Over the past decade, researches on follicular biology are focused on granulosa cells and their interaction with the oocyte. While, the theca cell has been somewhat forgotten as a necessary and vital part of the folliculogenesis and follicular developmental process in the mammalian ovary. Theca cells synthesize androgens required by the developing follicles, modulate angiogenesis and provide essential nutrients and steroid hormones along follicular developmental process. Their function is also enabled to contribute the process of cell apoptosis and follicular development via mediating the crosstalk with oocyte and granulosa cells. In this study, the advance in the function of theca cells in the mammalian ovary are reviewed to represent the crucial role of theca cells during follicular development and promote the research on the function of theca cells.

 Haploid embryonic stem (haES) cells contain one single set of chromosome and share some characteristics of normal embryonic stem cells, including the ability of self-renewal in vitro and forming multiple functional cells of all the tissues and organs. Recent studies demonstrate that haES cells are divided into parthenogenetic and androgenetic mouse embryos. Abundant data and evidence show that haES could be used as an ideal tool for genetic analyses. Here, we summary the recent progresses, potential applications, and some limitations of the mammalian haES. We also provide some viewpoints about haES combined with our study.

The aim of this study was to identify the active control area of the chicken Piwil1 gene core promoter, to predict the binding sites of the transcription factor, and to investigate the effect of the NF-Y binding sites on the chicken Piwil1 gene core promoter activity. A series of promoter deletion mutants were directly subcloned into pGL3-Basic vector. The transcriptional activity of the recombinant plasmids was detected by Dual-Glo^® Luciferase Assay System after transfecting COS-7 and GC-1 cells. In the deletion analysis of Piwil1 promoter, the core promoter essential for transcriptional activation was identified. With point mutations and Dual-Glo^® Luciferase Assay System，the effect of the NF-Y binding site on the chicken Piwil1 gene core promoter activity were identified. The activity of the -1 377--268 bp region in COS-7 and GC-1 cells were different, while the activity of -221-+1 bp region was almost disappeared. Data analysis demonstrated that there was the NF-Y binding site in -268--221 bp region. By point mutations, the activity was decreased significantly. Therefore, NF-Y site might has an important effect on Piwil1 gene promoter activity. NF-Y is an important regulatory element in Piwil1 core promoter. Furthermore, the results provide the foundation for analyzing the promoter activity and transcriptional regulation mechanism.

This study was performed to examine the haplotypes and copy number variation (CNV) of genome in chromosome 16 of Beijing-You chicken and their association with IgG level. A total of 995 chickens were genotyped using 60 K Beadchip and IgG level were measured at 80 d in serum. PennCNV, Haploview 4.1 and PHASE 2.0 were used to analyze CNV, linkage disequilibrium (LD) and haplotypes. The results represent two CNVs and two Blocks. In Block1, three haplotypes and six diplotypes were observed where IgG level of H1H2 was 1.45 times as H3H3 (P<0.05). In Block2, nine haplotypes and nine normal diplotypes were analyzed, but no significant difference were identified (P>0.05). Detection of copy number variations at GGA16（201 882-299 166 bp）was 1.25 times as that of the duplication of this region (P<0.05). Eight SNPs markers were chosen for analysis. The SNP markers Gga_rs16057310, Gga_rs15788237 and Gga_rs15788101 were significantly differed in IgG level among genotypes (P<0.05). In all, Block1, CNV2 (201 882-299 166 bp) and SNPs Gga_rs16057310, Gga_rs15788101, Gga_rs15788237 may be influenced on IgG level. The genes in these regions such as CD1b, TRIM27,ZNF692 and MHC could be related with chicken immunity function.

