Abstract:Objective
To study the effects of TRAIL and endostatin double genes in recombinant plasmid pshuttle-Egr1-shTRAIL-shES combined with X-ray irradiation on the proliferation,cell cycle procession and apoptosis in human vascular endothelial ECV304 cells.Methods Cells were divided into control,empty vector pshuttle,TRAIL single-gene plasmid pshuttle-Egr1-shTRAIL,endostatin single-gene plasmid pshuttle-Egr1-shES and TRAIL plus endostatin double-gene plasmid pshuttle-Egr1-shTRAIL-shES transfection groups.The cell transfection was done by lipofectamine-mediated method,and no transfection in control group.After plasmid transfection,the cells were exposed under X-ray irradiation at the doses of 0,0.1,0.5,1.0,2.0 and 5.0 Gy,  respectively.Then the time-course and dose-effect patterns of the TRAIL and endostatin protein expressions induced by irradiation were detected by ELISA,and the effects of TRAIL and/or endostatin single or double gene therapy combined with radiotherapy on the proliferation,cell cycle procession and apoptosis in ECV304 cells were detected by MTT assay,flow cytometry (FCM) with PI single-staining or/and Annexin Ⅴ double-staining.Results After 2.0 Gy X-ray irradiation,the expression levels of TRAIL and endostatin proteins in the supernatant of cultured ECV304 cells transfected with pshuttle-Egr1-shTRAIL-shES at other time-points were increased when compared with those at 0 h (P<0.01),and reached to the peak value at 24 and 12 h, respectively.Furthermore,their expression levels of TRAIL and endostatin protein induced by irradiation were increased significantly with the enlargement of radiation doses (P<0.05 or P<0.01).Under the exposure of X-ray irradiation,as compared with those in  control and pshuttle groups,the A490 values detected with MTT in pshuttle-Egr1-shTRAIL,pshuttle-Egr1-shES and pshuttle-Egr1-shTRAIL-shES groups were reduced in a time- and dose-dependent manner,and the apoptotic rates and the cell percentages in G^₂ + M phase were increased,while those in G₀/G₁ phase were declined.In particular,the cellular effects mentioned above in pshuttle-Egr1-shTRAIL-shES group were significantly higher than those in pshuttle-Egr1-shTRAIL and pshuttle-Egr1-shES groups(P<0.05 or P<0.01).Conclusion TRAIL and endostatin double-gene-therapy in combination with radiotherapy could inhibit cell proliferation,affect cell cycle procession and promote apoptosis in ECV304 cells,and its therapeutic efficacy is better than those of single gene-radiotherapy or radiotherapy.

Abstract:Objective
To investigate the influence of apocynum venetum extract (AVE) on angiotensin Ang Ⅱ-induced myocardial fibrosis in rats and to clarify its mechanism of anti-myocardial fibrosis.Methods The rat cardiac fibroblasts were pretreated with Ang Ⅱ,and then treated with different concentrations of AVE (0.8,0.4 and 0.2 mol•L^-1) or valsartan 10-6 mg•L^-1.The expressions of hydroxyproline,collagen Ⅰ,Ⅲ （ColⅠ,ColⅢ） were detected to identify the model of myocardial fibrosis.The expressions of  raf,ERK and c-fos mRNA were determined by RT-PCR.The expression of ERK protein was measured by Western blotting.Results The models of myocardial fibrosis were established successfully.Compared with AngⅡ group,the over expressions of  hydroxyproline,ColⅠ and ColⅢ in AVE groups were significantly decreased（P<0.05 or P<0.01）.Compared with control group,the expressions of raf,ERK and c-fos were increased obviously in Ang Ⅱ group;compared with Ang Ⅱ group,the expressions of raf,ERK and c-fos mRNA and ERK protein in AVE groups were decreased（P<0.05 or P<0.01）.Conclusion ERK pathway plays an important role in the occurrence and development of mycardiol fibrosis regulated by AVE.

Abstract:Objective
To investigate the effects of mesna or/ and in combination with ciprofloxacin on biofilm of Pseudomonas aeruginosa.Methods The broth microdilution method was performed to determine the minimal inhibitory concentration(MIC) of mesna and ciprofloxacin to PAO1,then a biofilm model of Pseudomonas aeruginosa in vitro was established,the biofilms were randomly divided into 3 groups: control group,low-dose mesna group and high-dose mesna group,the parameters　of biofilm structure were analyzed through pictures from CLSM with Image Structure Analyzer(ISA)software after the biofilms were treated with mesna;then the biofilms of Pseudomonas aeruginosa were randomly divided into: mesna group,CIP group,mesna+CIP group,they were treated with mesna,ciprofloxacin (alone and in combination),three dimensional images of the biofilms were obtained with Amira 5.2 software. Through a aboard design,different consistence  mesna (0,0.5,1.0，2.0 and 4.0 g•L^-1)were respectively combined with different consistence ciprofloxacin （0，1/2，1，2，4，8 MIC）,the number of bacteria in biofilms was measured by agar plate.Results The MIC values against PAO1 were 10 g•L^-1 for mesna and 0.12 mg•L^-1 for ciprofloxacin.The results of ISA showed that with the treatment of low-dose mesna,the biofilms  were decreased in thickness,average difusion distance(ADD)and textual entropy(TE) as compared with control group (P<0.01),however the areal porosity(AP) was increased(P<0.01),the same tendency of these parameters was observed in the high-dose group and the effects were more obvious(P<0.01); The images from CLSM showed that the  biofilms in mesna (2 g•L^-1) group were intensive and thicker,the masses of alive bacteria were found in the biofilms but no dead bacteria;ciprofloxacin(2 MIC) decreased the alive bacteria,the dead bacteria were increased relatively slightly;the biofilms were thinner and more scattered,plenty of bacteria were killed in the presence of ciprofloxacin (2 MIC) and mesna (2 g•L^-1).The single use of mesna at the concentration of 2 g•L^-1 could significantly decline the viable cells in biofilms,the individual use of ciprofloxacin at 2 MIC could significantly desease the total bacterial quantity;however,when 1 g•L^-1 mesna was combined with 1 MIC ciprofloxacin,the viable counts in biofilms were less than they were given alone.Conclusion Mesna can destroy the construction of biofilm of Pseudomonas aeruginosa and has the synergistic effect on bactericidal activity with ciprofloxacin.

Abstract:Objective
 To investigate the changes in the number of endothelial progenitor cells (EPCs) in whole blood,heart,brain and kidney tissues in rats at different growth stages,and to illustrate the relationship between the changes of the number of EPCs in rats and the diffrent growth stages.Methods The rats were divided into 0.5 months,2 months,8 months and 18 months groups,10 rats in each group.The blood was collected by abdominal aorta with EDTA-K2 anti-freezing when the rats were anesthetized by 2% pentobarbital sodium and centrifuged for collection of whole blood.Meanwhile,  the heart,brain and kidney tissues of the rats in each group were prepared as well as single cell suspension,and the EPCs were labeled with CD34 (FITC labeled) and CD133 (markers PE) double staining and  the changes in EPCs numbers in blood and suspension of single cell were detected by flow cytometry;the same places of heart,brain and kidney tissue microarray,and the tissue specific staining positive expressions of CD34 in EPCs were detected by immunohistochemistry.Results The number of EPCs detected by flow cytometry and immunohistochemical staining in blood and heart,brain,kidney tissues of the rats in 2 months and 8 months groups were significantly higher than those in  0.5 month and 18 months groups (P<0.05),but there were no significant differences between 2 months and 8 months groups(P>0.05).As well as 0.5 month and 18 months groups. Conclusion With the growth and development of rats,the number of EPCs is changed regularly in whole blood,heart,brain,kidney and other organs,and it may be closely related to the growth and development stages of rats.

