Ovaries are the most important reproductive organs in female animals.Understanding the molecular regulatory mechanisms of follicular development and hormone secretion is beneficial to improvement of livestock production，the diagnosis and treatment of human ovarian diseases.The miRNAs are small non-coding RNAs that are approximately 22 nucleotides (nt) in length.The majority of miRNAs are ubiquitous and highly conserved across species.The miRNAs regulate target genes via translational repression or degradation of target mRNA.Many studies indicated that miRNAs play critical roles in almost all ovarian biological processes，including folliculogenesis，follicle development，follicle atresia，luteal development and regression.The comprehensive investigation of ovarian miRNA will help to reveal the molecular mechanisms of ovarian functions post-transcriptionally.Herein，the advance in ovarian miRNA researches in recent years，especially in follicular development and ovarian diseases，are summarized.

Heat shock protein 70(Hsp70)，as molecular chaperones，plays an important role in protective effects and immune regulation on gastro-intestinal tract mucosa，however，its exact mechanism is still unknown.This paper review the expression and location of Hsp70 in gastro-intestinal tract mucosa，protective effects of Hsp70，especially the mechanism of immune regulation mediated by Hsp70.

The aim of this study was to detect CNV regions(CNVRs) from 150 Rongchang pig hybridization population using the Porcine SNP60 BeadChip.In this study，5 CNVRs (CNV regions) were identified across five autosomal chromosomes(chromosomes 1,7,8,13,14) and the predicted status for the CNVRs was 3 for deletion，2 for duplication.Among these 5 CNVRs，CNVR5 and CNVR1 were the novel found states，and CNVR2 and CNVR3 were reported by others but not had been validated.The CNVRs were validated by real time quantitative PCR.The results showed that CNVR2，CNVR3，CNVR5 had extensive polymorphism in different pig breeds.CNVR4 was the false positive states.Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that 46 protein-coding genes in verifying CNVRs possessed a great variety of molecular functions,such as olfactory receptor activity,ethanol metabolic process,neurological system development and fatty acid metabolism pathway.The result provide genetic variation materials for analyzing the genetic base of phenotypic variation between pig breeds.

The aim of this study was to determine and find the promoter core regulatory regions of Xuhuai goat’s c-Myc gene，and to investigate c-Myc gene regulation mechanism.According to the sequence of ovis aries c-Myc gene published in GenBank，the primers were designed and the different deletion fragments of 5′ flanking region of c-Myc gene promoter were amplified，and then were cloned into pEGFP-N1 and PGL3-Basic，then were transfected into gFF，COS7 and P19 cells and treated with TSA and NFAT1.SP1 transcription factor binding sites in the region of -402--249 bp was mutated，and the activity of dual-luciferase reporter gene was detected.The study showed that the region containing -1 334-+1 bp of c-Myc promoter had the maximal promoter activities，and sequence of -402-+1 bp had the basal promoter activities，further study showed that there were positive (the region of -1 334--971 bp，-587--147 bp) and negative (the region of -1 976--1 334 bp，-971--587 bp) regulatory domains，respectively.TSA and NFAT1 significantly enhanced the activity of c-Myc promoter.The activity was declined by the point mutation of SP1 binding site.By constructing the recombinant expression vectors containing different fragments of c-Myc gene promoter and comparing their activity，the core regulatory region of c-Myc gene promoter was identified.SP1 is an important regulatory element of c-Myc promoter core region，which lay a foundation for the further research on the mechanism of regulation and expression of c-Myc gene.

This experiment aimed to construct the eukaryotic expression vector of PLCζ，a preliminary study of its expression properties in progenitor cell line was conducted.PLCζ was amplified by PCR and cloned into vector pEGFP-N1，after correct identification，the recombinant plasmid pEGFP-N1-PLCζ was transfected to 293T cells and sheep fibroblasts.The expression and distribution of cell recombinant plasmid was observed under an inverted fluorescence microscope and laser scanning confocal microscope.After digestion and sequencing,the pEGFP-N1-PLCζ eukaryotic expression vector was successfully constructed，and its successful fusion expression with the green fluorescent protein was observed.This experiment realized the expression of pEGFP-N1-PLCζ in sheep fetal fibroblast cells，which will help us to study the activation of recombinant protein on the oocyte through nuclear transfer furtherly.

