To study the necessary of retinoic acid receptor α (RARα) for rat neuron function.
Methods Tissue digestion was used to isolate and cultivate the rat primary hippocampal neurons,and the adenovirus vector was used to  specifically silence the  RARα;Real-Time PCR was used to analyze the influence of silenced RARα in  retinoic acid(RA) receptors and the markers of nerve cells;live cell imaging analysis was performed to analyze the influence of the calcium excitability of neurons silenced RARα.Results The immunofluorescence results showed that  90% of the isolated cells expressed the  neuron marker neuron-specific enolase (NSE),the adenoviral transfection efficiency was up to 80%.The PCR results showed the expression of RARα in silenced RARα neuron was decreased by 75% (P<0.01),the other receptors were significantly decreased (P<0.01),but RARβ was significantly increased (P<0.05).The live cell calcium imaging results showed the calcium excitability in silent group was significantly reduced (P<0.05),however all-trans retinoic acid (ATRA) pretreatment for 24 h could significantly enhance the calcium excitability (P<0.01).Conclusion The absence of RARα can significantly reduce the neuron marker NSE expression of the primary hippocampal neurons,and significantly damage the neuronal calcium excitability.

bstract:Objective
To study the necessary of retinoic acid receptor α (RARα) for rat neuron function.
Methods Tissue digestion was used to isolate and cultivate the rat primary hippocampal neurons,and the adenovirus vector was used to  specifically silence the  RARα;Real-Time PCR was used to analyze the influence of silenced RARα in  retinoic acid(RA) receptors and the markers of nerve cells;live cell imaging analysis was performed to analyze the influence of the calcium excitability of neurons silenced RARα.Results The immunofluorescence results showed that  90% of the isolated cells expressed the  neuron marker neuron-specific enolase (NSE),the adenoviral transfection efficiency was up to 80%.The PCR results showed the expression of RARα in silenced RARα neuron was decreased by 75% (P<0.01),the other receptors were significantly decreased (P<0.01),but RARβ was significantly increased (P<0.05).The live cell calcium imaging results showed the calcium excitability in silent group was significantly reduced (P<0.05),however all-trans retinoic acid (ATRA) pretreatment for 24 h could significantly enhance the calcium excitability (P<0.01).Conclusion The absence of RARα can significantly reduce the neuron marker NSE expression of the primary hippocampal neurons,and significantly damage the neuronal calcium excitability.

Abstract:Objective To investigate the effects of recombinant plasmid pshuttle-Egr1-shTRAIL stable transfection in combination with X-ray irradiation on the TRAIL protein expression and the apoptosis in human SMMC7721 hepatoma cells. Methods The pshuttle-Egr1-shTRAIL packaged with liposome was stably transfected into SMMC7721 cells in vitro.The shTRAIL protein expressions were measured with ELISA assay,Annexin V-FITC kit was adopted to measure the apoptosis of pshuttle-Egr1-shTRAIL cells,and the changes in survival rate of SMMC7721 cells were measured　with cell cloning assay.Results The TRAIL protein expressions in pshuttle-Egr1-shTRAIL plus different doses of irradiation groups  were  significantly increased compared  with 0 Gy group（P< 0.001）.The percentage of apoptotic cells was significantly higher than that in 0 Gy group (P<0.05 or P<0.001),and the survival rate of SMMC7721 cells was decreased significantly (P<0.05 or P<0.001).Conclusion The pshuttle-Egr1-shTRAIL stable transfection in combination with irradiation can significantly induce the apoptosis of SMMC7721 tumor cells and inhibit the cell proliferation.

Abstract:Objective
To observe the changes of Th1,Th2 and and Th3/Tr1 cytokines IFN-γ,IL-10 and TGF-β after treated with ionizing radiation,and explore the radiationship between cell balance and radiation dose.Methods After whole-body irradiation with various doses （low dose:0, 0.075,0.100,0.200Gy;high dose: 0,1.000,2.000,4.000,6.000 Gy） for 16 h,ELISA was used to detect the conternts of IFN-γ (Th1-type cytokine),and IL-10 (Th2 type cytokine),TGF-β1(Th3/Tr1 type cytokine) in spleen and thymus.Results Compared with sham irradiation control group,the IFN-γ and TGF-β levels in spleen cells were decreased after  low-dose radiation (0.075-0.200 Gy),but there was no statistically significant difference（P>0.05）;however the IL-10 secretion was significantly lower than that in sham irradiation group (P<0.01,P<0.05).After  high-dose radiation (2.000-6.000 Gy),the IFN-γ,IL-10,and TGF-β levels in spleen cells were higher than those in sham irradiation group (P<0.01,P<0.05). The IFN-γ and IL-10 after low- or high-dose radiation expressed more than sham irradiation group (P<0.01,P<0.05).But the TGF-β level was lower than that in sham irradiation group after radiation at low doses (P<0.01),and it was significantly higher than that in sham irradiation group after exposured to high-dose radiation (P<0.01).Conclusion High and low-dose radiation may influence the secretion of Th3/Tr1 type cells regulatory factor TGF-β of mouse immune organs and paly a key role in different immune response effects  induced by different doses of radiation.

Abstract:Objective
To study the effect of ionizing radiation on the expression of hyaluronidase(HAase) in human fibroblasts and to discuss the role of HAase in the pathogenic mechanism of radiation-induced brachial plexus injury.Methods The fibroblasts were irradiated for 24 and 48 h respectively by X-ray with doses of 0.5,1.0,2.0,4.0 and 6.0 Gy,and sham irradiation group (0 Gy X-ray) was set up.The ultrastructural changes of fibroblasts were observed by transmission electron microscope,the expression changes of protein and mRNA of HAase were detected by ELISA and real-time PCR method.
Results After irradiated　for 48 h,the nuclear chromatin began to get together in 2.0 Gy group;the nucleus and chromosome fractured and the chromosome appeared empty bubble change in 4.0 Gy group;the chromatin was no longer apparent and the apoptosis change presented early in 6.0 Gy group.After irradiated for 24 and 48 h,with the increasing of　the radiation dose,the expression of HAase protein declined,there was significant difference compared with sham irradiation group(P<0.05).The expressions of HAase mRNA in 0.5 and 4.0 Gy groups were significantly higher than that in sham irradiation group(P<0.05),but the expressions of HAase mRNA in 1.0,2.0 and 6.0 Gy groups were significantly lower (P<0.05) at 24 h.The expression of HAase mRNA in 2.0 Gy,4.0 Gy and 6.0 Gy groups were significantly higher than that in sham irradiation group at 24  h(P<0.05),the expressions of HAase mRNA in 0.5 Gy and 1.0 Gy groups were significantly lower at 48 h (P<0.05).Conclusion The increase of the radiation dose could reduce the expression of HAase in human fibroblasts,and influence the occurrence of radiation-induced brachial plexus injury.