Abstract:Objective To explore the feasible of the expression of recombinant truncated human keratinocyte growth factor 1 (rhKGF1_d23)  in  insect cells and to provide a new method to product rhKGF1_d23.Methods The truncated gene of kgf1_d23 optimized  by insect prefer codon was amplified and subcloned into pFastBac vector.With the help of helper plasmid,the Tn7 section of pFastBac-kgf1_d23 was transposed to  the mini-attTn7 site of Bacmid DNA to obtain  the recombinant Bacmid-kgf1_d23.Bacmid-kgf1d23 shuttle vector was used to transfect sf-9 cells by lipofection kit to produce recombinant baculovirus P1.The  baculoviral stock was amplified and the virus titer was increased until P3 stock.The cells were broken by sonic and the lysate liquid and purified by heparin affinity chromatography and cation exchange; MTT method was used to detect the  rhKGF1_d23mitogen activity.Results  kgf1d23 gene was optimized by insect bias codon and the recombinant  pFastBac-kgfd23 and Bacmid-kgf1d23 had been successfully constructed.The SDS-PAGE results showed that  the rhKGF1D23 expressed mainly in cells and until 60 h post-infection,and the optimal harvest time was  in 84-96 h.Heparin affinity and SP cation exchange could remove 90% noisy proteins.The MTT results demonstrated that the purified rhKGF1_d23protein with 0.3-25.0 μg•L^-1  could significantly stimulate HaCat cells to proliferate in a dose-dependent manner.Conclusion The rhKGF1_d23expression is obtained in sf-9 cells with heparin binding characteristics,as well as immunogenic and cell biology activity.

Abstract:Objective
To explore the influence of multi-chamber piezoelectric pump on  the blood cells and coagulation system when it applied as blood driving pump. Methods Anticoagulation dog blood was utilized as circulating medium in the simulative circulatory system. There were three groups depended on the circulation time (0,1 and  2 h) and a control group. Hemoglobin concentration,white blood cell,red blood cell and platelet counts,hematocrit,free hemoglobin concentration were detected. The index of hemolysis (IH) of multi-chamber piezoelectric pump  was calculated as an important index of hemolysis damage. Results  Compared with control group,each index was not significantly changed in cycle 0 h group(P>0.05). Compared with control group,the  red blood cell count,white blood cell count,platelet count,hemoglobin concentration,hematocrit were decreased in cycle 1 h and 2 h groups（P<0.05).  Compared with control group,the  free hemoglobin concentration was increased,the fibrinogen concentration was decreased（P<0.05）.  Compared with cycle 1 h group,each index was not significantly changed in cycle 2 h group（P＞0.05）.The  IH of pump was calculated as 0.004 5,and it was significantly lower than international standard(＜0.1). Conclusion   Multi-chamber piezoelectric pump has good blood compatibility in vitro. It has advantages to apply as blood-driving pump.

Abstract:Objective
To observe and compare the photochemotherapeutic effects of BCPD-17 (hematoporphyrin derivative) and PSD-007 on osteosarcoma LM-8 transplanted in mice and to explore a novel photosenitizer for photodynamic therapy of  osteosarcoma Methods The tumor model was made by subcutaneous injection of osteosarcoma LM-8 on C3H mice.When the tumors reached about 6-8 mm in diameter,the tested mice were randomly divided into control group,PSD-007 group (5 mg•kg^-1),BCPD-17 groups (5 and 10 mg•kg^-1).After received intravenous injection of 5 or 10 mg•kg^-1 of PSD-007 and BCPD-17,respectively,the tumors were irradiated vertically with 630 nm or 670 nm laser (power density 200 mW•cm-2,energy density 240 J•cm^-2) for 20 min.The size of osteosarcoma was measured 7 d  after treatment.And all the tumors were extirpated,weighted and taken for pathological examination.The inhibitory rate of tumor was calculated.Results The results of pathohistological observation showed that in control group the size of mouse osteosarcoma was big and hyperchromatic with obvious nuclear atypia;in PDT treatment group,the turmor cells presented apoptosis,vacuolar degeneration and karyopycnosis,especially in BCPD-17 group. Compared with control group,the volumes and the weights of the tumors in PDT treatment groups were obviously decreased,there were significant differences(P<0.01).Compared with PSD-007 group,the inhibitory rates of tumor in BCPD-17 groups irradiated by the laser with wavelength 670 nm were increased,there were significant differences(P<0.01).Conclusion Both of the photosenitizers BCPD-17 and PSD-007 could obviously inhibit the growth of the tumors.Photosenitizer BCPD-17 has more anti-tumor effect.

Abstract:Objective
 To analyze the inhibitory effect of RNA interference (RNAi) on  the expression of IDH2 gene of  colon cancer cells HC７-8 and the influnce in the proliferation of colon cancer cells in vitro,and to provide a theoretical basis for colon cancer targeted therapy.Methods  The experiment was divided into  siRNA plasmid transfection,negative plasmid control and HCT-8 cells control groups.The recombinant plasmid which contained the siRNA of the IDH2 gene in the green fluorescence eukaryotic vector (PGPU6/GFP/Neo-IDH2) and the negative control were constructed and then they were transfected into colon cancer cells by using FuGENE HD Transfection Reagent. The expression of IDH2 gene mRNA was detected by real-time PCR. The proliferation abilities of colon cancer cells were detected by MTT.Results  Counted by fluorescence microscope,the transfection rate of cells was 74%. The inhibitory rate of IDH2 mRNA in  PGPU6/GFP/Neo-IDH2 group was 85.02%,which had significant difference compared with HCT-8 cells control group (P<0.05). Compared with neagtive control group,the proliferation ability of colon cancer cells was deceased by MTT assay (P<0.05).Conclusion Inhibition of the expression of IDH2 gene can infect the proliferation ability of colon cancer cells. It shows that IDH2 gene plays a role in the development of colon cancer and it may be beneficial in finding a new therapy for colon cancer.

Abstract:Objective
 To study the protective effect of panax quinquefolium 20s-protopanaxtriolsaponins (PQTS) on acute myocardial infarction in rats,and to clarify its mechanism. Methods The acute myocardial infarct model was prepared by left anterior descending  coronary occulusion for 24 h in openchest anesthetized rats. The rats were divided randomly into sham group,ischemia-reperfusion model group and PQTS 12.5,25.0， and 50.0 mg•kg-1 groups. All animals were administered one time everyday with intraperitoneal injection(i.p) for 3 d. The myocardial infarct size (MIS) was calculated. The activities of serum creatine hosphokinase(CK),lactate dehydrogenase (LDH),aspartate aminotransferase (AST) and superoxide dismutase(SOD),and the contents of malondialdehyde(MDA),nitric oxide(NO) and free fatty acid(FFA) were determined. Blood was collected to observe low shearing specific viscosity,middle shearing specific viscosity and high shearing specific viscosity of whole blood and plasma viscosity. At the same time,the platelet aggregation rate was also determined. Results Compared with model group,the MIS  in the  rats treated by PQTS (in  dosages of 12.5,25.0 and 50 mg•kg-1•d-1,i.p×3 d) were significantly reduced(P<0.05);the activities of serum CK,LDH and AST,and the contents of serum FFA and MDA were declined(P<0.05 or P<0.01);the activitie of SOD,and the content of NO were increased markedly(P<0.05). In addition,the low shearing specific viscosity,middle shearing specific viscosity and high shearing specific viscosity of whole blood and plasma viscosity as well as platelet aggregation rates were also declined significantly(P<0.05 or P<0.01). Conclusion PQTS has a significant protective effect on acute myocardial infarction,which may be related to increasing the activity of antioxidase in body,scavenging the damage of peroxidation from oxygen free radicals,correcting metabolic disorder of FFA in myocardial ischemia,and decreasing the viscosity of blood and plasma and function of the platelet aggregation etc.