The objective of this experiment was to study the effect of WY14643 on the development of early embryos and to explore the mechanisms of antioxidative damage capacity in early mouse embryos.The model of oxidative damage was established by H₂O₂ and added WY14643 to the medium of in vitro culture.H₂O₂ level in embryos were measured by DCHFDA .SOD activities and MDA contents were determined by using superoxide dismutase and malondialchehyche assay kit，respectively.The results showed that:(1)after different concentrations of WY14643 processing to 2 h，4-cell rate and blastocyst rate in the group of 0.1 μmol•L^-1 WY14643 treatment was significantly higher than other groups（P<0.05）;(2)after 0.1 μmol•L^-1 WY14643 with H₂O₂ co-culture，the embryo development rate was significantly higher than only H₂O₂ treatment group and control group（P<0.05）.(3)through DCHFDA fluorescence detection，the H₂O₂ level of WY14643 adding group was significantly lower than other groups（P<0.05）.(4)from the SOD activities and MDA contents detection，we found that SOD level of adding WY14643 group was significantly higher than other groups within each period（P<0.05）as opposite to the MDA level were significantly lower than other groups（P<0.05）.Those results indicated that WY14643 could improve the development of embryo by increasing activity of SOD enzymes to reduce lipid damage and then decrease the H₂O₂ level of the embryos in vitro culture.

After produce the transgenic animals，the primary task was important to detect integration sites of these animals.In this study，the integration sites were successfully cloned in two transgenic SCNT cattles with hLYZ gene by TAIL-PCR.The results showed that in two SCNT transgenic cattles，the exogenous genes integrated to cattle chromosome.The exogenous gene in the WS individual integrated to genomic clones (NW_003104566.1) on bovine chromosome 24 and that in the YW individual integrated to genomic clones (NW_003104440.1) on bovine chromosome 16，which located between LOC10030 and RGL1 gene.The deletions of 238 and 228 bp host genome were detected in WS and YW respectively.The consensus sequence for mammalian Topo I ( topoisomerase-I ) cleavage sites were found at 4 integration sequences (InS1-InS4).At the junction of exogenous gene and host genome (InS1 and InS3) in WS，homologous sequences of 1 nt were found.InS 2，InS 3 and InS 4 showed similar homologous sequences at the junction.

This study aimed to research the biological characteristics of retinal stem cells (RSCs) from bovine fetus.RSCs were isolated using physical method，and the surface markers of RSCs，Nestin，Chx-10 and Pax-6，were detected by immunofluorescence.The multi-lineage differentiation potential of RSCs was investigated and induced to differentiate into astrocytes，neurons and oligodendrocytes.The results showed that the fetal bovine RSCs not only expressed the special markers of stem cells but also induced to astrocytes，neurons and oligodendrocytes successfully.To sum up the RSCs isolated from fetal bovine have a high proliferation and multi-directional differentiation potential in vitro.Retinal stem cells for the future research provide the corresponding theoretical foundation.

The purposes of the study were to explore growth differentiation factor 9(GDF9) and its receptor position and expression in the adult dog testis.The expression and location of GDF9 and its receptor activin receptor-like kinase 5(ALK 5) in adult dog testis was detected by RT-PCR，immunohistochemistry and Western blot.The expression of protein GDF9 in testis of adult dogs can show that GDF9 is stage-specifically localized in the cytoplasm of round spermatids and pachytene spermatocytes of adult dog seminiferous epithelium.In particular，the expression of protein ALK5 in testis of adult dogs can show that ALK5 is localized in the cytoplasm of round spermatids.The study showed that GDF9 and its membrane receptor ALK5 were both expressed in adult dog testis.GDF9 is specifically secreted by the cytoplasm of round spermatids and pachytene spermatocytes.GDF9 regulates and sustains the connections between Sertoli cells，and between cells and germ cells by paracrine secretion.Also，GDF9 regulates the growth and migration of germ cells by autocrine secretion.