Abstract:Objective
To investigate the effects of ATRA on the proliferation and migration abilities and membranous distribution of Cx32 and Cx43 of colorectal cancer cells  Caco-2 and SW480 and to elucidate the potential mechanism.Methods The colorectal cancer cell lines Caco-2 and SW480 were divided into ATRA treatment groups and control groups.The Caco-2 and SW480 cells were treated with 1×10^-8，1×10^-7，1×10^-6，1×10^-5 and 1×10^-4 mol•L^-1 ATRA,and MTT assay was applied to detect the inhibitory rates of growth of Caco-2 and SW480 cells.After the SW480 cells were treated with the 1×10-5 and 1×10-4 mol•L^-1 ATRA,the wound healing assay was employed to determine the cell migration ability and flow cytometry was used to analyze membranous distribution of Cx 32 and Cx 43 in SW480 cells.Results Compared with control group,the growth inhibitory rates of Caco-2 and SW480 cells in ATRA groups after treated with 1×10-4 mol•L^-1 for 24,48 and 72 h were significantly increased（P＜0.01）;the migration abilities of SW480 cells were dramatically decreased after treated with 1×10^-5 mol•L^-1 ATRA for 24 and 48 h （P＜0.05）;the membranous distribution of Cx 32 and Cx 43 of SW480 cells was remarkably increased after treated with 1×10^-5 and 1×10^-4 mol•L^-1 ATRA for 24 and 48 h（P＜0.05）.Conclusion ATRA could increase the membranous distribution of Cx32 and Cx43 of colorectal cancer cells and inhibit the proliferation and migration abilities,which may be involved in the interaction between connexins and cytoplasmic proteins,as well as alteration of gap junctional intercellular communication.

Abstract:Objective
To check the induction of the expression of the genes related to the immune response of human hepatocytes at the early stage of RNA virus infection and to reveal the hepatocyte specific immune response.Methods The Sendai virus (SV) infected primary human hepatocytes were divided into  0,3 and 6 h groups, immunofluorescence (IF) assay was used to detect the location of IRF3 and IRF7.The SV infected primary human hepatocytes were divided  into 0,1,2 and 3 h groups,the  expressions of  IFNα1,IFNβ  and IFNλ3 and RIG-I in the hepatocytes at the early phase of infection  were detected by Real-time PCR.Results The IRF3 and IRF7 were detected in the cytoplasma before SV infection.The signal of IRF7 was detected in the nuclear of some cells after 3 h infection of SV,and the signals of both IRF3 and IRF7 were detected in the nuclear of some cells after 6 h infection.Compared with 0 h SV infection group,the  expressions of RIG-I, IFNβ  and IFNλ3  were significantly increased and  IFNα1 was slightly increased in 2 h SV infection  group （P<0.05）.Conclusion The nuclear translocation of the constitutively expressed IRF7 in hepatocytes occurs at the early stage of infection and is related to the induction of expression of the IFNs corresponding to the infection.It indicates that the hepatocytes have specific and fast immune response to the virus infection.

Abstract:Objective To investigate the morphological changes of cornea of rats and mice with type 2 diabetes mellitus induced with high-fat diet combined with low-dose STZ and clarify the characteristics of  the corneal injury in  rat and mouse models with diabetes and eye diseases.Methods Animals were divided into control groups and model groups.High-fat diet combined with intraperitoneal injection of low-dose STZ were used to induce type 2 diabetic rat and mouse  models.After 16/12 weeks,fasting blood glucose,free fatty acids and total cholesterol levels were measured with oxidase method and enzyme method.The histopathological changes of the cornea were analyzed by light microscope.Results Compared with control groups,there was no significant change in body weights of rats and mice in model groups (P>0.05);but the food intake,fasting blood glucose and lipid levels of rats and mice in model groups were significantly increased  (P<0.01).Compared with control groups,the corneal epithelium and stroma in model groups were obviously edema.Conclusion Both experimental diabetic models induced by high-fat diet and low dose of STZ in rats and mice have significant damage to the cornea after 16/12 weeks.

Abstract:Objective To study the activities of anti-duck hepatitis B virus (anti-DHBV) of three novel nucleoside analogues and their effects on histomorphology of duck liver,and to explore  novel potential anti-HBV agents.Methods 　The anti-DHBV activities were analyzed by fluorescent quantitative PCR in experimental groups with various doses of 030703,030605,030705 ( 2,10 and 50 mg·kg^-1 ) and adefovir dipioxil control group (10 mg·kg^-1).Compounds were taken by oral administration per day and last 30 d.The serum specimens were obtained at the time of before adminstration,15 d and 30 d after administration and 2 weeks after withdraw.The serum DHBV DNA was detected with fluorescence quantitative PCR.At the same time,DHBV DNA positive and negative groups were set up.The duck liver specimens were obtained after experiment.The  histomorphological changes of duck liver caused by three tested compounds were observed  under optic microscope.Results After administration with compound 030703,there were 3 ducks (60%) in high dose group,2 ducks (40%) in middle dose group,1 duck (20%) in low dose group,in which the DHBV DNA contents were decreased to 1/3 of primary contents;compared with adefovir dipioxil control group,the inhibitory effects in high dose group had significant difference (P<0.01),whereas the inhibitory effects in middle and low dose groups had no significant difference (P>0.05).After administration with compound 030605,there were 4 ducks (80%) in high dose group,3 ducks (60%) in middle dose group,1 duck (20%) in low dose group,in which the DHBV DNA contents were decreased to 1/3 of primary contents;compared with adefovir dipioxil control groups,the inhibitory effects in high dose group and middle group had significant difference (P<0.01),whereas the inhibitory effect in low dose group had no significant difference (P>0.05).After administration with compound 030705,there were 3 ducks(60%) in high dose group,2 ducks (40%) in middle dose group,0 duck (0%) in low dose group,in which the DHBV DNA contents were decreased to 1/3 of primary contents;compared with adefovir dipioxil control groups,the inhibitory effects in high dose group had significant difference ( P<0.01),whereas the inhibitory effects in middle and low groups had no significant difference ( P>0.05).Compared with adefovir dipioxil control group,the effects  of  all three test compounds(high,middle and low dose groups) on duck liver inflammation were similar to that in  adefovir dipioxil control group.Conclusion Three novel test compounds 030703,030605 and 030705 have obvious anti-DHBV activities and might improve duck liver inflammation to some extent.It has been suggested that all of these novel compounds be potential anti-HBV agents.

Abstract:Objective
 To construct pGEX-4T-1/SUMO3 recombinant plasmid in order to express the full length of small ubiquitin-like modifier 3 (SUMO3) in E.coli and purify the GST-SUMO3 fusion protein. Methods The DNA sequence of SUMO3 (full length: 312 bp) was amplified by PCR using the template plasmid pcDNA3.1/SUMO3 and was then cloned into the expression vector pGEX-4T-1.After identified by restriction enzyme digestion and sequencing,the recombinant clone was transformed into the competent expression cells of E.coli BL21.GST-SUMO3 fusion protein was induced by IPTG and further purified by Glutathione Sepharose 4B to obtain a fusion protein with molecular weight of 38　000.Results It was identified that the recombinant expression vector of pGEX-4T-1/SUMO3 contained a 312 bp insert fragment by BamHⅠ and XhoⅠ double digestion and the insert fragment showed exactly consistant sequence with SUMO3.The fusion protein of SUMO3 combined with GST was successfully expressed and purified with 0.1 mmol·L^-1 IPTG induction.Conclusion The SUMO3 protein combined with GST-tag is gained successfully,which provides the basis for the preparation of SUMO3 antibody and the further functional research of SUMO3.