Abstract:Objective
 To explore time window of inducing neuronal apoptosis successfully via building and evaluating oxygen glucose deprivation/reperfusion (OGD/R) model in vitro to mimic ischemia and reperfusion inducing apoptosis of SH-SY5Y cells,and to build a basis for revealing the potential mechanisms of neuronal apoptosis. Methods  SH-SY5Y cells were employed and divided into control group and experimental group treated with   OGD/R  for different time. The cell viability was calculated by the method of MTT method.The OGD10h/R groups in which the cell viability was decreased approximately 50% 24 h after reperfusion and  control groups were selected for further assay. With light microscope,fluorescence microscope and transmission electron microscope the variation of structure and morphous in cells at different time were observed. The changes in  lactate dehydrogenase (LDH) levels, malondialdehyde (MDA) contents,superoxide dismutase (SOD)  activities  were detected at different time of reperfusion. Flow cytometry was used to analyze the cell cycle distribution and the apoptotic rates at different time of reperfusion. Results  The MTT results showed that the cell viabilities were decreased significantly with the prolongation of OGD time. At reperfusion 24 h,the cell viability was (44.91±1.21)% in OGD10h/R groups. The cell viabilities in other different OGD groups had significant differences (P<0.05 or P<0.01). In OGD10h/R groups the inverted microscope and HE staining results showed that the cell densities were decreased following varied shape and deep blue pycnotic nuclei.The AO/EB fluorescence staining results showed the apoptotic body emerged.Under electron microscope,the cells showed heterochromatin fringe agglutinated in nucleus which were viewed as the appearance of apoptosis and autophagia. The level of LDH in culture fluid and the contents of intracytoplasm MDA at different reperfusion time were increased with the prolonged time and at reperfusion 24 h respectively reached the peak and platform phase,compared with control groups the differences were significant (P<0.01). The intracytoplasm SOD activities at different reperfusion time were declined at first and recovered later with the prolonged reperfusion time,compared with control groups the differences were significant (P<0.01). By  flow cytometry analysis on cell cycle phase at different reperfusion time,in OGD 10h/R groups with the prolonged reperfusion time,the percentage of the cells in G0/G1 phase was increased at first and was decreased later with its peak of (74.09±2.62)% at reperfusion 24 h. The percentage of the cells in G2/M phase reached its peak of (26.85±1.35)% at R 0 h and was declined gradually later. The percentage of the cells in S phase was declined gradually and reached the lowest level of （19.2±1.58）% at reperfusion 24 h. Compared with control groups at the same time,the differences were significant (P<0.05 or P<0.01). Conclusion OGD10h and reperfusion (OGD10h/R) to mimic cerebral ischemia and reperfusion in vitro is a reliable model that induces SH-SY5Y cells apoptosis,which provide a basis for the futher study on neuronal apoptotic mechanisms.

Abstract:Objective To investigate the interference effects of puerarin on multitude brain regions,which contains cortex,cerebellum,hippocampus and striatum of acute alcoholism rats,by detecting the contents of  superoxide dismutase(SOD),malondialdehyde (MDA) and glutamate (Glu) in rat brain and  the praxiological changes after administration with ethanol and puerarin.Methods Male SD rats were randomly divided into control group,alcoholism groups (high dose group,middle dose group and low dose group),and puerarin group（the rats were treated with middle dose alcoholism）.The contents of SOD,MDA,Glu  and the rodduration were detected.Results Compared with control group,the contents of SOD and Glu in alcoholism groups were reduced,while the MDA was increased.The contents of SOD and MDA changed in  dose-dependent manner.The rodduration in alcoholism groups was significantly shorter than that in control group (P<0.05 or P<0.01).Compared with alcoholism groups,the rodduration in puerarin group became longer,the contents of SOD and Glu were increased,the content of MDA was reduced(P<0.05 or  P<0.01),and the most significant changes were found at hippocampus among the four brain regions.Conclusion  Puerarin has interference effects on oxygen free radical and Glu in brain  of acute alcoholism rats,and it exhibites a regulatory effect on praxiology of acute alcoholism rats.

Abstract:Objective To discuss the relationships between  the serum creatine kinase (CK),lactate dehydrogenase (LDH) and skeletal muscle injury in rats after different load movement and to provide a theoretical basis for study on muscle injury with scientific and accurate judgement.Methods 70 Wistar male rats were divided into non-exercisegroup (control group), a bout of eccentric exercise group(AE) and 7 d  eccentric movement group(SE).Gastrocnemius were taken immediately，24 h and 7 d aftera bout of eccentric exercise or the last exercise, respectively.The changes of tissue structure and serumCK,LDH,malondialdehyde(MAD) and superoxide dismutase(SOD) activities were observed.Results The tissue structure of skeletal muscle of rats in AE group was damaged seriously  24 h after a bout of eccentric exercise;the tissue structure in SE group was also damaged seriously in the immediate time at the last exercise,showing the muscle fiber distortion, shrinkage,disappearance of cross striations and other changes.The activities of serum CK,LDH and MDA reached the peak in the immediate time after exercise in AE group,there were significant differences compared with control group  (P<0.001).The activity of serum SOD was increased significantly 24 h after exercise in AE groups compared with control group  (P<0.001).The activities of serum CK,LDH,MDA,and SOD in SE group were significantly increased in the immediate time  after the last exercise,and the differences between SE group and control group were significant (P<0.001).Conclusion The activities of serum CK,LDH and MAD of rats appears time course changes,and it is not consistent with the skeletal muscle tissue damage,and the changes of skeletal muscle injury has a certain delay phenomenon.The activity peaks of  serum CK,LDH,MAD,and SOD 7 d after eccentric exercise  are consistent with the skeletal muscle injury.LDH can be the indirect index to evaluate the repair degree of the skeletal muscle injury after eccentric exercise.

Abstract:Objective To investigate the effect of fluoxetine on the expression of 5-HT type 1A (5-HT1A) receptor in hippocampus tissue of posttraumatic stress disorder (PTSD) rats and to explore the mechanism of fluoxetine in the treatment of PTSD.Methods The male Wistar rats were randomly divided into control group,model group and fluoxetine group.The rat PTSD model was set up using SPS & S method.Hippocampus tissue was isolated and the expression of 5-HT1A receptor wasdetected with Western blotting and RT-PCR.Results The 5-HT1A receptor protein expressions in hippocampus tissue of rats in control group,model group,and flouxetine group were 1.18±0.12,0.39±0.05,and 0.71±0.09,respectively. The 5-HT1A mRNA expressions were 0.95±0.12,0.19±0.05,and 0.74±0.05 in three group,respectively. There were significant differences between model group,flouxetine group and control group (P<0.05).Compared with model group,the 5-HT1A mRNA in flouxetine group was increased (P<0.05).The index was inhibited after treatment with furoxetin. Conclusion Fluoxetine could inhibit the 5-HT1A receptor expression in hippocampus of PTSD rats.

Abstract:Objective To study the influence of the regulatory subunit of protein kinase CK2β in the early development of fertilized mouse eggs by the means of specific siRNA and to clarify the mechanism of regulation of early development of fertilized mouse eggs.Methods One-cell stage fertilized mouse eggs were collected after superovulation,and microinjected with TE buffer,scramble siRNA or CK2β siRNA,then the effect of siRNA  was examined by RT-PCR and Western blotting.After RNA interference,the morphological changes and cleavage rate of mouse embryos were observed under dissecting microscope,and the M-phase promoting factor（MPF）activity of fertilized mouse eggs were detected by autoradiography.Results Compared with TE buffer and scramble siRNA groups,the expression levels of CK2β mRNA and protein in CK2β siRNA group were significantly reduced（P<0.01）; the activity of MPF in CK2β siRNA group reached its peak value at 28.5 h after human chorionic gonadotropin（hCG) injection,about 30 min earlier than TE buffer and scramble siRNA groups.The embryos treated with CK2β siRNA entered the M phase at 28.5 h after hCG injection,and the cleavage rate was as high as 89.5% at 32.0h,markedly higher than those in TE buffer and scramble siRNA groups (P<0.01).Conclusion CK2β siRNA may promote the early development of fertilized mouse eggs via activating MPF activity ahead of time.

