Embryogenesis was a complex progress which is regulated by many transcriptional regulation factors. With the development of high-throughput sequencing technology, non-coding RNAs (ncRNAs) are found to play an important role in the embryogenesis, X chromosome inactivation, gender regulation and the development of animal brain. Firstly, miRNAs regulate the expression of genes related to embryogenesis through targeting the 3′ UTR of the targeted mRNAs. Secondly, lncRNAs mediate mammal X chromosome inactivation and insect X chromosome dosage compensation through interfering transcription or modifying chromatin. Thirdly, piRNAs can maintain the DNA integrity of the germ cell in the way of silencing transposon and regulating silkworm sex differentiation. Lastly, circRNAs can act as the spongy of miRNAs and play a great role in the development of mammal animal brain. Here, we elucidate the advances of ncRNAs through the regulation during animal embryogenesis, which will provide a reference for further study on the mechanism of ncRNA during the embryogenesis.

Veterinary drugs are widely used in the livestock breeding, and it plays a significant role in treating and preventing disease. However, misuse of veterinary drugs may lead to accumulate or remain in animal tissue or feed, such as liver and muscle, which could be harmful to human health. Therefore, the detection methods of veterinary drug residues are particularly important. This paper comprehensively discussed sample preparation of multi-residue of veterinary drugs in animal derived food and animal feed, mainly about the research of dozens or hundreds of multi-classes veterinary drugs. The development of sample preparation was further discussed.

The objective of this study was to investigate whether microRNAs play roles in puberty onset of the chicken. The Solexa deep sequencing was performed to analyze the miRNA expression profiles in the hypothalamus of hens before puberty onset (BPO) and after puberty onset (APO). The target genes prediction and functional analysis were also carried out through bioinformatics method. 374 conserved and 46 novel miRNAs were identified in the chicken hypothalamus. 144 conserved miRNAs were differentially expressed (reads>10, P<0.05) from BPO to APO transition. 2 013 putative genes were predicted as the targets of the 15 differentially expressed miRNAs (｜log2(fold-change)|>2.0, P<0.01). Functional analysis suggested that these 15 miRNAs played important roles in transcriptional regulation and signal transduction during puberty onset. The results suggest that miRNA as novel partner involve in chicken puberty onset. Considering the characteristics of miRNA functional conservation, the results will provide new theoretical base for studying the molecular regulation mechanism of animals puberty onset.

This study was conducted to obtain the sequence of goat FGF21 gene, elucidate the expression characteristics of FGF21 gene in different tissues, and detect the expression of FGF21 and FGFR during intramuscular preadipocyte differentiation. Jianzhou Big-eared goat was used as laboratory animal in this study. The goat intramuscular preadipocytes were obtained using collagenase II. RT-PCR was used to clone FGF21 gene. Real-time quantitative PCR (qPCR) was performed to detect the expression of FGF21 in different tissues and the expression levels of FGF21 and its receptors during goat intramuscular preadipocyte differentiation. The full-length sequence of goat FGF21 gene was 666 bp including a complete 630 bp open reading frame (ORF), encoding 209 amino acids (Accession No.: KT288195). FGF21 gene was expressed in all examined tissues including heart, liver, spleen, lung, kidney, adipose tissue and longissimus dorsi, and enriched in liver and adipose tissue. During the differentiation of goat intramuscular preadipocyte, the expression pattern of FGF21 gene was the same as that of FGFR1 and FGFR2, peaked at 2nd day, extremely higher than other days (P<0.01). The FGF21 mRNA was enriched in adipose tissue and peaked at 2nd day during goat intramuscular preadipocyte differentiation, indicating it might be a regulator in the early differentiation progress via FGFR1 or FGFR2. These results provide data for further elucidating the molecular mechanism of FGF21 gene in regulating goat IMF deposition.