Abstract:Objective To investigate the expressions of aquaporin 2（AQP2） and aquaprin 8（AQP8） in eutopic and ectopic endometrium of patients  with  endometriosis（EMs）and  to elucidate  possible impact of AQP2 and AQP8 on EMs. Methods 53 patients in our hospital were selected,among them 37 cases were EMs patients,they were divided into eutopic endometrium(Eu-Em) group and ectopic endometrium(Ec-En) group;15 healthy subjects were used as control(Co-Em) group.The AQP2 and AQP8 mRNA and protein expressions in endometrium tissues in   Eu-Em group,Ec-Em group  and  Co-Em group  were detectedwith real-time reverse-transcriptase polymerase chain reaction and Western blotting.Results Compared with Co-Em group,the AQP2 and AQP8 mRNA levels in Eu-Em group had no significant difference(P<0.05);the AQP2 mRNA level in Ec-Em group was significantly high than those in Co-Em and Eu-Em(P<0.05),the AQP8 mRNA level in Ec-Em group was significantly lower than those in Co-Em and Eu-Em group(P<0.05).Compared with Co-Em group,the AQP2 and AQP8 protein expressions in Eu-Em group had no significant difference(P>0.05),but the AQP2 and AQP8 protein expression levels in Ec-Em group were  significantly lower than those in Co-Em and Eu-Em groups(P<0.05).Conclusion AQP2 and AQP8 may play an important role in occurence and development of endometriosis.

Abstract:Objective To explore the effects of velvet antler polypeptide/ nano β-tricalcium phosphate（β-TCP）/ gelatin material on the osteogenetic differentiation in rabbit mesenchymal stem cells (MSCs),and clarify the mechanism.Methods  Cell cycle and proliferation were examined by flow cytometry and MTT assay in the third generation MSCs treated with 5 mg/g velvet antler polypeptide/20 % nano TCP/gelatin material for 48 h.The growth of MSCs cocultured with the material was  observed by scanning electron microscope.The morphology  of apoptotic MSCs was observed by AO∕EB fluorescence staining assay.The activity of alkaline phosphatase (ALP) was detected with automatic biochemistry analyzer,and the mineralized nodules  in MSCs were counted after 21 d treatment.Results  The material could significantly promote the proliferation of MSCs,the relative growth rate was 126.45 % after treated with the material for 48 h.The percentage of MSCs in the S phase was 45.84 %（7.96% in control）,and the apoptotic  rate was 2.08 %（13.68%  in control group）.The calcified nodules and the activity of ALP were increased after treatment with material for 21 d. MSCs could adhere to the surface of the material and go well with it.Conclusion The material has the effects on stimulating the proliferation,differentiation and mineralization of rabbit MSCs.

Abstract:Objective To construct the fusion gene eukaryotic expression vector pEGFP-N1-SUMO-3,and to study the expression and localization of GFP-SUMO-3 fusion protein in human prostate cancer cell line LNCaP.Methods The encoding region of SUMO-3 was obtained by PCR,and cloned into eukaryotic expression vector pEGFP-N1.Then the pEGFP-N1-SUMO-3 plasmid was transfected into LNCaP cells by liposome.The expression of the fusion protein was detected by Western blotting,and the subcellular localization  was observed by fluorescence microscope.Results The recombinant plasmid  pEGFP-N1-SUMO-3 was identified by BamHⅠ and XhoⅠ double digestion.As expected,it showed two bands of 4 700 and 312 bp.The recombinant plasmid also showed right sequence by the full length sequencing.The pEGFP-N1-SUMO-3 plasmid was transfected into prostate cancer LNCaP cells.A specific protein expression band at molecular weight of 39 000 was detected with  using GFP-antibody by Western blotting method.When observed by fluorescence micro
scope,the GFP-SUMO-3 fusion protein was mainly located in the nucleus of LNCaP cells.Conclusion The recombinant plasmid pEGFP-N1-SUMO-3 is successfully constructed,and GFP-SUMO-3 fusion protein is successfully expressed in mammalian cells and mainly located in the nucleus.

Abstract:Objective To construct the double interference vector targeting both PTTG and survivin gene and detect its silencing effects on PTTG and survivin gene in human glioma   U251 
cell line.Methods Two effective targeting sequences were chosen according to PTTG and survivin cDNA sequences in GenBank,then they were inserted into pGenesil-2.1 vector and pGenesil-2.1-PTTG and pGenesil-2.1-survivin plasmids were constructed. The recombinant plasmids were identified by restriction endonuclease and DNA sequencing. pGenesil-2.1-survivin and pGenesil-2.1-PTTG plasmids were digested by HindⅢ and BamHⅠ separately,long fragment of pGenesil-2.1-survivin plasmid and small fragment of pGenesil-2.1-PTTG (about 380 bp) were collected by 1% agarose gel electrophoresis.The double interference vector pGenesil-2.1-PTTG-survivin siRNA was constructed by uniting long fragment of pGenesil-2.1-srvivin plasmid and small fragment of pGenesil-2.1-PTTG.This plasmid was identified by restriction enzyme digestion and then its silencing effects on PTTG and surviving gene were detected. Results By restriction endonuclease and DNA sequencing,the eukyaryotic expression plasmid of pGenesil-2.1-PTTG,pGenesil-2.1-survivin and pGenesil-2.1-PTTG-survivin siRNA were constructed correctly.The result of RT-PCR showed that PTTG and survivin gene mRNA levels were decreased obviously after this plasmid was transfected into U251 cells for 48 h.Conclusion pGenesil-2.1-PTTG-survivin siRNA plasmid is constructed successfully.The PTTG and survivin gene mRNA levels in U251 cells are decreased effectively by transfecting this plasmid into U251 cells.

Abstract:Objective To observe the effect of mongolian milkvetch root on nitric oxide(NO)，interleukin 5(IL-5)，inflammatory cells in bronchoalveolar lavage fluid (BALF) and serum of bronchial asthmatic rats and clarify the efficacy of mongolian milkvetch root in treatment of asthma．Methods 60 Wistar rats were randomly divided into normal group，model group，budesonide suspension for inhalation intervention group and low,middle,and high dose mongolian milkvetch root intervention groups，10 rats in each group．The rat bronchial asthma models were established by egg protein(OVA) sensibilization and long-term inhalation provocation．The rats in mongolian milkvetch root intervention groups were lavaged with mongolian milkvetch root 2 mL (0.3 g/100g)，and the rats  in  budesonide suspension for inhalation intervention group were  inhalated with  budesonide　suspension  2 mL at 21 d after expreriment;the rats in normal group and model group  were lavaged with normal saline orally，once each day for 14 d.The behavior of rats was observed and  the changes of NO，IL-5，inflammatory cells in BALF and serum of  bronchial asthmatic rats in each group were detected.Results The model of asthma was set up successfully by observing the behavior of rats，then the behavior of the rats became better after treatment，but did not reach the normal state.The NO  and IL-5 levels in  BALF and serum in mongolian milkvetch root intervention groups were significantly reduced compared with model group (P<0.01)，and the percentages of the total inflammatory cells and eosinophils in BALF were significantly reduced (P<0.01).Conclusion Mongolian milkvetch root could inhibit the increasing of  IL-5 and NO in BALF in asthmatic models，and could significantly reduce activation and infiltration of airway inflammatory cells and eosinophils，thus reduce airway inflammation and asthma symptoms or avoid the onset of asthma to achieve the purpose of prevention and treatment of asthma.