The current study was conducted to investigate the effect of dietary phosphorus (P) levels on the true P digestibility (TPD) estimated by the regression method in soybean meal (SBM) for growing pigs.Forty-eight barrows with an average initial BW of (36.0±0.1) kg were allocated to 6 diets in a 2×3 factorial arrangement of 2 diet types (low P diet without corn supplementation or high P diet supplemented with 50.0% corn) and 3 SBM levels (13.0%，26.0% and 39.0%) by a randomized complete block design to study the effect of diet type (low P diet or high P diet) on the determination of TPD in SBM by the regression method.There were 8 replicates in a treatment group with 1 pig per replicate.The results showed that dietary P intake，fecal P output，digested P and retained P for growing pigs were increased linearly with increasing levels of SBM (P<0.01).Compared with the low P diet，pigs fed the high P diet increased dietary P intake，fecal P output，urinary P output，digested P and retained P (P<0.05).Using the regression method，the estimates of TPD in SBM for pigs fed the high P diet was 36.11%，which was lower than the determined value of 43.38% for pigs fed the low P diet (P<0.05).In conclusion，the estimated values of TPD in SBM were higher for pigs fed the semi-purified diet (low P diet) than that for pigs fed the practical diet (high P diet).Therefore，diet P levels should be considered when using the determined values of TPD in feed ingredients derived from pigs fed semi-purified diet by the regression method for diet formulation.

This experiment was conducted to investigate the effects of melamine on ruminal fermentation parameters in vitro.Melamine was supplemented at graded levels：0，2，10，50，250，1 250 mg•L^-1，respectively.Incubation mediums were taken after incubation of 0，1，2，4，6，8，12，24 and 48 h，respectively.Melamine，total fatty acid (TVFA)，acetic acid，propionic acid，butyric acid，valeric acid，isobutyric acid，isovaleric acid and ammonia nitrogen concentration were determined.The result showed that melamine supplementation resulted in no change in MEL degradation during 48 h incubation （P>0.05） but decreased dry matter degradability (DMD)（P<0.05）.With MEL supplementation at the level of 1 250 mg•L^-1，molar proportion of propionic acid decreased（P<0.05）while ratio of acetic acid to propionic acid increased（P<0.05）.Molar proportion of butyric acid increased（P<0.05）with MEL supplementation.Melamine showed an inhibitory effect on ruminal microbial fermentation and might not be effectively used by ruminal microbes.Therefore，MEL could not be used as a non-protein nitrogen source for feeding ruminant animals.

 The effects of dietary vitamins (vitamin E and beta-carotene) and zinc on semen quality，antioxidant indicators of Holstein bull in hot weather were analyzed.Sixty healthy and purebred Holstein bulls (BW=1 010± 80 kg) were randomly classified into 4 groups (n=15)：group Ⅰ：control group (basal diet); group II：basal diet +100 mg•kg^-1DMZn; group III：basal diet+antioxidant (300 mg•kg^-1DMvitamin E and 60 mg•kg^-1DMbeta-carotene) and group IV：basal diet +100 mg•kg^-1DMZn + vitamins (300 mg•kg^-1DMvitamin E and 60 mg•kg^-1DMbeta-carotene).The trial duration was 120 days.The results showed that vitamins and zinc could significantly increase the semen quality，vitality of fresh semen (group Ⅳ 0.649 vs groupⅠ0.575)，sperm density (group Ⅳ 15.16×10⁸ mL^-1 vs groupⅠ11.81×10⁸ mL^-1)，vitality of frozen semen (group Ⅳ 0.34 vs groupⅠ0.283) and acrosome integrity rate (group Ⅳ 45.2% vs groupⅠ41.8%) but decreased the sperm deformity rate significantly (group Ⅳ13.47% vs groupⅠ 16.81%).However，there was no significant effect on ejaculate volume (group Ⅳ11.56 mL vs groupⅠ10.33 mL).In addition，the serum Cu-Zn SOD activity could be significantly increased by the supplemental vitamins or vitamins + zinc（P＜0.01）; the individual or combination of supplemental vitamins and zinc significantly positively affected the serum T-AOC（P＜0.01）; and GSH-PX activity significantly （P＜0.01）; the serum MDA (P=0.053 5) concentration and hydroxyl radicals (P=0.069 8) were markedly reduced as the result of adding vitamins and zinc.Supplemental vitamins significantly increased the total SOD（P＜0.01）and T-AOC（P＜0.01）activities in seminal plasma; the Cu-Zn SOD activity （P＜0.05） in seminal plasma was increased by the addition of zinc.In conclusion，a diet with 300 mg•kg^-1DMof vitamin E，60 mg•kg^-1DMof beta-carotenoids and 100 mg•kg^-1DMZn could improve the semen quality in hot weather.

