Lipoprotein lipase (LPL) is produced by parenchymal cells from mainly adipocytes and myocytes, but it is involved in hydrolysing triglycerides in plasma lipoproteins at the capillary lumen. For decades, the mechanism by which LPL reaches its site of action in capillaries was unclear, but the problem was recently solved. Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 (GPIHBP1) picks up LPL from the interstitial spaces across endothelial cells to the capillary lumen. When GPIHBP1 is absent, LPL is mislocalized to the interstitial spaces, which lead to severe hypertriglyceridaemia. Some cases of hypertriglyceridaemia in humans are caused by Gpihbp1 mutations that interfere with the ability of GPIHBP1 to bind to LPL. Here, we review recent progress in understanding the role of GPIHBP1 in health and disease and discuss some remaining unresolved issues regarding the processing of triglyceride-rich lipoproteins.

 The aim of this study was to study the genetic diversity of PRLR gene in Hu sheep to discuss its effect on Hu sheep maternal behaviors and provide the genetics analysis method for evaluating sheep maternal behaviors. 81 parturition ewes were used to detect single nucleotide polymorphism of PRLR gene exon10 by using PCR-SSCP, in which 63 individuals were observed for maternal behaviors. The methods of normal distribution test, cluster analysis and nonparametric test were used to reveal the Hu sheep maternal behavior distribution rules and the difference of Hu sheep maternal behavior with various genotypes. The result showed that only the products amplified by primers P3 displayed polymorphisms. The genotypes were AA, AB and BB, and each genotype fitted Hardy-Weinberg equilibrium. The statistical data of licking, nursing, treading and refusing to feed lamb didn’t fit normal distribution. The results of nonparametric test with multiple independent samples showed that the difference of observations for licking, nursing and trampling of individuals with AA, AB and BB genotypes was extremely significant (P<0.01).No obvious difference was found in individuals with three genotypes for the behaviour of refusing to feed young sheep (P>0.05). As the result showed, the partial sequences of PRLR gene exon10 had polymorphisms and there was correlationship between PRLR gene and maternal behaviours. According to the preliminary inference, PRLR gene could be used as candidate gene affecting the Hu sheep maternal behavior.

The study aimed to screen the differentially expressed genes of skin tissue in fine wool sheep with different fiber diameter. Affymetrix GeneChip ® Bovine Genome Array and Agilent Sheep Oligo Microarray were used to screen differentially expressed genes of skin tissue in fine wool sheep with different fiber diameter. Significance Analysis of Microarray (SAM) methods was used to identify the differentially expressed genes. GO (Gene Ontology) and the pathway analyses were conducted on differentially expressed genes using a free web-based Molecular Annotation System 3.0 (MAS 3.0). A total of 267 genes were identified to be differentially expressed genes from 2 different Microarray, 106 were up-regulated expression genes and 161 were down-regulated expression genes. GO and Kegg analysis revealed that the expression levels of the most differentially expressed genes were related to cell proliferation，differentiation，apoptosis, protein biosynthesis, Wnt signaling pathway and Keratin constitution etc. Subsequently, Real-time PCR was performed to validate seven differentially expressed genes which were screened out by the microarray approach and sufficient consistency was observed by the two methods. The results suggest that the gene expression profiles of fine wool sheep with different fiber diameter are variant，their different expression may alter the formation and differentiation of skin follicle and wool fiber, and some important genes identified may be useful in further study on molecu1ar markers of fiber diameter in fine wool sheep.

 The effect of two mutations in Interleukin-8 (IL-8) gene for 588 Holstein cows from 72 bull families of Haifeng dairy cattle farm imported from Australia were investigated on 7 milking performances: tested day milk yield, milk fat percentage, milk protein percentage, 305 d corrected milk yield, 305 d milk fat yield, 305 d milk protein yield and somatic cell score (SCS). The aim was to explore molecular marker in the breeding of dairy cattle. The polymorphism of IL8 gene was detected by polymerase Chain reaction-single strand conformation polymorphism (PCR-SSCP) technique. The association between polymorphic sites of IL8 gene and milking performances , SCS was analyzed by the least square mean model. The results showed that a mutation from G to A was identified in the 5′ upstream of -180(G/A), the genotypic frequencies of GG, GA and AA were 0.226, 0.476 and 0.298, respectively. The gene frequencies of G and A were 0.464 and 0.536, respectively. No significant effects of mutation were observed on milking performances and SCS except on 305 d milk fat yield. The 305 d milk fat yield for GG genotype was higher than GA (P<0.05). A mutation from A to G was identified in 2 789 of IL8, and the genotypic frequencies of GG, GA and AA were 0.297, 0.506 and 0.197, respectively. The gene frequencies of G and A were 0.550 and 0.450. This mutation showed significant effects on tested day milk fat percentage, milk protein percentage, 305 d corrected milk yield and SCS. The AA genotype had greatly significant higher milk fat percentage than GG and GA genotypes (P<0.01). Milk protein percentage for AA was greatly significant higher than that GA (P<0.01). GG genotype had greatly significant higher SCS than GA and AA (P<0.01). 305 d corrected milk yield for GA was significant higher than AA (P<0.05). The polymorphisms of IL8 gene had a greatly genetic effects on milking traits and mastitis resistance, so this gene could be a useful genetic marker for Holstein cows imported from Australia.

