The genomes of seven goat populations were screened by using microsatellites as molecular markers, including Yichang White goat ,Matou goat, Xiangdong Black goat ,Fuqing goat ,Daiyun goat ,Huanghuai goat and Yangtse River Delta White goat .A total of 23 microsatellite markers were used and genetic diversity and genetic distance were also determined .The results showed that only 21 loci showed polymorphism in all population in 23 loci .Alleles were homozygotic in each population in BM0203, but there was genetic diversity among populations. Alleles were homozygotic in Xiangdong Black goat and Yichang White goat in BM6444 ,but more than one allele was detected in other populations. Average heterozygosity of all populations was 0.819 0 ,and the mean polymorphism information content (PIC) of all seven populations was 0.630 5~0.691 9 .A UPGMA dendrogram was constructed based on Nei’s standard genetic distance . Matou goat and Xiangdong Black goat were grouped at first, then Fuqing goat, Daiyun goat, Yangtse River Delta White goat, Hunghuai goat joined them respectively, Finally, Yichang White goat clusted with all above.

The genetic diversity of twenty-seven indigenous chicken breeds or strains in east China was detected with thirty microsatellite markers with high polymorphisms. The genetic parameters of mean polymorphic information content (PIC), mean heterozygosity (H), and Nei’s standard genetic distances (Ds) were calculated. The clustering dendrogram was obtained eventually based on Ds genetic distances of thirty microsatellite and UPGMA method. The results were as follows: the mean heterozygosity of Hetian chicken was the highest (0.687 0), and that of Langshan chicken was the lowest (0.617 0), and that in other breeds or strains was also more than 0.6, which reflected their rich diversity. The Ds genetic distances suggested the longer differentiation. UPGMA tree was obtained through analysis of Ds genetic distance, twentyseven indigenous chicken breed were clustered into ten groups based on the dendrogram. In the tree, Velutinate silky, Jining bairi, Hetian, Anyi wahui, Xianju, Jiangshan silky, Liyang, Jinhu silky, Lingkun, Yugan silky, Kangle, Xiaoshan, Langya, Huaibei ma and Huainan sanhuang were first grouped together; Baier and Pudong chicken had their own branch respectively; Chongren ma and Silky were fourth grouped; Langshan and Shouguang were fifth grouped; Luyuan, Dongxiang green eggshell and Wenshang barred chicken had their own branch respectively; Ningdu sanhuang and Luxi game chicken were ninth grouped; the last group was only Zhangzhou game chicken. According to those breeds or strains having their own characteristics and rich diversity based on the analysis of their genetic diversity, we should rationally explore their superiorities.

The objective of this study was to investigate tissue-specific expression of SGLT1 and GLUT2 mRNA in different intestine segments of Arbor Acre (AA) chicken by relative quantative RT-PCR. The results showed that the expression of SGLT1 mRNA decreased gradually along the downward intestine. SGLT1 mRNA abundance of duodenum, jejunum and ileum was 76.19% (P<0.01), 42.86%(P=0.06) and 38.10%(P=0.07) higher than that of colorectum, respectively. And SGLT1 mRNA expression of duodenum was higher 23.33%(P=0.18) and 27.59%(P=0.10) than that of jejunum and ileum, respectively. The GLUT2 mRNA expression of duodenum was close to that of jejunum, the difference was not significant (P>0.05). Single gene PCR results indicated the GLUT2 mRNA abundance of duodenum and jejunum were much more higher than that of ileum and colorectum. Further researches should be done on the physiological function of tissue-specific expression of SGLT1 and GLUT2 mRNA in chicken intestine.

The keratin family includes epithelial (soft) keratins and hair (hard) keratins, and can be divided into acidic typeⅠand basic to neutral typeⅡsubfamilies. A cDNA was library constructed with poly (A)⁺RNA from adult goat skin and 392 ESTs were analyzed. Based on sequence homology comparisons with the known typeⅠhair keratins sheep 8CⅠand hHa1, A fulllength cDNA that encoded goat hair keratin gHaⅠ(GenBank Accession No.AY510110. 1) could be identified, it derived 413 amino acid sequence comparisons with the sheep 8C1 type Ⅰ mRNA exhibits the highest homology (97.8%). In site hybridization with the gHa1 cRNA probe on skin revealed a strong expression in cortex of both primary and secondary hair follicles.