Abstract:Objective To investigate the protective effect of  cardiomyopeptidin(CMP) for injection preconditioning on the immature myocardial ischemia-reperfusion injury in young rats,and clarify its mechanism.Methods 30 young healthy SD rats were randomly divided into sham-operated group,model control group and CMP　group (n=10).The levels of serum lactic dehydrogenase （LDH）,creatine kinase MB（CK-MB）,troponin Tn-T were detected;the contents of superoxide dismutase (SOD),malondialdehyde (MDA),nitric oxide(NO),total nitric oxide synthase (TNOS),inducible nitric oxide synthase (iNOS)and endothelial nitric oxide synthase（eNOS）in myocardium tissue were measured;the expression levels of iNOS,eNOS and Caspase-3 protein in myocardial tissue were detected by immunohistochemical method.The expressions of iNOS,eNOS and Caspase-3 mRNA  in myocardial tissue were mea
sured by real-time fluorescence quantitative PCR.The changes of myocardial structures were observed with optical microscope and transmission electron microscope.The apoptosis of myocytes was observed by TUNEL.Results Compared with model control group,the levels of LDH,CK-MB,Tn-T in CMP group were decreased(P< );Compared with sham-operated group,the contents of MDA,SOD,TNOS,and iNOS were increased and the expression levels of iNOS and caspase-3 mRNA were significantly down-regulated in CMP group,but less than those in model control group（P<0.01 or P<0.05）.Several pieces of myocardial hemorrhage and inflammatory cell infiltration were observed in model control group,severe degeneration and necrosis of myocytes could be found,the apoptotic myocytes were significantly increased;while in CMP group,the myocardial pathological injuries were mitigated,only mild degeneration and necrosis of myocardial cells could be found,the vascular structure was near-complete,the number of apoptotic myocytes between sham operation group and control group.Conclusion Cardiomyopeptidin for injection preconditioning can protect immature myocardium against ischemia-reperfusion injury in young rats.The mechanism may be associated with reducing oxygen free radical (OFR) and NO production,inhibiting the  expressions of iNOS mRNA,Caspase-3 mRNA and Caspase-3 protein in myocytes,and reducing apoptosis.

Abstract:Objective To explore the mechanisms of DNA damage in rat H9c2 cells in vitro induced by different concentrations of cadmium chloride (CdCl2).Methods SCGE method was used to detect the DNA damage in rat H9c2 cells after exposed to 0,5,10,30,50 and 80 μmol?L-1 CdCl2 for 6,12 and 24 h.Western blotting was applied to observe the poly ADP-ribose polymerase (PARP) activity after 24 h exposure.Cytochrome-C (Cyt-C) and apoptosis inducing factor (AIF) expressions were measured by immunocytochemical method.Results With the increasing of CdCl2 concentration and exposure time,the DNA damage levels and rates of rat myocytes in 10,30,50 and 80  μmol?L-1 dose groups were increased after treated for 12 and 24 h.There were significant differences between various groups (P<0.05 or P<0.001).After treatment of CdCl2 for 24 h,the significant PARP cleavage fragment of 89 000 was found in 30,50 and 80  μmol/L dose groups.With the increasing of CdCl2 concentration,the Cyt-C and AIF protein expressions were significantly increased,there were significant differences between various groups (P<0.05 or P<0.001).Conclusion The toxic mechanisms induced by CdCl2 in rat H9c2 cells may be related to inducing the DNA damage and affecting the expressions of PARP,Cyt-C and AIF.There exists dose-dependent relationship.

Abstract:Objective To study the effects of ABM polysaccharide,flammulina velutipes polysaccharide and lentinan on the fermentation properties of lactobacillus acidophilus,and explore whether they can be considered as kinds of new prebiotics,and screen the best concentrations of the three fungi polysaccharides when they are used as prebiotics. Methods Fermented milk with lactobacillus acidophilus was studied by adding  different concerntrations of carbon sources. Experimental groups were added with ABM polysaccharide (0.2%,0.4%,0.6%,0.8%, and 1.0%）,flammulina velutipes polysaccharide (0.2%,0.4%,0.6%,0.8%, and 1.0%）and lentinan (0.4%,0.6%,0.8%,1.0%, and 1.2%）,positive control group was added with the traditional prebiotics inulin (1%,2%,3%,4%, and 5%）;the microbiological indexes,viscosities and pH values of  all  fermented milk samples were detected,and compared with negative control group.Results In the experiments of detecting microbial indicators,the viable count indexes were the largest when the ABM polyasccaride and flammulina velutipes polysaccharide concentrations were 0.6%,and the lentinan concentration was 0.8%,the results were 10.34,9.75 and 9.95 logcfu/mL;the viable counts of experimental groups were all larger than those in negative control group.In the experiments of detecting viscosity,the viscosity indexes were the largest when the ABM polyasccaride and lentinan concentrations were 0.8%,and flammulina velutipes polysaccharide concentration was 0.6%,the results were 3 800,3 300 and 2 800 mPa.s;the viscosities of experimental groups were all larger than those in  negative control group.In the experiments of detecting pH,the pH values were the lowest when the ABM polyasccaride and flammulina velutipes polysaccharide concentrations were 0.6%,and the lentinan concentration was 0.8%,the results were 4.74,5.12 and 5.08; the pH values of experimental groups were all lower than those in  negative control group.Conclusion ABM polysaccharide,flammulina velutipes polysaccharide and lentinan  have influences in the fermentation properties of lactobacillus acidophilus.All of them can reach the standard of being prebiotics.ABM polyasccaride is the best prebiotics in this study.

Abstract:Objective To investigate the effects of different molecule weight astragalus polysacharin isolated from the annual astragalus membra-neaceus(APSⅠ)on the expressions of pro-inflammatory and anti-inflammatory cytokine in lipopolysaccharides (LPS)-stimulated RAW264.7 cells,and explore the anti-inflammatory effects of APSⅠ.Methods The RAW264.7 cells were cultivated in vitro,RAW264.7 cells or LPS-induced (1 mg?L-1) RAW264.7 cells were stimulated by APS Ⅰ,then divided into control group,LPS model group,APS Ⅰ group,LPS + different molecule weight APS Ⅰgroups(APS Ⅰ-A,APSⅠ-B,APSⅠ-C).The effects of different molecule weight and different concentrations of APSⅠon the proliferation of RAW264.7 cells were examined by CCK-8 method. The content of NO in the RAW264.7 cells supernatant fluid was detected by means of nitrate reductase.The concentrations of TNF-α and IL-10 in the supernatant fluid was determined with ELISA.Results Compared with control group,the proliferation of RAW264.7 cells was inhibited significantly after treated with APS Ⅰalone (P<0.05 or P<0.01),the levels of NO and IL-10 expressions were increased apparently (P<0.05) while the secretion of TNF-α was decreased (P<0.05);Compared with LPS model group,APS Ⅰ combined with LPS significantly increased the proliferation activity of RAW264.7 cells in LPS + APS Ⅰ groups (P<0.05
 or P<0.01),the levels of  No and TNF -α expressions were decreased significantly (P<0.05 or P<0.01),and the secretion of IL-10 was increased notably (P<0.05 or P<0.01).The effects of APS varied by different molecular weights,the inhibition of APSⅠ-C on the expressions of NO and TNF-α was more obvious,which had a stronger promotion on the secretion of IL-10 than APSⅠ-A and APS- B.Conclusion APSⅠhas anti-inflammatory effect on the LPS-stimulated RAW264.7 cells,the anti-inflammatory activation of APSⅠvaries by different molecule weights.APSⅠwith small molecule weight has more significant activation.So the biological activity of APSⅠmay be associated with molecule weight.