The objective of this study was to investigate the effect of different sources of fiber on intestinal morphological development of Jilin White geese.Fifty-six geese were fed with diets containing corn straw and Leymus chinensis as the major source of fiber.The height of the villi，depth of the crypts，muscle thickness of the small intestine were measured on d 11，18，25，32，39，46 and 53 by the histological section technique.The results showed that jejunum villus height in geese fed corn straw and Leymus chinensis groups were greater than the other states of intestine.The absolute growth peak of the villus height and muscle thickness from d 11 to 25 were highest in geese fed corn straw and Leymus chinensis groups.The villus height，crypt depth of duodenum and jejunum were significantly higher in geese fed with Leymus chinensis than that in geese fed with corn straw (P＜0.01).Compared to the geese in Leymus chinensis group,those fed with corn straw had higher muscle thickness.The ratio of villus height to crypt depth positively correlated with the increasing age in different intestinal segments in corn straw and Leymus chinensis groups，and the geese in the Leymus chinensis group had higher ratio of villus height to crypt depth than that in corn straw group in the duodenum and jejunum (P＞0.05).The results indicate that，under this experiment condition，intestinal morphological development of Jilin White geese is significantly influenced by different sources of fiber from d 11 to 53，the development of duodenum and jejunum of geese fed with Leymus chinensis is better than those fed with corn straw.

The aim of this study was to establish a triplex for simultaneously detecting porcine epidemic diarrhea virus (PEDV)，porcine enterovirus-9 (PEV-9)，and porcine kobuvirus (PKV).Two pairs of special primers were designed according to the published sequences of PEDV，PKV in GenBank，one primers of PEV-9 were synthesized according to reference.A mixture of 3 paris of primers was used for amplification of viral nucleic acids，yielding 3 different amplicons with sizes of 681 bp，454 bp，and 313 bp for PEDV，PKV and PEV-9 respectively by optimizingthe reaction condition.Testing of the sensitivity of multiplex RT-PCR indicated that the lowest detection limits were 5.97 pg for PEDV，0.11 pg for PEV-9，0.74 pg for PKV，respectively.For the specificity test，all of pathogens in the negative controls were found no amplicons.A total of 46 specimens from piglets with acute diarrheas collected from Sichuan province were tested by triplex RT-PCR，and the positive rate were as follows：PEDV 76.1%，PKV 39.1%，PEV-9 73.9%，and the positive accordance rate between simplex and triplex PCR of PEDV，PEV-9，PKV were 100%，91%，and 94% respectively.In addition，the fact that three pathogens were detected simultaneously in one sample indicated mixed infection in the diarrhea disease.It was also suggested that the pathogenic mechanisms of three kinds of pathogens is somehow synergetic.This study showed that the triplex RT-PCR may be a useful tool for rapid and sensitive etiological diagnosis for acute viral diarrheas in piglets and provides an effective technical support for pathogenic molecular epidemiology investigation.Besides，it established the foundation that we can further explore the relationship between the traditional diarrhea pathogens and potential new intestinal pathogens.

In this study we constructed and identified the mini-genome of Duck Hepatitis A virus (DHAV), for analysis of the functional and structural properties of DHAV un-translated region (UTR).To construct a bicistronic of DHAV mini-genome (pRluc-fLuc)，the ORF of DHAV was replaced by firefly luciferase (fLuc) gene through enzyme digestion and the Renilla luciferase (Rluc) gene was inserted before 5′UTR.We also added the hammerhead ribozyme ahead Rluc gene and the hepatitis delta virus ribozyme behind 3′UTR.We transfected the pRluc-fLuc into DF-1 cells，the fLuc gene expression could be detected at 8 hours post transfected (h.p.t.) and with the levels peaking was at 24 h.p.t.The results showed that the DHAV mini-genome was constructed successfully and can be used for explore the role of UTR in virus transcription and translation.