In order to understand if the apoptosis of follicular cells and oocytes exists in the buffalo fetals ovary, and if does, what is the apoptotic characteristics, ovaries was chosen from a five-month-old fetal buffalo, the apoptotic characteristics of follicular cells and oocytes was studied by paraffin section, HE staining, light microscope and terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) technology. The results were shown that large number of apoptotic cells existed diffusedly on 5-month-old fetal ovary. Many apoptotic follicular cells and some apoptotic oocytes were distributed solely and evenly among a lot of preantrum follicles of ovary cortex. The main changes of apoptosis were cell shrinkage, nuclear condensed in histology, DNA terminal labeling appeared TUNEL positive. These results suggested that the apoptosis of ovarian follicular cells and oocytes was already existed in buffalo fetal ovary, and the apoptosis was the potential mechanism of arosing preantrum follicle atresia.

To improve the hatchability of introduced, high yield chickens at Tibet altitude，and ameliorate the egg performance and reproduction traits of the local Tibetan chicken，four mating combinations between the Recessive White and Tibetan chicken were employed in this study. The influences of the mating system and environmental factors on hatchability were investigated. The results showed that the fertility of the egg was significantly lower for those using Tibetan chicken as maternal parent than those using Recessive White as maternal parent (P<0.05), while hatchability was on the contrary (P<0.05). Especially during the late stage of incubation，the embryonic mortality of Recessive White was significantly higher than the Tibetan eggs by 7.95%. The hatchability of combination Ⅲ whose female parent is Tibetan chicken was higher than Ⅱ whose female parent is Recessive White by 20.44%，which further provided a theoretical basis for cross using the Tibetan chicken as maternal parent. Binary logistic regression analysis results also showed that the mating combinations and environmental factors were of the importance for the egg hatchability.

Two nitrogen-balance experiments were conducted to determine the optimum balance pattern among the four amino acids (lysine,methionine,tryptophan and threonine) in the diet on digestible basis for piglets (10kg) challenged with lipopolysaccharide or with saline as control.The method of amino acid deletion was adopted.All pigs were of a single genetic background and geographical site of origin.In each group fifteen Landrance×Rongchang castrated piglets were allotted randomly to one of five groups of dietary amino acid regimens. Positive control group was supplemented with four amino acids. One of the four amino acids was deleted by about 25% in the respective treatment groups. After the nitrogen-balance experiments,the slaughter technique was introduced to determine the ideal amino acid digestibility of the positive control by using chromium oxide(3g/kg) as the indicator. The results showed that the ideal amino acid pattern of Lys∶Met∶Thr∶Trp on the digestible basis was 100∶27∶29∶59 for 10-kg pigs under immunologic stress, and 100∶30∶21∶61 for piglets under normal condition.

Nine growing wethers of Inner Mongolia white cashmere goats (IMWCG) with the same father, fitted with permanent cannula at the rumen, T-shape cannula at the proximal duodenum and terminal ileum were used to investigate the effect of rumen-protected amino acid (RPAA) products on nitrogen metabolism in IMWCG. The experimental goats fed diet with the same nitrogen (CP 9.93%) level of a diet consisted of hay(70%), corn(19.65%) and soybean meal(4.5%), were divided into three group in a free random block design as follows: rumen protected amino acids group (group A), normal amino acids group(group B), and control group(group C). Total collection of feces and urine was performed every 24h for 5d. The results showed that supplemental rumenprotected methionine (RPMet) and rumenprotected lysine (RPLys) increased nitrogen intake (P<0.05), and retained nitrogen (P>0.05), and had a tendency to decrease urine urea nitrogen excretion (P>0.05). However, plasma free methionine and lysine concentrations were unaffected by supplemental with RPMet and RPLys.

