Abstract:Objective
To study the effects of the cotreatment of cadmium chloride (CdCl2) and Smac over-expression on proliferation and apoptosis of hepatocellular carcinoma cells,SMMC-7721,and provide an experimental basis for the Smac-based tumor gene therapy and the development of cadmium as the antihepatoma drug.Methods SMMC-7721 were divided into five groups,which were control,pcDNA3.1+,pcDNA3.1+-hSmac,single CdCl2-treated and CdCl2 plus Smac groups respectively. The cells in control group had no transfection or CdCl2 treatment,while those in pcDNA3.1+ and pcDNA3.1+-hSmac groups were transfected with null vector pcDNA3.1+ or pcDNA3.1+-hSmac respectively through lipofectamine-mediated method,and those in CdCl2-treated groups were exposed to CdCl2 at the concentration of 10,20 and 30  μmol·L^-1 respectively,and those in CdCl2 plus Smac groups were transfected with pcDNA3.1+-hSmac firstly,and then treated with CdCl2 after transfection for 24 h.MTT assay was used to determine the effect of Smac over-expression and CdCl2 on proliferation,and both AO/EB fluorescence staining and FCM with Annexin Ⅴ and PI double-staining methods for apoptosis.Results The  MTT results showed that compared with control group,except for  the null-vector pcDNA3.1+ group,the inhibitory rates in the rest groups were increased significantly (P<0.01 or P<0.001).In  single CdCl2-treated groups,the inhibitory rate was increased with the increasing of treatment doses,whereas,each Smac coupled group presented significantly higher inhibitory rate than the corresponding single CdCl2-treated group (P<0.001).AO/EB staining manifested that the  cells in control group were in good condition,while those in pcDNA3.1+-hSmac,30  μmol·L^-1 CdCl2 treatment,and CdCl2 plus Smac groups appeared the  morphological changes of apoptosis,especially in the last group.Furthermore,the results acquired by FCM showed that the apoptotic rate in pcDNA3.1+-hSmac group was increased significantly when compared with control and pcDNA3.1+ groups respectively (P<0.001).However,the apoptotic rate in CdCl2 pluse Smac group was the highest,which was significantly higher than that in single 30  μmol·L^-1 CdCl2-treated group (P<0.001).Conclusion CdCl2 has cytotoxicity to SMMC-7721 cells.It could inhibit cellular proliferation in a dose-dependent manner,and induce apoptosis.Smac over-expression  could inhibit cellular proliferation and slightly induce apoptosis independently.

Abstract:Objective
 To investigate the effects of antidepressant venlafaxine on the apoptotic cell percentage,death rate and cell cycle of the hippocampus neurons injured by glutamate(Glu) and demonstrate the neuroprotection mechanism of   venlafaxine.Methods The primary cultured hippocampal neurons were used  to set up Glu injured model.Three groups were included in the experiment: control group,Glu model group and venlafaxine administration group.The changes in apoptotic cell percentage,death rate and cell cycle of neurons were detected respectively by FCM before and after administration with venlafaxine.Results The apoptotic cell percentage in Glu model group was increasd compared with control group (P<0.01),but venlafaxine could reverse it.Compared with Glu model group,in venlafaxine administration group the    apoptosis was suppressed at low concentrations,the apoptotic cell percentage was decreased at 10  μmol·L^-1 venlafaxine （P<0.05),nevertheless,the   apoptosis was promoted at high concentrations,the apoptotic cell percentage was increased dramatically when treated with  100  μmol·L^-1 venlafaxine  (P<0.01).The  death rate of neurons in Glu model group was increased compared  with control group(P<0.05 ),the death rate of neurons  was reduced in venlafaxine administration group  compared with  Glu model group (P<0.05 or P<0.01).Compared with control group,the apoptotic cell percentage of G0/G1 phase in Glu model group was increased (P<0.001),the apoptotic cell percentage of S phase was decreased (P<0.01),the cell percentages of G_²+M phase   didn’t change significantly.The  apoptotic cell percentages in venlafaxine administration groups showed an decreasing trend at G₀/G₁ phase,which decresed to 64.5% and 50.8% of  that in Glu model groups when treated with  100 and 200  μmol·L^-1  venlafaxine (P<0.05).The apoptotic cell percentages of  S phase in venlafaxine administration groups were increased,which were 3.6,4.7 and 4.2 times of that in Glu model group when treated with 100,200 and 500  μmol·L^-1 venlafaxine respectively (P<0.01).Conclusion Venlafaxine can prevent the hippocampal neurons from the damage induced with Glu,elevate the neuron survival rate,reduce the LDH release,keep the cell membrane intact,and decrease the apoptotic cell percentage and death rate of neurons,which has the neuroprotection.

