanwhile acetic acid and propionic acid were decreased in different degree.Lactic acid contents were significantly increased (P < 0.05).The quality of stylo silage was poorer when it was silaged directly and the Flieg grades could be improved adding PFJ.In conclusion,adding PFJ could improve the fermentation quality and nutrient value of stylo silage and 4% PFJ was the proper application rate.

Effects of four kinds of diets on nutrient digestion and metabolism and blood biochemical indexes of one-year-old Yili horse were studied in this experiment.16 health horses(all males) with an average body weight of (227.38±16.48)kg and similar age (one-year old) were used in a staging and grouping experiment design.First stage had groupsⅠ and Ⅱ,second stage had groups Ⅲ and Ⅳ.The horses of group Ⅰ were fed with alfalfa 8 kg/d,pelleted concentrate 0 kg/d; The horses of group Ⅱ were fed with alfalfa 7 kg/d,pelleted concentrate 1 kg/d;The horses of group Ⅲ were fed with alfalfa 6.3 kg/d,pelleted concentrate 1.7 kg/d;The horses of group Ⅳ were fed with alfalfa 5.6 kg/d,pelleted concentrate 2.4 kg/d.Each stage lasted for 21 d (15 d pretrial period and 6 d trial period).The results showed that the digestibility of DM,OM,CP and P were increased with increasing of feed levels of pelleted concentrate,while the digestibility of NDF,ADF and Ca showed downtrend.DE and ME were significantly increased with increasing of feed levels of pelleted concentrate (P > 0.05).N retention and retention rate showed an increasing tendency,N retention and retention rate of groups Ⅲ and Ⅳ were extremely significant higher than those in group Ⅰ (P < 0.01).Ca retention and retention rate decreased with increasing of feed levels of pelleted concentrate,and Ca retention and retention rate of group Ⅰ were extremely significant higher than those in groups Ⅱ,Ⅲ and Ⅳ (P < 0.01).P retention was extremely significant increased with increasing of feed levels of pelleted concentrate (P < 0.01).There were no significant difference in the content of TP among all trial groups (P > 0.05),but the content of BUN in groups Ⅱ,Ⅲ and Ⅳ were lower than those in group Ⅰ (P > 0.05).Thus,comprehensive nutrient digestion and metabolism,blood biochemical indicators of four of kinds diets in one-year-old Yili horse,the requirement of crude protein,digestible protein,metabolizable energy,digestible energy were 14.28,9.16 g/(kg·BW) and 1.74,1.41 MJ/(kg·BW),respectively.

The aim of this test was to study muscle fiber characteristics and meat quality traits of Chinese Simmental cattle.Ten Chinese Simmental bulls were selected and longissimus dorsi,psoas major and semitendinosus were collected.Histological characteristics,pH,water holding capacity,shear force and texture profile analysis (TPA) were measured.The results showed that the diameter of these muscle fibers increased in the rank order type Ⅰ < ⅡA < ⅡB.The number and area percentage of type Ⅰ muscle fiber were highest in longissimus dorsi and that of ⅡB muscle fiber was highest in psoas major and semitendinosus.Longissimus dorsi and semitendinosus had lower pH,springiness,pressing lossing rate and higher shear force,hardness and resilience comparing with psoas major.In conclusion,muscle fiber characteristics could affect meat quality,especially textural properties of Chinese Simmental bulls.Increasing the percentage of type Ⅰ muscle fiber and decreasing ⅡB type could improve beef tenderness.

