Classical genetics suggests that the genetic information of the organism is determined by the genes, that is, only the change of DNA sequence can be inherited, but the epigenetic DNA modifications can't be inherited because of the cell reprograming. Recent studies have found that some of the epigenetic modification caused by environmental factors can also be passed to the offspring, and DNA methylation is one of the main mechanisms of epigenetic modification, which is a hot spot in the transgenerational inheritance field. Herein, we focus on the transgenerational inheritance of DNA methylation caused by environmental factors, which may provide some references for further study on transgenerational inheritance.

Wip1 (Wild-type p53-induced phosphatase1) is a member of PP2C (The type 2C family of protein phosphatases) family of phosphatases, which plays important roles in DNA damage repair and has attracted increasing attention as a oncogene. Wip1 knockout mice were viable, but exhibited reduced longevity, along with multiple-system abnormalities, including reproductive system, immune system, endocrine system and nervous system. The detailed phenotypes included male reproductive organ atrophy, male infertility, increased susceptibility to pathogens, diminished T-and B-cell function, insulin resistance, impaired body weight, declined fat mass,elevated anxiety-and depression-like behaviors and so on. The studies for the underlying molecular mechanisms further demonstrated that Wip1 played critical roles in a variety of physiological and pathological processes through dephosphorylating different substrate proteins. Therefore, in this review, we comprehensively discussed the phenotypes caused by Wip1 knockout in mouse as well as the possible underlying molecular mechanisms, which may provide information for further understanding the biological function of Wip1, and the possible mechanisms underlying the physiological and pathological processes in mammals.

The recombinant expression vector is vital in production of genetically modified animals, which includes complete exogenous gene expression cassette containing the target gene promoter, terminator and reporter gene. This paper briefly introduces the genetically modified animal production technology and counts the vector frequency commonly used in the production of genetically modified pigs, cattle and sheep during nearly 10 years. The statistical results show that promoter frequency of the genetically modified pigs from more to less are casein, CAG, CMV promoters, terminator frequency are rabbit β-globin poly A, casein poly A, SV40 poly A and BGH poly A; The promoter frequency in genetically modified sheep from more to less are casein, BLG and CMV promoters and terminator are casein poly A, BLG poly A, BGH poly A, SV40 poly A and rabbit β-globin poly A; The promoter frequency in genetically modified cattle from more to less are casein, CMV and human α-lactalbumin promoters and terminator are SV40 poly A, BGH poly A and casein poly A. Multiple detection and screening strategies aiming at the promoter and terminator are proposed according to the statistical results. This paper also analyses the possible problems in the future in genetically modified animal detection, this review will provide reference for regulatory perspective and detection of genetically modified animals.

Avian-origin reovirus, which is a kind of important poultry and waterfowl pathogen, can cause serious damage to the bursa of Fabricius, spleen and thymus of infected birds, resulting in immunosuppression and increased susceptibility to secondary diseases that greatly hinder the healthy development of global poultry and waterfowl industry. The innate immune response is the hotspot in the field of microbiology and immunology due to its earlier and quick non-specific response against pathogens. Adaptive immune response can eliminate the intruding pathogens,and defend against reinfection of the same pathogens, which provides immunological basis for the immunization of host. In this review, we highlight the current understanding of host innate immune response and adaptive immune response induced by avian-origin reovirus and the effects of such virus infection on the host immune system. This review will provide a basis for understanding the virus-host interaction and vaccine development.