Based on the previous studies, the aim of this study was to further improve the efficiency and accuracy of genotyping. Two high-throughput genotyping methods(Taqman and SNaPshot) for FecB detection in Chinese sheep breeds had been established in the present study. The results showed that the two methods not only reduced the cost but also saved the time and increased efficiency dramatically. The two methods had potential application value in large-scale molecular breeding introducing FecB. The detection results of 23 264 sheep including 10 Chinese indigenous sheep breeds, 3 crossbred breeds and some hybridized sheep populations collected since 2003 showed that the allele B frequencies in Small Tail Han and Hu sheep were much higher than other breeds. The allele B frequency in Small Tail Han sheep was 0.432-0.833. The allele B frequency in Luxi Blackhead sheep had been improved (up to 0.432) during upgrading and selection stage, while decreased during intercross stage. These results can provide reference data for improving sheep fecundity in sheep breeding program by introducing FecB gene.

The aim of this study was to understand 5′ regulatory region and promoter region features of FKBP6 gene, to provide evidence for exploring the mechanisms of differential expression of FKBP6 gene in yak and yak-cattle testicular tissue. The sequence of 5′ regulatory region of yak FKBP6 gene was obtained by cloning and sequencing, and was analyzed by bioinformatics methods. The core promoter region of FKBP6 gene was identified using Dual-luciferase assay system and the transcription factor binding sites related to spermatogenesis were predicted using bioinformatics software. The 5′ regulatory region of yak FKBP6 gene was 1 354 bp in length by cloning, sequencing and splicing, which shared the similarity of 99.71% with cattle. The 5′ regulatory region sequence of yak FKBP6 gene contained several potential promoter regions, typical CAAT-Box and CpG island, but no TATA-Box. The core promoter region of FKBP6 gene was located in －263-－167 nt of 5′ regulatory region of FKBP6 gene by luciferase activity analysis and contained transcription factor binding sites related to spermatogenesis including CAAT-Box, E-Box, CTCF and CREB. The identification of the FKBP6 gene core promoter and the transcription factor binding sites provide a basis for further research on the regulation of FKBP6 gene expression in yak testis.

The aim of this study was to investigate the impact of dominant effects on the predictive accuracy of genomic breeding values. Based on the dependency between additive and dominant effects using BayesA model, we proposed two submodels:1) the additive effect and dominant effect were independent of each other in BayesAD1 model; 2) the dominant coefficient and absolute values of additive effect were independent of each other in BayesAD2 model, in which the dominant coefficient followed normal distribution. Using simulated datasets, we compared the predictive accuracy of genomic estimated breeding value (GEBV) among the additive model (BayesAD0) and dominant models (BayesAD1 and BayesAD2). We further investigated the effect of number of QTLs (quantitative trait loci), size of full-sibs family and ratio of additive variance to dominant variance on predictive accuracy of GEBV. The results showed that the dominant models slowed down the declining of the predictive accuracy of GEBV in subsequent generations. Moreover, the predictive accuracy of GEBV increased as the ratio of dominant variance increase. When the ratio of additive variance to dominant variance reached 0.25, BayesAD2 was 20.3% and 28.4% higher than the accuracy of the BayesAD1 and BayesAD0, respectively. In addition, the size of full-sibs family affected the predictive accuracy of GEBV positively. And an increase in the number of QTL was accompanied by a reduction on the predictive accuracy of GEBV. These results indicate that a better prediction of genetic values is intended, when the dominant variance are large just as low-heritability traits.

To investigate the expression patterns of Kruppel like factor 2 (KLF2) and peroxisome proliferator activated receptor gamma (PPARγ) during the adipogenic redifferentiation of porcine dedifferentiated fat (DFAT) cells and lay a foundation for revealing the mechanism of porcine DFAT cell redifferentiation, we applied the “ceiling” culture to obtain porcine DFAT cells from mature adipocytes, detected the surface antigens of DFAT cells by flow cytometry, and then, identified the adipogenic differentiation of DFAT cells by methods of morphology, oil red O staining and real-time PCR. The results showed that the dedifferentiation sign was clearly observed, showing that the horn like protrusions were formed, large lipid droplets were decomposed into small lipid droplets, and lipid droplets were extracellularly discharged after porcine mature adipocytes were seeded on the third day, the lipid droplets were almost discharged in 9 day, and the bottom wall was covered with primary cells in 12 day. The positive rates of mesenchymal stem cell surface antigen CD29, CD44 and CD105 were 99.0%, 97.8% and 99.5%, respectively, while the percentages of hematopoietic stem cell surface antigen CD45 and CD34 were only 3.55% and 4.55%. Then, after 3 days of adipogenic induction of F3 generation DFAT cells, the spindle shaped cells became wider and shorter, and lipid droplets were observed in the cytoplasm. With the increase of induction time, the number and volume of lipid droplets increased, and the induction efficiency was more than 80% after 12 day induction. During the progress of adipogenic induction, the relative expression levels of KLF2 were 0.47±0.02, 0.35±0.07, 0.31±0.09 and 0.11±0.07, and PPARγ expression levels were 1.62±0.01, 2.03±0.04, 3.22±0.04 and 3.49±0.06 on the 2, 5, 10 and 15 days, respectively. In conclusion, porcine DFAT cells with high purity were successfully obtained, owning the mesenchymal stem cell properties and the adipogenic redifferentiation competence, and, KLF2 expression levels decreased, while PPARγ expression increased with the extension of induction time during adipogenic redifferentiation of DFAT cells, indicating that KLF2 might play an inhibitory role, while PPARγ could take on a positive role during the adipogenic redifferentiation of DFAT cells.