Abstract:Objective To study the influence of cyclosporin A （CsA）in the expression of macrophage inflammatory protein-1α (MIP-1α) mRNA in kidney tissue of  streptozotocin (STZ) - induced diabetic rats,and to explore the related mechanism.Methods Male SD rats were used to set up diabetes models with 50 mg/kg body weight by tailvein injection of STZ.The diabetic rats were randomly divided into 6 groups:CON (control group),DM(diabetic model),AMI(administrated with insulin), AML(administrated with CsA 1 mg/kg/d),AMM(administrated with CsA 4 mg/kg/d),AMH(administrated with CsA 8 mg/kg/d) groups(n=10).The expression levels were detected by RT-PCR.The MASSON positive staining areas and optical densities between various CsA groups were compared.Results Compared with  CON group,the expression level of  MIP-1α mRNA of rat lympthocyte in DM group was significantly increased(P<0.05 ).Compared with DM group,the MASSON positive staining areas and optical density values in CsA groups were significantly decreased(P<0.05).Conclusion The MIP-1α mRNA expression in kidney tissue of diabetic rats is increased and renal fibrosis is obvious.CsA intervention can decrease the MIP-1 mRNA expression in kidney tissue and reduce  renal fibrosis.

Abstract:Objective To study the effect of salidroside on rat renal ischemia reperfusion injury,and to clarify  its mechanism of action.Methods 54 healthy SD rats were randomly divided into nine groups:control group,ischemia-reperfusion groups by clamping 15,30,45 and 60 min,intraperitoneal injection of salidroside solution groups by clamping 15,30,45 and 60 min.The serum creatinine (Scr) in various groups were detected.The pathological changes of rat kidney tissues were observed by light microscope.The expressions of HSP72 and NF-κB in kidney tissue were determined by Western blotting and immunohistochemistry.Results Compared with ischemia-reperfusion groups,the levels of  Scr in salidroside solution groups(clamping 15,30 and 45 min) were significantly decreased (P<0.01),the  pathological damage was significantly reduced,and the expressions of HSP72 and NF-κB in kidney tissue were also decreased significantly.Compared with  control group,the  Scr levels in salidroside groups were increased slightly,especially in  clamping 60 min group,it was increased significantly(P<0.01),but in  clamping 15,30 and 45 min groups, the changes were not obvious.Conclusion Application of salidroside can significantly reduce renal ischemia-reperfusion injury in kidney tissue,the mechanisms may be related to  enhancing the antioxidant capacity of the kidneys,reducing inflammatory cell infiltration  and apoptosis in kidney tissue and inhibiting  the HSP72 and NF-κB expressions in kidney tissue.

Abstract:Objective To investigate the anti-fatigue effect of   lycium barbarum polysaccharide(LBP)  and 1,6-fructose diphosphates (FDP) and to provide scientific basis for improving training effectiveness and race performance.
Methods 140 mice were randomly divided into saline-negative control group,LBP group (20 mg/kg),FDP group (300 mg/kg) and the 2×2 analysis variance groups.The burden swimming mice were used as animal models.The swimming time,liver glycogen,muscle glycogen,lactate dehydrogenase (LDH),superoxide dismutase (SOD),blood lactate (LAC) and blood urea nitrogen (BUN) were detected.The synergy effect of LBP and FDp was analyzed.Results  Compared with negative control group,the loading swimming time in FDP and LBP groups was prolonged (P<0.01),which indicated that they played synergy effect on swimming time.The best doses of LBP and FDP were 20 mg/kg and 300 mg?kg-1(P<0.01).The activities of LDH and SOD were increased(P<0.01) and the levels of LAC and BUN were decreased in LBP group,and the liver glycogen and muscle glycogen of  mice were increased in FDP group(P<0.01).Conclusion FDP and LBP exhibits a synergy effect on anti-fatigue,its mechanism may be related to that FDP can increase the  liver glycogen and muscle glycogen reserve of mice  which can provide more energy,and LBP can accelerate the decomposition ofmetabolites,and make mice persist more time in endurance exercise.

Abstract:Objective To explore the dynamic expressions of apoptosis-related factors of Bcl-2,Bax and p53 in rats with cerebral ischemia-reperfusion injury,and to provide the experimental evidence for the neuroprotective mechanism in cerebral ischemia-reperfusion injury.Methods 52 adult male Sprague-Dawley rats were randomly divided into sham control group (n=12) and cerebral ischemia-reperfusion 2,3,6,8,12 h groups(n=8).Suture method was used to set up the middle cerebral artery occlusion (MCAO) and reperfusion model.The neuronal apoptosis of the MCAO brain tissues at different time points was detected by in situ terminal deoxynucleotidyl transferase labeling technique (TUNEL); the dynamic expressions of Bcl-2,Bax and p53 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry.Results The TUNEL staining results  showed that the apoptotic cells began to appear 2 h after reperfusion  in the surrounding area in ischemia-reperfusion group,and they were increased significantly at 6-12 h.The mRNA and protein expressions of Bcl-2,Bax and p53 tended to increase with the prolongaction of ischemia and reperfusion time.The cell morphology was relatively complete in the cells with high expression of Bcl-2 protein,and the cells with high expressions of Bax and p53 protein were severely damaged.The mRNA and protein expressions of Bcl-2 appeared earlier than Bax and p53.Conclusion Bcl-2 has a protective effect on nerve cells.Early regulation can promote the expression of Bcl-2 and reduce the expressions of Bax and p53,and it plays an important role in reducing the damage of nerve cells in the process of ischemia and reperfusion.

Abstract:Objective To evaluate the repair effect  of bone marrow mensenchymal stem cells(MSCs) transplantation  on   acute liver injury of rats and to explore the feasibility and validity.Methods The Wistar rats were treated with 10% CCl4 olive oil mixture by intraperitoneal injection to copy acute liver injury model.The model rats were randomly divided into treatment and control groups.Hochest33342-labelled rat bone marrow MSCs 0.5 mL (1×106 cells) were transplanted intothe rats by tail vein,portal vein and liver injection in treament groups,respectively.The same volume PBS was transfused into the rats in various control groups.On day 3 and 7 after transplantation,the liver function indexes of rats  were detected,the distribution of Hochest33342-labelled cells and the pathological changes  of liver tissue were observed by fluorescence microscopy at same time.Results Compared with control group,the levels of serum ALT and AST were decreased on day 3 and day 7 after transplantation in intraportal vein and intraliver injection groups (P<0.01),but there were no statistically significant differences in tail vein group (P>0.05);and they were decreased in intraliver injection group compared with intraportal vein group(P<0.05).The Hochest33342 labelled cells were not found in vein group and control groups at different time,but they  were found in intraportal vein and intraliver injection groups;the days were more,the cells were more.The pathological structure of liver displayed multiple necrosis of hepatocyte,diffuse degeneration of hepatocyte and infiltration of inflammatory cells in rats treated with CCl4 for 24h.The severity of pathological structure of liver wasn’t alleviated in tail vein group at different time after transplantation,but it was alleviated in intraportal vein and intraliver injection groups on day 3 and day 7;the improvement of pathohistology intraliver injection group was better than that in intraportal vein group.Conclusion Intraportal vein,intraliver injection transplantation of bone marrow MSCs can  ameliorate and repair liver function of rats with acute liver injury.The route of intraliver injection transplantation is better than intraportal vein transplantation.

