As a powerful tool，gene knockout technology has been widely used in biological and medical researches，and gradually utilized to other fields such as agriculture，livestock husbandry and veterinary science.Comparing to traditional homologous recombination，several novel techniques including RNA interference (RNAi)，zinc finger nuclease (ZFNs)，transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeat (CRISPR/Cas），which have been developed in recent years，can introduce delicate alteration and silencing in complex genomic DNA and RNA.These techniques are facilitating the disease prevention and trait improvement of big domestic animals.The improvement of these techniques and emergence of new techniques，and their application in domestic animals will push the development of livestock husbandry.This article is a brief overview focusing on the application of new gene knockout techniques include RNAi，ZFNs and TALENs in big domestic animals such as swine，bovine and caprine.

This study aimed to construct minicircle expression vector of AQP11 and observe the effect of transfection in Hela cells.AQP11 gene was amplified from the porcine intestinal mucosa and minicircle expression vector minicircle-AQP11 was constructed.The recombinant plasmid minicircle-AQP11 was transfected into Hela cells to investigate the expression of GFP by green fluorescent microscope.The sequencing results showed that the whole sequence of AQP11 gene containing a whole CDS was amplified successfully.Minicircle expression vector minicircle-AQP11 was constructed successfully.GFP was successfully expressed in Hela cells by the detection of bright green fluorescence for one week.Which provide new ideas for further elucidating the biochemical process and functional regulation of AQP11.

The present study was conducted to screen the anti-CSF virus-related genes and to analyze the genes expression under the different CSF-antibody levels in Landrace and Yorkshire.Agilent porcine 4×44K microarray was used to analyze and obtain the differentially expressed genes in four research groups:virus mimic PolyI:C transfected cells，DNA methytransferase inhibitor Aza-CdR transfected PK15 cells，PolyI:C and Aza-CdR transfected cells，mock cells.The differentially expressed genes were analyzed by cluster and gene ontology (GO).The results showed that significantly differentially expressed genes of myxovirus resistant （MX1） and the double-stranded-RNA-dependent protein kinase（PKR） mainly enriched in the biological process of defense response to virus.The expression levels of both genes in peripheral blood of Landrace and Yorkshire were detected under different CSF-antibody levels.The expression levels of two genes were significantly higher in both breeds with high antibody levels than that with non-immune levels (P<0.05).In addition，MX1 was highly expressed in Yorkshire peripheral blood than that in Landrace (P<0.01).Considering MX1 and PKR are important genes in interferon pathway，the results indicate that both genes can be served as potential candidate genes for defending CSFV infection.

Through investigating morphological changes and organ primordia formation of the porcine embryos，we want to lay the foundation for organgenesis study in pig，and for utilization of pigs as the model of human disease.Porcine embryos from day 7.0 to 18.5 postinsemination were collected to observe appearance of embryos by stereomicroscopy.Organ primordial of 14.0-18.5 d embryos were inquired using paraffin section and HE staining.The results showed that blastocysts had hatched from zona pellicuda at 7.0-7.5 d.The majority of conceptuses were filamentous，and the primitive streak emerged at 12.0-12.5 d.And embryos were in somite stage at 14.0-14.5 d.The data of tracking organ primordial were shown as follows.For renal formation，the pronephros appeared at 15.0-15.5 d，and the mesonephros emerged at 16.0-16.5 d and remained at 18.0-18.5 d.For brain development，the prosencephalon and the mesencephalon were clearly visible at 15.0-15.5 d.Four brain vesicles and telencephalon were observed at 18.0-18.5 d.For eyes formation，optic vesicle appeared at 15.0-15.5 d，and optic cup was showed at 18.0-18.5 d.For stomach and liver，the buds emerged at 16.0-16.5 d，and liver bud transformed into hepatic cord at 18.0-18.5 d.For heart generation，atrium and ventricle as well as valves were formed at 18.0-18.5 d.The results showed that bronchial tree was observed at 18.0-18.5 d.These results indicate that heart，liver，kidney，brain，lung，stomach，limb buds，etc.are beginning to form during 15.0-18.5 d.