This study aimed to explore the application of GBLUP and penalized regression methods in genomic selection of the hematological traits in pigs. We chose 13 hematological traits from the immune resource population collected by our laboratory as the analyzed objects. We used the genotyping data of Illumina PorcineSNP60 Genotyping Beadchip to conduct the genomic selection analysis, in which GBLUP and 3 penalized regression methods (ridge, lasso and elastic-net) were used based on additive model and additive-dominance model. The results showed that the accuracy of genomic selection was positively correlated with estimated values of chip heritabilities of traits. The results of cross-validation analysis showed that the MCV (mean corpuscular volume) had the highest prediction accuracy among 13 hematological traits. The prediction accuracy of additive model and additive-dominance model were different in different traits. In total trend, the prediction accuracy of the lasso and elastic-net regressions were lower than that of the ridge regression and GBLUP. But in a few traits, such as NE%, it was opposite. In conclusion, there is no optimal genomic prediction method that could be suitable for all traits, and we should consider the genetic characteristics of the target traits when choosing a genome evaluation method. This research provides important reference information for the practical application of genomic selection for immune traits in pigs.

The aim of this study was to clone the coding sequence of FoxO 4 gene of Congjiang Xiang pig and preliminarily explore the function of FoxO 4 gene. The relative expression level of FoxO 4 gene was detected by qRT-PCR in different tissues of Congjiang Xiang pig at the age of 6 month.The recombinant plasmid pEGFP-N3-FoxO4 was constructed by inserting the FoxO 4 gene coding sequence into pEGFP-N3, and then transiently transfected into the subcutaneous preadipocytes of Congjiang Xiang pig by liposome-mediated transfection.The expression levels of FoxO 4, PPARγ, LPL, FAS, ACC, ATGL, HSL in transfected cells were detected by qRT-PCR. The results showed that FoxO 4 gene had the highest expression level in adipose tissue. The constructed plasmid pEGFP-N3-FoxO4 was successfully expressed in subcutaneous preadipocytes. The expression level of LPL, FAS, ACC, ATGL and HSL genes were significantly up-regulated in the subcutaneous preadipocytes when compared to the control group.These results revealed that FoxO 4 gene had certain regulation on fat deposition and might be a key candidate gene for the pork quality, which will provide theoretical basis for further studying the mechanism of fat deposition.

The aim of this experiment was to study the influence of somatic cell nuclear transfer on IGF2-H19 locus methylation in cloned goat fibroblast cells. Goat ear fibroblast cells (GFC, control group) and cloned goat ear fibroblast cells (CFC, experimental group) were used as experimental materials. When cells in the 2 groups were cultured to passage 5, using cell counting method for cell growth curves, flow cytometer for cell apoptosis, real-time PCR for gene relative expression level and bisulfite sequencing PCR for the methylation status of differentially methylated region (DMR). The results showed that cell growth curves in the 2 groups were typical "S"-shaped, but apoptosis rate in CFC group was increased significantly(P<0.01) compared with that in GFC group. The expression level of Dnmt1 (P<0.01), Dnmt3b (P<0.01), Tet1 (P<0.05), Tet2 (P<0.05), H19 (P<0.05) and IGF2 (P<0.01) in CFC group were decreased significantly compared with that in GFC group. However, no statistical differences were found at the expression levels of Tet3、Dnmt3a and IGF2R between the 2 groups. Compared with GFC group, the methylation level of IGF2 DMR1 (74.1% vs. 57.8%, P<0.01) and DMR2 (76.8% vs. 40.0%, P<0.01) were decreased significantly, while the methylation level of IGF2-H19 imprinting control region (ICR) was increased significantly (68.8% vs. 84.0%, P<0.01) in CFC group. Somatic cell nuclear transfer resulted in aberrant methylation of IGF2-H19 locus by influencing Dnmts and Tets genes expression, which could affect genomic imprinting, and then influence the efficiency of serial recloning.