The immune effects of three different unmethylated CpG oligodeoxynucleotides (CpG ODN) on LaSota lived vaccine against Newcastle disease in chickens inoculated by intranasal and eyedrop route were investigated and the CpG ODNs were evaluated for their ability to activate B lymphocytes to induce HI antibody, promote T lymphocytes proliferation, induce macrophage nitric oxide (NO) production, and stimulate mRNA expression of IFN-γ,IL-6 and IL-1β in peripheral blood mononuclear cells (PBMC) in this study. After two successive immunizations, our results demonstrated that the birds in CpG ODN1-group produced a mean serum HI antibody titer of 8.2log2, a lymphoproliferative stimulation index (SI) of 9.836 and a NO production of 35.833 μmol/L. There existed an increment by 2log2 (P<0.05), 4.4(P<0.01), 27.6 μmol/L (P<0.01), respectively, compared with control group (inoculated with LaSota lived vaccine only). CpG ODN2 containing the GACGTT motif in group 2 showed a faint efficacy and there was no difference between this group and control group. However, deviation of flanking bases of CpG ODN3 in group 3 significantly reduced or diminished its stimulatory activity. Meanwhile, it was found that there were remarkable mRNA expression of IFN-γ，IL-6 and IL-1β in peripheral blood mononuclear cells of CpG ODN1-treated birds(P<0.05), in despite of CpG ODN3 induced a little higher level of mRNA expression of IFN-γ than that of CpG ODN1-group. These results indicate that CpG ODN1 is a potent LaSota lived vaccine adjuvant for stimulating both antibody and cell mediated immune responses in chickens.

Firstly,the mice were immuned with the expression plasmids by intramuscular injection. The first group was inoculated with pVAXLPH, the second group was inoculated with pVAXLPF, the third group was inoculated with pVAXLPH and pVAXLPF, the fourth group was inoculated with pVAXLPH, pVAXLPF and pVAXLPN, the fifth group was inoculated with empty vector DNA pVAX1 as negative control, and the sixth was inoculated with attenuated CDV vaccine as positive control. A total of three inoculations were performed at biweekly intervals. Indirect ELISA, neutralization and lymphocyte transformation assays were employed to determine the immunological response. As a result, the mice inoculated with these plasmids developed humoral and cellular immune responses against CDV. pVAXLPN did induce cell-mediated immunity, at high level. Tow weeks after the last injection ,the ELISA titers against CDV of the fourth group is 0.332, and SN titers against CDV of the fourth group is 1∶(4-16), the OD570 of lympho-spleencytes proliferation against CDV and ConA stimulation of the the fourth group is 0.384 and 0.356, respectively, the results of CD4⁺/CD8⁺ of the the fourth group is 2.05±0.38.Secondly, twenty 45-60 day-old CDV seronegative dogs were divided into four groups. All dogs in each group were immunized for 3 times. The first group was injected with a mixture of 200 μg of all the three plasmids containing the H, F and N gene, three times at biweekly intervals, the second group was injected with a mixture of 200 μg of all the three plasmids two times and with attenuated CDV vaccine once, the third group were injected with pVAX1 plasmid as negative control, and the fourth group was injected with attenuated CDV vaccine as positive control. Indirect ELISA, neutralization and lymphocyte transformation assays were employed to determine the immunological response. Two weeks after the last injection,the ELISA titers and SN titers against CDV of the first group is 0.296 and 1∶(8-32), respectively ;And the ELISA titers and SN titers against CDV of the second group is 0.433 and 1∶(16-64), respectively. As a result, the dogs inoculated with these plasmids developed humoral and cellular immune responses against CDV. The induced immune response in second group was higher than that in the first group.