The aim of this study was to clone the chicken Heme oxygenase 1 (HO-1) gene，predict its protein structure and function，and reveal its expression characteristics in tissues.In this study，Hailan White fowl (W-36) was used as experimental animal，the CDS region of chicken HO-1 gene was cloned by qRT-PCR，the protein structure and function were predicted by using bioinformatics，RT-PCR was used to analyze the expression characteristics in tissues.These results showed that the CDS of chicken HO-1 gene was 891 bp，encoding 296 amino acids.The conservation of its nucleotide and amino acid sequences were not high compared with other species.The protein of chicken HO-1 was transmembrane and non-secretory，it had a domain of the HO family，it is classified in EC1.14.99.3.It could bind to HEM，BLA，OXN，Q80，SUC，TRE and Na⁺ molecules，the HO-1 was expressed widely in tissues，and the expression level was higher in the brain，brain stem，artery，sciatic nerve，duodenum and small intestine.HO-1 protein is predicted that it can regulate the sugar and salt function，and is expressed widely in tissues，with important biological function in the circulatory system，nervous system and digestive system.

This experiment was conducted to explore the effect of combination of retinoic acid and Stra8 gene on male cESCs
differentiation.The sex of cESCs was determined by PCR to select only the male cESCs which divided into four groups，(1) both Stra8
transfected and RA-induced group，(2) only Stra8 transfected group，(3) only RA-induced group and (4) control group.Each group was repeated for three times.Cell morphology changes observations and qRT-PCR were performed to detect the effect of RA，Stra8 gene，and their combination on male cESCs differentiation.The results showed that there were small and round clones in the Stra8 transfected and RA-induced group in contrast to the other groups in which the cESCs were disintegrated.The expression quantity of stem cell marker genes was gradually declined in each group with extension of the incubation time.In the Transected Stra8 and RA induced group，the germ line related genes expressions were higher than that of the other groups (P<0.05) with the maximum amount at 6 d post induction and then gradually decreased.These results indicated that RA and Stra8 gene combination can promote the male cESCs differentiation to germ cell direction synactic that was proved by both significant cellular morphological changes and also the highly expressed germ cell marker genes.Through this induction system，it will not only establish a good foundation for germ cell induction and differentiation，but also provide basic idea for the late stage cESCs differentiation mechanisms.

In order to explore the ecological type difference between the same breeds in different areas，the ecological types of Jiaxian Red cattle in different areas was studied by the combination of microsatellite markers and morphological markers.The results showed that the 19 microsatellite loci were all with higher polymorphic and genetic parameters，which suggested that there was greater genetic variation within the breeds.The genetic parameters variance analysis indicated that the difference of genetic diversity was closely related to geographical distribution in each group.The Jiaxian Red cattle had the unique ecological types under the rich ecological conditions of Henan province.The Nei’s standard genetic distance and cluster analysis results showed that the genetic distance were minimum between the center areas and preservation field of Jiaxian Red cattle populations and had a close relationship，so the two populations were gathered together firstly，and then the two populations were aggregated with the edge region populations.The strong positive correlationship between geographic distance and genetic distance were found.The comparison between body size indexes indicated that the body size parameters were smaller in the conservation field population than that in the populations in central and edge areas，in which the central area was the biggest one.The geographical factors and production environment had an important influence on the body size indexes.The statistical results of hair color and angle type showed that each region had subtle differences in Jiaxian Red cattle populations in appearance，but all of populations had the intrinsic characteristics of the pure Jiaxian Red cattle.The results indicate that the phenotypic differences are consistent with molecular genetic basis of Jiaxian Red cattle in different areas，which prove the stability of the local ecological breeds.

This study aimed to clone bovine desmin gene promoter fragments of different lengths,and analyze its function.The bovine desmin gene promoter deletion sequences of different lengths were amplified by PCR,pGL3-desminpro dual luciferase expression vector transfected to skeletal muscle satellite cells and bovine fetal fibroblasts for activity detection was constructed.Meanwhile bovine MyoG and α-actin genes promoter expression vectors were transfected in order to compare the desmin,MyoG and α-actin genes promoter expression activity.The results showed that bovine desmin gene in the range of 132-2 106 bp promoter could start the expression of dual luciferase reporter gene at different degrees in bovine skeletal muscle satellite cells,and had muscle-specific.300 bp desmin gene promoter had the highest activity and higher than that of MyoG and α-actin genes promoter activity in bovine.Thus the 300 bp desmin gene promoter activity was higher and was adaptive to the muscle-specific promoter,which has laid an important foundation for the production of transgenic bovine.