 This experiment was conducted to study the expression patten of IGF-Ⅰgene regulating growth and development of sheep and correlation of its SNPs with growth traits in sheep. The IGF-1 gene expression change of IGF-Ⅰin brain,musle,skin,liver and heart were detected at six growth stages(15,60,105,150,195,240 days) by qRT-PCR approach and the correlation of SNPs of IGF-Ⅰgene with growth traits were analyzed by SSCP method in Liangshan Semi-wool sheep population. The results showed that the first inflection point of the expression of IGF-Ⅰgene in musle, skin, brain, liver and heart tissues was at 105 day and the second inflection point was at 195 day without the liver tissue. From 15 to 105 day, the expression of IGF-1 gene in musle, brain and liver tissues decreased at 60 day, and then increased at 105 day, however, it is not significantly different at 15 and 60 day, and increased significantly at 105 day in skin and heart tissues.After 105 day, the expression of IGF-Ⅰ were degraded in all of the tissues at 150 day,and the expression in liver tissue was decreased gradually from 105 to 240 day, but the expression in skin and brain tissues were higher at 195 day than that at 150 and 240 day, and no difference at among 150, 195 and 240 day in musle and heart tissues.The two SNPs which were detected with 587 Liangshan Semi-wool sheep were moderate polymorphism. The SNP of the P-1 primer affected significantly birth weight(P<0.05) and the SNP of the P-2 primer affected significantly weaning weight and the weaning daily gain(P<0.05). The results suggest that the expression of IGF-Ⅰin all tissues have the similar expression patterns,and strong correlation between SNPs of IGF-1 and confirmed the early growth traits provide theoretical basis for the growth regulation and can be applied to assisted selective breeding with the SNPs in Liangshan Semi-wool sheep.

This study aimed to clone ovine LPIN1 gene and to analyze its ontogenetic mRNA expression in seven adipose tissues of two breeds with distinct tail types, and based on which, to reveal the role of the gene in fat metabolism of sheep. RT-PCR technique was used to amplify LPIN1 gene. Bioinformatics methods were used to predict the physicochemical properties and domains in Lipin1 protein. Ninety six animals from Guangling Large Tailed and Small Tailed Han (48 for each breed with equal sex ratios) were slaughtered at 2, 4, 6, 8, 10 and 12 months of age, respectively. Adipose tissues were collected from great and small omental, mesenteric, retroperitoneal, subcutaneous, perirenal and tail fats to study the ontogenetic mRNA expression by real-time PCR. The results showed that the coding region was 2 688 bp in length, coding 895 amino acids. Lipin1 was a kind of unstable and hydrophilic protein with 10 potential glycosylation and 88 phosphorylation sites, but without transmembrane domain and signal peptide. N-and C-terminals contained two highly conserved regions(Lipin-N and LNS2 (Lipin/Ned1/Smp2)).  LPIN1 mRNA expressed in all adipose tissues studied, but the highest in perirenal fat and the lowest in mesenteric fats. Months of age, tissue and breed as well as their twoway interactions had significant influences on the LPIN1 mRNA expression. LPIN1 gene is significant in the genetic improvement of meat quality traits of sheep based on its direct participating in and indirect regulating fat metabolism.

In order to detect the association of UBA52 gene and sexual precocity in goats, female sexual precocious goat breed Jining Grey goat and female sexual late-maturing goat breed Liaoning Cashmere goat were selected. RT-PCR method was used to detect the expression profile of UBA52 gene in tissues at different developmental stages of the two breeds，and then real-time PCR technology was used to exam the mRNA expression level of UBA52 genes in hypothalamus, pituitary gland, ovary, uterus and adrenal gland. The result showed that UBA52 gene was widely expressed in the organizations at different developmental stages of the two breeds, and its expression level in the uterus was the highest.The results also showed that the expression level of UBA52 in pituitary gland in puberty was significantly lower than that in juvenile period of Jining Grey goat（P<0.01）, while the expression level in juvenile period Ⅱwas significantly higher than that in juvenile period Ⅰof Liaoning Cashmere goat, and its expression level in puberty was significantly lower than that in juvenile period Ⅱ（P<0.01）. The results indicated that the up-regulated expression of UBA52 gene was negatively correlated to goats estrus. So it was concluded that the up-regulation of UBA52 may reduce the secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) to suppress the occurrence of goat puberty.