This experiment was conducted to clone and identify the spliced variants of the TLR4 gene, and analyze the regulation of their gene expression in duck. Based on the sequence of TLR4 gene in GenBank(accession number:JN048668)，the primers were designed，and the TLR4 gene was obtained by the RT-PCR. The temporal and spatial expression pattern was detected by qRT-PCR to analysis the levels of transcript variants in different tissues by constructing the polyI:C infection duck model. One new transcript variants(TLR4-b) of duck TLR4 gene was found, and sequences analysis showed that it was an alternative exon between the first and the second exon. The tissue expression analysis showed that the expression of TLR4 was mainly wild type(TLR4-a). And the expression of TLR4-a mRNA always increased firstly, then decreased in lung and spleen after the stimulation of ployI:C, but the expression of TLR4-b had little change. The new splicing isoforms(TLR4-b) of TLR4 gene was cloned and identified sucessfully in duck. The expression of TLR4-a was higher than that of TLR4-b under normal conditions, but the expression level of TLR4-a gene firstly increased, then decreased after ployI:C stimulation. The results preliminary reveal the structure of TLR4 gene transcript variant in duck, and its function mainly depends on TLR4-a gene.

 To understand PRLR’s role in duck, a novel isoform with only one ligand binding region of duck prolactin receptor gene (PRLR) was identified by rapid amplification of cDNA ends (RACE) strategy. Sequence analysis showed that the isoform was 2 535 bp long, contained 431 bp 5′ untranslated region (5′-UTR), 1 884 bp coding sequence and 220 bp 3′ untranslated region (3′-UTR) and comprises 11 exons. The predicted PRLR contained 627 amino acids (aa) and comprises a signal peptide of 24 aa at N-terminus, thus, its mature protein was 603 aa long. Different from canonical avian PRLRs with two tandemly repeated ligand binding regions within the extracellular domain, the putative protein only contained one ligand binding region, similar to the vertebrate PRLRs.The ligand binding region contained 2 pairs of cysteine residues and a pentapeptide of 5 aa known as WS motif as well, the 2 typical features of highly conserved in the members of class 1 cytokine receptor superfamily. In the meanwhile, we investigated one point mutation and one single nucleotide polymorphism (SNP) which located in 5′- and 3′-UTRs of the duck prolactin receptor respectively, and the SNP (C2 391T) in the 3′-UTR was in an extreme departure from Hardy-Weinberg equilibrium in the duck population by Chi-square test.

 This experiment was conducted to study the effects of insulin on mRNA expression of genes related to milk protein and fat synthesis in bovine mammary cells cultured in vitro. Mammary epithelial cells were cultured for 24 h with the following hormone treatments: no hormones (NH, control), 50 ng·mL^-1 hydrocortisone + 200 ng·mL^-1 prolactin (FP) or 100 ng·mL^-1  insulin + 50 ng·mL^-1 hydrocortisone + 200 ng·mL^-1 prolactin (IFP). The relative expression of the related genes of milk protein and fat synthesis were measured by real-time PCR. The results showed that the mRNA relative abundance of β-casein (CSN2), κ-casein (CSN3), Acetyl-CoA carboxylase (ACACA), fatty acid synthase (FASN) and sterol regulatory element pinding protein1 (SREBP1) in IFP group were significantly up-regulated(P<0.05). In addition, signal transducers and activators of transduction 5(STAT5B) and ETS-related transcription factor Elf-5 (ELF5) in JAK2-STAT5 signal pathway, as well as Phosphoinositide-3-kinase (PI3K), protein kinase B(AKT1) and eukaryotic translation initiation factor 4E(EIF4E) in PI3K/Akt/mTOR siganal pathway in IFP group were significantly up-regulated (P<0.05), while the IFP hormone combinations had no effect on TSC1, TSC2 or RHEB transcription in AMPK singal pathway. The data demonstrated that insulin induced the mRNA expression of genes related to milk protein synthesis through JAK2-STAT5 and PI3K/Akt/mTOR signal pathway and the mRNA expression of genes related to milk fat synthesis through PI3K/Akt/mTOR and SREBP signal pathway in bovine mammary epithelial cells cultured in vitro, which indicated insulin, hydrocortisone and prolactin regulate the mRNA expression of genes related to milk protein and fat synthes together.