The experiment was conducted to investigate the effects of complex nutritional regulation additives on meat quality of grazing sheep in cold season.A single factor random block design was used,48 grazing ewes with 7 months old and the average body weight (28.01±1.70) kg were randomly divided into 4 groups:Group Ⅰ without supplement;Group Ⅱ with routine concentrate;Group Ⅲ with routine concentrate+complex formulation group 1 (0.30% malic acid,0.06% cysteamine,0.08% saccharicterpenin);Group Ⅳ with routine concentrate+complex formulation group 2 (0.60% malic acid,0.12% cysteamine,0.16% saccharicterpenin).And there were 3 repeats for each group with 4 grazing ewes in each repeat.At the end of experiment,the longissimus dorsi of ewes were collected to determine the content of meat nutrient and amino acids.The results showed that:① The crude fat content of group Ⅲ was higher than groupⅠ,Ⅱ and Ⅳ by 27.92% (P < 0.05),16.67% (P > 0.05) and 7.85% (P > 0.05),respectively.The crude protein content of group Ⅲ was higher than groupⅠ,Ⅱ and Ⅳ by 7.35% (P < 0.05),3.94% (P > 0.05) and 2.49% (P > 0.05),respectively.The shear force in three supplementary feeding groups were extremely significantly lower than that in the control group (P < 0.01) and compared with group Ⅱ,group Ⅲ was decreased by 3.98% (P < 0.05).② Compared with group Ⅰ and Ⅱ,the proline content of group Ⅳ was increased by 38.34% (P < 0.01) and 22.75% (P < 0.05),respectively.While the proline content of group Ⅲ was higher than groupⅠand Ⅱ by 19.63% (P > 0.05) and 6.15% (P > 0.05),respectively.Except for proline,other amino acids were not significant different among the four experimental groups (P > 0.05).In conclusion,in this trial,supplementing complex nutritional regulation additives had good effects on crude fat,crude protein and shear force and amino acids contents,also improved the flavor and nutritional value of grazing sheep in cold season.Taking into consideration various factors,the effect of group Ⅲ (supplementing routine concentrate+complex formulation group 1) was the most obvious,which had the practical guiding importance in improving the meat quality of grazing sheep in cold season.

The study was conducted to investigate the effect of VA and ZnMet on growth performance and part of antioxidant indices of weaning piglets.A total of 81 28-day-old three way cross(Duroc×Landrace×Yorkshine) healthy weaning piglets with an initial average body weight of (7.94±0.62) kg were randomly assigned into 9 groups with 3 replicates per group and 3 piglets per replicate.Factorial design of two factors and three levels was used in this study,the three levels of VA were 0,3 000 and 6 000 IU/kg,and the three levels of ZnMet were 0,300 and 400 mg/kg.The pre-trial lasted for 7 d and the trial lasted for 21 d.Piglets were weighed with an empty belly one by one on the 0,7 and 14 d of the test.Daily intake was recorded everyday.At the same time,the ADG and F/G were calculated.Three piglets without eating were randomly selected in each group for collecting serum samples at the age of 35 and 56 days.The results showed that:① compared with control group,supplementation of VA or ZnMet increased ADG significantly (P < 0.05),while had no influence on F/G (P > 0.05).In addition,ADG was significantly affected by the combination of VA and ZnMet (P < 0.05).VA and ZnMet had interaction effect on ADG,while F/G was not influenced by the combination of VA and ZnMet (P > 0.05).②At 35 days,compared with control group,both the activity of serum CuZn-SOD and GSH-Px increased significantly in experimental groups except the activity of serum GSH-Px in the group added with 300 mg/kg ZnMet;At 56 days,compared with control group,supplementation of ZnMet or VA could significantly improve serum CuZn-SOD activity of piglets (P < 0.05).Supplementation of both VA and ZnMet could significantly increase serum GSH-Px activity (P < 0.05),while supplementation of VA alone or ZnMet alone had no effect on GSH-Px activity (P > 0.05).The results indicated that supplementation of VA or ZnMet alone or the combination of them could promote the growth performance and antioxidant function of weaning piglets.

To study the effect of luteolin on blood indexes,liver and kidney in mice with acute mercury poisoning,28 mice were randomly divided into four groups:Control group (intraperitoneal injection 0.9% saline),luteolin group (lavage 100 mg/kg luteolin),mercuric chloride group (intraperitoneal injection 4 mg/kg mercury chloride) and mercury chloride+luteolin group (intraperitoneal injection 4 mg/kg mercuric chloride,lavage 100 mg/kg luteolin).The activities of ALT in serum,AST,CREA and BUN contents,blood WBC,RBC,HGB content and GSH and MDA contents of liver tissue were detected.Morphological changes of liver and kidney tissues were observed.The results showed that compared with the control group,the activities of ALT and AST of mercuric chloride group were extremely significantly increased (P < 0.01),serum CREA,BUN,blood WBC and liver tissue MDA contents were significantly increased (P < 0.05) while blood RBC,HGB and liver tissue GSH contents were significantly decreased (P < 0.05).Liver,kidney pathological changes were obviously.Compared with mercuric chloride group,the activities of ALT,AST in serum,CREA,BUN,blood WBC and liver tissue MDA contents of mercuric chloride+luteolin group were significantly decreased (P < 0.05),while blood RBC,HGB and liver tissue GSH contents were significantly elevated (P < 0.05).Liver,kidney pathological changes were attenuated obviously.The poisoning were characterized by inflammation and the occurrence of anemia when acute mercury poisoning occurred,liver and kidney showed different degrees of injury in mice.Luteolin could reduce the toxic effects of acute mercury poisoning on blood,liver and kidney.