The aim of this study was to study the feasibility of yolk antibody indicating the ALV infection status in SPF chickens instead of serum antibody.Eggs and serum samples were collected one to one correspondence from 40 23-weeks-old SPF chickens，which 21 chickens were positive to ALV-Ab and 19 chickens were negative.The egg yolk was diluted into 1∶150，1∶200，1∶300 respectively to compare the correlation of yolk antibody and serum antibody in the ELISA tests.76 SPF chickens with different ALV-Ab level were breed separately，eggs were collected once a week and serum was collected every three weeks from each chicken during the age of 25 to 34 weeks，totally 836 egg yolks and 304 serum samples were collected.They were tested the antibody response to ALV-Ab by ELISA detection kits.The yolk antibody correlated well with serum antibody when the egg yolk was diluted at the 1:300 dilution otherwise the false negative or false positives may arise.Antibody response to ALV-Ab in 836 egg yolks correlated well with 304 serum samples during the age of 25 to 34 weeks and the consistency rate is 94.7%.The yolk antibody can be as a good index to estimate the ALV infection in SPF chickens instead of serum antibody.

Capsular polysaccharide export protein (CPEP) gene of Haemophilus parasuis SC-1 was amplified by PCR and then was cloned into the pMD-19T vector.After being certified by sequence and digestion with BamHⅠand XhoⅠ respectively，the gene were cloned into the pET-32a(+) vector.The recombinant vector was transformed into BL21（DE3）and then induced by IPTG.The product of expression was studied by SDS-PAGE and Western blot，and then was purified.We further evaluated the immune responses and protective efficacy of purified CPEP in mice models.The results showed that the recombinant vector pET32a-CPEP was constructed successfully.The SDS-PAGE and the western blot results showed that the purified protein was 35 kDa and the recombinant CPEP possessed reactogenicity respectively.The protective capacity of the anti-CPEP antibodies was evaluated by the inoculation of highly virulent strain SC-1.Vaccinated animals had a delayed course of disease and 40% of the animals survived the lethal challenge.The partial protection achieved with the recombinant CPEP supports their potential as candidates to be included in future vaccine formulations against H.parasuis.

To study the population genetic diversity of T.multiceps isolated from Sichuan and the phylogentic relationships with other taenia species，the complete sequences of mitochondrial Cytochrome c oxidase subunit 1 (cox1) and Cytochrome b (Cyt b) genes of 20 Coenurus collected from goats (11 from brain，and 9 from intramuscular) in Sichuan were analyzed and phylogentic trees were reconstructed.The results showed that the complete lengths of mitochondrial genes sequenced in this study were 1 623 bp for cox1 gene and 1 068 bp for Cyt b gene.Variable sites were 59 and 9，respectively.The sequence divergence between the cox1 sequences of Coenurus isolated from Sichuan ranged from 0.1% to 2.5%，while differences in Cyt b sequences ranged from 0.1% to 0.7%.Seventeen haplotypes and 6 haplotypes were detected in cox1 and Cyt b，with the global haplotype diversity being 0.968 ± 0.033 (cox1) and 0.737 ± 0.068 (Cyt b)，the nucleotide diversities were 0.005 98 ± 0.001 33 for cox1 and 0.003 06 ± 0.000 79 for Cyt b，and the average genetic distances were 0.006 for cox1 and 0.003 for Cyt b，respectively.The phylogenetic trees inferred from both cox1 and Cyt b showed that all 20 Coenurus isolated from Sichuan were the species T.multiceps.A closer relationship was found between Sichuan isolates and Iran and Turkey isolates，and it was closer than that between Sichuan and Gansu，Guangzhou and Italy isolates.The results indicated that cox1 gene and Cyt b gene were suitable markers for detecting genetic diversity of T.multiceps，and the diversity of T.multiceps based on Cyt b gene was lower than cox1 gene.