The study focused on the effect of FSH and insulin on the proliferation of granulosa cells and the production of estradiol when the sheep follicle granulosa cells were cultured in serum-free medium. Granulosa cells were collected and treated with a range of concentration of FSH and insulin, and cultured for 168 h using long term method. Observed and counted under the microscope. Estradiol concentration was measured by double antibody ABC-ELISA. Results indicated that granulosa cell proliferation and estradiol production in the groups treated with FSH and insulin were superior to that in other groups treated alone. At 10 ng·mL^-1 concentration of insulin and 5.0 or 25.0 ng·mL^-1 concentration of FSH in medium, the number of granulosa cells were most and there is no significant difference between these two treated groups; at 10 ng·mL^-1 concentration of insulin and 5.0 ng·mL^-1 concentration of FSH in medium, Estradiol concentration was the highest (P<0.05) and the culture system was optimal. The multiplication and secretion of estradiol in sheep ovarian granulosa cells were induced by FSH. Insulin promotes the multiplication and estradiol secretion of FSH-induced sheep follicular granulosa cells in vitro. FSH and insulin stimulate the multiplication and estradiol secretion of sheep follicular granulose cells in a concentration-dependent manner.

 To study the localization of GHR and IGF-IR and the expression of GHR mRNA and IGF-IR mRNA in uterus of Jining Grey goats during postnatal development. Real-time flourescence quantitative PCR was used to detect the expression pattern of GHR mRNA and IGF-IR mRNA, and SABC method was applied to research the distribution of positive cells of GHR and IGF-IR in the uterus. GHR and IGF-IR were mainly localized in the cytoplasm of glandular epithelium, stromal cells, endothelium and smooth muscle cells of the uterus, occasionally in the nuclei. The changed trends on the positive cells of GHR and IGF-IR were similar with that on the expression of its mRNA. Although the expression of IGF-IR was less than that of GHR, the expression of GHR and IGF-IR both reached its peak on the puberty (D60) and the sexual maturity (D120)（P<0.01）, Then decreased and remained at a relatively stable level. GHR and IGF-IR play an important role in the growth of the uterus, and the puberty and the sexual maturity may be the fastest period of the uterus during postnatal development.

This experiment was conducted to study the quantitative relation model between the nutrient requirement and growth performance of Xiangcun Black pig at different growth phases. According to the uniform design U8*(8⁵), the experiment took crude protein(CP),lysine(Lys) and total phosphorus(TP) as three factors and set eight levels. The experiment was divided into 10-30,30-60,60-90 kg three growth phases based on physiological characteristics of Xiangcun Black pig. In each stage, forty eight Xiangcun Black barrows with approximate initial weight (10.08±0.49)，(30.07±0.73) and (59.96±0.83) kg were selected, which were randomly divided into eight treatment groups with six replicates per treatment group and one Xiangcun Black pig per replicate. The results showed that, except the TP requirement of Xiangcun Black pig in 30-60 and 60-90 kg stage, requirement of three nutrient factors was lower than recommended requirement of China Feeding Standard of Swine. The best growth performance of Xiangcun Black pig was obtained when the optimum combination of the three factors was CP 17.18%, Lys 0.90%,TP 0.56% at 10-30 kg stage with 0.48 kg·d^-1 for daily gain and 2.58 for feed conversion rate, CP 15.84%, Lys 0.79%,TP 0.52% at 30-60 kg stage with 0.68 kg·d^-1 for daily gain and 2.97 for feed conversion rate, CP 13.74%,Lys 0.68%,TP 0.47% at 60-90 kg stage with 0.77 kg·d^-1 for daily gain and 3.60 for feed conversion rate. The experiment also showed that CP was the most important factor to average daily gain(ADG) and Lys was the most important factor to feed conversion ratio(FCR). In conclusion, nutrient requirement of Xiangcun Black pig is different from other feeding standard of swine，and further research is needed for nutrients in addition to CP, Lys amd TP.