This research was conducted to establish a simple and effective method for isolation and purification of pig chorionic trophoblast cells. Single cell suspension was prepared from pig fullterm placentas by the method of trypsin-DNaseⅠdigestion and two density (35% and 45%, v/v) Percoll gradient centrifugation.Morphology observation,growth curve measurement, immunofluorescence and transmission electron microscopy (TEM) were applied to cells identification. The results showed that the cells were polygon or quasi-circular in shape, which exhibited epithelioid and paving-like spreading growth. After 48 hours culture, some cells started developing into syncytiotrophoblasts. Immunofluorescence assay showed that the proportion of Cytokeratin 7 positive cells was more than 91%, indicated the high purity of pig chorionic trophoblast cells. The results of TEM revealed that the cells had typical structural features of trophoblasts. These results indicated that our research could be used to obtain high-purity pig chorionic trophoblast cells simply and effectively by the method of trypsin-DNaseⅠdigestion and 2 density (35% and 45%, v/v) Percoll gradient centrifugation.

The melanin content of hair and distribution characteristics of mature melanocytes in skin of mink with different coat colors were analyzed and observed, which will provide theoretical basis for futher study on regulatory mechanism of mink coat color. The content of total melanin (TM), eumelanin (EM) and pheomelanin (PM) in hair, which were collected from Jinzhou black, Silver Blue and Jilin White mink in molting and pelting periods, respectively, were determined by microplate assay with sepia as standard sample. Toluidine blue, dopa and dopa with toluidine blue staining were conducted to observe the distribution of skin mature melanocytes. The results showed that the content of TM and PM from black hair in pelting period were 1.17 and 1.20 times (P<0.01) than that of molting period, and the value of EM was higher than that of molting period (P<0.05). There were not different for the content of TM, EM and PM in gray hair (P>0.05) between the two periods. The content of TM and PM from white hair in pelting period were 1.27 and 1.22 times (P<0.05) than that of molting period. Histological staining revealed that the mature melanocytes were distributed both in skin with black and gray hair. In the skin of black hair, a large number of pigment granules were distributed at epidemis and the top of hair follicle and a few positive staining band was in the outer root sheath and medulla. There was less dopa-positive band at epidemis and the top of hair follicle in mink skin with gray hair. Moreover, the positive staining of the outer root sheath in skin with gray hair was weaker than that of black. However, there was not obvious dopa-positive staining area in mink skin with white hair. Results of the present study indicated that the formation of gray and white hair were related with the content of PM. The mature melanocytes at the top of hair follicle might be the main cytological basis of hair pigmentation in mink.