The aim was to explore the efficience of in vitro fertilization and embryo development of the cattle-yak. The experiment was conducted to investigate the effects of in vitro maturation time, oocyte grades, different treatment methods of the semen, in vitro culture system and embryonic vitality and development after freezing and thawing on in vitro fertilization cattle×yak. The results showed that following the prolonging of in vitro maturation time, the extruding of the first polar body(Pb1) increased. And the Pb1 of grade A oocytes cultured in 24 h was significantly higher than that of B and C grade oocytes. But there was no significant difference between cattle oocytes and yak oocytes in same culture time and grade. Cleavage rate with BO fluid washing centrifugal process group is significantly higher than that of the Percoll group, but no significant differences on the blastocyst rate. Two kings of IVF cattle-yak embryos(cattle♀×yak♂vs. yak♀×cattle♂) co-culture with oviductal epithelial cells, cumulus cells and SOF medium and there was no significant difference in cleavage rate, but blastocyst rate and hatched blastocyst rate of co-cultured group were significantly higher than that of SOF medium group. The rate of normal morphology embryo (90.52%±5.94% vs. 91.53%±7.34%), survival rate within 24 h (61.45%±8.44% vs.64.72%±11.82%) or 48 h(44.81%±6.21% vs. 48.47%±10.04%) of the embryos after thawing were no significant difference. Taking 7 bovines including 4 yellow cattle and 3 cattle×yak as recipients, which were transplanted after natural estrus .The pregnancy rate was 57.14% and the length of average pregnancy was 258 days.

The research aimed to reveal the content and distribution rules of energy and protein in different body tissues for 20-35 kg Dorper and Thin-tailed Han crossbred ram lambs fed with different diet levels, and to establish some mathematical models to predict body composition and chemical composition by means of live weight in crossbred sheep. Twenty-one lambs were randomly assigned to three groups，one group fed ad libitum (AL), and the other two groups fed either 70% (IR70) or 40%(IR40) of ad libitum. Each group contained seven sheep and every sheep was a replicate. A slaughter trial was performed when the lambs in AL group reached 35 kg of live weight. Dry matter, crude protein, fat as well as gross energy content were measured in different tissues including bone, muscle, fat, blood+viscera, hide and fleece. In body composition, AL group lambs had the highest proportion of fat to moisture and showed significant difference to IR40 group (P<0.05), while the proportion of protein/fat was higher in IR40 group than that in AL group (P<0.05). No significant difference was found in the ratio of the muscle, blood+viscera or hide weight to fleece-free empty body weight (P>0.05), but the ratio of bone to fleece-free empty body weight was significant difference (P<0.05) among three treatments. Energy content (MJ·kg^-1) and protein content(%) in the tissues showed no significant difference (P>0.05) among three feeding levels. The ratio of dry matter or protein to fleece-free empty body weight showed no significant difference between the three groups (P>0.05), however, IR40 group lambs had a higher ratio in moisture and ash than that of AL and IR70 group lambs(P<0.05) . The feeding level significantly affected the proportion of crude protein, fat and moisture in Dorper and Thin-Tailed Han crossbred ram lambs between 20 to 35 kg of live weight, and a highly negative correlationship was found between fat and moisture. The parameters of E/GE and CP/GCP in different tissues were affected by the feeding levels but the impact was not consistent. The protein content was relatively stable while the fat content was easily affected by age and the feeding levels. There was a strong correlationship between empty body weight and different tissues and organs, and a strong correlationship was also found between fleece-free empty body weight and the main chemical composition in body.