Objective
To observe the effects of angiopoietin-2(Ang-2) on the expressions of Tie-2,PI3K,Akt gene and protein in the colon cancer cell line (HCT-8) and analyze the relationship between Ang-2 and PI3K/Akt.Methods Different  doses of Ang-2 （0,0.2,0.4,0.8,1.2,2.0， and 3.0 mg?L-1） were added into the DMEM medium of HCT-8 cell line and the effective stimulus concentration of Ang-2 on HCT-8 was selected by MTT.The cells were divided into four groups: normal control group (C group),serum-free culture medium group (SD group),Ang-2 group (SA2 group) and LY294002 group (SA2L group),the gene and protein expressions of Tie-2,PI3K and Akt were detected by RT-PCR and Western blotting.Results Different concentrations of exogenous Ang-2 showed various proliferation effects on the HCT-8 and the most effective dose of Ang-2 was 2 mg?L-1.Compared with SD group,the expressions of Tie-2,PI3K and Akt gene and protein in SA2 group were increased significantly (Tie-2: all P<0.01;PI3K: P<0.01 or P> 0.05;Akt: P<0.01);compared with SA2 group, the expressions of three genes and proteins in SA2L group were decreased significantly (Tie-2: P<0.05 or P<0.01;PI3K: P<0.05 or  P<0.01;Akt: P<0.01 and P<0.05). Conclusion Ang-2 can promote the proliferation of the colon cancer cells through the Ang-2/Tie-2/PI3K/Akt pathway which can be inhibited by LY294002.

Objective
To investigate the effects of growth differentiation factor 5 (GDF5) on the proliferation and migration of endothelial cells,and clarify the mechanism of its repair. Methods Vascular endothelial cells were cultivated with different concentrations (10,50,100  μg?L-1) of GDF5, and normal control group was set up at the same time.The cell proliferation was detected by method of CCK8 method. Three-dimensional cultivation was used to investigate the effect of GDF5 on the migration of endothelial cells.Results CCK8 experiment showed that GDF5  inhibited the proliferation of vascular endothelial cells;with the increasing of  GDF5 dose,the inhibitory rate was increased correspondingly even more than twice,there were significant differences between various groups(P<0.05).Three-dimensional experiment showed that GDF5  obviously promoted the movement of vascular endothelial cells, with the increasing of   GDF5 dose,the cell movement was significantly increased,there were significant differences between GDF5 groups and control group(P<0.05).Conclusion GDF5 can inhibit the  proliferation of endothelial cells and promote the migration of endothelial cells.

Objective To investigate the relationship between the renin-angiotensin system (RAS)and asthma,and provide the theoretical basis for pathogenesis and treatment of asthma.Methods The mouse asthma models were set up and divided into control group, model group,candesartan low-dose group, and candesartan high-dose group.The serum of mice was collected, the levels of serum angiotensin Ⅱ (AngⅡ) and angiotensin Ⅰ (AngⅠ) were detected;the changes of angiotensinogen (AGT),angiotensin-convertion enzyme (ACE),angiotensin Ⅱ type 1 receptor(AT₁R) and angiotensin Ⅱ type 2 receptor (AT₂R) in the RAS of the mouse lung tissue were delected by Western blotting and RT-PCR.Results The serum Ang Ⅱ level in model group was higher than that in control group(P<0.05),but the level of AngⅡ in candesartan groups had no significant changes compared with model group（P>0.05）.The serum Ang Ⅰlevels  in each group had no significant changes（P>0.05）.The expressions of AT1R and ACE protein in model group were markedly increased compared with control group(P<0.05),the AT1R expressions in candesartan groups were significantly decreased compared with model group(P<0.05),but ACE unchanged.There were no significant differences of the expressions of AT2R protein between various groups（P>0.05）.The ACE mRNA  expression in model group was increased significantly compared with control group(P<0.05),but there was no significant change in candesartan groups compared with model group（P>0.05）.The AT₁R mRNA expression in model group was significantly increased compared with control group(P<0.05),the AT₁R  expressions in candesartan groups were significantly reduced compared with model group(P<0.05).The AT₂R mRNA in model group was significantly decreased compared with model group(P<0.05),the expressions in candesartan groups were significantly increased compared with model group(P<0.05).The expression of AGT mRNA in each group had no significant change.Conclusion In the course of asthma of mice,RAS activation is involved in the pathogenesis of asthma,and the AT₁R blocker can reverse the changes of the expressions  of ACE,AT₁R and AT₂R.

Objective
To investigate the influence of filamin A(FLNa) on drug-i
nduced apoptosis of breast cancer cells and explore its mechanism.Methods MDA-MB-231/MB 231-KD(FLNa+/ FLNa－) cells were treated with different doses(2.5,5.0,10.0 and 20.0 μmol?L-1)MMC,AS2O3 and DDP,meanwhile control group was set up.The inhibitory rates of proliferation of breast cancer cells were detected with MTT method.MDA-MB-231/MB231-KD(FLNa+/FLNa－)cells were treated with different doses (2.5，5.0，10.0 and 20.0 μmol?L-1)MMC,meanwhile control group was set up.The apoptotic rates of breast cancer cells were determined with flow cytometry.Results Compared with control group,the inhibitory rates of proliferation of FLNa+ and FLNa－ cells in  2.5,5.0,10.0,20.0 μmol?L-1 MMC groups were significantly increased(P<0.05),and were higher than those in AS2O3 and DDP groups(P<0.05)；there  was significant difference between  two kinds of breast cancer cells，the inhibitory rate of FLNa－ cells was higher than that of FLNa+ cells (P<0.05).As the dose increasing, the number of apoptotic cells was increased,there were significant differences compared with control group(P<0.05);the inhibitory rate of FLNa－ cells was higher than that of FLNa＋ cells and there was significant difference (P<0.05).Conclusion The expression of FLNa can reduce the  drug-induced apoptosis of breast cancer cells.

Objective
To investigate the  inhibitory effect of proliferation of parthenolide(Par) on salivary gland adenoid cystic carcinoma cell line SACC-83 cells in vitro and clarify its related mechanism.Methods The SACC-83 cells were treated with different concentrations(5.0，7.5，15.0，30.0，40.0 and 80.0 mg?L-1) of Par in vitro,the proliferation inhibition of SACC-83 cells was detected by MTT colorimetry,the cell cycle distribution and apoptosis status were analyzed by flow cytometry,the expression of Livin protein was determined by Western blotting.Results The growth of SACC-83 cells in Par groups was inhibited effectively in concentration-　and time-dependent manner.The  apoptotic rates in  Par  groups were 14.7%±2.1%,30.5%±3.4% and 38.7%±2.7%, respectively, after treated with  7.5,15.0 and 30.0 mg?L-1 Par for 24 h, there were significant differences compared with  control group (3.2%±0.9%,all P<0.01).The expressions of Livin proteins in Par groups were significantly down-regulated,there were significant differences compared with control group(P<0.05);and there were significant differences between different doses of Par groups（P<0.01）.Conclusion Par could obviously inhibit the proliferation of SACC-83 cells.The expression of Livin  protein may be related to this effect.