30 dogs and 180 rats were divided into 6 groups randomly and respectively, then irradiated by pulse microwave with 5 different power densities ranged from 1.004 mW/cm² to 0.974 kW/cm² in different group. 12 months after irradiation, animal temperature, heart rate, electrocardiogram and serum myocardial enzyme spectrum were tested to study the changes of heart function，and the pathologic histological changes of myocardium were observed with paraffin slice dyed by HE,Pollak and PTAH respectively, and ultrastructure changes of rat myocardium were examined by electron microscope. The results showed that irradiation influenced the heart functions and damaged myocardial tissue structure although animal temperature and heart rate showed unapparent changes. Cardiogram showed that S-T wave lower and conduction retardation, and serum myocardial enzyme spectrum was disorder significantly stand by decreased activity. Irradiation by pulse microwave can injure structure including ultrastructure of myocardial cells, sinus, Purkinje cells and wall of blood vessels. Damage occured in whole heart and was severer in conduction system. These results indicated that heart is one of the susceptive organs exposed to electromagnetic fields. High power pulse microwave can induce morphological and functional injury of the heart and the related systems. The injuries showed the characteristics of acute, durative and repairable, and the damage degrees may correlate to irradiation dosage in a certain extent.

To investigate the apoptosis mechanism of hepatocyte induced by endotoxin(ET) and protective effect of Cation A (CA), the nitric oxide (NO) levels of liver tissue and the percentage of apoptosis of hepatocyte were measured 3，4，8，12 h post treatment respectively in the normal control group, ET group and CA protective group. The NO levels of all detected times were increased markedly in ET group (P<0.01), the percentage of apoptosis of hepatocyte displayed three periods: increased, decreased and increased, which were confirmed pathologically with obvious damage of cells and tissue of liver. While in CA group the above mentioned parameters and damage of cells and tissue of liver were reduced significantly (P<0.01, P<0.05). Results showed that NO might be involved in pathogenesis of endotoxic shock, primarily through the induction of hepatocytes apoptosis, and CA could be used in preventing or treating it by mechanism of effectively reducing ET-induced production of inflammatory factors and massive NO of liver.

Antimicrobial MICs of 30 pathogenic salmonella isolates from swine were determined by using plate dilution method. 11 antibiotics (belonging to aminoglycosides, tetracyclines, sulfonamides and chloramphenicols) were used. The results showed that twenty-eight strains (93.3%) of these salmonella isolates were resistant to at least one antimicrobial agent; resistance to the following antibiotics was common: tetracycline (83.3%), doxycycline (80%), sulfamethoxazole (80%), sulfamethoxazole timethoprim (76.7%), streptomycin (60%), kanamycin (56.7%) and chloramphenicol (56.7%). 25 pairs of primers were designed and antimicrobial resistance genes were amplified, sequenced and analyzed. A total of 13 different antimicrobial resistance genes conferring resistance to four categories of antimicrobials were identifed and the homologous rate of each gene was very high compared with the correspondence gene in GenBank (≥98.1%). At least one antimicrobial resistance gene was detected in 28 pathogenic salmonella isolates, accounting for 93.3% (28/30). The following antimicrobial resistance genes were commonly present: sulⅠ (76.7%), aph(3′)-Ⅱa (60%), tetC (60%), Cat1 (43.3%), tetA (40%) and aadA1 (36.7%). Antimicrobial phenotypes results were very consistent with the antimicrobial resistance genes (≥88%).

For investigating the effects of Shenpang acupoint-stimulation on reproductive endocrine, the changes of estrogen receptors immunoreactive (ER-IR) neurons after Shenpang acupoint-stimulation were studied by using the technique of immunohistochemistry SP. The ER-IR positive productions were detected in most nuclei of thalamus. In acupuncture group, a great number of ER-IR positive productions, with clear dendrite, existed in the nucleus paraventricularis thalami, nucleus ventralis lateralis thalami, nucleus ventralis medialis thalami, nucleus ventralis principalis thalami, nucleus centromedianus thalami, nucleus reticularis thalami, nucleus periventricularis thalami, and were stained strongly. In addition, the ER-IR positive production mainly located in plasma, nucleus and neurite, also existed in the cytoplasmic membrane. In contrast, a few of neurons existed in the above nuclei in control group, and the neurons were stained weakly. The expression of estrogen receptors in the above nuclei increased after the Shenpang acupoint-stimulation.