The aim of this experiment was to study the effects of electronic feeding station on stress response and feeding behavior of growing pigs. One hundred and twenty-two Large White growing pigs with initial weight of (28.2±3.41) kg were randomly allotted into station 15 group, common silo 15 group and station 8 group by pens. The pigs were stratified by sex. The 15 groups had 3 replications, and 8 group had 4 replications. Feeding period was 44 days. During the test period, the skin temperature of pigs was tested, and the saliva was collected to determine the cortisol concentration. At the same time, the feeding behavior was analyzed. The results showed that salivary cortisol level of pigs had no significant differences between the three treatments. The station significantly increased the pig skin temperature (P<0.05) on 2^nd day, and significantly decreased skin temperature (P<0.01) on 15^th day. Feed intake of pigs in station 8 group was slighter higher than that in station 15 group, and the gap of feed intake tended to be lager after 30 days. Intake times of pigs in station 8 group was lower than that in station 15 group, but the cumulative feeding time during a day was higher than that in station 15 group. Both station 8 and 15 group had feeding peaks at 09:00, 15:00 and 19:00，but the daily feeding behavior curve of pigs in station 15 group was more gentle.

The experiment was conducted to investigate the effects of ADF levels on growth performance, nitrogen metabolism, cellulase activity and caecum fermentation of Rex Rabbits from weaning to 3-month-old. Using a one-factor completely randomize design, two hundred weaned Rex Rabbits with similar body weight were randomly divided into 5 treatments(40 replicates per treatment,1 rabbit per replicate):feeding diets containing ADF 15%, 17%, 19%, 21%, 23%, respectively. The trial lasted for 7 days for adaptation, and 53 days for test, respectively. By animal breeding experiment, digestibility experiment and slaughtering experiment, indexes were determined. In the case of the initial body weight had no significant difference, the results showed that the levels of dietary ADF significantly affected the feed intake（ADI）and feed gain ratio(F/G) (P＜0.05); the dietary ADF levels significantly affected the nitrogen intake(IN) (P＜0.01) and fecal nitrogen（FN）(P＜0.05); the nitrogen utilization（RN/IN）and the bioavailability of nitrogen（RN/DN）reached to the maximum in C group; the levels of dietary ADF significantly affected NH3-N concentration and butyrate acid concentration (P＜0.01) in caecum;The activity of xylanase and salicin were significantly increased as the levels of ADF increase (P＜0.01). In conclusion, the suitable ADF levels was 19% for the Rex Rabbits from weaning to 3-month-old.

 To study the effect of STI on the generation of oxygen free radical in vivo at different growth stages of mice, 180 clean and male KM mice were selected and randomly divided into three groups. The first group as a control group was fed control diet, the second group(STI group) was fed control diet containing STI，the third group(VC group) was fed STI diet supplemented with vitamin C. After mice being fed 1, 2, 3, 4 and 5 week, serum and pancreas samples were collected to measure oxidative and antioxidative parameters. The results showed that, compared with the control group, the oxidant parameters significantly increased and the antioxidant parameters significantly decreased in serum and pancreas of STI group (P<0.05). In the whole growth periods, MDA content increased in the first two weeks, and achieved the maximum in the third week, then dropped. The ability of anti-superoxide anion firstly increased, then dropped, again increased and decreased. The ability of inhibiting hydroxy radical slowly increased. After VC being supplemented, the level of oxidation decreased, but its value was still significantly higher than that of control group (P<0.05). The activities of T-SOD in serum and pancreas dropped in the first two weeks, achieved the minimum in the third week, then increased. The activities of CAT in pancreas slowly increased . After VC being supplemented, the ability of antioxidant defenses in serum and pancreas were enhanced, but still lower than that of control group (P<0.05). The present study indicates that the effect of STI on free radical level in vivo of mice is related to growth periods, and this effect is the most significant in the third week. In addition, VC can interfere with the role of STI on the organism and effectively improve the status of oxidative stress.