Abstract:Objective
To construct the  CCNL1 eukaryotic expression vector pcfDNA3.1（+）-CCNL1,pVAXⅠ-CCNL1 and pEGFP-C1-CCNL1 and identify the  expressions of  CCNL1 genes in MDA-MB-231 cells.Methods The eukaryotic expression plasmid pcDNA3.1（+）-CCNL1,pVAX1-CCNL1 and pEGFP-C1-CCNL1 were transfected into the MDA-MB-231　cells by Fugene HD Transfection Reagent.After 48 h  transfection,the differential expressions of pcDNA3.1（+）-CCNL1,pVAX1-CCNL1 and pEGFP-C1-CCNL1 in  MDA-MB-231　cells were detected by real time PCR and Western blotting.Results After 48 h transfection,there was green fluorescence only in  pEGFP-C1-CCNL1 by  fluorescent microscope,not in pcDNA3.1（+）-CCNL1 and  pVAX1-CCNL1; the expressions of mRNA and protein from high to low were pcDNA3.1（+）-CCNL1,pVAX1-CCNL1 and pEGFP-C1-CCNL1 recombinant vector in MDA-MB-231 cells.Conclusion The recombinant vectors pcDNA3.1（+）-CCNL1,pVAX1-CCNL1 and pEGFP-CCNL1  are constructed succesfully.The expression of pcDNA3.1（+）-CCNL1  in  MDA-MB-231 cells is higher.

Abstract:Objective
To investigate the effect of P38MAPK on apoptosis of HSC-T6 cell stimulated by acetaldehyde and the change of P38 protein expression and to provide the basis for clinical treatment of liver fibrosis.Methods Control group(HSC-T6 plus 10% calf serum containing DMEM medium),aldehyde group(acetaldehyde was added based on   control group) and experiment groups(adding inhibitor of P38MAPK SB203580 based on  aldehyde　group,the final concentrations were 4,8,16  μmol·L^-1) were set up in this experiment.The apoptotic rate was detected by FCM,the expression of phosphorylated P38MAPK(P-P38MAPK) was determined by Western blotting,the change of caspase-3 protein level was observed by SABC.Results Compared with control group,the apoptotic rate and caspase-3 positive expression rate of HSC-T6 cells  in  aldehyde  group  were  significantly                     decreased (P<0.05 or P<0.01),the level of P-P38MAPK was increased(P<0.01).After administration of SB203580,the apoptotic rate was increased gradually(P<0.01),the expression of P-P38MAPK was decreased(P<0.01),the positive expression rate of caspase-3 was increased(P<0.01),and all in the does-dependent manner.Conclusion SB203580 could promot the apoptosis of HSC-T6 cells,and the occurrence of apoptosis could be related to phosphorylation of P38MAPK and activation of caspase-3.