The study aimed to carry out the comparison proteomic analysis of wapiti antler and bone based on Label-free. In this paper, the wapiti antlers and bone aged 130 d were selected as experimental material, then the protein components of antler and bone were analyzed by using the Label-free proteomics methods and bioinformatics methods. The results showed that a total of 1 138 proteins were identified, including 934 deer bone proteins and 835 antler proteins. Through the functional annotation of proteins successfully identified, deer bone proteins were specifically enriched in the process of low-density lipoprotein receptor particle metabolism, substantia nigra development, angiogenesis and regulation of cell apoptotic signaling pathway. The antler proteins were enriched in the peptide hormone processing, regulation of activated T cell proliferation and other biological processes. The proteins related to peptide hormone processing process were CPE, CPN1, ECE1, and the proteins related to regulation of activated T cell proliferation were IGF2, PRKAR1A, PYCARD. A total of 335 differentially expressed proteins (fold change>2, P<0.05) were screened out by SPSS software. Among them, 75 proteins were up-regulated in deer bone and 260 proteins were up-regulated in antlers. In addition, cathelicidin 1 and cathelicidin 5 were highly expressed in antlers, while cathelicidin 6 was highly expressed in the deer bone.The results fill the blank in the study of antler and deer bone protein components, and lay the foundation for the further study on the pharmacological active substances of antler and deer bones.

The objective of this study was to develop a software tool for exploring social genetic effects of some ecomomic traits of livestock and poultry. According to the previous selection theories of the social genetic effects and its optimal selection methods developed by our group, we developed a software system for simulating, predicting and optimizing the social genetic effects (SSPOS) using QT, Fortran and R etc., which was a tool for comprehensively analyzing the social genetic effect of economic traits in livestock population and visualizing the results using many enriched interfaces. We could effectively simulate selection progress of social genetic effects under different cases, rapidly predict the social genetic effects for some actual data by SSPOS, and obtain the optimal selection decision of social genetic effects for the target traits in breeding schemes. The results indicate that SSPOS is a useful tool to investigate the social behavior interaction of livestock and poultry, and can rapidly analyze social genetic effects of the economic traits.

The aim of this study was to find out signaling pathways and candidate genes that related to cold tolerance of Mongolia cattle by RNA-sequencing and bioinformatics analysis of Mongolia cattle skin tissues in winter and summer. Transcriptome sequencing skin tissues of adult Mongolia cattle in winter and summer were carried out by Ion Proton^TM Sequencer, respectively. And then data processing, reference genome alignment, gene differential expression analysis, GO and KEGG enrichment analysis and candidate gene screening were applied on the transcriptome data; Additionally, the reliability of the transcriptome data was detected by Real-time PCR. The results showed that 86 954 060 and 69 837 009 reads were obtained from winter and summer groups, respectively; The percentage of the uniquely mapped reads was 73.21% in winter group, and 75.55% in summer group. There were 182 DEGs between winter and summer groups, compared to summer group, 117 were up-regulated and 65 were down-regulated in winter group; Real-time PCR analysis showed that the transcriptome sequencing results were reliable; The results of GO analysis showed there were 21, 39 and 224 terms enriched in the cellular components, molecular functions and biological processes, respectively; KEGG analysis showed that the DEGs were significantly enriched in lipid metabolism, blood coagulation, tyrosine metabolism, steroid hormone biosynthesis and retinol metabolism related pathways; 8 candidate genes were screened out. In summary, lipid metabolism, blood coagulation and melanin synthesis related pathways and genes as well as hair synthesis related genes may play an important role in cold resistance of Mongolian cattle.

