Abstract:Objective
 To investigate the effect of chloride transport in nonspecific phagocytic process of human retinal pigment epithelial(RPE) cells which was treated with DNDS and bumetanide,and to discuss the mechanism for chloride transport’s effort in nonspecific phagocytic process.Methods  Human RPE cells(ARPE-19) were cultivated using millicell cell culture inserts system,and flow cytometry was performed on ARPE-19 cells that had been challenged with fluorescent latex beads.The bumetanide of 1,5,10,50,100,500,1　000 μmol·L^-1 and DNDS of 0.01，0.10，0.50,1.00,3.00,5.00,10.00 mmol·L^-1  were used in the experiment as chloride transport pathway blockers for 6 h,then flow cytometry was used to detect the phagocytic index.Results Compared with control group,different concentrations bumetanide didn’t affect the nonspecific phagocytic process of RPE cells（P>0.05）；when the basal membrane of RPE was treated with  0.01 and 0.10 mmol·L^-1 DNDS,there were no significant differences of phagocytic index compared with control group (P>0.05)；however when treated with 0.50, 1.00,3.00,5.00 and 10 mmol·L^-1 DNDS,the phagocytic indexes were significantly decreased (P<0.05 or P<0.01),and as the concentration increased,the phagocytic indexes were decreased gradually;when the apical membrane of RPE was treated with above concentrations of DNDS,there were no significant differences of phagocytic index compared with control group(P>0.05).Conclusion Apical membrane bumetanide-sensitive Na⁺-K⁺-2Cl^－cotransporter doesn’t participate in the nonspecific phagocytic process of RPE cells,basal membrane DNDS-sensitive Cl－/HCO₃^－ exchange and chloride channel play important roles in the nonspecific phagocytic process of RPE cells.

Abstract:Objective
To investigate the effect of the ultra-filtration extract mixture from Angelica Sinensis and Hedysarum Polybotrys on heat shock protein 70(Hsp70)expression in cardiomyocytes,and clarify its antioxidation on cardiomyocytes．Methods Myocardial cells from 1-3 d neonatal rats were cultivated in DF medium and the cellular injury was induced by H₂O₂（400 μmol•L^-1）．The ultra-filtration extract mixtures were given in three doses of 3.75，7.50， and 15.00  g•L^-1．The beating rates of cardiomyocytes were observed by phase-contrast microscope.The apoptotic rates of cardiomyocytes were measured by flow cytometry．The expressions of Hsp70 mRNA and protein in cardiomyocytes were measured by RT-PCR and Western blotting．Results Compared with normal control group,the beating rates of cardiomyocytes were significantly decreased (P<0.01) and the apoptotic rates of cardiomyocytes were significantly increased(P<0.01) in model group,and the expressions of Hsp70 mRNA and protein in cardiomyocytes were increased(P<0.05，P<0.01).Compared with model group,the beating rates of cardiomyocytes were significantly increased(P<0.01) and the apoptotic rates were significantly decreased (P<0.01) in each treatment group,and the expressions of Hsp70 mRNA and protein in cardiomyocytes were significantly increased（P<0.01，P<0.05）.Conclusion The ultra-filtration extract mixture has the protective effects on cardiomyocytes injured by H₂O₂ and can up-regulate the Hsp70 expression in cardiomyocytes．

Abstract:Objective
 To exploit the medicinal values of Scorzonera ruprechtiana Lipsch.et Krasch.through investigating its chemical constituents. Methods  The herbs of Scorzonera ruprechtiana were extracted with 70% ethanol,the extractives were purified with  macroporous resin and isolated with silica gel,and ODS column chromatography.NMR,UV,IR spectrometers were used to obtain spectroscopic data.
Results  According to the single spot in TLC,three flavonoid glycoside compounds and one sesquiterpenoid glycoside compound were isolated from the herbs of Scorzonera ruprechtiana,and their structures were identifited through physico-chemical constants and spectroscopic analysis.They were 5,7,3′,4′-tetrahydroxy-flavone 8-C-β-D-glucopyranoside (Ⅰ); 5,7,3′,4′-tetrahydroxy- flavone 6-C-β-D- glucopyranoside (Ⅱ); 5,7,4′-trihydroxy- flavone 6-C-β-D- xylofuranosyl-(1→2)-β-D- glucopyranoside (Ⅲ);3-O-β-D- glucopyranosyl-3 - hydroxy-1α,5α,7α H- guaia-4(15),10(14),11(13)- trien-12,6α-olide (Ⅳ).Conclusion  Three flavonoid glycoside compounds and one sesquiterpenoid glycoside compound are isolated,and the flavonoid glycosides are isolated from Scorzonera ruprechtiana Lipsch.et Krasch.and also from Scorzonera genus plants for the first time.

Abstract:Objective
To investigate the enhancement of hypoxia responsive element (HRE) on  killing effect of retrovirus mediated HSV-tk/gancyclovir (GCV) suicide gene system controlled by human telomerase reverse transcriptase (hTERT) promoter on   tumor  cells under hypoxia condition.Methods The recombinant retroviral vectors including the HRE hypoxia responsive element,the hTERT promoter and suicide gene tk based on retroviral vector pLNCL were constructed．The recombinant retroviral vector was transfected into PA317 cells for package by liposome transfection， 2 weeks after G418 selection，G418-resistant PA317 colonies were obtained and amplified．The supernatant of cell culture was harvested and used to infect NIH3T3 cells to measure the viral titer of recombinant retrovirus.The viral supernatant was used to infect lung cancer cells SPC-A1 and liver cancer cells Bel-7402.The positive clones were obtained by G418 selection.The experiment was divided into control group,SPC-A1/tk group and Bel-7402/tk group.Under the hypoxia (1%O2 ，94%N2) condition,GCV-mediated cell growth inhibition was determined with method of tetrazolium( MTT).FCM combined with PI and AnnexinV-FITC double pigmention methods were used to observe the apoptosis and cell cycle of two tumor  cell lines infected by the recombinant retrovirus.
Results As compared with control group, SPC-A1/tk and Bel-7402/tk showed high sensitivity to GCV in vitro under hypoxia condition,the apoptotic rate was  increased significantly (P<0.001).As compared with group of recombinant retrovirus with tk driven by the hTERT promoter,the survival rate in group of recombinant retrovirus with tk driven by HRE-modified hTERT promoter was decreased significantly (P<0.01),the apoptotic rate was  increased significantly (P<0.01),and both the cell populations at G₁ and G₀ phases were increased significantly (P<0.05),and the cell population at S phases was decreased significantly (P<0.05).Conclusion  HRE can enhance the  killing effect of HSV-tk/GCV suicide gene system controlled by the hTERT promoter on tumor  cells line under hypoxia condition.

Abstract:Objective
To establish the method to induce human bone mesenchymal stem cells(hMSCs) to differentiate into cardiomyocytes in vitro. Methods  hMSCs were isolated and purified from the bone marrow of human by density gradient centrifugation and adhering to the culture dishes.The fourth passage of MSCs was induced by 5-azacytidine(5-Aza) and bFGF for four weeks.The expression of Desmin,Troponin I and α-sarcomeric actin were detected by immunocytochemistry 3-4 weeks after induction.Results
hMSCs treated with 5-Aza and bFGF had long cytoplasmic process  one week  after induction,and had multiple branches  two weeks after induction,and they connected with adjoining cells forming myotube-like structures  four weeks after induction. The induced hMSCs were stained positively for Desmin,Troponin I and α-sarcomeric actin.Conclusion  5-Aza and bFGF can induce hMSCs to differentiate into cardiomyocytes in vitro.