Abstract:Objective
To investigate the expressions of both urotensin Ⅱ(UⅡ) and its receptor GPR14 in rat vascular smooth muscle cells (VSMC) under the intervention of urantide (UⅡ receptor antagonist),and clarify its possible mechanism.Methods The VSMC from rat thoracic aorta were cultivated in vitro with tissue explant method,and divided into normal control group (C group),UⅡ group (M group),positive control group (Flu group) and five intervention groups (10^-10,10^-9,10^-8,10^-7 and 10-6 mol·L^-1 urantide).The proliferation index(PI) and S-phase cell fraction(SPF) of  in vitro cultured VSMC were detected with MTT method and flow cytometry.Results Compared with C group,the VSMC proliferation was increased at each time point(P<0.01),and   PI and SPF were also increased significantly  (P<0.01);compared with M group, the VSMC proliferation in 10^-10－10^-6 mol·L^-1 urantide groups were significantly decreased(P<0.05 or P<0.01)，and PI and SPF were significantly reduced  (P<0.05 or P<0.01),among which 10-6 mol·L^-1 urautide group had the most significant effect. Conclusion Urantide can  block the mitogenic effect of  UⅡ on atherosclerosis(AS)  of the major disease-promoting cells VSMC,providing a new perspective and experimental basis for clinical application of urantide to treat AS.

Abstract:Objective
To investigate the best time of protective effect of puerarin on alcoholism rat hearts by  comparing the intervention  effect of puerarin on heart in rats with alcoholism at different administration time.Methods The male SD rats were randomly divided into control group,alcoholism group,and puerarin groups（treatment group，simultaneous administration group and prevention group).The contents of aspartate aminotransferase (AST),creatine phosphokinase (CPK),superoxide dismutase(SOD),malondialdehyde (MDA),and the activities of Ca²⁺-Mg²⁺-ATPase and  Na^+-K⁺-ATPase were detected by enzymic method and colorimetric method.Results Compared with alcoholism group,the content of SOD,and the activities of Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase in puerarin groups were increased,the contents of MDA,AST and CPK were reduced(P<0.05 or P<0.01).Compared among puerarin groups each other,the results were better for instantaneous administration group than the other two groups（P<0.05 or P<0.01）,besides the MDA in acute alcoholism group had no significant difference(P>0.05).Conclusion The puerarin has protective  effect on heart of rats with acute and chronic alcoholism,especially when it is used simultaneously with alcohol.

Objective To investigate the inhibitory effect of Codonopsis lanceolata total saponin(CLTS) on  human hepatoma HepG-2 cells and clarify its regulatory effect on apoptosis.Methods The HepG-2 cells were divided into 6 treatment groups(100,200,300,400,500， and 600 mg/LCLTS) and control group. The inhibitory rate of HepG-2 cell growth was detected by MTT assay and the  IC50 value was calculated after treated for 72 h. The HepG-2 cells were divided into 3 treatment groups(100,150 and 300 mg/L CLTS) and control group. The apoptotic rate  was determined  by flow cytometry,the activities of cspase 8,9 and 3 in carcinoma cells were determined by microplate reader. Results With the increasing of the concentration of CLTS,the inhibitory rate of HepG-2 cell growth was increased,the IC50 value was 239.623 mg/L,the different concentrations of CLTS inhibited the  HepG-2 cells in  dose- and time-dependent manner(P<0.01). The apoptotic rates in treatment groups were significantly higher than that in control group(P<0.01). The activities  of Caspase-8,9 and 3  were remarkably increased after treated with  CLTS  compared with  control group in a dose-dependent manner(P<0.01).Conclusion CLTS  could significantly inhibit the growth of HepG-2 cells.The induction of apoptosis through up-regulating caspase-8 and 9,then activating caspase-3 is probably one of its molecular mechanisms.

Objective
                   
To investigate the expressions  of gene associated with retinoid-interferon mortality-19 (GRIM-19) and its target gene product signal transducers and activator of transcription 3(STAT3)in human duodenum carcinomas,and study their effects in occurrence and development of duodenum carcinoma.  Methods 32 specimens of duodenum carcinomas,30 specimens of duodenal gastritis and ulcer tissue were included in this study.The expressions of GRIM-19 and STAT3 protein were determined by immunohistochemistry.Results The protein expression of GRIM-19 in duodenum carcinoma tissues were obviously lower than that in duodenal gastritis and ulcer tissues (P<0.05);furthermore,the protein expression of GRIM-19 was correlated to cell differentiation(P<0.01).The protein expression of STAT3 in duodenum carcinoma tissues were obviously higher than that in duodenal gastritis and ulcer tissues (P<0.01),and the expression of STAT3 was correlated to cell differentiation and lymphatic metastasis(P<0.01).The significantly negative correlation was found between the expression of GRIM-19 and STAT3 in duodenum carcinoma tissues(r=－0.470，P<0.01). Conclusion The low expression or absence of GRIM-19 may play an important role in the tumorigenesis of duodenum carcinoma.The low expression of GRIM-19 and the high expression of STAT3 co-exist in duodenum carcinoma tissues. 

Objective To explore the inhibitory effect of manumycin on proliferation of human myeloid leukemia cell line K562 and provide the theoretical basis for manumycin in treatment of leukemia.Methods Control group (K562 plus 10% fetal bovine serum containing IMDM medium) and manumycin group (the fatal concentrations were 2，4，8，16 μmol/L）were set up in this experiment and were treated for 24，48，72 h separately.Then the K562 cell growth was observed under microscope and the inhibitory rate was measured by MTT.Compared with control group,after 24 h,the survivin protein expressions in K562 cells treated with 4 and 16 μmol/L manumycin were examined by Western blotting method. Results The observation under microscope showed that the K562 cells in control group showed good growth state,well-distributed dense,fixed-size,clear outline and strong refraction.Compared with control group,the cells in 8 μmol/L manumycin group showed smaller size,thin dense,unfixed-size,unclear outline and weakened refraction;the cells in 16 μmol/L manumycin group showed less cell dense,anisocytosis,unclear cell outline and cell fragments.The MTT results showed that compared with control group,the inhibitory rate of K562 cells was raised with the drug concentration increasing and exposed time prolongation （P<0.05 or P<0.01），exhibiting a time-dose-effect relationship.The Western blotting results showed that compared with control group,after 24 h, the survivin protein expression in 16 μmol/L manumycin group was decreased.Conclusion Manumycin can stimulate apoptosis and inhibit tumor cell growth by down-regulating survivin protein expression and cure leukemia.

Objective  To study the effects of formaldehyde on memory and the content of neurotransmitter in the cerebral tissue of mice,and explore the effects of formaldehyde on central nervous system and its possible mechanism. Methods  48 Kunming mice were randomly divided into 4 groups：negative control,three experiment groups（low dose group:1/24 LC50,21.0 mg/m³;middle dose group: 1/12 LC50, 42.0 mg/m³;and high dose group:1/6 LC50,84.0 mg/m³）.The mice in experiment groups were exposed to formaldehyd by inspiration in a static total enclosure chamber for 12 weeks.At the first day of the 6th week,8th week and 12th week,the memory of mice was determined,and the neurotransmitters in the cerebral tissue were determined at the end of exposure to toxicants.Results   In  water maze test,at the 8th week,the   swimming time of achieving the goal of the mice in high dose group  was longer than that in control group（P<0.05）.At the  12th week,the swimming time of achieving the goal of the mice in middle dose group and high dose group was increased compared with control group（P<0.01）.And the swimming time of achieving the goal of the mice in high dose gr
oup increased compared with low dose group.（P<0.01）.In high dose  group,the swimming time of achieving the goal at the 12th week was increased compared with the 8th week（P<0.01）.The contents of Ach in cerebral tissue in experiment groups were lower than that in control group(P<0.01）,and the contents of Glu in cerebral tissue were higher than that in control group(P<0.01）.The content of NO in cerebral tissue of mice in each group had no significant difference(P>0.05).Conclusion Long-term exposure to formaldehyde can cause the decreasing of memory,decreasing of the content of Ach and increasing of the content of Glu in the cerebral tissue.It indicates that the change of  neurotransmitter contents  may be one of mechanisms of decreasing of memory in mice.