The objective of this study was to investigate the effect of fermented feed from cottonseed meal mixed substrate on chickens plasma metabolites via metabolomic technology based on high performance liquid chromatography-mass spectromentry (LC-MS). The data of plasma metabolites profile collected by LC-MS, which included four groups of control group (CG), Candida tropicalis group (CT), Saccharomyces cerevisiae group (SC) and compound fermentation group (CF), were transformed and the peak characterization was obtained by the commercial software automatically. The total difference of metabolites among CG, CT, SC and CF were found through partial least square-discriminant analysis (PLS-DA), and the differential metabolites were selected according to the value of variable importance in the projection（VIP>1.5）and P (P<0.05). The results showed that significant difference of plasma metabolites were found after adding the fermented feed to diets and the relative contents of these metabolites were all increased significantly (P<0.01) compared with the control group. These metabolites were mainly phosphatidylcholine (PC), phophatidylethanolamine (PE), cholesterol ester (CE), sphingomyelin (SM), diacylglycerol (DG) and triacylglycerol (TG) in CT; PC, CE, SM, DG, TG in SC; and PC, PE, DG in CF. The influence of microorganism fermented feed from cottonseed meal mixed substrate on chicken plasma metabolites mainly was revealed in the significant increase of lipid metabolites. The PC and DG were common difference metabolites in all treatment groups.

The aim of the study was to investigate the effects of berberine hydrochloride on lipid metabolism, endocrine and antioxidan function of meat rabbit. 128 New Zealand rabbits with 50-day-old and similar body weight were randomly allotted into four groups. One group was the control group and the others were the treating groups. There were thirty-two replicates in each group and one rabbits were used in per replicate. The experiment was designed by single factor random arrangement. The rabbits in the control group were fed with the basal diet. And the rabbits in the three treating groups were fed the basal diets with 10, 20 and 30 mg·kg^-1 berberine hydrochloride supplementation, respectively. The whole experiment lasted thirty-four days. The duration of the preliminary experiment was 7 d and the duration of the formal experiment was 27 d. The results showed that: with increased supplementation level of berberine, the concentration of ether extract, triglyceride and total cholesterol in longissimus dorsi and liver and the concentration of triglyceride, total cholesterol and low-density lipoprotein cholesterol in serum decreased, and the concentration of high-density lipoprotein cholesterol in serum and the activity of lipoprotein lipase and hepatic lipase in liver increased; every supplementation level of berberine increased the level of growth hormone, triiodothyronine and insulin in serum and decreased the level of glucagon and the concentration of glucose in serum (P<0.05 or P<0.01); the 20 and 30 mg·kg^-1 supplementation levels of berberine increased the level of tetraiodothyronine in serum(P<0.01); with increased supplementation level of berberine, the activity of total superoxide dismutase increased and the concentration of maleic dialdehyde decreased. The results suggest that: with the supplementation of berberine to the diet of meat rabbits, the lipid metabolism could be improved, the endocrine system could be regulated and controlled, and the antioxidan function could be enhanced.

The objective of this study was to construct the infectious molecular clone with molecular marker of subgroup J avian myeloid leukosis virus (ALV-J) strain SCGS-1. A full-length infectious clone of ALV-J (pUC-SCGS) was constructed by cloning and combining of three fragments using PCR method from SCGS-1. SalⅠ site was introduced on 4684nt of SCGS-1 by overlapping PCR to form another infectious clone and named pUC-△SCGS. The two plasmids, pUC-SCGS and pUC-△SCGS, were transfected into CEF, and the rescued viruses were detected by PCR, avian leukosis virus antigen test kit and indirect immunofluorescence assay (IFA). Digestion and sequence analysis revealed that the infectious clone pUC-SCGS and pUC-△SCGS were constructed correctly. PCR, ELISA test and IFA results showed that the 3rd and 4th generation of rescued virus were positive, while the controlled CEF were negative. Rescued virus and the virus with molecular marker of subgroup J avian myeloid leukosis virus were successfully constructed, named rSCGS-1 and r△SCGS-1.