In this study, the role of Gata6 in porcine induced pluripotent stem cell (iPS cell) and extraembryonic endoderm stem cell (XEN cell) isolation was analyzed deeply. The required genes were cloned using templates of cDNAs, which were reverse transcripted from the mRNAs of the fetal tissues of Nongdaxiang pigs, and the retroviral plasmids pMXs-pOct4, pMXs-pSox2, pMXs-pKlf4, pMXs-pMyc, pMXs-pGata6, pMXs-pLin28, pMXs-pTbx3 and pMXs-pNanog were constructed, then, porcine fetal fibroblasts were infected with different combinations of genes. The results showed that co-transfection of Gata6 with Oct4, Sox2, Klf4 and mMyc (G6OSKM) could promote early reprogramming efficiency. Gata6 replacement of Oct4, with Sox2, Klf4 and mMyc (G6SKM), also could induce porcine fetal fibroblasts to iPS cells, but the reprogramming efficiency decreased, compared with the control group of OSKM (Oct4, Sox2,Klf4 and mMyc). Over expression of Gata6 forced pig iPS cells to undergo differentiation at the late stage of reprogramming, and changed into XEN cell like cells morphologically. These cells could maintain the morphology of XEN cells in mouse XEN cell culture medium, express XEN cell marker genes such as Gata4, Gata6 and Sox17. Overexpression of Gata6 in porcine iPS cells obtained by OSKM 4 factors also produced XEN cell like cells. The results indicate that overexpression of Gata6 can generate porcine XEN cell like cells, can provide theoretical basis for the isolation and derivation of porcine XEN cells from normal embryos and the exploration of the mechanisms of porcine early embryo developmental regulation.

This research was carried out to study the effect of low protein level diets with balanced amino acids on intestinal antimicrobial peptide and microbial flora in fattening pigs. Twenty pigs (Duroc×Landrace×Yorkshire) weighed (60±2.5) kg were randomly assigned into 4 treatments:14.0% protein level diet (14.0% CP), 12.5% protein level diet supplemented with Glu, Lys, Met, Thr, Trp (12.5%CP), 11.0% protein level diet supplemented with Glu, Lys, Met, Thr, Trp (11.0% CP), 11.0% protein level diet supplemented with Glu, Lys, Met, Thr, Trp and BCAA(Val、Ile、Leu)(11.0% CP +BCAA). Each treatment had 5 replicates, each replicate had one pig. The trial was divided into 5-d pre-trial period and 30-d test period. The results showed as follows:(1) Reducing the dietary protein level led to lower concentration of urea nitrogen and uric acid(P<0.05); (2) Low protein level diets significantly reduced the level of β-defensin-2 in jejunum and ileum mucosa as well as mRNA expression levels of ANG1 and ANG4 (P<0.05). (3) Different protein levels of diets had significant effect on the partial amino acid compositions of the ileum and rectum microorganisms (P<0.05); (4)The Observed species of 12.5% CP group were increased significantly compared with 14.0% CP group (P<0.05). The Shannon, PD of 11.0% CP group and Shannon of 11.0% CP+BCAA group were decreased significantly (P<0.05) compared with 14.0% CP group. There was no significant difference in Bacteroidetes and Firmicutes abundance among 4 groups (P>0.05). The relative abundance of Acidobacteria and Gemmatimonadetes in 12.5% CP group and 11.0% CP group as well as the relative abundance of Gemmatimonadetes in 11.0% CP+BCAA group were decreased significantly compared with 14.0% CP group(P<0.05). The relative abundance of Proteobacteria in 14.0% CP group was higher than that in 12.5% CP group and 11.0% CP+BCAA group (P<0.05) and was not significantly different with 11.0% CP group (P>0.05), the relative abundance of Campylobacteraceae of 14.0% CP group was higher than that in other groups(P<0.05). The relative abundance of Coprococcus and Clostredium_sensu_stricto in 14.0% CP group were higher than that in other groups (P<0.05). The relative abundance of Lachnospiraceae FCS020 in 11.0% CP group and 11.0% CP+BCAA group were higher than that in 12.5% CP group and 14.0% CP group (P<0.05). Low protein level diets with balanced amino acids can improve nitrogen utilization and maintain the balance of the intestinal microbial flora. Low protein balanced diets with BCAA help to increase the amino acid balance and maintain intestinal immune function.