Microencapsulation provides a physical barrier to the Lactobacillus against the harsh environmental conditions and gastrointestinal passage thereby increasing the number of viable cells reaching the small intestine,and it is also beneficial to storage,processing and transportation.Spray drying is a common technology used in microencapsulation process because it is a continuous and rapid process with low cost and high reproducibility.However,it can cause a decrease in microorganism numbers and activity due to high inlet and exit air temperatures and evaporation rates during the process,which has strict requirement for Lactobacillus in the selection of wall materials and preparation parameters.This article reviewed the research progress of spray drying for microencapsulated Lactobacillus,which contained the selection of microcapsule wall material,technological parameter and the evaluation method of microencapsulated Lactobacillus.It could provide the theoretical basis of preparation for microencapsulated Lactobacillus and its industrial production.

The experiment was conducted to study the optimal supplementary feeding quantity of grazing beef cattle in spring in Inner Mongolia grassland.Seventy-five Simmental beef cattle with average weight of (281.19±17.15)kg were selected and divided into 5 groups (group A,B,C,D and E) with 15 replicates per group.Those cattle grazed optionally and supplemented with 0,0.8,1.7,2.5,3.4 kg/d concentrate,respectively,according to beef cattle weight of 0,0.3%,0.6%,0.9% and 1.2%.Pre-trial period was 15 d,the trial period was 60 d.The results showed that,with the increasing of concentrate supplementation,the daily gain of beef cattle was gradually significant increased (P < 0.05);Group D were 331.25%,130.00% and 38.00% higher than that of group A,B and C,respectively,the apparent digestibility of dry matter and crude protein were significantly higher than that of other groups (P < 0.05).The daily income of group D was 11.22 yuan and were 183.33%,102.53%,36.17%,20.52% significantly higher than that of group A,B,C and E (P < 0.05),the fattening income of group D was the highest.According to above results,the optimum adding levels was 2.5 kg/d in spring under the condition of grazing in Inner Mongolia.The higher daily gain and higher economic benefits could be obtained under the optimal supplementary feeding quantity.

This experiment was designed to study the complex of Lactobacillus rhamnosus fermented herbal and Bacillus subtilis on White Feather broiler immunity performance and impact of Escherichia coli infection.360 one-day-old broiler chickens were randomly divided into 3 groups with 4 replicates in each group and 30 chickens per replicate.The pretrial period lasted for 7 d,and the experiment lasted for 35 d.The chickens in the group Ⅰ(positive control group) and group Ⅱ(negative control group)were all only fed a basal diet,group Ⅲ was test group,by additive 1% fermented herbal preparations,groups Ⅱ and Ⅲ were intraperitoneal injection of 1 mL E.coli at 35 d,broiler mortality,immune organ index,cecalmicroflora content,immunoglobulin levels,IL-2 and IL-6 content were tested.The results showed that injection of E.coli caused massive death of chickens,group Ⅱtook up to 75.00%,it was significantly higher than groups Ⅰ and Ⅲ (P < 0.05),the mortality in group Ⅲ was significantly lower than group Ⅱ(P < 0.05),was only 23.33%.Injection of E.coli maked spleen index and thymus index of group Ⅱincreased significantly (P < 0.05),the spleen index and thymus index of groups Ⅰ and Ⅲ were no significant difference (P > 0.05).E.coli counts was significantly decreased after injectionin group Ⅲ (P < 0.05),but the number of intestinal Lactobacilli of group Ⅲ was significantly increased (P < 0.05),and inhibited the propagation of E.coli,the counts of E.coli in groups Ⅰ and Ⅲ were no significant difference (P > 0.05).At 42 d,the sIgA of the intestinal fluid in group Ⅲ were higher than that of groups Ⅰ and Ⅱ with 11.99% and 36.56%,respectively(P < 0.05).The serum IgG concentrations of group Ⅲ was higher than that of groupsⅠand Ⅱwith 14.68% and 28.15%,respectively(P < 0.05).At 42 d,the IL-2 content of group Ⅱ was the lowest,it was significantly lower than group Ⅲ(P < 0.05),the IL-6 of group Ⅲ was significantly lower than group Ⅱ(P < 0.05).