The aim of present study was to clone the CDS sequences of buffalo YY1 gene，and screen its effective shRNA fragments.The cDNA of buffalo fetal fibroblast(BFF) was used as the PCR template，the CDS full-length sequence of buffalo YY1 gene was amplified by using touch down RT-PCR methods.The purified CDS fragment was inserted into pMD18-T vector，and the positive plasmid was identified and sequenced.Fusion expression vector of buffalo YY1 gene was constructed by inserting the YY1 CDS fragment into pEGFP-N1 vector，named as pYY1-EGFP N1.The pYY1-EGFP-N1 plasmid was transfected into BFF cells by Lipofectamine^® LTX reagent.Two shRNA fragments targeting buffalo YY1 gene were designed and synthesized，their lentiviral expression vectors were constructed，named as pSicoR-GFP-YY1 shRNA1/2 respectively.YY1 recombinant plasmid (pYY1-EGFP-N1) and shRNA lentiviral expression plasmid (pSicoR-GFP-YY1 shRNA1/2) were co-transfected into 293T cells by using Lipo-LTX reagent at the ratio of 1:1 or 1:6，pSicoR-GFP and pSicoR-GFP-1864 plasmid were used as blank and negative control respectively.At 72 h after transfection，the cells were harvested，and the total RNA was extracted.The expression of YY1 gene in each transfected cells was detected by qRT-PCR and Western-blot methods. The results showed that the CDS sequence of cloned buffalo YY1 was 1 248 bp，the nucleotide homology of YY1 gene between bovine and buffalo was 99%.The constructed pYY1-EGFP-N1 plasmid could transiently express in BFF cells.The results of qRT-PCR showed that the two designed shRNAs could inhibit the expression of buffalo YY1 mRNA，the shRNA2 fragment had significantly higher inhibition effect than shRNA1 (93.64% v 50.77%) (P<0.05).The results of Western-blot analysis confirmed that the two shRNAs had inhibition effect on gene expression of buffalo YY1.The above results indicated that the CDS sequence of buffalo YY1 was obtained，two effective shRNA fragments targeting buffalo YY1 gene were selected，which laid foundation for further study the role of YY1 gene in buffalo embryo development.

The purpose of this study was to identify trans-vaccenic acid (trans-11 C18:1，t-VA) hydrogenating bacteria in vitro from rumen of dairy cows and to analyze biochemistry characteristics and phylogenetic position.The t-VA-hydrogenating bacterium was isolated from the rumen of Holstein dairy cows based on enrichment culture under anaerobic conditions.The strains with high degree of hydrogenation were screened.The biochemical map were detected by gas chromatograph and the biochemical characterization of RV was analyzed by VITEK 2 compact system.The 16S rDNA，sodA，rpoB and recN genes were sequenced and used for phylogenic tree building.The newly isolate bacteria named RV with t-VA hydrogenating efficiency of 82.1% was isolated.RV was strictly anaerobic，gram-staining-positive，globose or oval shaped，and had optimum growth performance at 39 ℃and pH 6.0-7.8.The carbon or energy sources of RV came from D-xylose，D-galactose，D-maltose，D-mannose，pullulan，sacchrose，D-trehalose，D-cellobiose，etc.It contained some enzyme activities (e.g alpha-galactosidase and arylamidase) and resistant phylotpye to bacitracin，novobiocin，polymixin B.The gram-positive biochemical map showed that the similarity of RV reached 99% to that of pig intestine Streptococcus.The sequence of 16S rDNA of RV had high identity to Streptococcus equinus and Streptococcus bovis (99%-100%).The sequence of the funtional genes(sodA，rpoB and recN) had the hightest homology to S.bovis(99%，99% and 98%).One t-VA hydrogenating bacteria (a strain from S.bovis) named RV is isolated from rumen.To our known，this is the first study to find that S.bovis has t-VA hydrogenating activity.