This experiment was conducted to study the effect of portal ammonia on urea cycle and gluconeogenesis in liver of pig. Eight pigs (Duroc×Landrace×Yorkshire), with the average weight of 20 kg, were randomly assigned to two treatments with 4 pigs in each treatment. Pigs were surgically implanted with catheter in the portal vein and hepatic vein. Then 65 and 35 mmol•L^-1 of ammonium chloride (NH₄Cl) were infused into liver by portal catheter. Serum samples were collected to analyze metabolites based on gas chromatography-mass spectrometry metabolomics. Liver samples were collected to examine gene expression and enzyme activity in urea cycle and gluconeogenesis. The results showed that, compared with low concentration NH₄Cl, the infusion of high concentration NH₄Cl significantly increased the content of urea, glucose and glucose-6-phosphate(P<0.05), the content of alanine, glutamate and aspartate were significantly decreased (P<0.05); addtionally, high concentration NH₄Cl significantly upregulated the expression of carbamoyl-phosphate synthase 1 (CPS1), ornithine carbamoyl-transferase (OTC), arginase 1 (Arg1), phosphoenolpyruvate carboxykinase (PCK) and glucose-6-phosphatase (G6PC), meanwhile the corresponding enzyme activities increased 50.77%, 44.47%, 41.24%, 46.99% and 54.57%（P<0.05）, respectively. These results indicated that, with the ammonia load increased into liver, the urea cycle was increased and the additional nitrogen was supplied by amino acid metabolism (alanine, aspartate, glutamate), of which the carbon skeleton were used for hepatic gluconeogenesis.

This study was conducted to investigate the effects of the combination of Glucagon-like peptide-2 (GLP-2) and dipeptidyl peptidase-Ⅳ (DPP-Ⅳ) inhibitors with different concentrations on cell proliferation, metabolism and enzyme activity of primarily intestinal epithelial cells cultured in vitro from 28-day-old weaned piglets, and aimed to explore the method of promoting GLP-2 effects. The experiment, taking 28-day-old weaned piglets’ intestinal epithelial cells as a model, studied the effects of DPP-Ⅳ inhibitors (KR62436) with different concentrations on the cell proliferation and metabolism. The 2×3 factorial design was adopted to study the effects of combination of GLP-2 (1×10^-10, 1×10^-9 and 1×10^-8 mol•L^-1) and KR62436 (0 and 1×10^-11 mol•L^-1) on cell proliferation, metabolism and apoptosis. The results were as follows: there was no significant effect of DPP-Ⅳ with 1×10^-11 mol•L^-1 on cell number, MTT OD, cell protein retention, total cell protein level, LDH activity, CK activity and Na⁺, K⁺-ATPase activity (P>0.05). Compared with the control group, the DPP-Ⅳ group with 1×10^-10 mol•L^-1 showed lower MTT OD value (P<0.05), and higher LDH and CK activity (P<0.05). In the groups that treated with different concentrations of GLP-2 and low concentrations of KR62436, with the GLP-2 doses increasing, cell number, MTT OD value, cell protein retention, total cell protein level and Na⁺, K⁺-ATPase activity increased significantly (P<0.05), and LDH and CK activity decreased significantly (P<0.05). KR62436 could significantly promote GLP-2 effects. The results indicate that the low concentration DPP-Ⅳ inhibitors have no significant effect on cell growth, while the high concentration DPP-Ⅳ inhibitor have a negative effect on cell metabolism and integrity. Combination of GLP-2 and low concentrations DPP-Ⅳ inhibitors may have favorable effects on intestinal epithelial cell proliferation, metabolism and integrity than GLP-2 alone.

The present study was conducted to measure greenhouse gases (CO₂, N₂O and CH₄) emissions and investigate emission characteristics of broilers using respiratory chambers. Three hundred Arbor Acres broilers (1-day-old) with good health and similar weight were randomly allotted into 3 respiratory chambers, and reared on the net until 42-day-old. A continual monitoring system for ventilation rate and the greenhouse gases concentration were used in this research to determine the 3 gases(CO₂, N₂O and CH₄) emissions. The results showed as follows: the CO₂, N₂O and CH₄ emissions for broilers were (31.40±11.45) g•d^-1•bird^-1, (9.13±0.43) mg•d^-1•bird^-1 and (0.40±0.15) mg•d^-1•bird^-1, respectively. Total CO₂, N₂O and CH₄ emissions from 1-day-old to 42-day-old were estimated: (1 318.94±480.91) g•bird^-1, (374.01±17.68) mg•bird^-1 and (16.67±6.17) mg•bird^-1, respectively. The daily emissions of CO₂ and N₂O increased with the growth of age of broilers, and then remained stable after 24-day-old and 27-day-old, respectively. The CH₄ emissions were relatively lower, and could be ignored when assessing the effect of greenhouse gases on environment.