The effects of PCV2 on NF-κB signal were studied to explore the modulation mechanism of PCV2 on release and changes of pro-inflammatory factors from lymphocytes in PCV2 infected piglets. Five conventional post-weaning piglets free of PCV2 and PRRSV antibody and antigen were chosen as the experiment animals. The cells isolated from spleens with pinhead of injectors were distributed into experimental group and control group. The lymphocytes of experimental group were incubated with PCV2 at 1 × 10⁵ TCID₅₀, control group was incubated with equal volume of 1640. The PCV2-inoculated and mock-inoculated lymphocytes at 0, 6, 12, and 24 h post-inoculation (HPI) were collected respectively. NF-κB was translocated into nuclear by confocal laser scanning microscope. The protein levels of NF-κB/P65 and phosphorylation-IκBα were detected by Western Blot and NF-κB DNA binding activity was detected by electrophoretic mobility shift assays (EMSA). The result of confocal laser scanning microscope showed that nuclear translocation of NF-κB/P65 after lymphocytes incubation with PCV2 increased gradually with the increased incubation time. The NF-κB/P65 proteins increased significantly (P<0.05) in PCV2-inoculated lymphocytes compared with controls at the same time. The results of NF-κB DNA binding activity revealed the same changes with NF-κB/P65 protein. The protein levels of phosphorylation-IκBα in plasma was highest at 24HPI, and the levels in experimental group were significant increasing (P<0.05) compared with control group at 6 HPI, 12HPI and 24 HPI. All the results demonstrated that PCV2 can activate the NF-κB signaling pathway through the degradation of IκB phosphorylation, and the signal could play an important role in modulating the expression of the some pro-inflammation cytokines.

 This experiment was conducted to study the initial secretive features of specific IgA antibodies against P36, P46 and P97R1 proteins produced in the respiratory tract after the pigs were affected with Mycoplasma hyopneumoniae (Mhp). Total seven pigs were infected with Mhp strain. The nasal swabs were collected at the 1st, 2nd, 4th, 6th, 8th, 12th, 16th and 21st day post inoculation (dpi) to detect specific IgA titers against P36, P46, P97R1 proteins by the three established indirect enzyme-linked immunosorbent assays (ELISA). The IgA titers were normalized by the total protein concentration of nasal swab samples. The results showed that levels of three sorts of specific IgA start to rise at the 6th dpi and reach the peak at the 12th dpi. The high levels could be kept until to end of assay (the 21st dpi). The secretive features of three specific IgA antibodies were consistent as well as the secretive amounts on the same day (P>0.05). In conclusion, P36, P46 and P97R1 proteins could induce the specific IgA antibody in the respiratory tract during the early stage of infection with the same trends and secretive amounts.

This experiment was conducted to develop the infection model of Mycoplasma hyopneumoniae in vitro and study the adhesion characteristics by indirect immunofluorescence method in different cell lines. Several Mhp strains which include field strains (XLW-1, XLW-2, XLW-3, WX), international standard strain 232 and the attenuated strain (168) were selected, and infected Pig Kidney15 （PK15）. The results showed that all the Mhp whole bacteria strains could adhere to PK15 cell lines with different adhesion efficiency. And the order of adhesion ability of Mhp was field strain XLW-1, WX, the attenuated strain (168), international standard strain 232, field strain XLW-2, XLW-3. The field strain XLW-1 had the best adhesion ability, so it was selected as test strain to study the adhesion ability to other cell lines, such as the original generated cells of pig tracheal epithelial, porcine alveolar epithelial cells (SJPL), porcine alveolar macrophages (3D4/31) and rhesus monkey kidney cells (Marc145). XLW-1 strains can adhere to different cell lines in vitro, but the different adhesion capacity also showed on the different cell lines. XLW-1 strain has good adhesion ability to pig tracheal epithelial cells in primary separation, the SJPL cell lines and 3D4/31 cell lines, while very weak ability to the non-swine cell lines, Marc145 cell line. In conclusion, Mycoplasma hyopneumoniae adhesion model was developed on different cell lines, and different Mhp strains showed different adhesion ability on the same cell lines, as well as the same strain showed the different adhesion on different cell lines.