This research aimed to study the mechanism of orally administered probiotics Escherichia coli（E.coli） Nissle 1917 regulating intestine barrier in weaned piglets challenged or unchallenged by enterotoxigenic E.coli Abbottstown.Twenty piglets with an average live body weight of (6.35±0.58) kg randomly assigned to 4 treatments:(1) Weaned piglets were provided a based diet (CON);(2) Weaned piglets were orally administered probiotics E.coli Nissle 1917 in the test (EcN);(3) Weaned piglets were challenged by enterotoxigenic E.coli Abbottstown in the pre-test (EcA);(4) Weaned piglets were challenged by enterotoxigenic E.coli Abbottstown in the pre-test and orally administered probiotics E.coli Nissle 1917 (EcN+EcA) in the test.Each treatment had one repeat and one repeat had one piglet.This experiment had a 3 d pre-test and 21 d test.Weaned piglets in the EcA and EcN+EcA groups were challenged by 5×10⁹ enterotoxigenic E.coli Abbottstown in the pre-test (EcA).Weaned piglets in the EcN and EcN+EcA groups were daily orally administered probiotics 1×10¹⁰ E.coli Nissle 1917 in the test.The results showed as follows:Compared with the CON group,orally administered 1×10¹⁰ probiotics E.coli Nissle 1917 could improve growth performance (P<0.05),decrease diarrhea ratio,increase the concentration of occludin (P<0.05),decrease the concentration of calprotectin (P<0.05),down-regulate the mRNA expression levels of Protein kinase R (PKR),Toll-like receptor-2 (TLR-2) and eukaryotic initiation factor-5A (eIF-5A) (P<0.05),and up-regulate the mRNA expression levels of Endothelial growth factor (EGF),Hepatocyte growth factor (HGF),Insulin-like growth factor (IGF-I),Trefoil peptide factor-3 (TFF-3) (P<0.05) in jejunal mucous membrane in weaned piglets unchallenged by enterotoxigenic E.coli Abbottstown;Compared with the EcA group,orally administered 1×10¹⁰ probiotics E.coli Nissle 1917 could improve growth performance (P<0.05),decrease diarrhea ratio,decrease serum concentration of DAO (P<0.05) and NO (P<0.05),increase the
concentration of occludin (P<0.05),decreased the concentration of calprotectin (P<0.05),down-regulate the mRNA expression levels of PKR,eIF-5A and TLR-2 (P<0.05),and up-regulate the mRNA expression levels of IGF-I,HGF,TFF-3 and EGF (P<0.05) in jejunal mucous membrane in weaned piglets challenged by enterotoxigenic E.coli Abbottstown.These results indicate that orally administered 1×10¹⁰ probiotics E.coli Nissle 1917 can protect greatly the intestinal barrier of early weanling piglets challenged or unchallenged by enterotoxigenic E.coli Abbottstown.

The aim of this study was to determine the role of gN gene of pseudorabies virus (PRV). The PRVs rPRV-gE^-/TK^- and Bartha K61 were selected as parental strains, and mutants with gN gene deletion were constructed by homologous recombination. PCR amplification and sequence analysis showed that the mutants with gN gene deletion (rPRV-gE^-/TK^-/gN^- and rPRV-Ba-gN^-) were constructed successfully. Compared with parental strain, growth speed of rPRV-gE^-/TK^-/gN^- was slightly slow in the early stage and its titer was not affected on PK15 cells, while both growth speed and titer of rPRV-Ba-gN^- were significantly lower on PK 15 cells. The LD₅₀ of both strains rPRV-gE^-/TK^-/gN^- and rPRV-gE^-/TK^- were more than 1×10⁶TCID₅₀, the virulence of rPRV-Ba-gN^- was attenuated compared with the parental strain Bartha K61. The two gN deletion mutants could provide higher antibody level and protection against virulent PRV challenge than the parental strains. The deletion of gN gene in
PRV may contribute to decreased virulence and increased immune protection.