The experiment was conducted from March 2015 until August 2015 at Bayan Dai cattle farm,Xinjiang Xin Sheng Yuan company in Yili.To study the influence of light music (cello music,violin music and bass music) on neurotransmitters levels and milk performance of dairy cows,36 health Holstein cows with similar body weight,age,parity and milk yield were randomly divided into two groups,control group and test group,each group with 18 heads.All these cows were managed at the same conditions.The test had three sections,each section lasted for 40 days.Every 20 days the daily yield was determined,and the blood and milk samples were collected.The results showed that all these three instrumental music could promote milk yield and the milk yield of violin music group were significantly higher than that of the control group (P < 0.05).Violin music significantly increased Gly level and reduced NPY level (P < 0.05).Bass music could significantly reduce NPY level (P < 0.05).The results suggested that violin music could be used as the environment music in dairy cows production.

The study was undertaken to study the inhibitory effect and mechanism of the water extracts of Holly Bark.Minimal inhibitory concentration (MIC) was tested through twice micro-dilution method and minimal bactericidal concentration (MBC) was tested by agar plate dilution counting method (SDA)of the water extracts of Holly Bark on producing extended spectrum β-lactamases (ESBLs) bacteria,then the bacteriostatic mechanism of the water extracts of Holly Bark through bacteria curve was tested,the cell ultrastrueture was observed,permeabilty changes of cell wall and membrane,nucleic acid synthesis inhibitory experiment were researched.The results showed that the water extracts of Holly Bark could affect bacteria curve compared with the blank control,destroy the cells seriously,permeability of both cell membrane and wall increased,extravasation of cytoplasm appeared,the content of AKP and soluble protein in the bacterial liquid increased,E.coli DNA fluorescence intensity weakened obviously.Research showed that the bacteriostatic mechanism of the water extracts of Holly Bark was based on its roles about changes on the cell wall and membrance permeability,inhibitting nucleic synthesis of bacterial.

The study was aimed to observe the effects of Codonopsis pilosula polysaccharides(CPPS) and selenium-Codonopsis pilosula polysaccharides₅(sCPPS₅) on the immune functions of immunosuppressive mice induced by cydophosphamide (CY).96 mice were randomly divided into 8 groups:Normal control group,CY model group,CY+CPPS (low,middle and high dose groups) and CY+sCPPS₅ (low,middle and high dose groups).Mice in normal control group and CY model group were lavaged with normal saline,the mice in other groups were lavaged with CPPS and sCPPS₅ by 0.2,0.4 and 0.6 mg/g,respectively.Administration was carried out once a day for 10 successive days.On the 11th day,mice were manufactured immunosuppressed by a three-day intraperitoneal injection of 0.04 mg/g CY,except for the normal control group.After administrated with drugs for 24 h,the immunological indexes of the mice were detected.The results showed that spleen and thymus indexes of CPPS and sCPPS₅ groups were significantly higher than that of CY group (P < 0.05).The A_{570 nm} when stimulated with ConA and LPS,serum IgG and IgM contents of CPPS and sCPPS₅ groups were significantly higher than that of CY group (P < 0.05) and sCPPS₅ possessed better efficacy than CPPS.In conclusion,CPPS and sCPPS₅ could strengthen immune activity of the immunosuppressive mice,but sCPPS₅ was better than CPPS and selenylation modification could improve the immune activity of polysaccharide.

The test was conducted to study the pharmacokinetics of compound traditional Chinese medicine preparation for locoweed detoxication in rats.After oral drug preparations,the absorption circumstances of active ingredients in vivo were detected by ultraviolet spectrophotometry and then the relative pharmacokinetic parameters were calculated based on identifying the effective components.The results showed that a suitable ultraviolet spectrum analytical method was established for 8 compounds at first.It had shown that the effective components were absorbed into the blood quickly after oral administration,and the maximum plasma concentration of each composition was got after about 1 h,indicating that the efficacy developed rapidly.The metabolic rates of Codonopsis pilosula saponin,polysaccharide and ferulic acid were fast,while the action time of the metabolites of glycyrrhizic acid,saikosaponin,astraloside and prim-O-glucosylc-imifugin were longer.The results suggested that pharmacolinetic process of this medicine preparation in rats basically conformed with two compartment model (expect the hesperidin).The determination method was simple,and had high accuracy and stability,as well as suitable for the pharmacokinetic study.