Recombinant baculovirus containing M and sM gene of transmissible gastroenteritis virus (TGEV) were constructed using Bac-To-Bac Baculovirus expression system, and recombinant M, sM proteins were expressed by infecting the sf9 cell with recombinant baculovirus. Firstly, M gene and sM gene of TGEV were amplified, then were cloned and inserted into donor plasmid pFastBacTMDual to obtain recombinant donor plamid, pFastBac^TMDual-M and pFastBac^TMDual-M-sM. Plasmid pFastBac^TMDual-M and pFastBacTMDual-M-sM were transformed into E. coli DH10Bac respectively. Recombinant baculovirus rBac-M and rBac-M-sM were obtained by transfection of the sf9 cells with Bacmid-M and Bacmid-M-sM. Western blot and indirect immunofluorescence assay (IFA) were conducted, and the results showed that recombinant M protein and sM protein was expressed successfully, two proteins had a positive reaction with antiserum. In vitro experiments of virus-like particles (VLP) assembly of TGEV was performed by infection of insect cell with rBac-M and rBac-M-sM. It is showed that the expressed M proteins alone and co-expression of M and sM proteins in insect cell Sf9 infected by recombinant baculovirus could form VLPs morphologically similar to virion of TGEV. Diameter of VLP is variable from 61 nm to 101 nm. This study provides the foundation for researching the VLP assembly of TGEV.

The aim of current study was to identify the genotype and genetic variation of Echinococcus granulosus in the Tibetan plateau of China, and provide the basic molecular data needed for studies of the molecular diagnosis, epidemiology, prevention and control of echinococcus diseases. The whole mitochondrial ND5 gene sequences of 43 isolates from the Tibetan plateau were analyzed. We found out that 42 isolates were belonged to E. granulosus G1 genotype, while one isolate from Tibetan sheep was identified as E. granulosus G6 genotype according to phylogenetic tree. There were 43 mutation sites in G1 isolates, which were separated in 27 haplotypes. The haplotype diversity and nuclear diversity were 0.940±0.028 and 0.001 93±0.001 14, respectively; And mean genetic distance between haplotypes was 0.002 6. Parsimony network showed a radialized expansion from a main founder haplotype H₉. The analysis of mismatch distribution showed a unimodal structure. Additionally, significant negative neutrality indices were detected. All these results suggested that the main genotype of E. granulosus in the Tibetan plateau was G1 with low genetic variability.

The aim of this study was to establish the culture method of piglet oral mucosal epithelial cells in vitro. Piglet buccal cells were cultured and subcultured with the direct explant technique, trypsin digestion, combined method of DispaseⅡand trypsin digestion, and DispaseⅡ combined with explant technique. Morphological characteristics were observed under the light microscope and transmission electron microscope (TEM). For second cultures, the expression of cytokeratin was detected by indirect immunofluorescence, cell growth curve was measured by MTT method，cell cycle distribution was evaluated by flow cytometry. The results showed that in the same culture medium the explant technique and enzymatic method were not successful, the cells obtained by the method of combination of dispaseⅡand trypsin digestion were difficult to attach and grow, but the cells obtained by the combination of DispaseⅡ and explant technique proliferated quickly, and were of high purity, which can be passaged continuously till to the 10-11 generations, and the cells were identified with the characteristics of oral mucosal epithelial cells. The culture method of piglet oral mucosal epithelial cells in vitro has been established successfully, which may provide ideal research material for study the immortalization of oral mucosal epithelial cells, swine foot-and-mouth disease (FMD) pathogenesis and its vaccine.