This study was aimed to assess the influences of avian reovirus (ARV) infection on expression of innate immune genes in peripheral blood lymphocytes of SPF chickens.The quantitative real-time PCR assays was performed to determine the transcriptional levels of TLR3, TLR7, TLR21, MDA5, IPS-1, IRF-3, IFN-α, IFN-β, IFN-γ, IFITM3, Mx1 and OASL in 0 h, 12 h, 1, 2, 3, 5 and 7 days post infection (dpi) in peripheral blood lymphocytes of SPF chickens infected with ARV S1133. These results showed that the mRNA transcriptional levels of TLR3 and TLR21 were significantly reduced within 7 dpi （P<0.05 or P<0.01）; the mRNA transcriptional levels of TLR7 and MDA5 were obviously promoted and specially reached the summit maximum values on 7 dpi and 1 dpi, respectively (P<0.05 or P<0.01). The mRNA level of IPS-1 was down-regulated within 7 dpi. Meanwhile, the mRNA level of IRF-3 was up-regulated during the whole transcrption course, and IFN-α, IFN-β and IFN-γ mRNA levels were significantly increased and reached the peak at 2-3 dpi. The mRNA levels of IFITM3, Mx1 and OASL were significantly up-regulated and peaked at 3 dpi (P<0.01). Therefore, ARV infection is able to modulate the increased mRNA transcription of TLR7, MDA5, IRF-3, IFN-α, IFN-β, IFN-γ, IFITM3, Mx1 and OASL, and reduce the mRNA transcription of TLR3, TLR21 and IPS-1, which would contribute to further understand ARV pathogenesis and host innate immune responses to ARV infection.

In order to investigate the role of semen of yellow feather grandparent roosters in avian leukosis (AL) eradication process，a, b, c three different breeds of yellow feather grandparent roosters were chosen to be sampled. ALV p27 antigen ELISA was used in direct detection of cloacal swabs, while anti-coagulated blood and semen sample were used to isolate and detect exogenous ALV based on DF-1 cells. Upon comparative analysis, the results showed that the virus could be isolated from semen, while the result of ALV p27 antigen ELISA S/P value of cloacal swabs and the virus isolation of plasma were negative. Three results couldn’t completely correspond respectively. Individual AL deteetion of cloacal swabs, plasma or semen could lead to missed detection. Therefore in AL eradication of yellow feather breeds, semen as the test sample is feasible and should not be neglected, multiple samples should be considered in order to improve the detection rate and speed up the purification process.

The aim of the present study was to explore the influence of nucleocapsid protein (NP) to the proliferation of duck enteritis virus (DEV). We designed and constructed pSilencer-DEV-NP according to the DEV NP gene sequence in GenBank. After these pSilencers were transfected into duck embryo fibroblast (DEF) cells in three different ways, the expression of pSilencer DEV-NP to DEV proliferation in DEF cells was analyzed by fluorescence microscope and FQ-PCR. The results showed that the green fluorescence was observed by microscope in DEF cells transfected with pSilencer-DEV-NP-l, pSilencer-DEV-NP-2, pSilencer-DEV-NP-3 and pSilencer-DEV-NP-4, respectively. The silencing efficiency of pSilencer-DEV-NP-1 to 4 on DEV proliferation were 81.10%, 72.69%, 77.35% and 67.69%, respectively. FQ-PCR was used to amplify and calculate. The results showed that pSilencer-DEV-NP-1 could silence the DEV proliferation in DEF cells by three transfection ways, but the silencing efficiency was different. The silencing effect of pSilencer-DEV-NP-1 was the best at 60 h after transfection and its efficiency was 69.20%. The second highest efficiency occurred in the way of simultaneously DEV infection and pSilencer transfection, the best situation at 48 h was 63.79%. The worst efficiency was 52.58% at 24 h after the operation with the method of pSilencer-DEV-NP-1 transfection after DEV infection. These results suggest that the nucleocapsid protein has a certain effect on DEV proliferation, which lays a theoretical foundation for clarification of the mechanism of DEV replication and proliferation.