 In order to study the effect of the Bacillus probiotics on blood cells and tissue structure of broilers, 480 newly hatched AA chicken were randomly divided into control group, antibiotic group, experiment Ⅰ and Ⅱ groups. The control group was fed with basal diet, and the antibiotic group was fed with 50 mg·kg^－1 chlortetracycline in the basal diet, and the experimental Ⅰ and Ⅱ groups were respectively fed with 200 and 400 mg·kg^－1 the Bacillus probiotics in the basal diet for 42 days. The results showed: compared with the control group, the quantity of red cells and the ratio of basophilic granulocytes and monocytes were significantly increased in 21- and 42-day- old broilers (P<0.05), and the weight and organ index of thymus and bursal of the 42-day-old broilers were respectively increased by 46.60% and 38.23%, 66.67% and 47.62% (P<0.05); The weight and organ index of bursal were significantly increased by 35.40% and 36.92%, 88.03% and 153.97% in the 21- and 42-day-old broilers (P<0.05), and the weight and organ index of thymus of the 42-day-old broilers were significantly increased by 46.27% and 28.13% in experiment Ⅰ group (P<0.05); the quantity of white cells was obviously decreased, and the weight and organ index of spleen were also obviously decreased in the antibiotic group at the 42 d (P<0.05). Compared the tissue structure of immune organs in experiment Ⅰand Ⅱ group with the control group, the quantity of spleen node were increased, and periarterial lymphatic sheath were thickened; the thymic cortex were also thickened, and the quantity of thymic corpuscle were decreased; the bursal node were greatened with tightly rank lymphocytes, and which change were more obvious in the experiment Ⅰthan the experiment Ⅱ group. But the tissue structure of immune organs showed different degree atrophy in the antibiotic group. The results suggested that addition of 200 mg·kg^－1 the Bacillus probiotics are better to improve development of immune organs and strengthen immune function than that of 400 mg·kg^－1 the Bacillus probiotics.

Interleukin-10 (IL-10) has been recently identified as a multifunctional cytokine because of its close link with immune regulation and anti-inflammatory responses. The aim of this study was to clone and express rabbit IL-10 gene in order to prepare the polyclonal antibody against recombinant IL-10. The IL-10 gene of New Zealand white rabbit was amplified by RT-PCR from spleen total RNA, then was sub-cloned into prokaryotic expression vector pET-32a and expressed in E. coli BL21 (DE3) host cells. Fusion protein was purified and immunized into guinea pigs so as to prepare polyclonal antibody. Finally, the sensitivity and specificity of the polyclonal antibody were detected through indirect ELISA and Western blot, respectively. As a result, rabbit IL-10 gene was cloned, and the obtained 537 bp fragment had 99% identities to the previously identified rabbit IL-10 gene (NM_001082045) at nucleotide levels; recombinant vector pET-IL-10 was successfully constructed; SDS-PAGE electrophoresis showed that the fusion protein was well expressed; the anti-IL-10 polyclonal antibody showed high sensitivity (1:56 000) and specificity. The above results indicate that IL-10 gene has been successfully cloned and expressed, and the guinea pig anti-rabbit IL-10 polyclonal antibody has been prepared by immunization with the purified recombinant IL-10 protein, which lays the foundation for further studies on rabbit IL-10 gene functions.

The present study was undertaken to study the inhibitory effect and mechanism of α-terpineol on Escherichia coli, to provide a theoretical basis for the development of new drugs. Minimal inhibitory concentration (MIC) were determined by the same ratio dilution method with broth medium and minimal bactericidal concentration (MBC) by SDA of α-terpineol on Escherichia coli. Permeability change experiment of cell wall and membrane, nucleic acid synthesis inhibitory experiment, SDS-polyacrylamide gel electrophoresis and endotoxin inhibitory experiment, were study in the study. Results were as follows: MIC and MBC of α-terpineol on Escherichia coli are all 0.78 μL·mL^-1; Permeability of cell wall and membrane increasing; DNA fluorescence intensity is reduced and bacterial protein bands become light obviously; Endotoxin decomposed and show no typical pyrogen reaction in experimential group. It is suggested that the bacteriostatic mechanism of α-terpineol is based on its roles about changing the cell wall and membrance permeability, inhibite synthesis of nucleic acid and proteins and anti-endotoxin activity.