Objective To study the subtypes of  50 cases of  tonsillar malignant  lymphoma and  the expressions  of Ki-67,bcl-6 and CD10 and explore  their bilolgical significance.Methods By  means of  histomorphology and  immunohistochemistry,50 cases of  primary tonsillar biopsy were reviewed  according to the  new classification of WHO(2008). Results There were 37  cases of   follicular  lymphoma(FL), 9 cases of diffuse large B-cell lymphoma(DLBCL)，2 cases of Burkitt’s lymphoma(BL) and 2 cases of peripheral T-cell lymphoma(PTCL）,16 cases of  FL grade 1 and 2.  The expression of  Ki-67 was in 25%（4/16）  of  the  indolent  lymphoma,and  58.06%（18/31） of the aggressive lymphoma.The Ki-67  expression showed significant difference between different grades of malignancy (P<0.05).The variable expression of bcl-6 was   in  88%(44/50) of  NHL of tonsil.Coexpression of bcl-2+ and bcl-6+  in 40%（8/20）of the low grade FL，61.89%（13/21） of  the high grade FL and 77.78%（7/9）of DLBCL.The CD10 expression showed statistical significance between genders(P<0.05).Conclusion All the cases of tonsillar malignant lymphoma are non-Hodgkins lymphoma.The most histological subtypes is FL,in turn DLBCL,BL and PTCL.There is a rising tendency of Ki-67  expression from low grade FL to aggressive DLBCL.The expressions of bcl-6 vary  in  different  subtypes  such  as  FL，DLBCL，BL  and  PTCL.There  is  an increase trend of bcl-2+ and bcl-6+ coexprssions in low grade FL,high grade FL and DLBCL in sequence.

Objective To study the expressions of Epstein-Barr virus(EBV) and Bcl-2 protein in Hodgkin and Reed-Sternberg(HRS) cells of Hodgkin lymphoma(HL)  in young adults and clarify the relationship between them. Methods The clinical and pathological data of 48 patients with HL between 16 and 36 years old were collected and reviewed.The immunohistochemical method was employed to detect the expressions of CD30,CD15,CD45,LMP-1 and Bcl-2 in HRS cells of young patients with HL.In situ hybridization at tissue chip was used to detect the expression of EBV encoded small RNAs (EBER1 and EBER2).Statistical analysis was done to investigate the association between EBV and Bcl-2 expression.Results According to WHO 2008 classification criteria,no nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) was found in this group,all the cases were classical Hodgkin lymphoma (CHL),among which mixed cellularity classical Hodgkin lymphoma (MCCHL) accounted for 42.67%(20/48),nodular sclerosis classical Hodgkin lymphoma (NSCHL)  27.08%(13/48),lymphocyte-rich classical Hodgkin lymphoma (LRCHL)  25.00% (12/48),lymphocyte-depleted classical Hodgkin lymphoma (LDCHL)  5.25%(3/48).The positive rate of EBV  was 25.00% in all the 48 cases,and 66.67% (2/3),30.00%（6/20），25.00% (3/12),7.69% (1/13) in LDCHL,MCCHL,LRCHL and NSCHL,respectively;but not associated with gender,age and histological subtype of patients（P>0.05）. The Bcl-2 protein was expressed in 64.58% of the 48 cases,its positive rates were 84.62%（11/13），65.00%（13/20），50.00%（6/12），and 33.33%（1/3）in NSCHL,MCCHL,LRCHL,and LDCHL,respectively;and not relevant to gender,age,EBV  expression and histological subtype of patients（P>0.05）.Conclusion MCCHL is predominant in the four CHL subtypes.The positive rates of EBV  and Bcl-2 protein expression are  not related to gender,age and histological subtype of patients,and no relationship is found between EBV and Bcl-2 protein expression.

Objective To study the correlation between the expression of Galectin-3 and Bcl-2,Mmp-2 and K-ras in laryngeal squamous carcinoma(LSCC) and the relationship between Galetin-3 expression and  clinicopathological characteristics.Methods The expressions of Galectin-3,Bcl-2,Mmp-2 and K-ras  in LSCC tissues of 32 patients,normal mucosa tissues of 32 cases,and polyp of vocal cord tissues of 10 cases were detected by immunohistochemistry staining.The relationship between  their expressions and the pathological characteristics of LSCC,clinical staging and lymph node metastasis in LSCC was analyzed.The correlation between the expression of Galectin-3 in LSCC tissue and Bcl-2,Mmp-2 and K-ras expressions in LSCC tissue was analyzed.Results The differences in the  expressions of Galectin-3,Bcl-2,Mmp-2 and K-ras between  normal mucosa tissues,polyp of vocal cord tissues and LSCC tissues were significant  （P<0.05）.The expressions of Galectin-3 and Bcl-2 were related to pathological grade of LSCC（P<0.05）,the expression of Bcl-2 was related to clinical stages and lymph node metastasis of LSCC（P<0.05）.The expression of Galectin-3 had positive correlation with  Bcl-2,Mmp-2, and K-ras expressions in LSCC tissue(r₁＝0.993，r_２＝0.989，r_３＝0.992).Conclusion The high expressions of Galectin-3,Mmp-2 and K-ras are closely related to the occurrence and development of the LSCC.The expression of Galectin-3 is positively related to the expressions of Bcl-2,Mmp-2 and K-ras.Co-expression of Galectin-3,Bcl-2,Mmp-2 and K-ras are able to promote the pathogenesis of LSCC.Galectin-3 is possible to become a new target gene of LSCC gene theray.

Objective  To investigate the influence of terbutaline in combination with inhalation of budesonide  on the β2-adrenergic receptor， T lymphocyte subgroup and eosinophile granulocyte (EOS) and the improvement effect on lung function in the acute attack asthmatic patients.Methods 56 patients with moderate and severe acute attack asthma were selected and randomly divided into control group and  treatment group（n＝28）.The patients in control group were treated with  common method such as doxofylline and the patients in treatment group were treated with an additional treatment by applying combination inhalation: budesonide + terbutaline,two times a day;the β2ADR,T lymphocyte  subgroup,EOS, and lung function were detected before and after  treatment.Results There was no change  in the level of  β2AR in peripheral blood of patients after treatment in  two groups(P>0.05);CD4%,CD4/CD8 and EOS counts　of the patients  were obviously reduced(P<0.05),PEF and FEV1/FVC were obviously increased(P<0.05) after treatment  in two groups,the effect was  obvious in  treatment group compared with control group(P<0.05).Conclusion The combination inhalation has the effect of improving asthmatic patient in acute attack period and achieving clinical control in the long term.