Glycyl-tRNA synthetase is an important member of aminoacyl-tRNA synthetase family.However,the knowledge about expression and regulation mechanism of GlyRS in the development of bovine mammary gland is limited.To reveal the relationship among GlyRS and bovine mammary gland development and lactation,EdU immunofluorescence and flow cytometry were used to analyze the impact of GlyRS on the number of DNA and the influence of the cell cycle of BMEC,and the expression of Cyclin D1 of BMEC by Real-time PCR and Western blotting.The results showed that the DNA synthesis of BMEC,the numbers of cells in S and G2/M phase,and Cyclin D1 expression levels in BMEC were all increased after BMEC was transfected with a GlyRS overexpression vector,whereas all of the above indexes were reduced after BMEC was transfected with a shRNA targeting against GlyRS.This study revealed that GlyRS accelerates cell cycle transformation,increases the number of DNA synthesis by upregulating the expression of Cyclin D1,thereby promoting BMEC proliferation.

To build up the method of isolating and culturing the dairy endometrial epithelial cells for further study, mechanical method and enzymatic isolation method were used in this study.0.1% pronase was used to soak uterine horns for 16 to 20 h at 4 ℃, and a surgical knife was used to scrape cells from dairy endoemtrium. The dairy endometrial epithelial cells were cultured by trypsin digestion method. The cultured cells were identified by keratin antibody through immunohistochemistry, and the proliferation of the third generation were measured by MTT assay.The results showed that morphology of the cultured cells were similar to epithelia cells by phase-contrast microscope. In addition, the eighth generation still maintained the similar morphology and growth state comparing to the primary generation cells, and the cytokeratin-positive cells ration was more than 80%. The methods used in this study were suitable for culturing the endometrial epithelial cells with highly quantity, activity and purity, it could be popularized in related labs.

The aim of the experiment was to study the effect of alcohol extract of Aster tataricus L.f.on the contraction of isolated guinea pig tracheal smooth muscle.The guinea pig tracheal smooth muscle was prepared by isolated tracheal thermostatic perfusion method.The effects of Aster tataricus L.f.on tracheal smooth muscle under the resting state and contraction stimulated by acetylcholine (Ach),histamine (His),CaCl₂,Ca²⁺ release and influx in cells without calcium were measured by BL-420F biological function experiment systems.The results showed that Aster tataricus L.f.could induce the contraction of isolated trachea at resting state at a low concentrations of 0.002 to 0.008 g/mL and exerted a relaxation on the isolated trachea when at the higher concentrations of 0.008 to 0.196 g/mL.All of the three concentrations of Aster tataricus L.f.could inhibit the tension stimulated by Ach,His and CaCl₂ which were antispasmodic agents,led to the non-parallelled rightward moving of cumulative concentration-response curve and the decline of maximal responses.The suppressive effect on His was the strongest,followed by Ach and CaCl₂,and the suppression could counteract the contraction induced by extracellular Ca²⁺influx significantly,and they were all expressed concentration-dependent manner.These results indicated the alcohol extract of Aster tataricus L.f.could relax tracheal smooth muscle by acting on M-receptor and histamine receptor and blocking Ca²⁺ passage,thus inhibiting extracellular Ca²⁺influx.

The study was aimed to establish the method of simultaneous detection of four kinds of tetracyclines,including oxytraoycline,tetracycline,chlortetracycline and doxyeycline in eggs by high performance liquid chromatography (HPLC).The eggs samples were detected with HPLC after through the procedure of extraction of 0.1 mol/L Na₂EDTA-Mcllvaine buffered solution,purification of Oasis MAX solid-phase extraction column,gradient elution of taking acetonitrile-0.01 mol/mL trifluoroacetic acid as mobile phase,separation with reverse liquid chromatography.The results showed that the calibration curves of four kinds of tetracyclines were linear correlated in the concentration range of 30 to 1 000 ng/g which the limits of detection was 10 ng/g and the limit of quantification was 50 ng/g.The average recoveries were 67.3% to 81.4%,relative standard deviations of intraday was 2.7% to 14.3%,relative standard deviations of inter-day was 0.9% to 5.0%.This mothod established in this study could simultaneously detect oxytetracycline,tetracycline,chlortetracycline and doxycycline drug residues in egg and it had high sensitivity,accuracy and precision.The method was rapid and simple to operate.