Abstract:Objective Through inspecting in vitro fibrillogenesis of the young and the old rat tail tendon collagen,as well as comparing the morphological changes  of the lumbar cancellous bone collagen,to study the influence of  age  in fibrillogenesis.Methods The tail tendon(5 g)  of  2-week and  20-month rats,was obtained,and the tail collagen  was  extracted by the dilute acetic acid.The  extracted collagen was freezed-dried in low temperature.15 mg tail collagen were take from 2-month-old and 20-month-old rats.Each wss divided into three groups,5 mg in each group,and the collagen was melted in 10 mmol/Lacetic acid solution  at 4℃,the titanium metal screws were infiltrated into the solution.After adding buffer the  fibrillogenesis was taken in 37℃.The screws were picked up separately at 2,4  and 8 h,and the  morphological changes of  collagen of the surface of the screws were observed under  scanning electron microscope(SEM).10 2-week rats and 10 20-month rats were sacrificed to remove the L3 vertebral body,the regular specimen were made.The specimens were repeatedly washed by using physiological saline,dehydrated,and  sprayed gold,and then observed under SEM.Results The tail  collagen of rats in two groups did  not take fibrillogenesis in the screw surface at 2 h.At  4 h,the tail collagen   of   20-month-old rats   was gathered into the irregular and massive collagen polymer,no collagen fibers wre formatted  in surface of the screw.The tail collagen of 2-week-old rats turned into 0.07-0.20  μm  collagenousfibres.At 8 h,the tail collagen of  20-month-old rats  in the surface of the screw polymerized into the  different size collagen polymers,no formation of collagen fibers.The  tail collagen of 2-week-old rats  formated 0.65-1.35 μm collagen fibers.The SEM results showed that the  collagen of 2-week rat trabecular bone cross-linked into uniform collagen fibers.The fibers were closely arranged,and the orientation was consistent. The widths of the collagen fibrils of 20-month-old rat trabecular bone were not consistent. The arrangement of the fibrils was loose,sparse and disordered,and the orientations of the oblique collagen fibers between them were unorganized.Conclusion With the increaes of age,the fibrillogenesis capacity of type Ⅰ collagen gradually reduce,and lead to the morphological changes of bone collagen fibers,which may be very important reasons of  senile osteoporosis and increased fracture risk.

Abstract:Objective To establish an ovarian cancer cells with high expression of LPA1 by transfecting LPA1 genes into the ovarian carcer cell lines and to study thebiological characteristics of the transfected cells and to study the effects of lysophosphatidic acid (LPA) on the expressions and activities of MMP-2 and MMP-9 inovarian cancer cells with stable expression LPA1 genes.Methods The pcDNA3.1-LPA1 was transfected into ovarian cancer cells line (SKOV3 and 3AO)with lipofectaminereagent.Control and vector groups were set up.Transfection and expression were confirmed by the means of RT-PCR.The biological characteristics of the transfected cellswith expression LPA receptor genes were studied by the observation of their growth and proliferative period.The effects of LPA on the of MMP-2 and MMP-9 protein expressions and activities in transfected ovarian cells were analyzed by gelatin zymography.Results It was confirmed that the LPA1 genes had been stably integrated into 3AO and SKOV3 cell lines.The growth characteristics of transfected cells were not different before and after transfectation.The high expression of LPA1 gene was revealed by the means of RT-PCR.The expression levels of LPA1 mRNA in transfected cells （SKOV3-LPA1 and 3AO-LPA1） were significantly higher than those in control transfected cells and an empty vector groups,there were significant differences (P<0.05).The activities and protein expressions of MMPs in the cells  transfected with LPA1 receptor genes did not increase,compared with control and empty vector groups (P>0.05).Conclusion The ovarian carcer cells with high expression of LPA1 gene in vitro have been established.LPA1 plays a role as negative growth receptor in the process of LPA-induced MMPs invasion and metastasis of ovarian carcer cells.

Abstract:Objective To observe the effect of fluoxetine on phosphorylated microtubule-associated protein-2 (pMAP-2) expression in amygdala of depression model rats,and to provide the experimental basis for the study of efficacy mechanism of fluoxetine.Methods Male Wistar rats were randomly divided into control group,model group and fluoxetine group.The depression animal model was produced by giving the rats chronic unpredicted mild stress.The depressional behavior was examined using sucrose preference test,tail-suspension test and Morris water maze.The expressions of pMAP-2 protein and mRNA were detected using Western blotting and RT-PCR,respectively.Results The changes in body weight (%) and percentage of sucrose preference of rats in model group and fluoxetine group were significantly lower,while the tail-suspension immobility and the excape latency time were higher than those in control group (P<0.05).These indexes were reversed by fluoxetine.The expressions of  pMAP-2 protein and mRNA in model group and fluoxetine group were higher than those in control group (P<0.05),while the indexes were decreased after treatment with fluoxetine (P<0.05).Conclusion Fluoxetine can  reverse the expression increasing of pMAP-2 in amygdala neurons of depression model rats.

Abstract:Objective  To observe the effect of salmeterol(SLM) on inflammatory factors in bronchial veolar lavage fluid(BALF) of rats with bronchial asthma,and to investigate the therapeutic mechanism of salmeterol on bronchial asthma.Methods  48 SD rats were divided into 4 groups randomly:blank control group,model group,SLM group and inderal +SLM group.BALF of rats in various groups was collected.Then the eosinophil(EOS) was counted under light microscope,the apoptotic index of EOS was detected by TUNEL,and the levels of interleukin-5(IL-5)　 and Eotaxin were detected by ELISA.The pathological changes of lung tissues were observed by HE staining.Results  The levels of EOS,Eotaxin and  IL-5 in BALF of rats in model group and inderal +SLM group were all higher than those in blank control group (P<0.01).The apoptotic indexes in model group and inderal +SLM group were lower than that in blank control group(P<0.01).The levels of EOS,Eotaxin and IL-5 in BALF of rats in SLM group were all lower than those in model group(P<0.01).The apoptotic index in SLM group was higher than that in model group  (P<0.01).Inflammation was significant in lung tissues in model group and inderal +SLM group.Inflammation was mild in lung tissues in  SLM group.Conclusion The therapeutic mechanism of SLM on bronchial asthma may be related to lowering the number of EOS by the way of adrenoceptor-β2 to depress Eotaxin and IL-5 in BALF of asthma rats.

Abstract:Objective  To study the iron metabolism and iron nutrition state in obesity rats,and to provide the evidence for the prevention and control of iron malnutrition induced by obesity.Methods  27 male Wistar rats were divided into control group (n=10)and obesity group (n=17),the rats were fed with ordinary feed,high fat diet,respectively for 12 weeks,then they were killed by taking the eye blood.The hemoglobin,red cell count,hematocrit,serum iron,total iron binding capacity,serum transferrin saturation,hepcidin,transferring and ferritin of rats were determined.Results Compared with control group,the red cell count and hematocrit value in obesity group were increased,the hemoglobin content was decreased,in which the red blood cell number compared with control group showed significant differences(P<0.05).The serum iron,iron saturation values in obesity group were lower,the serum total iron binding capacity was increased,including serum iron,the iron saturation value compared with control group showed significant differences(P<0.05).The ferritin,transferrin and hepcidin values in obesity group were reduced.Wherein,the hepcidin and ferritin values compared with control group had significant difference (P<0.05).Conclusion Obesity can lead to iron malnutrition and the changes of iron metabolism indices.The obesity subjects should pay attention to  iron deficiency.

Abstract:Objective To explore the association between single nucleotide polymorphisms (SNPs) at rs2308765 and rs9269186 sites of HLA-DRB1 gene and hypertensive nephropathy, and to provide theroretical basis for study on genetic mechanism of hypertensive nephropathy. Methods Case-control study method was used and 97 cases were collected in the study. PCR-LDR technique was used to detect the SNPs of HLA-DRB1 in 45 patients with hypertensive nephropathy and 52 controls. The goodness-of-fit χ2test was used for Hardy-Weinberg equilibrium test in case and control groups. The differences of genotypic frequency and allelic frequency between case and control groups were test by χ2 test.Results There were G/T polymorphisms in  rs2308765 sites,two genotypes  G/G and G/T were found.The differences of genotype and allele distributions in rs2308765 between case and control were not significant (P>0.05). There were C/G polymorphisms in rs9269186 sites,three genotypes  C/C G/G and C/G were found.The differences of genotype and allele distributions in rs9269186 between case and control groups  were significant (P<0.05). Conclusion The correlation between SNPs at rs9269186 and hypertensive nephropathy is found, but not rs2308765.