Objective To study the association of four SNPs and haplotypes of Tim-1 gene with rheumatoid arthritis (RA) in Ningxia Hui population and provide theoretical basis early prevention of RA.Methods Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP),specific sequence primer-polymerase chain reaction (SSP-PCR) genotyping method were carried out to examine the －1637A>G,－1454 G>A，－416 G>C and －232 A>G polymorphic sites of Tim-1 gene,including 104 healthy individuals and 108 RA patients in Ningxia Hui population.Results The genotypic  frequencies at －1637,－232 sites and allelic frequency at －1637 site of Tim-1 gene had extremly significant difference (P<0.01) between RA patients and non-RA controls.The frequency of allele A in RA patients was significantly higher than that in non-RA controls (P<0.01) at －1637 site.The frequencies of the two haplotypes AGCA (OR 9.611,95%CI 3.13－29.52),and AGCG (OR 4.361,95%CI 2.12－8.96) in RA patients were higher than those in non-RA controls (P<0.01).The frequencies of the three haplotypes GGCA (OR 0.374，95%CI 0.22－0.64),GGCG (OR 0.199，95%CI 0.08－0.49) and GAGA (OR 0.023，95%CI 0.002－0.260) in RA patients were lower than those in non-RA controls (P<0.01).Conclusion －1637,－1454,－416,and －232 sites of Tim-1 gene have variations.－1637A>G polymorphisms of Tim-1 gene may be strongly associated with RA susceptibility in Ningxia Hui population.The haplotypes AGCA and AGCG  are  associated with RA.The haplotypes GGCA,GGCG,and GAGA have protective effects on  RA.

Objective To study the influence of activities of daily living (ADL)system intervention on the movement function and quality of life of stroke patients,and clarify its improvement on the quality of life of the patients.Methods The patients with stroke who fit the criteria were selected.According to unbalance index minimum principle,100 patients were randomly divided into control group (n=50)and intervention group (n=50).The patients in two groups received conventional therapy and nursing in Department of Neurology.Based on this,the patients in intervention group were given ADL system intervention.After 6 weeks intervention, the quality of life and their movement function in two groups were compared.Results The FMA of  patients in two  groups when they were selected had no  significant difference (P>0.05).After 6 weeks system intervention, the FMA in two  groups were increased, there was significant difference between intervention group and  control group  (P<0.05); the quality of life of each dimension and the overall quality of life of patients in two groups before intervention had  no statistically significant differences (P>0.05).After  6 weeks system intervention,except memory, thinking and communication,the physical dimensions,emotion,ability,mobile ability,hand function,participating in dimension and the overall quality of life of patients in intervention group were  better than those in control group (P<0.01). Conclusion ADL system intervention is helpful to the movement function   and  quality of life of  stroke patients.

Objective
To measure the acetabular morphology of Chinese on CT three-dimensional (3D) reconstruction image,and provide the evidence on the prevention and treatment of hip diseases.Methods 96 cases (192 sides) of adult hip CT scans were reconstructed,the acetabular index (AA),center-edge(CE),ACE angle,anteversion angle(AVA)，abduction angle(ABA),and vertical diameter(SID) were measured.Results The 　total acetabular index was (8.78 ± 5.34) °,of which male was (7.84 ± 5.55) °and female was (9.60 ± 5.06)°. The total CE was (33.59 ± 5.91) °,of which male was(34.55 ± 6.03) ° and female was(32.78 ± 5.70) °.The total ACE angle was (29.01 ± 5.65) °,of which male was (28.02 ± 5.94) °and female was (29.80 ± 5.30)°.The total AVA was (20.92 ± 5.55)°,of which male was (20.48 ± 5.08)°and female was (21.25 ± 5.89)°.The total ABA  was (51.27 ± 4.16) °,of which  male was (51.71 ± 4.37)°and female was (50.89 ± 3.96)°.The total SID was(53.79 ± 3.92) mm,of which male was (56.55 ± 2.64) mm and female was 〖JP2〗(51.46 ± 3.25) mm.Of the above data,there were statistical differences in the acetabular index,CE angle,ACE angle 〖JP〗and acetabular diameter between men and women(P<0.05),Chinese and foreigners(P<0.05).While there was no statistical difference between the left and right sides(P>0.05). Conclusion There are differences in acetabular morphology between men and women,Chinese and foreigners.Compared with simply using overseas data,it is better to study morphological parameters of native acetabulaum to instruct the preoperative preparation and operation of national total hip arthroplasty surgery.And it is meaningful to design national parameters.

Objective
To reveal the geographical distributing rule and supply a scientific basis for unifying the reference value standard of left atrial diameter (LAD) of Chinese presenile women in various region of China. Methods The LAD reference values were collected from 4 943 healthy presenile women in 154 areas of China. The correlation between the measured values of LAD and the geographical factors was analyzed with correlation analysis method.Applying the method of regression analysis and principal components analysis,one best regression equation was inferred by using T-test.Results The reference value of LAD was correlated with the geographical environment.The linear regression equation deduced from principal components analysis was:Y=30.53－0.001 104X₁　+0.001 060X₂－0.129 5X₃－0.043 92X₄－0.000 912 3X₅+0.103 5X₆+0.899 3X₇;　in the above equation,Ywas normal reference value of Chinese presenile women left atrial diameter (mm),X₁ was altitude,X₂ was annual sunshine duration（h）,X₃ was annual average temperature （℃）,X₄ was annual mean relative humidity（%）,X₅ was annual precipitation amount（mm）,X₆ was annual range of air temperature（℃）,X₇ was annual average wind speed (m·s^-1),the geography distribution figures of Chinese presenile women LAD reference values were exactly inserted with methods of Kriging.Conclusion If the geographical factors of a particular area are obtained,the LAD reference value of Chinese presenile women in this area can be calculated using this model and the normal LAD reference values of Chinese presenile women anywhere in China can be obtained from geography distribution figure.

Objective To explore the conditions of preparation technology of Sanku dropping pills and provide the basis for further development of traditional Chinese medicine in treatment of  coronary heart disease and angina pectoris.  Methods Time limit of dissolution,weight variation and appearance(length pellets,hardness,color uniformity) were used as evaluation indexes of the test,the single factor investigation method was adopt to select the variety of the matrix and the dropping distance;on the basis of orthogonal test,the good dropping conditions about the proportion of two types of matrix，matrix with the medicine and the temperature of liquid medicine were selected.Results The best pill formatting process was that the dimethyl silicon oil was used as refrigerant,the ratio of PEG 4000 to PEG6000 was 1∶6,the ratio of medicine to matrix was 1∶2,the dripping temperature was 80℃,the dropping distance was 5 cm,the dropping speed was 18－20 drops·min^-1.Conclusion The optimized moulding process results is stable and feasible,suitable for mass production 

Objective
To establish rat model fit to study cerebral vasospasm(CVS) induced by subarachnoid hemorrhage(SAH).Methods Wistar rats  were divided into sham operation group  (injected with of 0.3 mL saline into the magnum pool), SAH model group (injected with 0.3 mL no anticoagulation   autologous blood into the cisterna magna),SAH  nimodipine treatment group (injected introperitoneally with 0.2 mg·kg-¹ nimodipine 30 min after modeling and every day after operation).The neurological deficits of rats and incidence of decreased food intake were observed and  the largest rat basilar artery blood flow velocity (Vmba) was detected with transcranial Doppler(TCD) at the 1st,7th,14th and 21th after operation.Results Compared with sham operation group,the basilar artery blood flow,nerve dysfunction and decreased food intake (including 2-4) in SAH model group were consistent with the occurrence of schedule,mainly  increased at  the 1st day,peaked on day 7,decreased on  day 14,normal on  day 21,and the mortality rate was 0. Compared with SAH model  group,  the basilar artery blood flow velocity in treatment group was decreased, the incidence of nerve dysfunction and decreased food intake were decreased  also .Conclusion Through the second injection of blood into cisterna magna, a rat model closing to clinic and fiting for research on CVS induced by SAH is established,and nimodipine can significantly reduce the degree of CVS after SAH.