For optimizing in vitro maturation system of bovine oocytes,we firstly examined the influence of four different hormonal regimes(FSH+LH,HMG,FSH+LH+E₂ and HMG+E₂) on oocyte maturation rates.Then we studied the effects of epidermal growth factor (EGF) in the above defined medium on bovine oocyte maturation,in vitro development and quality of parthenogenetic embryos.The cell apoptotic index of parthenogenetic blastocysts was detected by TUNEL.No significant difference was observed in maturation rates in four groups supplemented with different hormones.However,human menopausal gonadotropin (HMG) provided steady maturation results in replicates.Maturation of oocytes was promoted by supplementation with 17β-estradiol (E₂).Combination of HMG and E₂ gave rise to steady and efficient mature results.The presence of EGF at 30 ng/mL concentration significantly increased maturation rate and blastocyst rate and reduced apoptotic cells in parthenogenetic blastocysts.Therefore,the optimal oocyte maturation solution could be supplemented with 0.075 IU/mL HMG,1 μg/mL E₂ and 30 ng/mL EGF.

Toll-like receptors (TLRs) is the main category of pattern recognition receptors.It is not only expressed in classical immune cells,but also expressed in the ovary and the genital tract of a variety of mammals.It plays an important role in ovarian activities.Biological functions of TLRs in the ovulation processes is the main research content in the field of reproductive immunology because that ovulation is a core event in ovarian activities and the key to determine the success or failure of reproductive.This review will concentrate on expression and distribution of TLRs in mammalian ovary,regulation mechanisms of TLRs expression and function,and the functions and significance of the ovulation process.Then we analyze briefly its possible functions in ovulation-related diseases.We want to provide a reference in research areas of reproductive immunology.

Mink suspected infection aleutian mink disease virus (ADV) from mink breeding areas in Liaoning province were tested with CIEP method.The mink with antibody to ADV were selected and culled.Liver,spleen,kidney and mesenteric lymph node samples were taken for pathological examination and the viruses were observed under electron microscope.The grinded tissue fluid filter was added penicillin and treptomycin and inoculated into CRFK cells and passaged by 6 times for virus isolation.And cells cultures were identified as ADV by PCR.Then they were inoculated into healthy mink.Three days later,the mink showed clinical signs,which including the loss of appetite,anemia,hair dull,antifeedant and binge drinking.Some minks showed neurological symptoms,manifested symptoms of convulsions,cramps,staggering gait,ataxia,or hind limb paralysis and died.The virus strains isolated and identified were named as the ADV-LN.

In order to provide a scientific prevention and treatment of porcine contagious pleuropneumia (PCP) disease in a pig farm of Tianjin,pigs with suspected PCP were carried on clinical comprehensive diagnosis and histopathological observations.A systematic analysis of isolated strain were conducted by laboratory diagnostic methods included bacterial isolation and culture,microscopic examination,biochemical tests,satellite phenomenon examination,PCR detection,animal pathogenicity experiments and drug sensitive tests.The most sick pigs were dyspnea,and the typical results of pathological anatomy and histopathological observations were fibrinous pneumonia.The gram-negative and polymorphous bacilli could be seen under the microscopy,its biochemical characteristics and satellite phenomenon were consistent with those of Actinobacillus pleuropneumoniae(APP).The amplified result of APP APX Ⅳ A gene was positive by specific primers.The isolated strain could be lethal to mice and it was highly sensitive to cefotaxime and florfenicol.Combined with the results of laboratory detection and the clinical comprehensive diagnosis,it was confirmed the isolated strain was APP.