Abstract:Objective
To examine the effects  of C-type natriuretic peptide (CNP) on the atrial dynamics and cylic nucleotides level in the atrial myocytes in rabbits and explore its mechanism of action. Methods  Atria were obtained from New Zealand white rabbits, and the experiments were performed using a  perfused beating atrial model. Briefly, the heart was removed from rabbit which had been anesthetized and the left atria was  dissected free, and then it was fixed in perfused beating atrial model. The effects of CNP (10, 30, or 300 nmol•L^-1) on atrial pulse pressure and atrial stroke volume were analyzed, and cAMP efflux and cGMP levels which were dealed with  30 nmol•L^-1 CNP were measured by radioimmnoassay . Results ①CNP (10, 30,and 300 nmol•L^-1) significantly decreased the atrial pulse pressure and atrial stroke volume in perfused beating atria compared with control group（P<0.05 or P<0.01）;these effects showed dose-dependent manner, and the maximum inhibitory rates in  300 nmol•L^-1  CNP group were 74.8% and 76.9%, respectively. ② 30 nmol•L^-1 CNP  significantly decreased the atrial pulse pressure and atrial stroke volume in perfused beating atria compared with control group（P<0.01）, and the cGMP level in the atrial myocytes was significantly increased, about 10.2 times,compared with control group(P<0.01), but the cAMP level didn’t  change (P>0.05).Conclusion CNP can inhibit the atrial dynamics,its mechanism may be related to the increasing of cGMP level through CNP-GC-cGMP signal pathway in rabbit atria. 

Abstract:Objective
To isolate and cultivate human bone marrow mesenchymal stem cells (hMSCs) in vitro,and observe the  expression of alternative lengthening telomere related promyelocytic leukemia protein (PML) in hMSCs,in order to evaluate its role in self-renewal and division growth processes of hMSCs.Methods hMSCs were isolated from bone marrow by Percoll density gradient centrifugation.Immunocytochemistry staining was used to detect the expression rate of PML body in hMSCs.Results There were no visible expressions of PML body in passage 1,3 and 5 of cultured hMSCs.There was positive expression of PML body in passage 7 hMSCs,there were brown particles in nuclei of hMSCs,the positive rate was (16.1±0.6）%.There were also positive expressions of PML body in passage 9 and 11  hMSCs,the positive rates were (16.8±2.6)% and (45.8±9.5)%, respectively;there were significant differences between passage 9,11 and passage 7  hMSCs（P＜0.05）.Conclusion Following long-time culture of hMSCs in vitro,ALT will appear in hMSCs to promote the lengthening of telomere,and maintain the ability of proliferation and self-renewal of hMSCs.

Abstract:Objective
To construct the recombinant vectors that coexpress wild type p53(wt-p53) and siRNA-mdm2 for gene therapy in androgen independent prostate cancer .Methods The subcloning,T-Acloning,and PCR technique were used to construct the recombinant vectors,named pcDNA3.1-p53,siRNA-mdm2,p53/siRNA-mdm2, and siRNA-scramble .The recombinant expression vectors mentioned above were transfected into PC-3 cells,the expression levels of the mdm2 siRNA and p53 after transfection were detected by the semi-quantitative RT-PCR analysis and Western blotting analysis.MTT assay was used to detect the  proliferation inhibition  of PC-3 cells after transfection.Results The recombinant plasmids containing both wt-p53 and siRNA-mdm2 were successfully constructed,and confirmed by DNA sequence analysis.After transfected for 48 h, the expression of GFP was observed.The expression level of wt-p53 was increased,and the expression of mdm2 was decreased detected by RT-PCR and Western blotting.MTT assay showed that the inhibitory rate of  proliferation of PC-3 cells was 40.4% after transfection.Conclusion The recombinant plasmids containing both wt-p53 and siRNA-mdm2 are successfully constructed,they can remarkablely inhibit the proliferation of PC-3 cells.

Abstract:Objective
To express and purify the recombinant BCG heat shock protein 70 (BCG HSP70),and remove endotoxin from it.Methods E.coli. BL21(DE3) containing the recombinant plasmid of pET28a/HSP70 (pET28a/HSP70/BL21) was induced with IPTG in 10 L fermentor,and the expression of BCG HSP70 was detected by SDS-PAGE. Then the bacteria were disrupted by sonication. BCG HSP70 was purified by successive applications of Nikel-affinity chromatography on Sepharose 4B,TritonX-114 washing,gel filtration on Sephadex G-25 and ion-exchange chromatography on Q-Sepharose FF.The purified BCG HSP70 was identified by SDS-PAGE.The concentration of the purified BCG HSP70 was detected by Lowry assay.And the purified BCG HSP70 was incubated in the 37 ℃ water bath for 0 to 4 h to analyze its stability.Endotoxin in the purified proteins was determined by Limulus amebocyte lysate assay.Results 96 g wet weight of bacterial pellet was obtained after 3 h of induction of pET28a/HSP70/BL21 in the fermentor,and the protein with relative molecular mass of 70 000 approximately accounted for about 29.6% of the total bacterial protein.The protein with relative molecular mass of 70 000 was purified to 96.5% purity,and the concentration of the purified protein was 1.2 g•Ｌ
^-1.When incubated in the 37 ℃ water bath for 4 h,the BCG HSP70 accounted for approximately 95.1% of the total protein.Endotoxin in the purified protein was less than 0.01 EU?•μg^-1.Conclusion The recombinant BCG HSP70 is expressed and purified successfully,the endotoxin in the purified BCG HSP70 is removed effectively.

Abstract:Objective
To obtain recombinant enzyme donor (ED) and enzyme acceptor (EA) fragments of  β-galactosidase in E.coli and establish new cloned enzyme donor immunoassays (CEDIA) for clinical use. Methods The encoding sequences of ED and EA fragments were amplified with pSV-β- galactosidase control vector as template and inserted into pGEM-Teasy vector,the target gene was chosen by double digestion,PCR and sequencing,then ED and EA fragments were inserted into the expression vector pET20b+. The competent cells of host strain of BL21 were transformed by the recombinant plasmid.The expression of the target protein was induced with IPTG and purified by Ni2+-NTA agarose column.The β-galactosidae with activity is formed.
Results The cloned fragments of ED and EA were 100% consistent with that of pSV-β-galactosidase control vector.The expression vector pET20b-ED and pET20b-EA were constructed and expressed.The target protein was purified by Ni2+ -NTA agarose column.The expressed fusion-protein ED fragment was 14 000 and EA fragment was 116 000 in SDS-PAGE as expected.Conclusion ED and EA proteins are prepared successfully and β-galactosidase with activity is formed.

Abstract:Objective
To identify the antimicrobial resistance and the mutations in acrAB-tolC efflux pump and marOR regulated genes in Shigella,then analyze their relations.Methods One hundred and fifty-nine clinical isolates of Shigella were collected.Susceptibility tests of tetracycline( TE),chloramphenicol(C),ampicillin(Am),gentamycin(GM),norfloxacin (NOR) and selectrin(SMZ-TMP) and organic-solvent tolerance test were performed in all isolates.The acrA,acrB,tolC and marOR genes of these isolates were amplified by polymerase chain reaction (PCR) respectively.The PCR products were digested by restriction endonuclease Taq Ⅰ or Hinf Ⅰ,then analyzed by single strand conformation polymorphism (SSCP).Results In 159 clinical isolates,137 multi-drug strains and 4 sensitive strains were discovered.The rate of multi-drug resistance was 86.2%. 122 strains were organic-solvent tolerance and the rate of organic-solvent tolerance was 76.7%. The marOR and acrAB-tolC genes were found in all strains and the lack of the four genes were not found. SSCP analysis revealed that the rates of mutations in marOR,acrA,acrB and tolC genes were 17.4%,5.8%,3.9%， and 2.6%,respectively.Conclusion  The Shigella spp.has high multi-drug resistance rate and organic-solvent tolerance rate,and the mutation rate of marOR gene is high too.