Abstract:Objective  To evaluate the diagnostic value of the Montreal Cognitive Assessment (MoCA) in mild cognitive impairment (MCI) by using 25/26 cut-off value with Meta-analyss. Methods PubMed, Medline, VIP, CNKI and WANFANG databases (from January 1 2006 to December 31 2011) were searched to collect studies which evaluated the diagnostic value of MoCA in MCI. The statistical information and quality of science were assessed and classified. The data was analyzed using Meta-Disc1.4 software.The diagnostic value of MoCA in MCI was evaluated by the pooled sensitivity, specificity,and the likelihood ratio. Results Thirteen literatures were collected including 2 in English and 11 in Chinese, and 9 684 subjects were included in the review which were grouped with 6 859 MCI patients and 2 825 cognitive normal individuals, all diagnosed by gold standard. Heterogeneity test showed that the heterogeneity was existed among the study. By using random effects models toanalyze the data, the value of the weighted sensitivity was 0.98(95% CI：0.97-0.98), the specificity was 0.55(95% CI：0.53-0.57),the positive likelihood ratio was 3.67(95% CI：2.67-5.05),and the  negative likelihood ratio was 0.0(95% CI：0.03-0.18). Conclusion The diagnostic ability of MoCA is good. High sensitivity and low specificity are found by using 25/26 as cut-off value. MoCA could be used as a screening test for MCI screening in Chinese population. Considering its low specificity,more detailed researches should be performed to localize this tool, and focus on its cut-off value establishment.

Abstract:Objective To observe the influence of azelnidipine  in clinical efficacy and serum levels of high sensitive Ｃ-reactive　protein (hs-CRP),superoxide dismutase(SOD) and 6-ketone prostaglandins 　F1α　 (6-keto-PGF1α) in  patients with hypertension and to clarify  the effectiveness of azelnidipine on realeasing blood pressure and suppressing the development of atherosclerosis (AS).Methods 48 patients with hypertension were randomly divided into azelnidipine group(n=24) and amlodipine besylate group(n=24).After two-week drug clearance，the patients in two group receieved azelnidipine (8-16 mg/d
) or amlodipine besylate (5-10 mg/d) for 8 weeks.The blood pressure and the serum levels of hs-CRP,SOD and 6-keto-PGF1α before and after treatment were detected and compared. Results The systotlic blood pressure(SBP) and diastolic blood pressure(DBP) of the patients in two groups were significantly decreased,the effective rates in azelnidipine group and amlodipine besylate group were 87.5 and 91.7%，there was no significant difference between two groups(P>0.05).There was no significant difference of blood biochemisal indexes of the patients between two groups before and after treatment(P>0.05).There were no significant differences of plasma hs-CRP,SOD,and 6-keto-PGF1α of the patients between two groups before and after treatment(P>0.05).Compared with before treatment,the hs-CRP levels of the patients in two groups were significantly decreased(P<0.05),the SOD levels were increased(P<0.05);the 6-keto-PGF1α level of the patients in azelnidipine group after treatment was increased(P<0.05),but there was no change in amlodipine besylate group(P>0.05).Conclusion Azelnidipine can significantly decrease the blood pressure. It inhibit the formation and development of atherosclerosis.

Abstract:Objective To explore the reliability and validation of intensity of self-guided brisk walking in simple obesity patients and to provide evidence for formulating special exercise prescription. Methods  One hundred simple obesity patients (BMI>28, obese group, O group) and 100 normal weight adults (BMI<24, normal weight group, NW group) performed two tests of 2-km brisk walking on alternate days at a self-guided pace (Test1 and Test2). The average pace, heart rate (HR),percentage of maximal heart rate (%HRmax) and ratings of perceived exertion (RPE) were measured during each test;the retest reliability of brisk walking was conducted by correlation test in all indexes of Test1 and Test2;criterion-related validation was analyzed through the threshold (60%-90%HRmax) recommended for improvements in cardiovascular fitness by American College of Sports Medicine (ACSM). Results There were no significant differences of all indexes within groups between Test1 and Test2 (all P>0.05) and correlation coefficients were higher than 0.9, suggesting high test-retest reliability. The pace was lowered (all P<0.01), HR was higher (all P<0.01), %HRmax was higher (all P<0.01) and there was no significant difference of RPE (P>0.05) in O group compared with NW group in Test1 and Test2. The relative exercise intensity in O group (72%HRmax) reached the threshold recommended by ACSM and criterion-related validation r=0.731 (P<0.01), but in NW group (59%HRmax) the threshold was lowered and criterion-related validation r=0.351 (P>0.05). Conclusion  Exercise intensity of self-guided brisk walking is suitable for simple obesity patients but not for normal weight individuals. RPE may be a valuable indicator for simple obesity patients to monitor exercise intensity of brisk walking.

Abstract:Objective To explore the effects of 8-week exercise prescription intervention on aerobic capacity, body composition, blood lipid and C-reactive protein (CRP) and to provideevidence for guiding exercise prescription in obese adolescents. Methods Sixty male obese adolescents were randomly divided into control group (CG) and exercise group(EG). Both two groups participated in physical education lessons (twice per week, 40 min per session), but only EG participated in additional 2 sessions per week of 60-75 min per session (8 weeks) of exercise prescription intervention. The resting heart rate (RHR), blood pressure, maximal oxygen uptake (VO2max), body composition, blood lipid and  CRP were determined before and after test. Results There were no significant differences in the indexes mentioned above between two groups before test. At the end of test, the RHR, systolic blood pressure and serum CRP were reduced (P<0.05 or P<0.01), while the VO2max and lean body mass (LBM) were elevated (P<0.05 or P<0.01) in EG compared with pre-value, but all the indexes in CG showed no significant change(all P>0.05). The RHR, systolic blood pressure and serum CRP were lower (all P<0.05), while VO2max and LBM were higher (all P<0.01) in EG than those in CG after test.Conclusion Eight weeks exercise prescription can effectively improve aerobic capacity, increase LBM and reduce cardiovascular diseases risk level in obese adolescents.

Abstract:Objective To investigate the relationships between transforming growth factor beta 1(TGF-β1),  polymorphism of human papilloma virus( HPV16) E6 gene and cervical carcinoma, and to clarify the possible mechanism of TGF-β1 in cervical carcinoma. Methods Fresh cervical tissues of hospitalized surgical patients between May 2007 to January 2009 were collected and divided into three groups, 17 cases of  cervical intraepithelial neoplasia Ⅰ(CIN Ⅰ);17 cases of CIN Ⅱ-Ⅲ;39 cases of cervical squamous cell carcinoma. The immunohistochemical method was used to observe the TGF-β1 protein expression in cervical tissues,and the direct PCR method was performed to test the genotype of HPV16 E6, finally the relationships between them and the occurrence and development of cervical carcinoma were analyzed. Results  The positive rates of TGF-β1 in CIN,CINⅡ-Ⅲ and  cervical squamous cell carcinoma group were 11.8% ,35.3% and 64.3%, respectively, and there were significant differences between various groups (P<0.01 ).The expression of TGF-β1 in cervical carcinoma  patients of stage ⅡB to ⅣA (88.9%) were significantly higher than that of stage ⅠB and ⅡA（76.7%）(P<0.01). The detection rates of HPV16　E6 in CINⅠ, CINⅡ-Ⅲ and cervical squamous cell carcioma group were 11.8%, 41.2% and 82.1%,there were significant differences between various groups(P<0.01). The positive rate of HPV16 E6 gene was positively related with the development of cervical carcinoma （r=0.592 ，P=0.001). By DNA sequencing, DNAman program was used to analyze the data, totally 14 sequences were the same as the standard strain，and the most common mutant site was D25E（42.5%). The TGF-β1 positive rates in HPV16 E6 non-mutant patients and in HPV16E6 D25E patients were 50.0% and 92.9%, respectively (P<0.05).Conclusion The occurrence and development of cervical carcinoma has something to do with TGF-β1 expression, and the HPV16　E6 polymorphism may be associated with the TGF-β1 which promote to growth and invasion of cancer.