The aims of this study were to investigate the effects of different starch types on intestinal digestion and absorption, serum biochemical indices and hormone levels of finishing pigs. Ninety barrows ((68±2.0) kg) were randomly allocated to 3 diet treatments with 5 replicates and 6 pigs per replicate. Purified waxy maize starch (WMS), nonwaxy maize starch (NMS) and pea starch (PS) were used as the only dietary starch sources to formulate experimental diets (the amylose to amylopectin ratio were 0.07, 0.19 and 0.28, respectively). At the end of 28 days of treatments, two pigs per replicate(near to the average body weight of the replicate) were weighed, slaughtered and the samples of jejunum mucosa and blood were collected, and the target traits were detected. The results showed that:1) Dietary starch types had no significant effect on the activity of digestive enzymes in jejunum mucosa of finishing pigs (P>0.05). 2) Compared with the WMS diet, the PS diet significantly down-regulated the mRNA expression of taste receptor type 1 member 2(T1R2), taste receptor type 1 member 3(T1R3) (P<0.05), whereas significantly up-regulated the mRNA expression of solute carrier family 1 member 5 (SLC1A5) (P<0.05); the NMS diet significantly up-regulated the mRNA expression of solute carrier family 7 member 7 (SLC7A7) (P<0.05); PS and NMS diets significantly down-regulated the mRNA expression of sodium/glucose cotransporter 1 (SGLT1) and glucose transporter 2 (GLUT2) (P<0.05). 3) In the serum biochemical indices, no significant effect was observed in the plasma glucose content among groups (P>0.05). Compared with the WMS diet, PS and NMS diets significantly reduced the plasma free fatty acid content (FFA) (P<0.05). 4) No significant effect was observed in the serum hormones levels among groups (P>0.05). It could be concluded that finishing pigs fed with high amylose to amylopectin ratio diet could down-regulate the mRNA expression of sweet taste receptors and glucose transporters, up-regulate the mRNA expression of some amino acid transporters, and then decrease the absorption of glucose, be beneficial to the absorption of amino acids.

This study assessed the effects of diets with different non-fiber carbohydrate (NFC)/neutral detergent fiber (NDF) levels on rumen fermentation parameters and bacterial community of male calves. Thirty 2-3 month-old healthy weaned male calves with (94.38±0.25)kg were randomly divided into 2 groups with 15 calves each, and were fed 2 types of total mixed ratio diet with the same crude protein level and different NFC/NDF ratios (1.35(A) and 0.80(D)), respectively. The experiment lasted for 105 d, with 15 d of adaptation period and 90 d of formal period. Six calves in each group were selected to collect rumen fluid through oral for 2 hours after morning feeding on the 120, 150 and 180 d. The pH, volatile fatty acids (VFA) and NH₃-N concentration and rumen microbial diversity of the rumen fluid samples were determined. The results showed that:1) The dietary NFC/NDF levels had no significant effect on rumen fluid pH and total VFA concentration (P>0.05); 2) Acetate, NH₃-N concentration and acetate/propionate ratio in group A were significantly lower than that in group D (P<0.05), propionate content in group A were significantly higher than that in group D (P<0.05); 3) The microbial diversity parameters of rumen fluid of calves in group A were significantly lower than that in group D (P<0.05); 4) At the phylum level, bacteriodetes proportion accounting for the total number of sequences of calves in group A was significantly higher than that in group D (P<0.05), Christensenellaceae_R-7 and Prevotellaceae_UCG-003 proportions accounting for the total number of sequences of calves in group D was significantly higher at the genus level than that in group A (P<0.05); 5) With the age increasing, the diversity indices except for Shannon index, were significantly different between the 2 groups (P<0.05), but there was no significant difference in rumen microbial composition between the 2 groups(P>0.05). In summary, the dietary NFC/NDF levels significantly affect the species and abundance of the rumen microorganisms, and the level of genus of microorganisms, finally affect the yield of acetate and propionate.