Abstract:Objective
 To construct the eukaryotic expression vector and express in mouse liver cancer cell Hepa1-6,and observe reproductive activity and tumorigenesis ability of transfected cells.Methods The MAGE-1 total length sequence was amplified from SMMC-7721 by PCR and  linked to pcDNA3 to construct pcDNA3-MAGE-1.The expression vector was transfected into Hepa1-6 cells by lipofectamine,and then the positive clones were screened by G418.The expressions of mRNA and protein in positive clones were detected by RT-PCR and Western blotting.Cell growth and proliferation in Hepa1-6 cells and Hepa1-6- MAGE-1 cells were detected by MTT,then they were inoculated in the right back subcutaneous of mice.Results  pcDNA3-MAGE-1 was constructed and transfected into Hepa1-6 cells,the expressions of MAGE-1 mRNA and protein were detected in Hepa1-6-MAGE-1 cells by RT-PCR and Western blotting,there  was no statistical difference in cell proliferation between two groups.The mice were all infected with tumor in two groups,and there was no difference in tumor size.
Conclusion The eukaryotic expression vector pcDNA3-MAGE-1 is successfully constructed,and the Hepa1-6 cell line which can stably express human MAGE-1 gene is established with good proliferation and tumorigenesis ability.

Abstract:Objective
To observe the effect of diterpenoid B derived from Rabdosia excise (DB) on the cell line of cervical cancer (Hela) and its related mechanism.Methods Hela cells at logarithmic growth phase were divided into control group and 0.5,1.0,2.0,4.0,8.0,16, 0 and 32.0 mg•L^-1 DB experimental groups,MTT  assay was used to detect the  inhibitory rate of cell growth at 24 and 48 h,and the morphologic changes of cells were observed by phase contrast microscope. RT-PCR was exploited to observe the mRNA expressions of p53,p21 and CDK2 of Hela cells.Immunocytochemical  staining was utilized to detect the protein expressions of P53,P21 and CDK2.Results  MTT result  showed that the inhibitory rate of cell growth at 24 h in 2.0,4.0 and 8.0 mg?mL-1 DB groups were 18.18%,28.89%,and 32.84%;the inhibitory rates  of cell  growth  at 48 h in  2.0,4.0,and 8.0  μg•mL^-1DB groups were 26.64%,47.03%, and 51.90%;DB  inhibited Hela cell growth in a dose- and time- dependent manner（P<0.05 or P<0.01）.Hela cells after treated with DB  manifested retarded proliferation,and became round and smaller in size,with some cells floating;the RT-PCR  and Western blotting results showed that after  treated with DB (2.0,4.0,8.0  μg•mL^-1) for 24 and 48 h,the mRNA and protein expressions of P53 and P21 in DB  groups were significantly higher than those in control group（P<0.01）,meanwhile the mRNA and protein levels of CDK2 were lower than those in control group（P<0.01）.Conclusion  DB can inhibit the  proliferation of Hela cells,its mechanism  may be related to  up-regulating the expressions of P53 and P21,down-regulating the expression of  CDK2,and  furthermore influencing the transition from G1 to S phase.

Abstract:Objective To investigate the inhibitory effects and possible anticarcinogenic mechanism of celecoxib on the proliferation of Lewis lung carcinoma cells in vitro and in vivo.Methods The proliferation of Lewis lung carcinoma cells　treated with different doses (50,100 and 200  μmol?L-1) of celecoxib for 12,24,48 and 72 h,was measured by MTT assay.The contents  of arachidonic acid and prostaglandin E2 in the culture supernatants at 24 h were measured by the methods of RP-HPLC and PGE2-specific ELISA respectively.Twenty C57BL/6 mice receiving tumor implantation were divided randomly into control group and treatment(200 mg•kg^-1) groups. All the groups were gavaged continuously with normal saline or celecoxib on the third day after implantation.The mice were killed on the 15th day,and the volume and weight of the tumor tissues were calculated.Results Compared with control group,50,100 and 200 μmol•L^-1 celecoxib inhibited the proliferation of Lewis cells (P<0.05),the inhibitory rate was  increased with the dose and time of treatment.The IC50 value of 72 h was (134.06±2.97)  μmol•L^-1.Compared with control group,celecoxib had no effect on the arachidonic acid content,but depressed the prostaglandin E2 level in the culture supernatants in 50  μmol•L^-1 treatment group (P<0.05). In 100 and 200  μmol•L^-1　 treatment groups,celecoxib  significantly increased the content of arachidonic acid and depressed the prostaglandin E2 level in the culture supernatants with the  increasing of the  dose  of celecoxib (P<0.01). The solid tumor volume and weight in treatment group were lower than those in control group (P<0.05), and the tumor inhiitory rate in treatment group was 50%.Conclusion Celecoxib can inhibit the growth of Lewis lung carcinoma cells in vitro and in vivo.The anticarcinogenic effect of celecoxib depends on not only increasing the prodcution of arachidonic acid,but also inhibiting the synthesis of prostaglandin E2 in Lewis lung carcinoma treated with celecoxib.

Abstract:Objective To label anti- c-erbB-2 monoclonal antibody  with ¹³¹I,and investigate its inhibitory effect on the growth of ovarian cancer in mude mice.Methods Anti-c-erbB2 monoclonal antibody was combined with ¹³¹I to produce radioimmunological targeting drug.The nude mice which had been inoculated subcutaneously with ovarian cancer SKOV3 cells and the diameter of tumor mass was about 1.0 cm were randomly divided into 3 groups:G₁,G₂ and control groups.The mice in G₁　and G₂ groups were treated with 11.1　MBq and 3.7　MBq anti-c-erbB2 monoclonal antibody labeled with ¹³¹Irespectively.The mice in control group was treated with mIgG labeled with ¹³¹I.The size and weight of tumor were determined once every week during 6 weeks after administration.Results Compared with  control group, the  size and weight of tumor in G₁ and G₂ groups were significantly decreased,and the inhibitory rate was increased(P<0.05). The size and weight of tumor in G₁　group were lower than those in G2 group(P<0.05),and the inhibirory rate in G1 group  was higher than that in G₂ group(P<0.05) .Conclusion Anti-c-erbB-2 monoclonal antibody labeled with radionuclide ¹³¹Icould inhibit the growth of ovarian cancer in nude mice and the effect of 11.1 MBq c-erbB-2 monoclonal antibody labeled with ¹³¹Iis better than 3.7 MBq  ¹³¹I-c-erbB-2.

Abstract:Objective
To screen the high chitinase-producing fungi from the samples in the environment,and investigate the inhibitory effect  of the enzymes from the strains on  the Candida  for the development of antifungal drug.Methods  Congo red-chitin plates and classical  DNS method were used to screen the high chitinase-producing strains,and  the filter paper containing the chitinase was used to detect the inhibitory effect of the enzymes.Results 6 chitinase-producing strains were screened from 47 strains.  2 high chitinase-producing strains（Aspergillus fumigatus JLC 50134 and Trichoderma sp.JLC 31235）were obtained. The enzyme from JLC 50134 had better inhibitory effect  on Candida albicans and Candida tropicalis,and the individual diameters of the inhibition zone were 4-5 cm and about 2 cm;while the enzyme from JLC 31235 had better inhibitory effect on Candida krusei and Candida glabrata than JLC 50134,and the individual diameters of the inhibition zone were about 2 cm and 1 cm. Conclusion High chitinase-producing strains (JLC 50134 and JLC 31235) are obtained ,and they have different inhibitiory effects on Candida.