Objective To investigate the association between the single polymorphisms(SNPs) of MMP-9 gene C1562T,R279Q and coronary heart disease (CHD). Methods 258 patients diagnosed with CHD and 183 healthy contols were selected. The serum levels of MMP-9 were determinated using enzyme-linked immunosorbent assay (ELISA). PCR-RFLP analysis was applied to detect the C1562T and R279Q SNPs of MMP-9 gene. Statistic
al soft ware SPSS 17.0 was used to analyze the frequency distribution of alleles and genotypes. Results The serum MMP-9 level  in CHD group was significantly higher than that in  control group（P<0.05）. The C1562T genotype frequencies of CT+TT genotypes and T allele were 24.4% and 11.8%  in CHD group and 13.8% and 7.3% in control group, respectively, and there were significant differences  between  two groups (P<0.05). The R279Q genotype frequencies of AG+GG genotypes and G allele were 60.2% and 41.7%  in CHD group and 68.0% and 41.5%  in control group, respectively, and there were no  significant differences between  two groups(
P>0.05). Conclusion The  MMP-9 level in patients with CHD  is increased and the polymorphisms in MMP-9 gene C1562T associates with the susceptibility to the CHD,but there is  no association between R279Q polymorphism and CHD.

Abstract:Objective To investigate the expression of zinc finger E-box binding homeobox 1(ZEB1) in lung squamous cell carcinoma,and  to analyze the relationship between the ZEB1 mRNA and protein expression levels and invasion and metastasisof lung squamous cell carcinoma. Methods 45 patients with lung squamous cell carcinoma performed surgical operation excision were chosen. According to the distance from the tumor tissue, lung squamous cell carcinoma tissue and adjacent normal lung tissue were selected, RT-PCR and Western blotting were used to detect the ZEB1 mRNA and protein expressions in lung squamous cell carcinoma and adjacent normal lung tissues, and the relationship between the clinical pathological features of lung squamous cell carcinoma and ZEB1 mRNA and protein expressions were analyzed. ZEB1 specific siRNA vector was constructed, then NCI-H2170 cells were infected, meanwhile untransfected group, control group and ZEB1siRNA transfected group were set up. Western blotting was performed to detect the interference effect, and then through the Transwell chamber the changes of NCI-H2170 cell invasion ability were observed. Results The positive mRNA expression rate of ZEB1 in lung squamous cell carcinoma tissues was significantly higher than that in normal lung tissue (P<0.05). The positive mRNA expression rates in the carcinoma tissues from the patients with positive lymph node or distant metastasis were significantly higher than those from the  patients without lymph node or distant metastasis(P<0.05). However, the expression of ZEB1 had no correlation with gender, age, tumor size, or tumor differentiation level of the patients with lung squamous cell carcinoma(P>0.05). The protein expression of ZEB1 was similar to the mRNA expression of ZEB1. The protein expression of ZEB1 in lung squamous cell carcinoma tissue was significantly higher than that in normal lung tissue (P<0.05). The protein expression level of ZEB1 in the carcinoma tissue from the patients with positive lymph nodes or distant metastasis was significantly higher than that from the patients without lymph nodes or distant metastasis (P<0.05). Reducing the protein expression of ZEB1 in NCI-H2170 cells, the number of cells that passed through the Transwell chamber was significantly lower than that in non-transfected control group as well as transfected control group (P<0.05). Conclusion ZEB1 displays a high expression in lung squamous cell carcinoma  tissue, and their expression changes are associated with the pathological features of lung squamous cell carcinoma;By RNA interference effectively silencing ZEB1 gene in NCI-H2170 cells, knockdown of ZEB1 could significantly reduce the invasion and metastasis ability of NCI-H2170 cells, suggesting that ZEB1 play an important role in lung carcinogenesis and development, and inhibition of ZEB1 expression might become a kind of method for treating lung cancer.

Objective To study the influence of different cements and margin designs on marginal microleakage in metal crowns in order to guide clinical design, operation, and to provide basis for  reducing microleakage in clinic.
Methods  45 extracted human teeth were randomly divided into 3 groups. Teeth were prepared to the different margin finish lines, respectively chamfer, shoulder and bevel, and cast metal crowns. The crowns in each group were randomly subdivided into 3 subgroups according to the cements:Rely X Unicem resin adhesive, 3M resin modified glass ionomer cement, and polycarboxylate cement. After thermocycling experiment,the teeth were dyed and developed,and the crowns were cut along the long axis of the teeth, then all specimens were evaluated under the microscope, and were graded using SPSS 17.0 software for statistical analysis on the degree of microleakage. Results With the same finish line, the resin adhesive demonstrated the least marginal microleakage,and polycarboxylate cement was associated with severe marginal microleakage(P<0.05);With the same cement, chamfer and bevel had no significant difference in microleakage(P>0.05),and they were both less than shoulder(P<0.05). Conclusion Resin adhesive luting system which is the first selection in clinic is better th
an glass ionomer cement and polycarboxylate cement;the chamfer and bevel which are the appropriate finish lines for decreasing microleakage are better than shoulder.

Objective To detect the expressions of the matrix metalloproteinase 2(MMP-2) matrix metalloproteinase 9(MMP-9) vascular endothelial growth factor(VEGF）,Flms like tyrosine kinase (Flk-1) and multiple tumor suppressor 1 (P16) in human glioma invasive tissues,and to  elaborate the correlation of the MMP-2,MMP-9,VEGF,Flk-1 and P16 with the glioma invasion.Methods The expressions of MMP-2,MMP-9,VEGF,Flk-1 and P16 were detected through in situ hybridization and immunohistochemistry in 33  cases of glioma invasive tissues.Results
 The positive rates of MMP-2 and MMP-9 in Ⅰ-Ⅱ　grade and Ⅲ-Ⅳgrade glioma invasive tissues were 20.00%,61.11％ and 13.33％,55.56%. The positive rates of VEGF and Flk-1 were 26.67％,66.67％and 20.00％,7
2.22％(P<0.01),respecttively；the P16 deletion frequencies were 13.33％ and 77.78％,there was significant difference(P<0.01).Conclusion The necessary condition in occurrence of glioma invision is the basilar membrane degraded by the matrix metalloproteinases and the proliferative activity of vascular endothelial cells.
 The descended expression rate for P16 reflects the biological behavior of glioma invasive tissue.

Objective  To investigate the association between habitual physical activity and onset of  postmenopausal osteoporosis (PMOP). Methods The hospital-based case-control study included 67 cases of PMOP (case group) and 67 healthy
women (control group). The women verbally answered the International Physical Activity Questionnaire for assessment of physical activity levels. This instrument evaluated the frequency and duration of activities performed during transportation
, at work, at home or in leisure time-over the course of recent one year. The risk of disease was represented by using odds ratio and 95% confidence interval.Results The physical activities in total and transportation, leisure-time were different between case group and control group（P< 0.01）. The physical activities in occupational, housework did not show significant difference between case group and control group（P>0.05). The physical activity in transportation significantly decreased the risk of PMOP (OR=0.223, 95% CI:0.058－0.856). The results demonstrated that moderate walking exercise in postmenopausal women appeared to be effective way to decrease the risk of PMOP (OR=0.064, 95%CI:0.007-0.563). Conclusion   The weight-bearing physical activities including weight-bearing physical exercise can be likely to decrease PMOP risk.