To develop genetic engineering living-vector vaccine of Bovine viral diarrhea-mucosal disease virus (BVDV)， E₂ gene of BVDV Changchun184 strain was selected and the complete sequence was analyzed，then the epitopes of E₂ protein were predicted by bioinformatics software.Six pairs of primers were designed according to E₂ gene sequence，and the epitopes coding regions (1-297，1-345，1-374，45-297，45-345，45-374) of E₂ gene were amplified by PCR and cloned into pMV361 vector.PCR，restriction endonuclease digestion and sequence analysis proved that the 6 fragments were inserted into the expected position.It was indicated that the six prokaryotic expression plasmids were constructed.The positive recombinant plasmids were electro-transformed into BCG for expression at 45 ℃.The SDS-PAGE and Western blot results indicated that the recombinant proteins could react with polyclonal antibody against BVDV.Our study provides a basis for the further studies on genetic engineering living-vector vaccine of BVDV.

This experiment was conducted to study the pathogenicity of very virulent MDV strain rMd5 between three different local chicken lines (Ji-ning-bai-ri, Wen-shang-lu-hua, Shou-guang-hei) in China. rMd5 strain was inoculated to chickens by intraperitoneal injection, 1-day-old chicken with 500 PFU and 7-day-old chicken with 1 500 PFU. The feeding trial lasted for 90 days. rMd5 causes specific lesions of Marek’s disease in three different local chicken lines, but the degree of the pathogenicity differs significantly: the mortality and oncogenicity of Shou-guang-hei are significantly higher than the other two chicken lines, which is almost similar to SPF chicken. In addition, the transmission capacity of rMd5 in Shou-guang-hei is the highest; By contrast, the resistance of Ji-ning-bai-ri and Wen-shang-lu-hua lines to rMd5 infection and disease are greater. Therefore, Ji-ning-bai-ri and Wen-shang-lu-hua can be candidate lines to screen for genetic resistance genes and study genetic resistance mechanism.

In June of 2011，sick chickens from a commercial flock at Zibo in shandong Province were sent to our lab for inspection.The flock size is 80 000.Main species breed in the farm were Black-bone silk fowl (BSF)，Ma-chicken and Hy-Line chicken.The BSF and Ma-chicken were reared in the same house， 4 000 each.The chickens were 360-day-old， and showed emaciation， listlessness，lethargy，pale comb.The mortality rate was about 10%.Hy-Line chickens in another house were not infected.Six of sick chickens were tested by gross，histopathology，blood smears observation and RT-PCR.The sequences of isolated viruses were analyzed by software.Necropsy observation showed enlargement of the liver and kidney，fibrosarcoma in chest，intestinal atrophy and liquefaction，soybean size of tumor nodules in pancreas.Histopathology observation showed that myelocytoma presented in the liver and arterial，and aneurysm existed in the leg.Fibrosarcoma appeared on the pectorales in BSF.Erythroblastoma of liver，spongy hemangioma of lungs were showed in Ma-chickens.Blood smear observation showed the occurrence of myeloid cell-like tumor cells，the number of neutrophils was significantly increased.Pathogen detection showed that ALV p27 antigen and antibody were both positive.The two kinds of chickens were both ALV-J positive in detection of RT-PCR.One and two strains of ALV-J were isolated from BSF and Ma-chicken，respectively.Comparing with the reference strains，the homologies of gp85 protein amino acid sequence were ranged from 82.3%-91.1%，suggesting greater genetic variability.It indicated that different host cross infection might happen and promote the rapid evolution of virus when mixed chickens infected with ALV-J.

The objective of the present study was to investigate the distribution of fimbrial genes in some avian Salmonella serotypes and the expression differences of fimbrial genes under different bacterial culture conditions.Seventeen kinds of fimbrial operon genes in Salmonella pullorum，Salmonella gallinarum，Salmonella typhimurium and Salmonella enteritidis were detected by PCR，and expression of fimbrial genes in the above Salmonella strains under three different culture conditions which include static broth，shaken broth and static plate were detected by
RT-PCR.Results showed that the four Salmonella serotypes had different fimbrial genotypes，10 of which were conservative，and the distribution of the other 7 fimbrial genes was different.RT-PCR detection results showed that the expression of fimbrial genes in different
serotypes cultured under three conditions had some differences，as the expression of fimbrial genes in Salmonella pullorum and Salmonella enteritidis cultured on static broth was more than that of others (P<0.05)，while the difference in Salmonella gallinarum and Salmonella
typhimurium was not significant (P>0.05).The results revealed that the four Salmonella serotypes had different fimbrial genotypes and the bacterial culture condition significantly impacted the expression of fimbrial genes in Salmonella pullorum and Salmonella enteritidis.