Abstract:Objective
To observe the effect of Ciwujianoside B(C-B) on apoptosis induced by hydrogen peroxide (H₂O₂) in cardiomyocytes and the expressions of its related regulating proteins,and clanify  the inhibitory effect of C-B on apoptosis induced by injury of oxidative stress in cardiomyocytes.Methods The neonatal rat cardiomyocytes were primary cultivated and the cells which beat autonomously well were randomly divided into normal control group,H2O2 group,100  mg•L^-1 C-B group and 200  mg•L^-1 C-B group.The cardiomyocytes were exposed to 200  μmol•L^-1H₂O₂ for 120 min to induce apoptosis.The morphological changes of cardiomyocytes were observed by microscope,the survival rate was detected by MTT.The apoptotic rate was analyzed by  flow cytometry labelled with AnnexinⅤ-FITC. The activity of Caspase-3 was detected by spectrophotometric method.The expressions of Caspase-3,PARP,Bcl-2 and Bax proteins were determined by immunoblotting.Results Compared with normal control group,the injuried cardiomyocytes were seen and the survival rate was decreased significantly (P<0.001),the apoptotic rate and Caspase-3 activity were increased (P<0.001 or P<0.01);the expressions of Caspase-3,Bax and the degradation of PARP were raised,and the expression of Bcl-2 was decreased in 200  μmol•L^-1H₂O₂ group.Compared with 200  μmol•L^-1H₂O₂ group,the injuried changes of cardiomyocytes were reduced,the survival rates were significantly increased (P<0.05 or P<0.01),the apoptotic rates were decreased (P<0.001);the activities of Caspase-3 were decreased significantly  (P<0.05 or P<0.01);the expressions of Caspase-3 and Bax were decreased,the degradation of PARP was attenuated,and the expressions of Bcl-2 were increased in 100 and 200  mg•L^-1 C-B groups.Conclusion C-B can obviously inhibit the apoptosis induced by H₂O₂ in cardiomyocytes,its mechanism may be related to the regulation of the expressions of the apoptosis-related  proteins Caspase-3,Bax and Bcl-2.

Abstract:Objective
To investigate the expression level of glucose regulating protein 78(GRP78) in SH-SY5Y cells in the Parkinson’s disease(PD)  model induced by dopamine(DA)，and provide the theoretical basis for PD．Methods PD models  induced by DA in SH-SY5Y cells were established.The cells in experimental group were treated with DA,and the cells in control group were treated with  equal volume of 0.2% vitamin C.The cells in two groups were incubated for 24 h, the viability of SH-SY5Y cells was assayed by Trypan blue staining,Hoechst 33342 fluorescence staining was used to estimate the apoptotic percentage．The proteins of SHSY5Y cells treated with or without DA were extracted 24 h after incubation，and then the maps of the proteins were established by 2D-DIGE(differential gel electrophoresis,DIGE).The altered protein spots were identified with MALDI-TOF MS and database searching.Results The survial rate of SH-SY5Y cells  in experimental group(53.2%) was lower than that in control group(100%)(P< 0.05)．The Hoechst fluorescence staining result showed that the nuclei of SH-SY5Y cells in exprerimental group appeared karyopyknosis and fragmentation which was consistent with nuclear morphology.A high expression protein spot in experimental group was confirmed as GRP78 by MALDI-TOF MS． Conclusion  DA can induce the apoptosis of SH-SY5Y cells,and the expression of GRP78 is increased in the process of apoptosis．The results demonstrate that the increase of GRP78 may be involved in the pathogenesis of PD．

Abstract:Objective
To establish E.coli strain and express human fibroblast growth factor 21(FGF21),and   provide experimental foundation for development of  new drugs for treatment of  type 2 diabetes.Methods Human FGF21 gene fragments were obtanined by PCR.After the FGF21 was fused with SUMO (small ubiquitin-related modifier) by PCR,the fused gene was expressed in E.coli BL21(Rosetta).By inducing high levels of expression with IPTG,the soluble proteins were obtanined.The fused protein was purified by DEAE Sepharose and Ni-NTA affinity chromatography.Once cleaved from SUMO,the purity of human FGF21 by SDS-PAGE was shown to be higher than 95%.Results  Acquired gene fragments of hFGF21 were identified by digestion and DNA sequencing with the human FGF21 reported in GenBank (Accession no.NM019113 ).The recombinant vector of pET20b-SUMO-FGF21 was constructed successfully.SDS-PAGE result proved that SUMO-FGF21 fusion protein with a relative molecular mass of about 19 401 was expressed.Western blotting result showed that the expressed products had specific reaction with anti-human FGF21 polyclonal antibody.Conclusion The engineering E.coli strain expressing human FGF21 fusion protein is successfully established and the purified protein is obtained.

Abstract:Objective
To extract and purify anti-tumor peptide of tumor chalone T42 peptide from gene engineering bacteria pTYB2-T42 and  identify its biological activity,and  provide the experimental basis for the research of its function mechanism and the clinical treatment of tumors.Methods The pTYB2-T42 in E.coli was cultivated,and fusion protein was produced by IPTG,T42 peptide was extracted by the method of affinity chromatograph,DTT was removed by molecular sieve G15 and then T42 peptide was identified by Trincine SDS-PAGE.The influence of T42 peptide on the cell activity was detected by MTT.The apoptosis of T42 peptide was detected by AO/EB.Results  The recombinant expression vector coding tumstatin T42 peptide was correct in size,its relative molecular mass was about 5 000.The result of MTT showed that the proliferation of growth of Hepg2 cells was inhibited by T42 peptide,and the survival rate of Hepg2 cells was decreased with the increase of T42 peptide dose,and IC₅₀ was 30 mmol•L^-1.The result of AO/EB indicated that the Hepg2 cells appeared typical characteristics of apoptosis after treated with T42 peptide for 24 h.Conclusion  T42 peptide evidently inhibit the proliferation of growth of  Hepg2 cells  in a dose-dependent manner,and it can promote the apoptosis of Hepg2 cells.

Abstract:Objective To investigate the association between herpes virus infection and primary graft failure（PGF）after penetrating keratoplasty（PKP）in naive rabbits. Methods Animal model of herpes virus latency was established in rabbit eyes.PKP was performed in only one eye in two groups.In group A,seven naive rabbit eyes received latent donor,while seven latent rabbit eyes received naive donor in group B.The cornea was observed with slit lamp at 3,5,7,14 and 28 d after PKP respectively.Cornea swabs  were taken and detected for DNA of four viruses,including herpes simplex type 1 （HSV-1）,herpes simplex virus type 2 ( HSV-2),cytomegalovirus (CMV) and varicella-zoster virus (VZV) by multiplex PCR.Results Two eyes in group A (2/5,28.6%) were suffered PGF at 5 and 28 d after PKP, respectively.HSV-1 and VZV DNAs were detected by multiplex PCR from cornea swabs,grafts,rims and trigeminal ganglions (TGs),but HSV-2 and CMV were not detected.The cornea graft of remain 5 eyes in group A (5/7,71.4%) and 7 eyes in group B （7/7，100%） were clear until 28 d after PKP.The multiplex PCR results of cornea swabs were negative. Conclusion The herpes virus infection of cornea donor is the main factor of PGF after PKP.Compare to HSV-2 and CMV,the rabbit cornea is more easily infected by HSV-1 and VZV.The latent viruses in donor can be reactivated by PKP,and PGF is probably caused by VZV besides HSV-1.

Abstract:Objective
Through studying the impact of donor age on the human periodontal ligament cells（PDLC）of primary culture,to find the best donor age of the human PDLC of primary culture,and provid a theoretical basis for convenient,quick acquisition of a large number of human PDLC.Methods 68 adolescent(12-28 years old) health  premolar were collected,which were extracted for the reason of orthodontic in Department of Oral and Maxillofacial Surgery.Then the teeth were divided into three groups according to donor age,namely 12-14 years old group,15-18　 years old group and  19-28 years old group.The primary cells were isolated from human periodontal ligament and cultivated by tissue explant culture technique,then passaged when the cells completely covered the bottom of culture flasks.Morphological analysis and immunocytochemical analysis were used to identify the cell lineage.Results Human PDLC of primary culture were obtained with tissue cultivation,the success rates in various   groups were different,The success rates in  12-14　years old group,15-18 years old group and  19-28　years old group were 66.67%,8.57%,and  0,respectively. The immunohistochemical result showed that when the cells were cultivated to passage 3 in 12-14 years old and 15-18 years old  groups,they were positive for vimentin staining,and negative for keratin staining,and they were proved to be tissue-derived mesoderm.Conclusion Donor age has a significant effect on the success rate of human PDLC and 12-14 　years old is the best donor age.