To get native porcine Ficolin α, Bac-to-Bac baculovirus expression system was used. Firstly, rBacmid-Ficolin α was constructed, and then transfected into sf9 insect cells to obtain recombinant baculovirus rBV-Ficolin α. The optimized expression conditions were detected by Western blot and analyzed by gray level using Image J software. Followed by purification, antiviral activity of target protein was assessed in vitro. Western blot results showed that the expression level of Ficolin α in transfected sf 9 insect cells was higher at the infection (MOI) of 7.5 when they were harvested at 96 h, and porcine Ficolin α had the antivirus activity against porcine reproductive and respiratory syndrome virus (PRRSV). In conclusion, porcine Ficolin α was successfully expressed in sf9 insect cells and showed strongly anti-PRRSV activity. This study provided a material basis for the study of antiviral activity and its mechanism of porcine Ficolin α.

This experiment was conducted to study the conservation and variation of P46 gene and P97R1R2 gene of three Mycoplasmal pneumonia of swine vaccine strains, e.g.168, RM48 and Z, from domestic production. The DNA of the four vaccine manufacturer products were extracted. The target gene fragments amplified by PCR were sequenced. And the nucleotides sequence and the deduced amino acids sequences were analyzed by DNAStar software, which compared with four international standard strains.The attenuated vaccine strain 168 showed 100% homology of P46 nucleotides sequence with vaccine strain Z and RM48, while 99.1% of that with strain J and 232, 98.9% of that with strain 7422 and 7448. As for P97R1R2 gene, strain Z and RM48 showed completely same identity, and 97.5% of identity with strain 168. The homology among vaccine strains, strain J, 232, 7448 and strain 7422 was lower, only from 90.1% to 93.6%. The major differences of the vaccine strains occurred in the repeat numbers of P97R1 amino acid sequence, which is benefit for differentiation of the attenuated vaccine strains.

The aim of this study was to characterize the dUTPase gene of Taenia multiceps and analyze its enzymatic activity. The recombinant Tm-dUTPase was expressed in prokaryotic system and the enzymatic activity was measured; Western blotting analysis and immunofluorescence localization were also performed. Sequence analysis showed that the Tm-dUTPase gene contains a 447 bp open reading frame and the mature polypeptide consists of 148 amino acid residues, with a predicted molecular weight of 16.39 kDa and PI of 6.263; the deduced amino acid of Tm-dUTPase shares 97% and 91% similarity with dUTPase from Cysticercus cellulosae and Taenia pisiformis respectively. The recombinant Tm-dUTPase was expressed as a soluble protein, and the purified protein has a concentration of 0.907 mg·mL^-1. The Western blotting showed that the recombinant Tm-dUTPase could be recognized by the serum of goat naturally infected with Coenurus cerebralis, indicating a relatively high immunoreactivity. The immunolocalization showed that the native Tm-dUTPase mainly distributed to the parenchymatous zone of the adult parasite, and widely distributed to the protoscolexes and out layer of the cystic wall of Coenurus cerebralis. The enzyme activity of Tm-dUTPase was 986.29 U·mg^-1, and the activity could be inhibited by EDTA and enhanced by Mg²⁺, Zn²⁺ and Cu²⁺. These results would greatly benefit the further study of the biological function of Tm-dUTPase.

In order to investigate species and genus distribution of yeast on hairs of giant panda and their pathogenicity on mice, thirty-seven hair samples of giant panda were collected and 45 isolates of yeast were isolated from the samples with traditional microbial isolation method. PCR amplification on the strains using the universal fungi primer of ITS1 and ITS4 and phylogenetic analysis were performed. And the 45 isolates belonged to 12 species. Drug sensitive qualitative test of 12 species of yeast were researched using 5 kinds of clinical commonly antifungal medicines by the Danish Rosco disc diffusion method. The results indicated that 12 species of yeast were isolated, including Trichosporon cutaneum, Trichosporon moniliiforme, Trichosporon interdigitale, Trichosporon akiyoshidainum, Trichosporon asteroids, Trichosporon brassicae, Trichosporon guehoae, Trichosporon jirovecii, Trichosporon montevideense, Trichosporon sp., Candida catenulate and Cryptococcus fragicola. Most of the yeast strains were sensitive to 5 kinds of antifungal drugs. C. fragicola was resistant to ketoconazole, C. catenulate and T. interdigitale were resistant to amphotericin B, C. catenulate and T. jirovecii were resistant to itraconazole, Trichosporon sp. was resistant to fluconazole,C. catenulate, T. interdigitale, T. guehoae and T. cutaneum were resistant to nystati. T. cutaneum, T. interdigitale, T. asteroids, C. fragicola were pathogenic to mice, and other isolates of yeasts were opportunistic pathogenicity. Various yeast species exist in surface of giant panda, and they have different drug sensitivity to fungicides. These results have greatly enriched the understanding of the yeast on hair of giant panda,will provide a reference for the dermatomycoses diagnosis and treatment of giant panda.