Brucella，Chlamydia psittaci and Coxiella burnetii are three important zoonotic pathogens known to infect dairy cows.The aim of this study was to develop a multiplex PCR method for the simultaneous detection of them.Three genus-specific genes sequences were selected according to the GenBank，including Bp26 of Brucella spp., 23S rRNA of Chlamydia psittaci and IS1111a of Coxiella burnetii spp.And three specific primers pairs were designed respectively.After optimizing PCR reactions and amplification conditions，the multiplex method successfully discriminated Brucella，Chlamydia psittaci and Coxiella burnetii of dairy cows.The method had better specificity and repeatability，the detection sensitivity of simplex method for the three genes both reached 3.1×10² copies·reaction^－1，the detection sensitivity of multiplex method for these reached 3.1×10³ copies·reaction^－1.Using this multiplex method to detect 172 samples including bloods，serums，placentas and milk of different abortion cattle，53 Brucella positive samples，2 Chlamydia psittaci and 9 Coxiella burnetii positive samples were detected out，2 co-infection of Brucella and Chlamydia psittaci and 2 co-infection of Brucella and Coxiella burnetii positive samples were detected out else，no co-infection positive samples of the three pathogens was detected out.The above results indicated this method can be used for simultaneously，rapidly and sensitively detecting Brucella，Chlamydia psittaci and Coxiella burnetii of dairy cows.

The aim of this study was to investigate the role of the scavenger receptors on dendritic cells in presentation of recombinant VP1-VP4 of foot-and-mouth disease virus (FMDV) to T cells in vitro. The prokaryotic expression vector of pET-32a-VP1-VP4 was constructed and the VP1-VP4 protein was expressed and purified. Bone marrow-derived dendritic cells (BMDCs) pretreated with scavenger receptor inhibitors were pulsed with recombinant VP1-VP4 antigen and then cocultured with lymph node T cells. The culture supernatants were harvested at different time points for
determining IFN-γ contents with ELISA. The results showed that the IFN-γ levels of experimental groups were significantly higher than that of control group at different time points (P＜0.01). Our data indicate that scavenger receptors on BMDCs are able to recognize recombinant VP1-VP4 protein and play a negative regulatory role in antigen presentation of BMDCs to initiate lymph node T cell responses.

Histological and Immunohistochemical techniques were used to observe the distribution and quantity of immunocytes，T lymphocytes and its subpopulation，B lymphocytes，Mononuclear/macrophage cells and APCs，then to analyze the immune function and establishment of the crop on histological and cellular level in chicken.The results showed：The lymphoid-tissues are located in the lamina propria at the junction of the crop and the cervical and thoracic parts of the esophagus.A significant count of CD3⁺，CD4⁺，CD8⁺ T lymphocytes are evident in the diffused lymphoid tissues and the interfollicular areas around germinal center where Bu-1b⁺ cells are located in.Parts of CD3⁺，CD8⁺ cells and MHC-Ⅱ⁺ cells are infiltrated to the epitheliums of the mucus and the mucosal glands，which are transformed to the lymphoepithelium；Ma⁺ cells are slightly distributed in the Lamina propria submucosa muscular layer of mucosa and MHC-Ⅱ⁺ emerged in all of the tissues.The results showed that as the first storage place of food in chicken，the crop could be responsible for Moisturizing and softening food through mucus from mucus-producing oesophageal glands in lamina propria；as well as posses histological and cellular capacity of attendant mucosal immunity.The crop is an important part of gut-associated lymphoid tissue (GALT).