This study was conducted to investigate the distribution of P-gp and the transcription level of mdr1 mRNA in liver, kidney and intestines of healthy piglets. Five crossbred pigs (Large white × Landrace × Duroc) of approximately 9 weeks of age with an average body weight of 20 kg were used. The localization of P-gp in healthy piglets was studied by immunohistochemisty method. And real-time RT-PCR method was used with gapdh as house-keeping gene to detect the transcription level of mdr1 mRNA. Immunocytochemical method using specific monoclonal and polyclonal antibodies against P-gp confirm high expression in liver, intestines and kidney of the porcine. In the porcine liver, only the apical membrane of the epithelial cells in the bile ducts was positive. In the kidney, immunoreactivity is obvious in the proximal and distal tubules. In the intestines, P-gp was localized in the apical membranes of the enterocytes and the epithelial cells of intestinal gland. The transcription level of mdr1 mRNA ranked from high to low in all tissues was ileum, colon, liver, jejunum, duodenum, rectum, cecum and kidney. The mdr1 mRNA transcription level of ileum was obviously higher than those of cecum and kidney (P=0.005, P=0.001) and significantly higher than those of duodenum and rectum (P=0.017, P=0.014). Also the transcription level of mdr1 mRNA of kidney was significantly lower than those of jejunum, colon and liver respectively (P=0.046, P=0.018, P=0.030). The results indicate that P-gp could be detected in all test tissues，but the mdr1 transcription level is different. The high expression level of P-gp in ileum and liver could play an important role in drug disposition in body and induce drug-drug interaction in clinic.

The aim of the present study was to find out the effects of drinking boron on osteogenesis of tibias of ostrich chicks. Sixteen ostrich chicks were randomly assigned to four groups. The ostrich chicks were fed with 0 (group A), 100 (group B), 200 (group C), and 400 mg·L^－1(group D) of additional boron in water for 30 d. At the end of experiment time, the ostrich chicks were slaughtered and their top quarter of tibias were obtained, then the samples were made into paraffin sections, observed and photographed under Olympus DHP microphotograph. The results showed as follows: 1) The width, the density of trabeculae area and the marrow cavity increased significantly in others three groups, especially in group C, compared with the Group A; 2) Perimeter, area, and relative volume of trabeculae increased significantly and the maximum were occurred in group C. There were a very large number of osteoblasts and osteoclasts on the surface of trabeculae in group C. The trabecular separating degree in Group C was the minimum and significantly lower than in Group A and Group B, but had no significant difference compared with Group D; 3) In Group D, chondrocytes became focally vacuolated gradually in calcified cartilage matrix area, nuclear appeared pyknotic phenomenon, finally degradation and death appeared. These results indicated that the additional boron supplemented in drinking could promote obviously tibias osteogenesis of African Ostrich Chicks, and 200 mg·L^-1 supplement boron in the drinking water appeared to be the most beneficial.

To study effect of ginsenoside Rb1 on oxygen free radicals in acute lung injury of mice induced by A/swine/HeBei/012/2008/swine influenza virus (H9N2 SIV), 120 six to eight weeks old BALB/c mice were randomly divided into three groups with forty in each．The mice in control group were inoculated intranaslly with an equivalent dilution of noninfectious allantoic. And that of acute lung injury group (ALI group) and Ginsenoside Rb1 group (G-Rb1 group) both were inoculated intranasally with H9N2 SIV diluted in sterile saline，and meanwhile，animals of G-Rb1 group were treated with Ginsenoside Rb1 (10 mg·kg^－1) by intraperitoneal injection continuously for up to seven days．The clinical signs and body losses were observed in eight infected mice of each group．At the same time，at the indicated time points after infection，lung histopathology was observed and the activity of T-SOD，inhibition ability of OH·, MDA and NO content of mouse lungs were detected．The results showed that mice of ALI group appeared depression，ruffled fur，feed intake reduction and weight loss post first 2 days of infection．Furthermore，pulmonary edema，hemorrhage，and a number of inflammatory cells exuding from the alveolar were observed in lungs of infected mice，however，mice organs in the control group showed no abnormality．For mice in G-Rb1 group, clinical symptoms were significantly improved, while survival time was delayed and mortality was decreased．On the 4^th, 6^th and 8^th day after infection：the inhibition ability of OH· and T-SOD activity were significantly reduced (P<0.05) in ALI group and G-Rb1 group mice compared with that of control group mice，but the indexes of G-Rb1 group were significantly higher than that of ALI group (P<0.05)；The contents of NO and MDA were increased significantly (P<0.01) in ALI group and G-Rb1 group mice compared with that of control group mice, but these contents of G-Rb1 group was significantly lower than that of ALI group (P<0.01). Results indicate that G-Rb1 could scavenge free radicals in acute lung injury lung induced by H9N2 SIV in mice，and G-Rb1 is helpful to attenuate the lung injury induced by oxygen radicals.