In order to select probiotic strains with probiotic potential isolated from Tibetan sheep,fresh excreta of the healthy female Tibetan sheep was processed with 90 ℃ water-curing treatment for 5 min,and Bacillus with resistance were screened.The morphological,physiological and biochemical characteristics and 16S rDNA sequence of the Bacillus were comprehensively analyzed and identified.Biochemical characteristics were studied by using antimicrobial test,acid resisting tests and bile salt resisting tests,hydrolytic enzyme test and other methods.The results showed that the Bacillus SWUN-G43 isolated from fresh excreta of Tibetan sheep had a significant antibacterial effect on E.coli and Salmonella.Through analysis and identification,the Bacillus SWUN-G43 was Bacillus pumilus.Its survival rate was 50.43% under the environment of pH 2.0 and 44.35% under the environment of 0.3% bile salt.It could produce protease and cellulase and was sensitive to 12 kinds of commonly used antibiotics.By studying,the Bacillus pumilus(SWUN-G43) had a certain degree of resistance and potential probiotic ability.Besides,the strain didn't contain R-plasmid and would not cause horizontal transmission of antibiotic resistance in the intestinal microecosystem.It also had the potential to be developed as a micro ecological additive.

In order to understand the prevalence of pig viral diseases in Guangxi province,and provide evidence for disease prevention programs,the reseach about investigation of Guangxi swine infectious diseases was conducted.Total of 410 samples were collected form Guangxi in the 1 January 2013 to 31 December 2014.The porcine reproductive and respiratory syndrome virus (PRRSV),porcine circovirus (PCV),hog cholera virus (CSFV),pseudorabies virus (PRV),porcine epidemic diarrhea virus (PEDV),transmissible gastroenteritis of pigs virus (TGEV),porcine rotavirus (PORV),swine influenza virus (SIV) were detected by PCR and RT-PCR.The average infection rate were 35.12%,18.54%,1.17%,0.98%,10.00%,2.44%,0 and 1.22%,respectively.PRRSV and PCV-2 mixed infection rate was 6.83%.The PRRSV had high infection rates in the autumn and winter,and the average infection rate were 36.67%,45.31% and 63.64%,48.78%,respectively.The PCV-2 had high infection rates in the spring,summer and winter,respectively,which was 44.44%,25.00% and 11.29%,19.35% and 39.39%,14.63%.PRRSV and PCV-2 were the main pathogens in mixed infection.They were mixed infected each other or with CSFV,PEDV,PRV,SIV,HPS and SS.The prevention and control of PRRSV and PCV-2 will be the key point in the future in Guangxi province.

To study the characteristics of 2 low hemagglutinating activity H9N2 avian influenza virus (AIV) strains.2 strains were identified by HA/HI,electron microscope observation,HA and NA sequencing.The virus characteristics were studied by morphology,chemistry and biology.2 isolates were identified as H9N2 AIV.The observations demonstrated that the virions expressed various shapes of spherical,oval,elongated forms and polymorphism with diameters in a range of 80 to 120 nm.Hemagglutinin(HA) of the 2 strains was medium-stable to heat,and they were heat-resistant under 50 ℃.2 isolates could agglutinate cock and human O type red blood cells.The agglutination did not appear on the original isolates,but recovered with the passage of virus gradually.2 isolates did not agglutinate pig,cattle,sheep,rabbit and horse red blood cells.2 strains were not sensitive to the ether and chloroform,and only one isolate was sensitive to acids.2 strains were not fatal to the chicken embryo,and the EID₅₀ were both 1×10^-7/0.2 mL.The small White mice showed no clinical symptoms and no deaths after being inoculated with the viruses via respiratory route in our experiment.Serum antibody HI titers of the survived mice were measured after inoculated 14 d,and the antibody of H9N2 AIV were positive.Some tolerant H9N2 strains existed in live animal markets of Shanghai area.

In order to develop subunit vaccine of mink enteritis virus,the immunogenicity of mink parvovirus protein NS1 and VP2 had been evaluated.Two pairs of primers were designed,and the full-length NS1 and VP2 genes had been amplificated,and then prokaryotic expression vector pET-32a-NS1,PET-32a-VP2 were constructed.After the analysis of SDS-PAGE and Western blotting,target proteins had been purified by His-Bind affinity chromatography.The immunogenicity of purified protein NS1 and VP2 were evaluated by serum ELISA testing,after inoculated BALB/c mouse.The results showed that the molecular mass of NS1 and VP2 protein were 83 and 67 ku by SDS-PAGF and Western blotting;Although both target protein NS1 and VP2 had the ability to induce BALB/c mouse to produce anti-MEV specific antibodies,the level of antibodies induced by the protein VP2 was higher than protein NS1.Mink parvovirus protein VP2 was more suitable for the development of subunit vaccine.