Cell entry of virus is an indispensable process in virus life cycle. However, the mechanism of classical swine fever virus (CSFV) cell entry is still undiscovered. Here, CSFV entry into ST cells by clathrin-mediated endocytosis was tested. Chemical inhibitor chlorpromazine and Dynasore and shRNA were used to disturb the clathrin-mediated endocytosis. The cell entry efficiency of CSFV was greatly decline when the function of clathrin and dynamin-2 were blocked. By using NH₄Cl and down-regulation of Rab5 and Rab7 gene, CSFV infection was inhibited. These results showed that the CSFV can entry into susceptible cells by using clathrin-mediated endocytosis pathway and the early endosome and late endosome were used for CSFV infection. This research provides new information to understand the CSFV infection.

Caprine parainfluenza virus type 3 (CPIV) can cause respiratory disease in goats, which could cause extensive morbidity and mortality when co-infected with Mycoplasma, bacteria and other pathogens. In this study, an indirect ELISA detection method based on CPIV3 N protein was established to monitor the disease. Primers were designed to amplify the N gene of CPIV3 JS2013 strain. Target gene was cloned into prokaryotic expression plasmid pET32a(+), to construct the recombinant plasmid pET32a-N, and then pET32a-N was transformed into Escherichia coli BL21 competent cells. Large amount expression of recombinant N protein was induced by IPTG. By optimizing the reaction conditions, an indirect ELISA was established. Clinical sera were tested by indirect ELISA and the results were compared with HI test. The recombinant plasmid was constructed successfully, and the expression of protein was identified by SDS-PAGE and Western blot, which confirmed that the protein was expressed correctly and had good immunogenicity and specificity. The purified protein was used as antigen and the indirect ELISA reaction conditions were optimized as follows: the coating antigen concentration was 1 μg•mL^-1, serum dilution was 1:200, serum reaction time was 60 min, HRP-labeled secondary antibody dilution was 1:6 000, reaction time of secondary antibody was 30 min, and substrate chromogenic time was 10 min. By testing of 138 clinical serum samples with the indirect ELISA and HI test, the coincidence rate between these two methods was 88.41%. The indirect ELISA established in this study was sensitive, accurate, which will be suitable and useful for the detection of clinical samples and large scale serological surveys.

In order to specify the Ghrelin expression and localization inside sika deer, we adopted the reverse transcriptase-polymerase chain reaction (RT-PCR), real-time fluorescent quantitative PCR (RT-qPCR ) and immunohistochemical technique to test the relative expression quantity of Ghrelin mRNA and Ghrelin protein location distribution in the esophagus, rumen, reticulum, omasum, abomasum, duodenum, jejunum, ileum, caecum, colon, liver, pancreas, thyroid, anterior pituitary, lung, kidney, spleen, ovarian and other organs of the sika deer. RT-qPCR results showed: Ghrelin mRNA was expressed in these organs, and the expression quantity in the abomasum was significantly higher than that of other organs (P<0.05); Immunohistochemical staining result showed that Ghrelin protein was expressed in these organs, most widely distributed in the digestive tract. The specific expression pattern of Ghrelin mRNA and Ghrelin protein inside Sika deer revealed that the new molecule may have extensive biological effect in the growth and development process of sika deer, and existed as an important adjusting mode especially in the aspect of gastrointestinal function.

In order to explore the effect of Lianmei extract on antioxidant index，gastrointestinal regulatory enzymes and duodenal apoptosis gene expression of heat and humidity diarrhea swine.Established heat and humidity environment artificially，and the swine fed E.coli to induce heat and humidity diarrhea model.The diarrhea swine were randomly divided into model group，positive drug group and Lianmei extract with high-，middle-and low dose groups，swine in the control group were fed at normal environment.Detected antioxidant enzymes，gastrointestinal regulatory enzymes，liver ATP enzymes vitality and duodenum apoptosis gene Caspase-3，-8，-9 mRNA expression changes for analysing the treatment mechanism of Lianmei extract.The results showed that the swine treated by Lianmei extract，The serum content of T-SOD and GSH-Px enzyme activity increased，LDH and MDA reduced，compared with model group，the difference was extremely significant (P<0.01).Acetylcholinesterase，amylase，liver Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase vitality significantly increased (P<0.05).Growth hormone，thyroid hormone T3 secretion improved significantly (P<0.05)，Glucocorticoids secretion decreased significantly (P<0.05).Meanwhile, duodenal apoptosis genes of swine recoveryed at different degree.Only the middle dose group Caspase-8 mRNA transcription decreased significantly to 2.62 times of that from the control (P<0.01).High- and middle dose group Caspase-3 mRNA transcription were reduced to 5.82 and 3.29 times of that from the control，compared with the model group，the difference was significant (P<0.05) or extremely significant (P<0.01).However，Caspase-9 mRNA transcription was significantly reduced (P<0.05) at high-，middle，low dose group and the amount of reduction was proportional to the concentration of the drug.The results indicated that Lianmei extract could achieve therapeutic heat and humidity diarrhea swine by enhancing diarrhea pigs anti-oxidative damage，improving gastrointestinal digestion and absorption capacity，promoting the secretion of growth hormone and reducing the expression of apoptosis gene.