Abstract:Objective
To build an airway remodeling model of bronchial asthma in mouse  with  house dust mite extract (DHM)，and to investigate the dose-effect and time-effect relationship.Methods Ninety  SPF  BALB/c  mice  were randomly divided into control group,low dose DHM group and high dose DHM group.Each group was randomly divided into three subgroups:three-week,six-week and nine-week groups.The mice was challenged with intranasal drip after sensitized with subcutaneous and peritoneal injection of DHM with differet doses.All mice were underwent pulmonary lavage in 24 h after the last challenge.the total inflammation cell count,eosinophil count in BALF,the IL-4 level and total serum IgE level were detected,and the internal perimeter (Pi),the wall thickness(d) and wall area(WA) of the bronchi were measured by computer image analytical system.Results In both high dose DHM and low dose DHM groups,the total inflammation cell counts and eosinophil counts,the levels of IL-4 and serum total IgE levels were significantly increased (P<0.05);the d/Pi and WA/Pi were statistically increased (P<0.05) compared with control group.The airway inflammation was increased significantly since the third week,and there was incremental change of morphological parameters in  high dose DHM group during 9 weeks.Compared with low dose DHM group,in high dose DHM group,the total inflammation cell and eosinophil counts were increased in 3 weeks( P<0.05),the levels of IL-4 and IgE were  increased in 3,6 and 9 weeks (P<0.05), while the d/Pi and WA/Pi had no significant change.
Conclusion DHM can induce airway remodeling model of bronchial asthma in mouse successfully,and the airway remodeling changes obviously with the prolongation of exposured time,and there is significant dose- and time-effect in the study.

Abstract:Objective To determine the protective effect of cardiopulmonary bypass(CPB)  with autologous lung as oxygenator on CPB-relative inflammatory response.Methods Twelve adult mongrel dogs were randomly divided into control group and experimental group.CPB using a membrane oxygenator (control group) or using the autologous lung (experimental group) for gas exchange was performed for 120 min in an alternating series of 12 mongrel dogs with 90 min heart arrest by crystalloid cardioplegia and 30 min reperfusion.The blood samples were collected at the same time point of pre-operation(T1)，60 min of CPB(T2)，and 1 h (T3),2 h (T4) after CPB．The plasma levels of IL-6，IL-10,TNF-α were detected with ELISA．  Results The plasma levels of IL-6，IL-10 and TNF-α of dogs in each group were significantly increased at T2,T3 and T4(P<0.01) compared with before CPB.The plasma levels of IL-6 and TNF-α in experimental group were significantly lower than those in control group at T2,T3 and T4(P<0.01).The plasma levels of IL-10 in experimental group were significantly higher than those in control group at T2,T3 and T4(P<0.05).Conclusion Extracorporeal circulation with autologous lung as oxygenator could reduce the increased amplitude of the plasma levels of TNF-α and IL-6，whereas enhance the increased amplitude of the plasma IL-10 level.The results indicate that extracorporeal circulation with autologous lung as oxygenator can reduce  the CPB-relative inflammatory respones.

Abstract:Objective To  compare the changes of   myocardial morphology,myocardial enzymology and hemorheology  between the acute myocardial infarction models  with blood stasis and the simple acute myocardial infarction models in rats  and elucidate the difference between the acute myocardial infarction models in rats with blood stasis and the simple acute myocardial infarction models in evaluation of pharmacodynamics.Methods Wistar rats were randomly divided into  control,acute blood stasis model,acute myocardial infarction sham operation,acute myocardial infarction model and  acute myocardial infarction model with blood stasis groups.The acute myocardial infarction models under the status of the acute blood stasis in rats were built to determine the parameters of myocardial infarct size (MIS) and the activities of serum aspartate aminotransferase (AST),creatine phosphokinase (CK) and lactate dehydrogenase (LDH),at the same time,platelet agglutinate rate (PAR),platelet aggregation(PAG),erythrocyte sedimentation rate(ESR),hematocrit(HCT) and the length,the dry weight,the wet weight of the thromb in vitro were tested,the parameters of thrombelastogram were also be detected.Results ①When the acute myocardial infarction models with blood stasis in rats and the simple acute myocardial infarction models were comparatively studied,the results showed that there were no obvious differences between them in MIS,the activities of CK,AST and LDH,and PAR and PAG（P>0.05）;② The increasing extent of the ESR of the acute myocardial infarction models with blood stasis in rats was much higher than that of the simple acute myocardial infarction models（P<0.05）,but there was no obvious difference between them in HCT（P>0.05）;③ The dry weight and the wet weight of the thromb in vitro of the acute myocardial infarction models with blood stasis in rats were  much heavier those  of the simple acute myocardial infarction models and the acute blood stasis syndrome models in rats（P<0.05 or P<0.01）,but there was no obvious difference between them in the length of thromb in vitro（P>0.05）;④ The increasing extent of the values of r,k and the reducing extent of the value of ma in the acute myocardial infarction models with blood stasis in rats were significantly higher than those in the simple acute myocardial infarction models（P<0.05）,but there was no obvious difference between the acute myocardial infarction models with blood stasis and the acute blood stasis syndrome models in rats（P>0.05).Conclusion The obvious differences are showed in the parameters of blood sedimentation and the weight of the thromb in vitro and the thrombelastogram,but there are no obvious differences in the indexes of MIS,the myocardium enzyme in  serum,HCT and the index of  function of the blood plate when the acute myocardial infarction models with blood stasis and the simple acute myocardial infarction models are comparatively studied.The results  show that it is  defective to evaluate pharmacodynamics of traditional Chinese drug only with  simple acute myocardial infarction models.

Abstract:Objective  To study the anti-tumor effect of recombinant human MUC1-MBP and mouse Muc1-MBP protein vaccines,and  provide the basis for clinical application of tumor vaccine.Methods The mice were divided three groups and respectively immunized with PBS  (negative control),human MUC1-MBP and mouse Muc1-MBP fusion protein.The cross reaction between anti-human MUC1 and anti-mouse Muc1 antibody was detected by ELISA.CTL-specific cytotoxicities of mice induced by MUC1 and Muc1 were detected by LDH.The anti-tumor effects of human MUC1 and mouse Muc1 were observed by establishing mice models of the lung cancer metastasis and the subcutaneous human breast cancer.
Results The cross reaction between the antibody of human MUC1 and the antigen of mouse Muc1 was stronger than that between the antibody of mouse Muc1 and antigen of human MUC1.The cytotoxicities of CTL from immunized mice by human MUC1-MBP and mouse Muc1-MBP to the MCF-7 cells were (48.7±7.1)% and (54.6±7.8)% or to LLC1 cells were (61.9 ±10.2)% and (43.5±8.4)%,respectively.The inhibitory rates of lung tumor nodules  of Lewis lung cancer in human MUC1-MBP and mouse Muc1-MBP groups were 94.4% and 68.5%,respectively.The average tumor volumes in human MUC1-MBP,mouse Muc1-MBP and PBS groups were  (23.5±15.7),(56.5±46.7) and (142.8±70.2) mm3,respectively,in the animal models of human breast cancer. The masses of tumor were obviously suppressed in MUC1-MBP group (P<0.01) and Muc1-MBP group (P<0.05) compared with   control group.Conclusion Human MUC1 and mouse Muc1 have common epitope of B and T cell and the human MUC1 could induce the stronger cross reaction.Human MUC1-MBP has significant anti-tumor effects than that of  mouse Muc1-MBP,and it  might be used to develop the protein vaccine against human carcinoma.