The objective of this study was to investigate harmful gases concentrations of dairy cow houses in winter in the chill region of Hebei province.Three styles of dairy cow houses and four styles of calf houses in Bashang plateau in Hebei province were used in this study to determine NH₃ and CO₂ concentrations.The results showed that the CO₂ concentrations were extremely significantly higher in well-closed dairy cow houses than that with exercise yards (P < 0.01).And the CO₂ concentration in the evening was higher than that in the morning and at noon,reaching up to 4 481.7 mg/m³(dairy cow house 1).The CO₂ concentrations in all calf houses were high in the morning and evening and low at noon.Besides,the CO₂concentrations in all dairy cow houses and calf houses were extremely significantly or significantly higher than that in the net roads or exercise yards (P < 0.01;P < 0.05).The NH₃ concentration of three dairy cow houses showed a sharp rise trend after 19:00 and the dairy cow house 3 only detected the low concentration of NH₃ which ranged from 0 to 0.36 mg/m³.The NH₃ concentration of the calve houses were changing from 0.71 to 5.28 mg/m³.In conclusion,the harmful gas concentration related to the building types and structure that CO₂ and NH₃ concentration were higher when the cowsheds were sealed,even exceeded standard badly.To reduce the harmful gas content in the house,the ventilation should be taken into account to improve the air quality.

The economic traits of livestock has been significantly improved due to long-term under natural and artificial selection,and the specific variation characterizations emerged from the selected genome regions.As time goes on,the some polymorphism frequency of gene has dropped or disappeared,and keep in a group contains a single haploid type of multiple genes.This frequency variation of gene in specific region on the genomes is called the signatures of selection.Identifying signatures of selection can provide a straightforward insight into the mechaism of domestication and further uncover the casual genes related to the phenotypic variation.The high density SNP chips and large scale resequencing technology have been successfully applied to genomic selection signature in livestock breeds.The methods to detect selection signatures can be classed into three categories:Site frequency spectrum based methods,haplotyped based methods and population differentiation based methods.In the present article,we summarized the methods of selection signature detection and development and application of genomic selection signature methods in livestock.It will provide useful information for researchers working with breeding and evolutionary biology.

In order to establish the quality detection method of Qingying oral liquid,Flos Lonicerae,Fructus Forsythiae,Rhizoma Coptidis,Radix Salviae Miltiorrhiae,Raidix Scrophulariae and Ophiopogon jɑponicus in the prescription were identified by TLC,and the method of HPLC for the determination of chlorogenic acid was studied.The results showed that the TLC identification was highly specific,the spot was clear,and there was no interference in negative sample.The concentration of chlorogenic acid had a good linear relationship with peak area in the range of 8 to 160 μg/mL.The average recovery rate was 97.86%,and the RSD was 1.2%.In conclusion,the detection method established in this study was simple,accurate,specific and reproducible,which could effectively test the quality of the Qingying oral liquid.

To investigate the predicted regression model for beef carcass yield and percentage of Liaoyu White cattle,60 cattles were slaughtered and determined 10 indicators about beef carcass yield and percentage,including carcass weight (X₁),the 5th rib thickness (X₂),product of 5th to 6th rib eye width and height (X₃),5th to 6th rib eye area (X₄),the 12th rib thickness (X₅),product of 12th to 13th rib eye width and height (X₆),12th to 13th rib eye area (X₇),the weight of kidney,heart and diaphragm fat (X₈),hind leg circumference (X₉) and thigh muscle depth (X₁₀).Then correlation and regression analysis between 10 indicators and beef carcass yield and percentage were taken using SPSS 19.0.The results showed that the correlation were the highest between X₁ and beef carcass yield,X₈ and beef carcass yield percentage.Besides,there were two credible regression model for beef carcass yield of Liaoyu White cattle:① Beef carcass yield (product of 12th rib eye width and height)=0.809X₁+0.158X₆+0.857X₈-13.967 (R²=0.986);② Beef carcass yield (12th rib eye area)=0.779X₁+0.318X₇+0.933X₈-17.779 (R²=0.990).