Objective To investigate the expressions of hepatitis B virus X protein (HBx) and carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) in HBV-related hepatocellular carcinoma tissue and to explore the relationship between the two genes and occurrence,development and invasion of hepatocelular carcinoma.
Methods  The expressions of HBx and CEACAM1 in 81 cases of HBV-related hepatocellular carcinoma tissues were detected by immunohistochemistry method,and the relationship with clinicopathological features was analyzed. The expressions of CEACAM1 in the human normal hepatocyte cell line QZG, hepatocellular carcinoma HepG2 and HepG2-X(HepG2 cells transfected with HBx) were detected by Western blotting. Results The positive rates of HBx and CEACAM1 expressions in 81 cases of HBV-related hepatocellular carcinoma tissues were 74.07%(60/81) and 71.60%(58/81)， respectively. The expressions of HBx and CEACAM1 were correlated with portal vein invasion,nodal metastasis and TNM staging(P<0.05 or P<0.01). The expression of HBx was negatively correlated with CEACAM1（rs=－0.310，P<0.05）.The CEACAM1 protein was significantly  decreased in HepG2-X compared with HepG2 and human normal hepatocyte cell line QZG.Conclusion The high expression of HBx and low expression of CEACAM1 are correlated with the invasion and metastasis of HBV-related hepatocelular carcinoma,and HBx may promote HBV related hepatocarcinogenesis and development by       inhibition CEACAM1.

Objective  To analyze the correlation between the bladder weight (BW) and intravesical prostatic protrusion (IPP) and the degree of bladder outlet obstruction（BOO） in  the patients with benign prostatic hyperplasia (BPH) and to explore the clinical value of transabdominal ultrasonography in judgement of BOO in the patients with BPH. Methods Urodynamic examination was performed in 124 BPH patients from March 2009 to January 2011.

124 patients were divided into 2 groups (non-obstruction and obstruction). The  BW  and IPP were measured by transabdominal ultrasonography,the correlation between BW,IPP and the parameters reflecting BOO was analyzed.  Results  The BW values in  obstruction group and  non-obstruction group were (77.47±44.34) g and (57.47±23.10) g. There was significant difference in BW between  obstruction group and  non-obstruction group(P<0.05). The IPP values in   obstruction group and non-obstruction group were (11.05±9.65) mm and

(7.67±7.99) mm. There was significant difference in IPP between obstruction group and non- obstruction group(P<0.05). The BW value of all the patients with BPH was negatively correlated with Qmax(r=－0.177,P<0.05) and positively  correlated with PdetQmax (P<0.05,r=0.201). The IPP value was negatively correlated with Qmax(r=－0.403,P<0.01) and positively correlated with PdetQmax (r=0.192,P<0.01). Conclusion
The BW and IPP are highly correlated with BOO ,and evaluating BW and IPP with ultrasound is a reliable method to determine the degree of BOO.

Objective
 To explore the extent of lymph node dissection for upper and middle thoracic esophageal cancer by comparing the efficacy of three-field lymphadenectomy(3-FL) and two-field lymphadenectomy(2-FL). Methods 119 patients    who were treated surgically with upper and middle thoracic esophageal squamous carcinoma from 2008 to 2010 were retrospectively reviewed. 46 patients were treated by 3-FL(3-FL group), and the remain 73 patients were treated by 2-FL(2-FL group). Results The lymph node metastasis rate in 3-FL group(58.7%) was much higher
 than that in 2-FL group (39.7%), the difference was significant(P<0.05).The cervical lymph node metastasis rate in 3-FL group was 21.7%. The upper mediastinal lymph node metastasis rate in 3-FL group(28.2%) was much higher than that in 2-FL group(11.0%)(P<0.05). The surgical morbidity in 3-FL group(39.1%) was much higher than that in 2-FL group(16.4%, P<0.01), the main difference was the rate of recurrent laryngeal nerve palsy. The one-year survival rate of the patients in 3-FL group(95.6%) was higher than that in 2-FL group(83.5%)(P<0.01).
Conclusion Compared with 2-FL, 3-FL can dissect the cervical and upper mediastinal lymph nodes more thoroughly, reduce the local recurrence rate, and improve the rate of survival. The patients with upper and middle thoracic esophageal squamous carcinoma without metastasis should be operated by 3-FL.

Abstract:Objective To detect the rate of Chlamydia infection of ocular adnexal lymphoproliferative diseases and to investigate the relativity between ocular adnexal lymphoproliferative diseases and   Chlamydial  infection.
Methods 118　patients with  ocular adnexal lymphoproliferative diseases were selected as experiment group, and 46 cases among them  with mucosa associated lymphoid tissue(MALT) lymphoma,41 cases of  hyperplasia of reactive lymphoid tissue,31 cases of other types of lymphoma. 20 cases of samples with ocular adnexal non- lymphoproliferative
diseases were selected as    control group,including 17 cases of vascular tumor and 3 cases of  thyroid associated ophthalmopathy.Then  the method of immunofluorescence was used to detect  the Chlamydia in two groups.It was positive when it showed over ten inclusionbodies in the vision of a high-rate magnification(×1 000).Otherwise it was negative.
Results 80 cases of ocular adnexal lymphoproliferative diseases in 118 cases had positive chlamydia infection(67.79%),while there was0 case  among  20 cases of ocular adnexal non- lymphoproliferative diseases in  control group(χ2=32.262, P<0.001).There was no significant differecnce of pasitive rate of Chlamydia infection between MALT lymphoma patients and non-MALT lymphoma patients(P>0.05). Conclusion There might be a relationship between common ocular adnexal lymphoproliferative diseases and    Chlamydia infection,especially the MALT lymphoma and the hyperplasia of reactive lymphoid tissue  have higher rates of   Chlamydia infection.The detection and treatment of Chlamydia infection provide a new direction of research for the diagnosis and treatment of  ocular adnexal lymphoproliferative diseases.

Objective
To investigate the health condition of the old people in the urban and rural communities of Changchun city,and to  analyze its influencing factors and provide evidence for elderly  to prevent related diseases and  the government to make care strategies. Methods ADL scale, SF-36 scale and self-designed questionnaire were used to investigate 600 old people from Guilin Road community in urban , Jingyue Development community in the urban and rural junction and Jiangjia community in rural by grading and systematic sampling. Some physical examinations were performed at the same time. SPSS 13.0 software  was adopt to deal with the data and compare the conditions of the three communities. Results The morbidity of chronic diseases in old people was 77.14%, and they were 87.36%, 83.61% and 71.22% in urban, urban and rural junction and rural communities. The morbidities of various chronic diseases were as follow:heart disease 51.43%, hypertension 36.96%, arthr itis 34.11%, cerebrovascular diseases 28.04%, respiratory diseases 23.04%, cataract 8.04%, malignant tumors 3.92%, Parkinson’s disease 1.42%, disability 0.53%. The old people who had at least two kinds of chronic diseases occupied 67.30%. The downtown community had higer morbidities of hypertension, heart disease, cerebr ovascular diseases and arthritis than the other two communities(P<0.001). Conclusion  The old people of Changchun communities has higher morbidity of chronic diseases. The medical workers in community health service center should provide more health guidance to improve the old people’s health.

Objective To examine the various factors that affect determination of astragaloside Ⅳ in Baoyuanqingxue granules,elucidate the extraction method and chromatographic conditions of determination of astragaloside Ⅳ, and to establish an effective method for determination of astragaloside Ⅳ. Methods High performance liquid chromatography(HPLC) method was used.The SB-C18 column (Agilent，ZORBAX，4.6 mm×150 mm，5 μm)was adopted,the mobile phase was acetonitrile-warer(36∶64) at the flow rate of 1.00 mL?min-1 with UV detection at 203 nm,the volume of injection was 20 μL and the column temperature was 30℃. Results  The resolution between astragaloside Ⅳ and other peaks was good. The calibration curve was linear in the range of 0.010 03-0.200 60　g?L-1（r=0.999 9).The average recovery(n=6) of astragaloside Ⅳ was 98.4% with RSD of 0.8%. Conclusion  The HPLC-UV method to detect the cotent of astragaloside Ⅳ in Baoyuanqingxue granules is simple and accurate. It can be used for quality control of astragaloside Ⅳ in Baoyuanqingxue granules.