The aim of this study was to prepare antiserum against the goose CD4 protein.Specific primers for goose CD4 gene were designed according to public Mallard duck reference sequence (AF378701) in GenBank.Then we obtained Northeast White Goose CD4 gene by RT-PCR successfully.According to goose CD4 ORF sequence，specific primers were further designed and goose CD4 extracellular region gene was cloned.We produced recombinant goose CD4 extracellular region protein in prokaryotic expression system (pET-32a（+）/Rosetta(DE3)pLysS).The recombinant protein was induced by IPTG and was purified by Ni-NTA column affinity chromatography.As immunogen，the purified recombinant protein prepared rabbit anti-goose CD4 extracellular region serum.I-ELISA，Western blot and Flow cytometric analysis showed that antiserum could identify recombinant goose CD4 extracellular region protein and peripheral blood T lymphocytes respectively.Indirect immunofluorescence test confirmed the purified antiserum could identify goose CD4 extracellular protein by eukaryotic expression system specifically.The above results show that antiserum can be used as detection reagent to detect goose CD4⁺ T lymphocytes.

Six 30-day-old piglets (post-weaned) were selected as experiment animals，which were free of porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) antibody and antigen as detected by ELISA and RT-PCR.The alveolar macrophages (AMs) isolated aseptically were divided into four groups，i.e.，control group，molecular myeloid differentiation 88 (MyD88) interference group (interference group)，PCV2 infection group (PCV2 group) and PCV2 infection-MyD88 interference group (PCV2-interference group)，and cultured in vitro.The MyD88 gene was knocked down by siRNA (small interfering RNA).After PCV2 infection，the cells and the cultural supernatants were collected at 0，6，12，24 and 48 h respectively.The gene-silencing efficiency of siRNA was detected by fluorescence quantitative PCR.The intracellular expression of MyD88 protein was detected by Western blot.The dynamic changes of IL-1β，IL-6 and IL-10 in the supernatants were assayed by ELISA.The results showed that siRNA could lead to 78% of the MyD88 gene silencing，and significantly affected the expression of MyD88 protein (P<0.01).The amount of MyD88 expressed in AMs in PCV2 group increased significantly after 6 h (P <0.05)，which was significantly higher compared to that in PCV2-interference group (P <0.01) at 12，24 and 48 h.The yield of IL-1β secreted in AMs at 6 h infected with PCV2，was notably higher compared with those in control group (P<0.01) and PCV2-interference group (P<0.05).The level of the secreted IL-6 in the PCV2 group was significantly higher than those in the control group (P<0.01) and PCV2-interference group (P<0.05) at 6 and 12 h.The yield of IL-10 secreted in AMs significantly increased (P<0.01) at 12，24 and 48 h infected with PCV2; the yield of IL-10 in PCV2-interference group significantly decreased (P<0.01) at 24 and 48 h.The results indicated that the infection of PCV2 contributed to different changes of the secretion of IL-1β，IL-6 and IL-10 in PAMs cultured in vitro，where MyD88 was involved as an important regulator.

An assay of SYBR Green Ⅰ real-time fluorescent quantitative RT-PCR was established to investigate the expression pattern of chicken PD-l，PD-L1 and PD-L2  genes at different stages after IBDV infection.The specific primers were designed according to the sequences of PD-l，PD-L1 and PD-L2  from GenBank to amplify the objective genes for corresponding plasmids construction，and then they were used as quantitative template to construct the standard curve and melting curve for detection sensitivity，specificity and repeatability.The established SYBRS Green Ⅰ real-time fluorescent quantitative PCR (Rea1 time FQ-PCR) assay worked well with a good coefficient correlation(R²>0.99) while the template concentration was from 1×10¹-1×10⁹ copies·μL^-1，the sensitivity was sufficient to detect at least 10¹ copies of the samples.The relative transcription levels of chicken PD-l，PD-L1 and PD-L2 mRNA in PBMCs from chicken were detected on the 3rd，5th，7th and 10th day after IBDV infection by the developed Rea1 time FQ-PCR assay.At the same time，the semi quantitative RT-PCR and IBDV rapid test strip for detection were carried out to analyze the viral loads in bursal tissue.Real-time RT-PCR analysis show that the expression of PD-1 increased significantly (P<0.05) in the 7th day after infection，the expression of PD-L1 and PD-L2  increased significantly (P<0.01,P<0.05) in the 3rd day after infection and the IBDV loads in bursal tissue are highest in the 5th days after infection.The established assay was able to detect the expression of the three genes in PBMC.Our results revealing the pattern of chicken PD-l，PD-L1 and PD-L2 genes at different stages after IBDV infection，which showed that the gene transcription increased positively correlated with IBDV loads，and PD-L1 , PD-L2 increasing transcription precede PD-1.