To investigate the antibacteria mechanism of Pulsatilla decoction, the Transwell system was used to establish the model of leukocyte migration induced by Lipopolysaccharide (LPS). The lysozyme concentrations and activity of the polymorph nuclear neutrophils (PMNs), which could pass through the rat intestinal mucosa microvascular endothelial cells (RIMECs) and not, were tested by the means of Agarose rocket electrophoresis and Fluorescence. The results showed that among the transendothelial migration of PMNs, the lysozyme concentrations of the “PD+LPS” group (compared with the control group) and the LPS group (compared with the PD group) significantly increased (P＜0.01), and their lysozyme activity also increased significantly (P＜0.05). There is no significant difference in the groups among the no transendothelial PMNs (P＞0.05). The results indicated that one of the antibacterial mechanism of PD is closely related with the protection against LPS and activation of transendotelial
migration of PMNs lysozyme.

To master the distribution regularity of Rex rabbit’s weight in different ages and seasons，and understand season’s impact on growth of Rex rabbits at different ages，growth data of Rex rabbits was allotted into 12×8 order matrix(season×age)，curve surface fitting tool of matlab2010a was adopted，multiple comparison between weights in different seasons was carried out.The results showed as follows：the obvious pit of weight distribution located between July to October and 91- to 180-day-old，difference between extreme weights at this stage achieved significant level (P<0.05)，and the difference of weight was up to 500 g approximately.And then，ridgy area of distribution was located in February to April and approximate 200-day-old，weight distribution present twin-peaks prior to approximate 56-day-old，and then single peak ；weight distribution of Rex rabbit born in different seasons demonstrated that rabbits born in summer gained maximal weight during 91 to 210 days，minimum weight was gained by rabbits born in the spring；daily gain surface distribution showed twin-peaks，which included endogenous growth and the compensation growth，peaks presented saddle-shape，the bottom of saddle was located in the summer.The first peak was presented in August，and the second peak was presented in June.The results indicate that the summer is the main season affecting the growth of Rex rabbits，and the suppression role almost involve in the whole age stages，winter mainly inhibit growth of rabbits before
56-day-old；rabbits own classical compensation growth ability，and the more serious impact of season on the first growth peak，the stronger compensation growth；Bivariate polynomial five times，which display better fitting effect，is suitable for growth prediction and exploration of the growth rule.

 This experiment was conducted to detect existence of GnRHR in caudal mesenteric ganglion (CMG) of female goat，and discuss the possible functional role of GnRHR in regulating reproductive activity through endocrine and autonomic nerve pattern.Immunohistochemical SP method was used to observe the distribution characteristics of GnRHR in CMG.The results showed that all the neurons in CMG were responded positively to GnRHR.The membrane and whole cytoplasm of neurons were strong positive stained with brown and the neurite was middle positive stained with yellow to GnRHR.Weak GnRHR immunoreactivity products distributed on passing nerve fibers and satellite cells.Image analysis documented a significant difference of GnRHR expression between neurons and non-neurons (P＜0.01).The results suggest that CMG’s neurons are the major targets of GnRH，which implies GnRH may regulate the function of reproductive organs through the GnRHR distributed in CMG.Therefore GnRHR may be a “transfer station” to coordinate reproductive activity through endocrine and autonomic nerve pattern.

In order to investigate the transcription of c-myc mRNA and iNOS mRNA in central nervous system in rats anesthetized with the Combined
Anaesthetic for Miniature Pigs (named as XFM), 30 rats were divided randomly into two groups: control group and XFM group. XFM group consisted of four subgroups: M₁ subgroup (rats received XFM intraperitoneally and then waiting the disappearance of right reflection); M₂ subgroup (rats received XFM intraperitoneally and then waiting for 1 h after the disappearance of right reflection); M₃ subgroup (rats received XFM intraperitoneally and then waiting the recovery of right reflection) and M₄ subgroup (rats received XFM intraperitoneally and then waiting for 1 h after the recovery of right reflection). The brain tissues were taken when reaching each time point. The transcription of c-myc mRNA and iNOS mRNA in central nervous system was detected with real time PCR. The results show that the transcription of c-myc mRNA in cerebral cortex and thalamus decreased significantly after receiving XFM (P＜0.01); The transcription of c-myc mRNA in cerebellum, hippocampus and brain stem increased significantly (P＜0.01). The transcription of iNOS mRNA in all brain regions decreased significantly after receiving XFM (P＜0.01). These results indicated that XFM could affect the transcription of c-myc mRNA and iNOS mRNA in central nervous system. This might be one of the anaesthetic mechanisms for XFM.