Abstract:Objective  To observe the effect of Tongxinluo on the expreeions of plasminogen activator inhibitor type-1 (PAI-1) and vascular cell adhension molecule-1 (VCAM-1) in rabbits with atherosclerosis(AS),and explore the possible mechanism.Methods Twenty four rabbits were randomly divided into control,cholesterol diet,and Tongxinluo groups.The levels of serum cholesterol(TC),triacylglycerol(TG),low density lipoprotein(LDL),PAI-1 and VCAM-1 were measured before and after treatment.The aortas were harvested for pathologic morphological observation.Immunohistochemistry analysis was used to evaluate the positive percentages of  PAI-1 and VCAM-1.Results There were no differences of TC,TG,LDL,PAI-1 and VCAM-1 between three groups before treatment （P>0.05）. 14 weeks after treatment,the TC,TG,LDL,PAI-1 and VCAM-1 levels in cholesterol diet group and Tongxinluo group were significantly higher than those in control group(P<0.01),while the TC,TG,LDL,PAI-1and VCAM-1 levels in Tongxinluo groups were signficantly lower than those in cholesterol diet group(P<0.01). The expressions of  PAI-1 and VCAM-1 in aorta in cholesterol diet and Tongxinluo group were higher than those in control group(P<0.01),and the expressions of PAI-1 and VCAM-1     in Tongxinluo group  were lower than those in cholesterol group(P<0.01).Conclusion PAI-1and VCAM-1 over express   in   AS rabbits,while inhibiting the expressions of PAI-1 and VCAM is likely to be one of anti-AS  mechanism of  Tongxinluo.

Abstract:Objective  To investigate the effect of purine nucleotides compensation on the levels of dopamine (DA) and norepinephrine( NE) in brain tissue in rats and its possible mechanisims for heroin addiction. Methods Sixty five male Wistar rats were randomly divided into five groups (n=13):control group(C),heroin group (H),heroin plus AMP and GMP group (HAG),heroin plus AMP group (HA),heroin plus GMP group (HG).The levels of DA and NE in brain tissue were examined by fluorospectrophotometry.The activity of tyrosine hydroxylase (TH) in brain tissue was detected by ELISA.The TH expression was determined by real-time PCR. The ultrastructure changes of VTA were observed by transmission electron microscopy.Results The levels of DA,NE,TH and the expression level of TH in brain tissue in H group were significantly decreased compared with C group (P< 0.05).While in HAG group,the DA,NE and TH levels were increased when compared with H group (P< 0.05). In HA group,the TH level was increased when compared with H group (P< 0.05).In HAG,HA and HG groups,the  TH expression levels  were increased when compared with H group (P< 0.05).Under electron microscope,the impairment in HAG,HA,and HG groups was less than that in H group.Neuropil swelled and medullary sheath was detached in H group.Autosome was increased in HAG,HA and HG groups.Conclusion Purine nucleotides compensation can partly alleviate the impairment which caused by heroin and increase the contents of DA and NE in brain tissue in heroin dependent rats.

Abstract:Objective  To observe the inhibitory effect of 3,4,5-hydroxy benzoic acid (TBA) on H₂₂ transplanted tumor in mice and  its influence in survival time and antioxidant capacity in vivo of H₂₂ tumor bearing mice .Methods H₂₂  tumor bearing mouse models were established.The H₂₂ tumor bearing mice were randomly divided into  control group,positive control group,TBA low dose group,TBA middle dose group,and TBA high dose group.There were 10 mice in each group.0.5% sodium carboxymethyl cellulose solution 2 mL·10 g^-1 was administered by oral in control group.CTX 20 mg·kg^-1 was injected intraperitoneally in positive control group.TBA was administered by oral in TBA low (0.13 g·kg^-1),middle(0.25 g·kg^-1) and high(0.50 g·kg^-1)dose groups.The H₂₂  tumor bearing mice were administered once a day at fixed time and  lasted  for 10 d.Then,the tumor  weight,the survival time,and  the SOD activity,MDA and GSH levels in serum in each group were detected.Results Compared with control group,the average tumor weights were  reduced in TBA middle dose group and CTX group (P<0.05)，there was no significant difference between other groups;compared with control group,the mean survival time in TBA middle dose group and CTX group  were longer (P<0.05),there was no significant difference between other groups;compared with control group,the SOD activities  in TBA middle and high dose groups were significantly increased(P<0.05);the MDA level  in TBA middle dose group was significantly decreased(P<0.05);the GSH levels were in TBA low,middle,high dose groups were significantly increased(P<0.05). Conclusion  TBA could inhibit the growth of H₂₂  tumor  and prolong the survival time of tumor bearing mice by oral administration. TBA could clear the free radicals in tumor bearing mice and up-regulate the  oxidative stress status of body.

Abstract:Objective  To observe the injury of skeletal muscle and apoptosis following limb ischemia reperfusion (LIR) in rats and investigate the protective mechanism of endothelin-A receptor antagonist BQ123 on the injury of skeletal muscle following LIR. Methods  Male Wistar rats were randomly divided into three groups (n = 8)：control group,IR group,BQ123+IR group.The contents of lactate dehydrogenase (LDH),creatine phosphate kinase (CK),malondialehyde (MDA)，initric oxide (NO),endothelin-1 (ET-1),and NO/ET-1 in plasma and the values of MDA,myeloperoxide(MPO),NO,ET-1,NO/ET-1,the percentage of DNA Chain% in  gastrocnemius tissues were measured.Immunohistochemistry (IHC) method was used to detect the expression of Caspase-3 in gastrocnemius tissues.Apoptosis was examined by means of TdT-mediated dUTP nick end labeling(TUNEL).Results  Compared with  IR  group,the levels of LDH,CK,MDA，ET-1 in plasma  in BQ123+IR group were decreased（P<0.01); the content of NO in plasma and the percentage  of NO/ET-1  were increased（P<0.01）,but the expression of Caspase-3 was obviously decreased （P<0.01）;the contents of MDA,MPO,ET-1 in gastroenemius tissues were decreased(P<0.01),the NO level,NO/ET-1,and the percentage of DNA chain were increased(P<0.05 or P<0.01);the expression of Caspase-3 in gastrocnemius tissues was signficantly decreased,and the apoptotic index (AI) in BQ123+IR group was also decreased （P<0.01）.Conclusion The endothelin-A receptor antagonist BQ123 hase protective effects on skeletal muscle  injury and apoptosis following  LIR in rats.

Abstract:Objective To observe the protective effects of ATP and panax ginseng triol set on the denerved muscle atrophy and motor end plate,and provide basis for the selection of operation opportunity in patients with never injury.Methods Adult male Wistar rats were used. The model of denervated gastrocnemius muscle was established by cutting off the sciatic nerve.The rats were randomly divided into control group,ATP group,low dose panax ginseng triol set (25 mg·kg^-1) group and high dose panax ginseng triol set (50 mg·kg^-1) group. During four weeks after operation, ATP was injected into gastrocnemius and the panax ginseng triol set was injected into belly. The muscle wet weight，diameter of myocytes，cross section area of myocytes and the morphological changes of motor end plate were measured 2,4,6,8,12,16,20, and 24 weeks after operation.  Results  After nerve injury, the muscle wet weight，diameter of myocytes，cross section area of myocytes were decreased. During 16 weeks after nerve injury, there were significant differences between ATP group , low dose panax ginseng triol set group, high dose panax ginseng triol set group and  control group(P＜0.01). At 20 weeks after nerve injury, there was no difference between ATP group and control group, but there were differences between low dose panax ginseng triol set group, high dose panax ginseng triol set group and  control group(P＜0.05). At 24 weeks after nerve injury, there were  no differences between  four groups. There were no significant changes of motor end plate during  4 weeks after nerve injury. Four weeks later, the motor end plate lose its normal morphous and changed into neurofilaments. Until 16 weeks after operation，the neurofilaments began to decrease. The number of neurofilaments in ATP and panax ginseng triol set groups was more than that in control group.  Conclusion  The denerved muscle atrophy and motor end plate cataplasia could be deferred by using ATP and panax ginseng triol set.