 Our research was conducted to observe the effect of metallothionein (MT) on cow heat stress splenic lymphocytes apoptosis, and study MT on the regulation of the thermo-oxidative stress-induced apoptosis/necrosis and mitochondrial membrane potential (ΔΨm). The splenic lymphocytes apoptosis phenomena were observed by using fluorescence microscope. The apoptosis and necrosis rates were assayed for adopting Annexin V-FITC with flow cytometer. The ΔΨm of cells was assayed with flow cytometer through loading JC-1. Results were as follows: MT only has inhibitory effect on the apoptosis induced by heat stress, and when MT was 5.25-8.25 μg·mL^－1, the apoptosis can be promoted lowly. The necrocytosis can also be promoted lowly by the lower MT when the concentration was 2.25 μg·mL^－1. And the ΔΨm value can be increased more by MT, but the high stress of cell require higher concentration of MT, and the normal cell require moderate concentration (3.75-6.75 μg·mL^－1), cell can not be protected enough by the other concentration. These findings indicate that the 1 hour culture in 43℃ of cow spleen lymphocytes can improve mitochondrial membrane potential, as well as the necrosis rate. Nevertheless, mitochondrial membrane voltage can be improved, and necrosis caused by heat stress could be restrained because of adding MT in cell culture medium. Therefore, It has more practical meaning for adopting adding MT to improve the integrity of cellular mitochondria than high fever, at the same time, it also shows that the mitochondrial pathway is one of signal transduction pathway which MT regulating apoptosis/necrosis.

To understand the pathogenesis of facial eczema of grazing sheep, the clinical disease process, pathological anatomy and histology, serum liver function parameters of facial eczema of grazing sheep in northern slope of Tianshan Mountain were investigated and determined. The fungal types isolated from 100 sick sheep, 100 healthy sheep and 5 dead lambs were analyzed by statistics. The results indicated that onset of the diseases was limited on certain areas only, and the disease most likely attacked young fine-wool sheep with the age less than 6 months old. The clinical manifestations were characterized by inflammational edema in face (ears and eyelids) and mandibular area. Postmortem examination of dead lambs showed enlargement of liver with yellow white patchs of necrotic lesion and tuberous sclerosis and fibrosis on section. Histologic examination of liver showed extravasated blood, severe lesion of liver cells and bile duct, and fatty degeneration. GGT levels of the disease animals were very high and varying from 94.93 to 260.78 U·L^-1, but the GGT in normal was 16.47- 61.59 U·L^-1. While the activities of ALT and total bilirubin were elevated and AST was normal in affected sheep. ALT and total bilirubin level was higher than normal (P<0.01), and the AST activity of sick sheep and lamb was not significantly different with healthy sheep (P>0.05). 205 experimental sheep had fungal infections; Pithomyces, Alternaria, Fusarium and Aspergillus isolated from diseased goats had higher separation rate than that of the others. The Pithomyces separation rate was highest. Others such as yeast, Rhizopus and Mucor were also separated in all sheep, but the separation rates were low. The results revealed that the facial eczema was hepatogenic light allergic eczema caused by hepatic dysfunction and hepatonecrosis in grazing sheep in the area, and the pathogen may be Pithomyces, Alternaria, Fusarium and Aspergillus.