In order to explore whether the superior cervical ganglion (SCG) of female goats was equipped with the conditions for the action of progesterone，thereby speculating that whether progesterone was involved in neural regulation of endocrine processes of target organs，we used immunohistochemical SP method to detect the distribution of progesterone receptor (PR) in SCG of female goat.The result showed that PR-immunoreactive-products were widely distributed in the SCG of female goat，and different levels of staining existed in neurons，Schwann cells，satellite cells and nerve fibers.All the neurons were PR positive，of which the cell membrane and cytoplasm was brown as moderate positive，and nuclei was brown as strong positive.Schwann cells，satellite and nerve fibers also had yellow，weak positive PR-positive-product.Image analysis showed that PR in the nerve cells was extremely higher (P＜0.01) than other non-nerve cells.The result proved that the neurons in the SCG were the one of main target cells of progesterone，implying that progesterone may act on the sympathetic postganglionic neurons of the SCG，and thus participates in the activities of the target organ dominated by the SCG，and PR in the SCG might act as a network node to coordinate the endocrine regulation of progesterone and neuroregulation of autonomic nerve on the activities of the target organ.

By using nanotechnology，ginsenoside(GS) and levamisole hydrochloride(LH) were combined with the carrier nanoemulsion to prepare compound GS and LH nanoemulsion(GS-LH-NE).Then its composition and acute toxicity were evaluated.The optimum blank nanoemulsion，which was used to prepare GS-LH-NE，was screened out through pseudo-ternary phase diagram.The enhancement effect of nanoemulsion containing different content of drug on the immunologic function of immunosuppressive mice were studied through splenic lymphocytes proliferation test，and the nanoemulsion with a better immunologic enhancement effect was screened out for ascertaining the composition of GS-LH-NE.The morphology of GS-LH-NE was observed using transmission electron microscope; the particle size was measured using laser particle size analyzer; the gavage acute toxicity of GS-LH-NE was measured using mice.The ascertained composition of GS-LH-NE was Solutol^®HS-15/glycerol/PEG400/IPM/distilled water(with the mass ratio of 25.33:10.14:2.53:6:56)，in which the content of GS and LH were both 30 mg·mL^－1 and 30 mg·mL^－1，respectively.GS-LH-NE was a yellow transparent oil-in-water nanoemulsion，which had spherical droplets with the particle size of 23.08 nm and PDI of 0.237，and its gavage LD₅₀ was 402.85 mg·mL^－1.GS-LH-NE is successfully prepared and provides the possibility to develop immunologic enhancer through nanotechnology.

In order to construct the model of heat accumulation of intestinal type of Qi-fen Syndrome，the rats were feed with the hot medicine and the rat wastes，then were injected with the inactive E.coli，and the clinic symptoms such as the in taking，drinking，body weight and body temperature were determined，even the intestinal organizational structure，and the number of intestinal IEL and GC in rat were studied.According to the clinic symptoms，in taking，drinking，body temperature of the rats，the results showed that that the methods of feeding with the hot medicine，wastes and injecting the inactive E.coli to the rats could make the model of heat accumulation of intestinal type of Qi-fen Syndrome in successfully when the rats were injected with the inactive E.coli 5 hours later.The changes of intestinal structure in rat injected with the inactive E.coli 5 hours later were significantly，and the numbers of the IEL and GC in the duodenum，jejunum and ileum decreased in significantly.Therefore，the model of heat accumulation of intestinal type of Qi-fen Syndrome in rat can be made in successfully after the rats were injected with the inactive E.coli 5 hours later.