Abstract:Objective To investigate the effects of Shugan Jieyu Panacea (SJP) on behavior and the levels of ACTH in plasma and T3,T4,TSH,rT3 in serum in depression model rats and explore the mechanism.Methods The model rats were lonely fed and received chronic moderate intensive unpredictable stimulation.Normal  control group,depressed model group,high dosage SJP group,middle dosage SJP group,low dosage SJP group and fluoxetine group were set up.Defferent drugs were used in various groups for 21 d,then the body mass,sugar consumption and the behavior changes of the rats were determined and the levels of ACTH in plasma and T3,T4,TSH,rT3 in serum were detected with radioimmunoassay. Results Compared with normal group,the body mass was  decreased（P<0.01）,the scores of movement and sugar consumption were decreased（P<0.01）in model group. Compared with model group ,the body mass was increased（P<0.01）,the scores of movement and sugar consumption were increased（P<0.01）,the levels of ACTH,T4,rT3 markedly decreased （P<0.05 or P<0.01） and the level of T3 was increased（P<0.05） in high,middle,low dosage SJP groups after treatment. At the same time,there was  no obvious difference between  SJP groups and  fluoxetine groups.Conclusion SJP can significantly improve the depression in rats,its mechanism may be connected with adjusting the function of HPAA and HPTA.

Abstract:Objective To observe the the protective effect of combination of low dose fosinopril and erbasartan  on experimental myocardial infarction in rats and its mechanism.Methods Wistar rats were randomly divided into sham operation,model,fosinopril (20 mg·kg^-1),erbasartan (100 mg·kg^-1) and  fosinopril + erbasartan(10 mg·kg^-1+50 mg·kg^-1) groups.The acute myocardial infarction models were established with ligation of left anterior descending coronary artery in rats except  sham operation group. The effects of fosinopril and erbasartan and  combination of fosinopril and erbasartan on myocardial infarct size (MIS) and the activities of serum creatine phosphokinase (CK),lactate dehydrogenase (LDH) and aspartate aminotransferase (AST),the contents of plasma endothelin (ET) and angiotensinⅡ(AngⅡ), the level of myocardial free fatty acid (FFA) in ischemic and non-ischemic area,and the viscosities of blood and plasma were determined.Results Compared with  model group,the MIS in  sham operation,fosinopril,erbasartan and  fosinopril + erbasartan groups was significantly reduced (P<0.05　 or P<0.01),the activities of serum CK,LDH and AST were significantly decreased (P<0.05 or P<0.01),the contents of plasma ET and the levels of myocardial FFA in ischemic area were significantly decreased (P<0.05),and the viscosities of blood and plasma were also significantly decreased (P<0.05),but the contents  of plasma AngⅡ and the levels of myocardial FFA in non-ischemic area had no significant changes.Conclusion Low dose fosinopril combined with  erbasartan  has protective effect on acute myocardial infarction in rats,just as high dose fosinopril or erbasartan,by reducing damage of FFA and ET on myocardium,improving myocardial metabolism,correcting state of bloodstream in myocardial ischemia and preventing thrombosis etc.

Abstract:Objective To investigate the relationship between PON1 rs2299262 htSNP polymorphism and cerebral infarction in Chinese northern Han population,and to illustrate whether rs2299262 is a genetic marker of large artery atherosclerotic infarction.Methods A case-control study was performed including 529 cerebral infarction patients and 305 normal subjects.In addition,the cerebral infarction patients were sub-grouped into 304 large artery atherosclerotic infarction patients and 225 lacunar infarction patients based on TOAST standard.PON1 rs2299262 genotypes were determined by PCR-RFLP.Results Compared with control group,the rs2299262 T allelic frequency had significant difference in large artery atherosclerotic infarction group(P<0.05).There were no significant differences of the distribution of rs2299262 T allelic frequency between case and control groups （P>0.05）.When they were divided by gender,there was no difference of the distribution of PON1 rs2299262 allelic frequency and genotypic frequency between the subgroups and control group （P>0.05）.Conclusion The patients with PON1 rs2299262 T allele have a higher risk of large artery atherosclerotic infarction than without it.PON1 gene rs2299262 is probably a genetic marker of large artery atherosclerotic infarction with high risk.

Abstract:Objective To investigate the curvature radius of distal femur and hypocondyle by imageological methods and compare the differences between them and gender differences between male and female,and provide morphological basis for design of knee joint prosthesis.Methods  69 cases of normal knee undergoing magnetic resonance imaging in knee extensor 180 ° position were selected in sagittal images,the anterioposterior dimension (AP),rd and rh were measured and the medial curvature index(MCI),lateral curvature index(LCI),distal condyle curvature index (DCI) and hypocondyle curvature index(HCI) were used as the curvature standards of distal femur and the femoral posterior condylar.Results The rd of male medial condyle was (32.58±4.84)mm,rh was (19.04±3.33)mm;rd of male lateral condyle was (32.82±3.95)mm,rh was (19.38±2.80). The rd of female medial condyle was (29.40±4.15)mm,rh was (15.99±2.38)mm;the rd of female lateral condyle was (29.30±3.16)mm,rh was (16.41±2.01)mm. The data showed statistical difference in MCI and LCI between male and female (P<0.05).There was statistical difference between female DCI and HCI (P<0.05).The result of  two-variable linear regression analysis showed the linear regression between AP,rh and rd (P<0.001).Conclusion There are statistical differences in distal femur and hypocondyle curvature between male and female.The curvature of female is larger,especially the medial hypocondyle.It is suggested that the asymmetry curvature design should be taken for the prosthesis’s hypocondyle part,that is,the curvature of medial distal condyle should be increased to form a continuous radius design with the medial hypocondyle which has larger curvature.

Abstract:Objective To investigate the lumbar vertebrae parameters of deer,sheep and human via anatomy appearance and  curvature  in animal models,and explore the posibility of deer and sheep as vertebrae model.Methods 30 fresh  samples  of  deer,sheep,human vertebrae were chosen.SPSS 17.0 software was used to statistically analyze the data get from  sliding caliper and CT.Results The  forward,middle,behind  heights of human vertebrae were 25.76,21.08 and 26.3 mm,in deer they   were 37.75,41.10 and 42.72 mm,in sheep they were  31.24,32.70 and 33.78 mm.The  upper,middle,inferior diameters of  human vertebrae were  49.91,44.26, and 52.85 mm,in deer they were  28.04,23.51 and 32.82 mm,in sheep they were  24.52,21.44 and 28.12 mm.The  upper,middle,inferior radius of human vertebrae were  36.42,32.43 and 36.48 mm,in deer they were  23.07,16.71 and 21.09 mm,in sheep they were  16.92,12.96 and 16.87 mm. The vertebral foramens in coronal radius and sagittal radius in human were 23.40 and 15.62 mm,in deer they were  16.10 and 12.31 mm,in sheep they  were  14.05 and 9.13 mm.The pedicle of vertebral arch in heigh and width in  human were  15.72 and 13.78 mm,in deer they were  15.72 and 13.78 mm,in sheep they were  24.97 and 6.80 mm.The  upper,inferior, middle curvature radius in  human were  2.76,23.78 and 19.98 mm,in deer they were  22.26,23.27 and 32.91 mm,in sheep tyey were  15.37,16.89 and 27.48 mm. The curvature radius  coronal and sagittal in  human were  22.83 and 39.57 mm,in deer they were  27.48 and 21.49 mm,in sheep they were  21.70 and 24.03 mm.Conclusion The vertebrae and vertebral foramen of deer have similarity at length,width,curvature,lamina of vertebral arch with human being.The height of vertebrae,pedicle of vertebral arch,radius of curvature of sheep have similarity with human being.The vertebrae of deer and sheep have similarity at morphometry with human being,the vertebrae of deer is better used as a model at pedicle of vertebral arch,the vertebrae of sheep is better used as a model at vertebrae lateral wall.