The aim of this study was to establish a simple, efficient and inexpensive strategy for expression and purification of Non-taged nanobodies. Chose nanobodies of porcine circovirus type 2 (PCV2) Cap protein as the target protein, three plasmids with N-terminus fusion tags pHSIE-Nb(His-Sumo-Intein）, pHSIE-intein-Nb（His-Intein）and pET32a-intein-Nb（His-Trx-Intein）were constructed and their effects were compared. The recombinant three plasmids were transformed into E. coil BL21(+) and induced by IPTG respectively. The expression and solubility of target proteins were tested under different conditions, and the favorable tags were selected. Recombinant proteins were purified by Ni-NTA column chromatography, and tags were removed by Intein as self-cleavaged, then target protein were prepared, based on which a double antibody sandwich ELISA method was developed. All these three different tagged nanobodies could be successfully expressed. A series of experiments lead to the finding that the placement of His-Trx-Intein before the Nb is the best strategy in availing the soluble expression of the tagged protein. After on-column cleavage was preceded at pH7.0，being incubated at 25 ℃ for 24 h, the recombinant un-tagged Nb were released. The results of double antibody sandwich ELISA showed that the Nb protein could react specifically with cap protein, and there was no significant difference (P>0.05) between positive control group and nanobody group, which suggested that this method of preparation of nanobodies was successful. An efficient expression and simple purification method of preparation of nanobodies was successfully established, which was efficient soluble expression, and simple purification, and no tagged active nanobodies could be prepared. The study provides a technical support for the further theoretical research and production in nanobodies.

The aim of this study was to study antibacterial effects of Jin-Ying-Huang-Gui Injection (JYHGI), a Chinese herbal formula, on Staphylococcus aureus (S. aureus)．JYHGI was prepared by routine method. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of S. aureus were detected with tube double dilution method in vitro. The mice model was prepared by vena caudalis injection of S. aureus to carry antibacterial experiment in vivo. In addition, we also studied the influence of JYHGI on ultrastructure and the specific activity of succinate dehydrogenase (SDH) of S. aureus. Antibacterial activity showed that the MIC and MBC of JYHGI to S. aureus were 31.2, 62.5 mg·mL^－1, respectively. Mortality of mice in JYHGI could significantly decrease mice mortality caused by S. aureus (P<0.05). The JYHGI treated cell wall of S. aureus were covered with a large number of floc, the cells seemed to be collapsed-like, some cell wall even ruptured. The SDH specific activity of JYHGI group were significantly lower than that of control group (P<0.01). It is concluded that the inhibition mechanism is that JYHGI could damage the cell wall and inhibit the enzyme system of S.aureus.

For comparing ELISA and IFA in the detection of J subgroup avian leukosis virus, five different dilution of NX0101 TCID₅₀ virus were inoculated to DF1 cell and cultured. In six different stages (1, 2, 3, 5, 7 and 9 days culturation), ELISA was applied to detect P27 antigen within supernatant and IFA was conducted to detect the infected cells respectively. Using more than 10^1.625TCID₅₀ of the inoculation dose, IFA achieves identification of all the infected cells at 2 days and 3 days post inoculation, but the results from ELISA is relatively lag of 2-4 days for confirmation. Inoculating less than 10^1.625TCID₅₀ of virus, IFA detect successfully the infected cells while ELISA still not detect by 7th day. The results suggest that IFA is more accurate and effective than commercial ELISA kit, ELISA have obvious hysteresis in comparison to IFA.

This experiment was conducted to study the pathogenicity of the H5N2 avian influenza virus isolates from wild bird and investigate its pathogenic and occurring mechanism. Two-week-old SPF duck (Shaoxing sheldrake) was infected by an isolate of LPAIV H5N2 subtype as natural infection model in this study. The results showed that virus can propagate more effectively in digestive tract. Despite of absent of obvious clinical symptoms, the infected SPF duck shed virus for a longer time. The virus has the infection ability by direct contact. The antibody against HI can be detected at 7-14 days after infection, and the antibody level reached its peak at the third week after infection. Besides, this virus can propagate with higher level only in caecum tonsil and bursa of fabricius, while the tissue damage occurred in bursa of fabricius, lung, thymus and pancreas in varying degrees. The results indicated that duckling plays a key role during AIV spread，whether it (even wild duck) cause by certain differences between the laboratory imitation nature and true nature infection, it remains to be further explored.