The aim of this study was to investigate MHC expression and localization in uterus and placenta of pregnancy rat, and to study the possible mechanism of immunomodulation classical class Ⅰmajor histocompatibility complex (RT1-A) antigens and non-classical classⅡ major histocompaibility complex (RT1-DM) antigens influenced by exogenous TGF-β1 (Transforming Growth Factor-β1). All of these parameters were examined by western blot and immunohistochemistry. The results showed that the expression of RT1-A and RT1-DM protein were examined in uterus and placenta during every stage of gestation. The expression of RT1-A protein was increasing gradually at uterus during all stage of gestation (D4-D19). The expression of RT1-DM protein was increasing at uterus from early-stage to mid-stage of gestation and decreasing in late-stage. In placenta, the expression of RT1-A, RT1-DM were decreasing gradually during mid-stage to late-stage of gestation. TGF-β1 treatment increased expression of RT1-A and RT1-DM in pre-implantation stage and decreased them in implantation stage (D6-D9) and late at uterus. During late-stage gestation (D19) TGF-β1 treatment significantly inhibited the expression of RT1-A and RT1-DM protein at uterus (P<0.01). The expression of RT1-A and RT1-DM was up-regulated during mid gestation significantly (P<0.05, P<0.01). Immunohistochemistry showed that TGF-β1 did not affect the localization of the two molecules: in early pregnancy RT1-DM mainly localized on uterine luminal epithelium, glandular epithelium, blood vessels, myometrium. RT1-A mainly localized on decidual blood vessels luminal epithelium, glandular epithelium, dicidualbasalis. During mid and late pregnancy, RT1-A and RT1-DM mainly localized on labyrinthine zones, spongiotrophoblast, glandular epithelium, uterus blood vessels. These results indicated that the expression of MHC antigens and TGF-β1 in maternal-fetal interface which may be helpful for constructing an appropriate micro-immunoenvironment of fetal parturition during normal pregnancy.

 Endometritis is one of the three severe diseases in dairy cows, which has serious impact on the development of dairy industry. Gram-positive bacteria are the major pathogens of endometritis. Lipoteichoic acid (LTA) is important component of the cell wall of Gram-positive bacteria. Toll-like receptor 2 (TLR2) is the main receptor of innate immune system, which recognizes pathogenic microorganisms and plays significant role in innate immune reaction. TLR2 has close relationship with endometritis caused by Gram-positive bacteria. In this study, mice were used as model, and uterus infusion was taken with different concentration of LTA. Meanwhile, endometrial epithelia cell culture was stimulated with different concentration of LTA in vitro. Pathological histological changes of endometrial tissue in mice were observed, and expression level of TLR2 and inflammatory cytokine TNF-α, IL-6 in endometrial tissue of mice was analyzed. The results could provide some basis for inflammatory signal pathway mediated by TLR2. The results indicated that infusion of different concentration of LTA led to acute endometritis, and inflammatory cells increased in endometrial tissues. Expression of TLR2, TNF-α, IL-6 mRNA in endometrial tissues and epithelial cells increased significantly after different concentrations of LTA stimulation. The expression level changed depending on LTA concentration. Low concentration led to high level expression, however, high concentration decreased expression level.

In the current study, a new method for detecting the sequence difference of duck Chromo-helicase-DNA-Binding 1 (CHD1) genes on Z and W sex chromosomes was established through amplifying DNA fragment length polymorphism, which aimed to solve some technical problems, such as the sex identification with nondamage, the misclassific rate of artificial sex identification during embryonic period and the injury coming from the rectal identification during neonatal period. In this study, an improved gender identification method was developed by DNA amplified fragment length ploymorphism between CHD1-Z and CHD1-W genes of birds. The gender identification PCR primers HPF/HPR were used, and the PCR product was cloned, sequenced and blasted; then the effectiveness and accuracy of this method had been explored by examples. 2% agarose gels were used to easily distinguish the 495 bp CHD-Z and the 351 bp CHD-W PCR amplicons, and the female (ZW) displayed two visible bands, whereas only a single band was found in the male (ZZ). The results indicate that the new detecting method for detecting the sequence difference of duck CHD1 genes is visual and reliable, and the molecular marker provided by this study is highly efficient and precise in the sex molecular biological identification of Chinese native domestic duck breeds.

In this study, an unknown virus was isolated from rectal swabs of cattle with severe diarrhea and hemorrhagic dysentery in Beijing. The unknown virus was identified as non-enveloped particles about 30 nm in diameter and containing RNA genome by physicochemical characteristics analysis. All of four layers which were purified by cesium chloride density gradient centrifugation were amplified with random polymerase chain reaction (PCR), and the products from layer 2 ranged mainly from 500 to 1 500 bp, showing comet tail-shaped entrainment phenomena, were purified and sequenced. The sequence blast results showed that the virus shared high gene homology with bovine enterovirus type 2. Therefore, specific primers of bovine enterovirus type 2 was used further to identify the virus, and the unknown virus was finally verified exactly belonging to BEV2 (93% nucleotide identity). Two healthy adult cattle were inoculated with the virus as experiment animals, but no clinical symptoms appeared, although antigen and serum antibody were positive after challenge.