Abstract:Objective To assess the efficacy and safety of quetiapine and haloperidol in the treatment of schizophrenia and the differences in adverse effects of them．Methods Meta analysis  on  the 14 papers about efficacy and safety of quetiapine and haloperidol in the treatment of schizophrenia was performed,and  the total effective rate,cure ratative,obvious effective rate and adverse effects were evaluated.Results ①Haloperidol and quetiapine had similar total effective rate (P=0.76);②there was significant difference of curative rate between haloperidol and quetiapine in treatment of schizophrenia(P=0.04);③Compared with haloperidol,quetiapine had a higher obvious effective rate(P=0.006);④the  incidence rates of akathisia,myotonia,dry mouth etc in quetiapine group were lower than those in haloperidol group (P<0.05).Conclusion Quetiapine is better than haloperidol on the cure rate and obvious effective rate in the treatment of schizophrenia．Further,quetiapine has less extrapyramidal reactions.

Abstract:Objective To investigate the dissociative effect of amikacin sulfate on platelet aggregation in patients with EDTA-dependent pseudothrombocytopenia (EDTA-PTCP),and develop an effective method for accurate platelet counting in this kind of samples. Methods Amikacin sulfate was added or not to the EDTA-K2 anticoagulated tube before collecting venous blood in an infrequent patient with EDTA-PTCP,and the platelet counts and other blood cell parameters at different time after blood withdrawal were examined by automatic cytoanalyzer. Amikacin was added at different time after collecting EDTA-K2 anticoagulated blood samples,and dissociative effect of amikacin on the platelet aggregation and the platelet morphological changes on the blood smear were observed.Results The platelet counts of EDTA-PTCP blood sample without amikacin were gradually reduced from 117×10⁹ L^-1 to 41×109 L^-1 with extending time (0.5-4.0 h) after blood withdrawal.In the blood sample pre-supplemented with amikacin,the platelet counts were within range of the normal reference values,and the platelet counts were relatively stable within 4 h at room temperature after blood drawn.When amikacin was added within 30 min after blood sampling,the platelet counts （186×10⁹ L^-1 -191×10⁹ L^-1 ）were within the  range of the normal reference values,the obvious platelet dissociation on the blood smear were found.When amikacin was added over 30 min after blood drawn,the platelet counts were gradually reduced from 168×10⁹ L^-1  at 40 min after standing  to 42×10⁹ L^-1 at 4 h after standing,and smear examination revealed significantly the platelet aggregation.Conclusion Amikacin sulfate is added either before blood withdrawal or within 30 min after blood sampling,the platelet counts of the patient with EDTA-PTCP examined by automatic cytoanalyzer are accurate and the determination of other blood cell parameters is not influenced.

Abstract:Objective To investigate the effectiveness and safety of administration of parecoxib-sodium on the patients with acute slightly  head injury for postoperative analgesia.  Methods  60 patients with slightly head injury,ASAⅠorⅡ,aged 18-78 years,scheduled for acute neurosurgical operation were randomly divided into two groups（n=30）:parecoxib-sodium group received intravenous parecoxib -sodium 40 mg immediately after operation and control group received intravenous equivalence saline at the same time.The tramadol hydrochloride consumption at 24 h after operation,the number of unsatisfied demand and the number of successfully delivered doses were observed,and the intensity of pain was measured using VAS scores;the adverse effects,the patients’ global evaluation of the postoperative analgesia and blood coagulation function before and after administration were recorded and compared between two groups. Results The tramadol hydrochloride consumption at 24 h after operation,the percentage of the patients who needed postoperative analgesia,and the VAS scores of the patients after operation in parecoxib-sodium group were significantly lower than those in control group(P<0.05).At the same time,the percentage of the patients in parecoxib-sodium group who considered the postoperative analgesia satisfied was higher than that in control group(P<0.05). And the percentage of patients’ adverse effects in parecoxib-sodium group was lower than that in control group(P<0.05).There was no significant difference in coagulation between parecoxib-sodium group and control group(P>0.05). Conclusion Parecoxib 40 mg given in vein after neurosurgical operation can reduce the tramadol hydrochloride consumption and improve postoperative analgesia in patients with acute slightly head injury．

Abstract:Objective To evaluate the efficacy and safety of vandenafil combined with mental intervention in aged male kidney transplant recipients with erectile dysfunction(ED).Methods        
One hundred and twenty male patients(aged more than fifty years)who had received kidney transplantations at least one year before and whose serum creatinine was under 150 μmol·L^-1 were selected randomly in the study.Their sexual function was assessed with IIEF questionnaire,and those with ED were treated with oral vardenafil combined with mental intervention for six months.The efficacy was assessed with IIEF.Its safety was appraised by measuring serum creatinine levels,creatinine clearances,liver function and concentrations of CSA and FK506.Results Seventy-eight aged men with ED received oral vardenafil combined with mental intervention for six months.IIEF score was  improved from 11.2±4.2 (before treatment)to 23.3±4.6(after treatment)（P＜0.05）;but renal function,liver function, FK506 and CSA concentrations remained unchanged.There were no significant adverse effects. Conclusion Oral vardenafil combined with mental intervention is effective and safe  for the treatment of ED in aged male kidney transplant recipients.

Abstract:Objective To   study  the levels of the population-based urinary polycyclic aromatic hydrocarbons (PAHs) metabolites,  1-hydroxypyrene glucuronide (1-OHPG) and 2-naphthol,  in polluted cities with different levels of PAHs,and to clarify the feasibility of 1-OHPG and 2-naphthol as an evaluation of PAHs pollution and health hazards on the population.Methods 60 students and their parents from the three cities (Datong,Changchun and Kunming) were chosen via cluster random sampling method to fill in questionnaire and  their urinary  1-OHPG and 2-naphthol levels were measured. Results The levels of urinary 1-OHPG in students （10.94 μmol·mol^-1±6.58  μmol·mol^-1 creatinine）and parents （5.87 μmol·mol^-1±6.87  μmol·mol^-1 creatinine）in Datong were  higher than those in Changchun (students:6.33 μmol?mol-1±8.54  μmol?mol-1 creatinine;parents:4.60 μmol·mol^-1±4.08  μmol·mol^-1 creatinine）and Kunming (students:1.90 ·mol^-1±3.57  μmol·mol^-1 creatinine;parents:1.62 μmol·mol^-1±2.44  μmol·mol^-1 creatinine）（P<0.01）.The levels of urinary 1-OHPG in students and parents in Changchun were higher than those in Kunming(P<0.01).Moreover,in the  children and their  parents,there were  no significant differences of the urinary 2-naphthol levels 　between three cities（P>0.05）. Conclusion The urinary 1-OHPG level could proper
ly represent the pollution condition of PAHs.It could be used as a biomarker to evaluate the population harm of PAHs pollution.

Abstract:Objective To illustrate the genotypes and distribution of the extended-spectrmu β-lactamases (ESBLs) of gram-negative bacterial isolates from several hospitals in Changchun,Jilin,China and to provide the reliable basis for antibiotic application. Methods Drug-resistant genes associated with ESBLs in gram-negative bacterial isolates were amplified by PCR. PCR products were sequenced and compared with the genotypes reported in  GenBank.Results  68 strains of 165 isolates exhibited drug resistance such as  TEM-type (12.12%),CTX-M-1 type (4.24%),SHV-type (3.64%) and CTX-M-9 type (3.03%).The multi-drug resistant genes including SHV + TEM-type,TEM + CTX-M-1 type,TEM + CTX-M-9 type,TEM + CTX-M-1 + CTX-M-9 type and SHV + TEM + CTX-M-1 + CTX-M-9 type were also detected. The positive PCR amplification products were  randomly selected and  sequenced,the SHV-11 subtype,TEM-1 subtype,CTX-M-3 subtype and the CTX-M-15 subtype,CTX-M-14 and CTX-M-18 subtype were obtained. Conclusion The genotypes of ESBLs in gram-negative bacterial  isolates mainly include TEM-1 subtype,CTX-M-14 subtype,CTX-M-18 subtype,SHV-11 in Changchun area.