To identify the sequence character and antibacterial activity of holin from phage Bp7, holin gene of phage Bp7 was amplified by PCR and amino acid sequence was compared and analyzed. Recombined plasmid pCold-holin was constructed and transferred into E. coli BL21 to express holin protein. The best induction conditions were explored. The protein was purified by affinity chromatography and its antibacterial activity was determined. The results showed that the holin of phage Bp7 contained a transmembrane region and belonged to Phage-holin-T family. The soluble protein was expressed in E. coli BL21. Turbidity test of E. coli BL21 showed that the expression of holin protein could significantly inhibit the proliferation of E. coli BL21, decrease the number of cells in 4 h, also the cells showed a agglutinative form. The results indicate that holin of phage Bp7 has antibacterial activity, which is helpful to further study of the antibacterial mechanism of holin.

As the specific energy regulator, the function of ghrelin was widely studied in the energy metabolism and reproduction regulation, but its exact regulatory mechanism in the estrous cycle of mice has still not been elucidated. In this study, by intraperitoneal injection of ghrelin to mice, we investigated its effect on feed intake and serum estradiol concentrations (competitive immunoassay electrochemistry luminescence detection) of mice during different stages of estrous cycle, the co-localization between estrogen receptor α (ERα) and ghrelin which existed in arcuate nucleus (ARC) were also studied by immunofluorescence. In the results, ghrelin (50 μg·kg^-1) enhanced the feed intake of mice in protesters, in two hours post injection (P<0.05); ghrelin suppressed the concentration of estrogen in serum (P<0.01), and there was no significant change in other estrous cycle. Moreover, ghrelin enhanced mice neuron activation (P<0.05 or P<0.01), and ghrelin directly act on ERα neuron in ARC nucleus in whole estrous cycle, and highest co-expression rate was occurred in the proestrus. Thus, we hypothesis that ghrelin has the ability to direct acting on ERα neurons in mice hypothalamus to effect on the concentration of estrogen in serum, and finally mediate mice food behavior in mice protesters.

This experiment was conducted to investigate the histological characteristics and age-related changes in yak kidney. Newborn (1-7 days old, n=5), adult (3-6 years old, n=5) and old (7-10 years old, n=5) yaks were selected as the subjects, and histological and statistical methods were used to observe the changes in yak kidney. The relevant data of kidney units was analyzed and measured by the image analysis software. Histological observations showed that yak kidneys consisted of the surface of the membrane and the internal substance. The malpighian corpuscles were composed of glomeruli and renal capsules. The wall of the renal capsule was rich in collagen fibers and the basal membrane of malpighian corpuscles was PAS positive. The kidney tubules included proximal tubules and distal tubules, which are rich in collagen fibers and had a positive PAS on the epithelium basal membrane. The results of measuring showed that the number of glomeruli per unit field (2.3×10⁶ μm²) decreased with age; while the number of glomerular cells and the size of glomerular nuclei increased with age; the differences was in the maximum cross-sectional area of glomeruli and malpighian corpuscle were the largest in adult, followed by the differences in the aged, with those in the newborn being the smallest. In addition, with the age increasing, the trend of changes in the number of tubule epithelial cells in the kidney cortex and medulla per unit field (2.3×10⁴ μm²), the size of proximal tubule and distal tubule epithelial nuclei were the same as the number of glomeruli and cells in glomeruli, the size of glomeruli nuclei, and the maximum cross-sectional area changes of glomeruli and malpighian corpuscle. The results suggested that the histological structures of yak kidney changed with the increase of age. Moreover, it is presumed that the contributing factors to this change might include glomeruli sclerosis and reduction of substantial cells, however, further empirical investigation is required to be conclusive.