The objective of this study was to investigate the changes of EGFL7, VEGF and VEGFR2 in lung of broilers with ascites syndrome and analyze the influence of compound traditional Chinese medicine for endothelium derived multiplication factor on AS. A total of 275 8-day-old Ross broilers were randomly divided into five groups, blank group bred with basal feed, model group bred with mixed feed (9-11 ℃, added 3% lard and 4% fish meal to basal feed, added 0.12% NaCl to drinking water), compound traditional Chinese medicine high, medium and low dose group bred with mixed feed and respectively given a dose of 2, 1, 0.5 mL•kg^-1 in drinking water. Using real-time fluorescence quantitative PCR and ELISA to detect the content of EGFL7, VEGF and VEGFR2 in lung tissue. Compared with blank group, ascites heart index in model group were increased significantly (P<0.01), and relative expression levels of EFGL7, VEGF, VEGFR2 mRNA and protein expression levels in lung of broilers in model group were increased significantly (P<0.01 or P<0.05); Compared with model group, compound traditional Chinese medicine high and medium dose group were able to improve clinical symptoms (mental state, ascitic fluid and hydropericardium), compound traditional Chinese medicine could effectively down regulate gene and protein expression levels of EGFL7, VEGF and VEGFR2 in lung of broilers (P<0.01 or P<0.05). The study showed the increases of gene and protein expression levels of EGFL7, VEGF and VEGFR2 in lung of broilers participated in the pathogenesis of AS. Compound traditional Chinese medicine can effectively prevent and treat of ascites syndrome by down regulate gene and protein expression levels of EGFL7, VEGF and VEGFR2 in lung of broilers to protect vascular endothelium.

In order to explore the correlation between adiponectin，leptin，visfatin in placenta and calf birth weight.56 naturally born healthy Chinese Holstein cows were used，in which，cows given birth to calf that were less than 40 kg were included in group A；between 40-45 kg were included in group B；more than 45 kg were included in group C.The placentas were surgically collected immediately after birth.RT-PCR and ELISA were used for evaluation of adiponectin，leptin，and visfatin in placenta，the correlation between adiponectin，leptin，visfatin in placenta and calf birth weight were analyzed.The results showed that，as the calf birth weight increased，the adiponectin and visfatin mRNA and protein expression levels increased，while leptin reduced first and then increased.The expression level of leptin mRNA of group C was significantly higher than that of group B (P<0.05). The difference among group A，B and C were not significant(P>0.05). Adiponectin and visfatin of group A，B and C were not significantly different (P>0.05). The expression level of adiponectin protein in group A and group B had no significant difference (P>0.05).The expression level of adiponectin in group C was significantly higher than that of group A and group B (P<0.05)，the expression levels of leptin and visfatin in group A，B and C had no significant differences(P>0.05). The mRNA and protein level of adiponectin and leptin in placenta were positively correlated with calf birth weight(P<0.01).The mRNA and protein level of visfatin in placenta with calf birth weight had no significant correlation(P>0.05).The mRNA and protein expression level of adiponectin，leptin and visfatin in 3 groups had no significant correlation respectively(P>0.05).The results demonstrated that adiponectin，leptin and visfatin were expressed in placenta of cows，which played a role in promoting of calf intrauterine growth cooperatively，adiponectin and leptin had great influence on calf birth weight，visfatin had little influence on calf birth weight.The study provided reference for studying mechanism between adiponectin，leptin，visfatin in placenta of cow and calf birth weight in the future.