To investigate the pharmacokinetics of Compound HuangBo Granule in mice, the effective component-berberine was detected as a representative index.Pharmacokinetics of orally administration of Compound HuangBo Granule at a dose of 25 g/kg (10.95 mg/kg berberine) were investigated . Serum concentration of berberine were measured using HPLC and concentration-time data were analyzed with pharmaceutical kinetics software. The results showed that the serum profiles of the drug's effective component-berberine were best described by a two compartment open model, the pharmacokinetics equation is C=17.8e^{-0.859 81t}+14.51 e^{-0.636 1t}.The main parameters were as follows: t_1/2α(0.81±0.07) h, t_1/2β(5.25±2.96) h, t_1/2Ka(0.76±0.05) h, AUC_0→∞(3.389 3±0.4３7 4) mg·L^-1·h, CL/F (3.281 8±0.446 3) L/h, V_^d/F(13.65±22.38) L/kg, T_peak(1.617 19±0.056 5０) h, C_max(1.149 98±0.056 04) μg/mＬ.

Young Hailan chicken are injected with Epimedium-proplis mixture twice on 3th day and 10th day respectively. The blood samples were taken at 7,14,21 and 28 days old. The measurement indexes of cell immunity contain transformation level of lymphoid cell, the formation rate of Erose garland and the content of lysozyme in serum. The result shows that Epimedium-proplis mixture can obviously increase the conversion level of lymphoid cell and the formation rate of E-rose garland of young chicken , so it is an effective insulation for single nuclear-monocyte system of young chicken,and has long-lasting effective period and good effection. Theory and experiment roots were provided by this test for Epimedium-Proplis mixture used as immunity enhancer in practice.

Here, we presented a Chlamydiaceae-specific 23S rRNA-based real-time PCR assay for simultaneous detection and quantitation of four members of Chlamydiaceae family, C. trachomatis, C. psittaci, C. pneumoniae and C. pecorum, using SYBR Green and Lightcycler^TM. The assay was characterized using plasmid constructs of the bacteria and verified on standard strains of all four species of the Chlamydiaceae and a large cohort of clinical samples collected from human and animals by comparison with fluorescence immunohistochemistry method. The results showed that the present real-time PCR assay was of high specificity and sensitivity. It was capable of detecting as few as 250 fg of chlamydial DNA (equivalent to 10^-1 IFU) and was applicable to both liquid cultures and clinical samples. This assay may therefore offer a rapid, economic and reliable method for screening of the chlamydiaceae pathogens.

Chicken interleukin 18 mature protein gene was amplified from the recombinant plasmid named pT-mChIL18 by polymerase chain reaction(PCR) and cloned into pcDNA3.1/V5-His-TOPO. The recombinant plasmid was transfected into chicken embryonic fibroblasts(CEF) with liposome. 24，48，72 hours after transfection, the expression of mChIL-18 were successfully tested by indirect immunofluorescence assay(IFA) with the antiserum of mChIL-18 expressed in E.coli.

The LppQ N-terminal gene of Mycoplasma mycoide subsp. mycoides SC was amplified from MmmSC HVRI X strain by PCR using the primers designed according to the reported MmmSC LppQ sequence. The products of PCR were cloned into the pUC18 and pUC19 vector respectively. Mutagenesis was performed in a one-step overlap extension PCR. Sequence analysis confirmed the successful mutation from TGA to TGG in amino acid 66 in the LppQ N-terminal gene. The fragments amplified by PCR containing the mutation site were subcloned into the pET32a expression vector. Restriction enzyme, PCR and sequencing analysis indicated that the recombinant expression plasmid pET32a-LppQ was successfully constructed, which may provide a basis of expression of the LppQ N-terminal gene in vitro.

The voltage-gated K⁺channels dynamical indices，resting membrane potential and potassium currents of bronchial smooth muscle cells(BSMC) were investigated by cell patch clamp technique. The experimental BSMCs of dogs were divided into control group,CD group,Tetramethylpyazine group,Baicalin group,Astraglus membranaceus group and Berberine group. The results indicated that the activation of Kv channel from CD group was significantly inhibited. This inhibitation was counteracted by Tetramethylpyazine, Baicalin and Astraglus membranaceus, which made the Kv channel of BSMC open. The most powerful one of four Chinese herbal medicinal ingredients was Baicalin. Baicalin increased the activation of the Kv channel of BSMC from CD dogs significantly and extended bronchial smooth muscle cells.