The experiment was conducted to study effects of heat stress on Hsc70 import-export nuclear and apoptosis in H9c2 cardiac myocytes. H9c2 cells were exposed to 42℃ as the heat stress model, inhibited expression of Hsc70 was performed by transfecting Hsc70 siRNA, the expression of Hsc70 in the cytoplasm and nucleus were detected by Western blot, LDH concentration of cell culture was detected by ELISA, and apoptosis was detected by Annexin V-FITC/PI double staining. The results showed that:in normal H9c2 cardiomyocytes, the level of Hsc70 in cytoplasm was high and that in nucleus was low, the level of Hsp72 in cytoplasm and nucleus was very low. There was no significant difference in the expression of Hsc70 after heat stress. The expression of Hsc70 in nucleus was significantly increased after heat stress 30 and 100 min (P<0.01), and decreased after heat stress 240 min. After heat stress, the expression of Hsp72 was significantly increased (P<0.01 or P<0.05). After Hsc70 inhibited, the level of cytosolic Hsc70 was significantly decreased, and Hsc70 import nucleus decreased significantly after heat stress. After Hsc70 inhibited, there was no significant effect on the expression of Hsp72 in cytoplasm and nucleus. Compared with the heat stress group, the expression of LDH was increased in the heat stress+Hsc70 siRNA group, and there was a significant difference between the two groups after heat stress 100 min (P<0.05). After Hsc70 inhibited, H9c2 cells were more prone to apoptosis after heat stress, and there was a significant difference between the two groups in heat stress 30 and 100 min (P<0.05). In conclusion, heat stress can cause Hsc70 import and export nuclear, and the low level of Hsc70 nucleus can cause of H9c2 cells damage aggravated and the rate of apoptosis increased after heat stress.

In order to investigate the region of apoptosis and the expression of genes related to apoptosis in cartilage cells of Tibial Dyschondroplasia (TD) in broiler chickens, TD model was established, in which eighty AA broiler chickens were divided into two groups; control group and experiment group (given thiram 100 mg·kg^-1 in feed). The morphology of chondrocytes and chondrocytes apoptosis in tibial growth plate of broilers were determined by HE (hematoxylin and eosin) staining and TUNEL technology. Real-time PCR and immunohistochemistry respectively were used to detect the mRNA transcription level of apoptosis-related genes in chondrocytes and protein expression levels. The HE staining result revealed that obvious cartilage plug formed and gradually bigger in the tibial growth plate after day 1, 2, 4 and 6 in TD broilers induced by thiram; and the abnormity of chondrocyte also increased. The TUNEL technology result showed that apoptosis levels of the chondrocytes significantly increased in cartilage lesion parts, especially in the hypertrophic zone. The Real-time PCR results showed that the mRNA transcription level of Bax was up-regulated from day 1 to 6 significantly (P<0.01). The Caspase-3 gene transcription significantly increased on day 1, 2 and 4 in TD broiler tibial growth plate (P<0.01). The transcription of anti-apoptosis gene BAG-1 was significantly decreased on day 1 and 2 in TD broiler tibial growth plate (P<0.01). The results of immunohistochemistry showed that the expression of Bax protein was significantly higher in the treated group than in the control group on day 1 and 4. It is concluded that the apoptosis of chondrocytes in TD broilers growth plate was related to the enhancement of pro-apoptotic gene expression and decreased of anti-apoptotic gene expression.