Abstract:Objective
To establish rat model of smoking-dosage cadmium and  study the effect of smoking-dosage cadmium on myocardial injury in rats,and to explore its molecular mechanism.Methods The rat models of smoking-dosage cadium were established as model group,control group was set up at the same time.The rats in 20,36 and 52 weeks were killed,then the contents of cadmium in heart and the changes of heart organ coefficient in rats were investigated.The contents of creatine kinase(CK),lactate dehydrogenase(LDH),glutathion(GSH),malonaldehyde(MDA) and activity of superoxide anion dismutase(SOD) in myocardial tissues were detected.The histopathological changes of myocardium were observed by light microscope with HE staining.RT-PCR was performed to examine the caspase-3,bcl-2 and bax mRNA expressions.Western blotting was used to determine the caspase-3,bax,caspase-9 and Fas protein expressions.Results Compared with control group,the contents of cadmium in model group were increased in 36 and 52 weeks（P<0.05）,the organ coefficient of heart in model group was significantly decreased in 52 weeks（P<0.05）,and the contents of CK and LDH in model group were increased obviously in 20,36 and 52 weeks（P<0.05）,and the activities of SOD in model group were decreased in 36 and 52 weeks（P<0.05）,the contents of GSH in model group were decreased in 20,36 and 52 weeks（P<0.05）,the contents of MDA in model group were increased in 20,36 and 52 weeks（P<0.05）.The myocardial cell disorder and focal hyperplasia of connective tissue were observed in model group by light microscope with HE staining.The results of  RT-PCR and Western blotting showed that  the expressions of caspase-3,bax and Fas were increased（P<0.05） and the expression of bcl-2 was decreased （P<0.05）.Conclusion Smoking-dosage cadmium can do harm to rat heart through  decreasing the cardiac enzymes and oxidative stress,at last result in the apoptosis of myocardial cells.

Abstract:Objective
 To study the features of Bifidobacterium breve carrying endostatin（ES）and interferon γ（IFNγ）and its anti-tumor effect   in mice with lung cancer.Methods The target genes ES and IFNγ were transferred into Bifidobacterium breve by plasmids and identified,then the Bifidobacterium breve marked by 3H-TdR were given into lung-cancer-bearing mice by caudal vein injection and by gavage.The experiment included control,Bifidobacterium breve without plasmid (B-b),Bifidobacterium breve with empty plasmid (B-b-pNZ44), Bifidobacterium breve with endostatin (B-b-pNZ44-ssEndostatin),Bifidobacterium breve with IFNγ (B-b- pNZ44- IFNγ) and Bifidobacterium breve conbination (B-b-pNZ44-ssEndostatin + B-b-pNZ44-IFNγ) groups.The distribution of  the Bifidobacterium breve,the changes of tumor size,the activity of splenic cytotoxic T cell (CTL),natural killer cell (NK),tumor necrosis factor-α (TNF-α) secretion activity and intratumoral micro-vessel density(IMVD) were detected.Results The Bifidobacterium breve carrying ES and IFNγ were successfully constructed,and accumulated inside of the tumor.The radioactivities of different organs were significantly decreased from the 3rd day compared with the 1st day(P<0.05);the radioactivities of tumor tissues were significantly increased with the prolongation of time from the 3rd day compared with the 1st day(P<0.05).The administration of Bifidobacterium breve carrying ES and IFNγ surpressed the growth of tumor,enhanced the activities of CTL,NK and the TNFα secretion activity and reduced the IMVD.Conclusion Bifidobacterium breve carrying ES and IFNγ can targetly implant in  tumor and have anti-tumor effect by immune enhancement and vessel growth inhibition.

Abstract:Objective
To observe the protective effect of adrenomedullin(ADM)  on the myocardial injury following limb ischemia reperfusion(LIR) in rats and its mechanism.Methods 30 Wistar rats were randomly divided into control group,LIR group and ADM+LIR group(n=10).The plasma and myocardial tissue concentrations of endothelin-1(ET-1) were determined by using radioimmunoassay.The levels of creatine kinase(CK),creatine kinase MB isoenzyme(CK-MB) in plasma and malondialdehyde(MDA),xanthine oxidate(XOD),superoxide dismutase(SOD) in myocardial tissue were measured in each group.The pathological changes in the myocardium were observed.Immunohistochemistry technique was used to determine the expression of ET-1 in myocardium.Results Compared with control group,the levels of ET-1,CK,CK-MB in plasma and ET-1,MDA,XOD  myocardial tissue were increased(P<0.01),and the content of SOD in LIR group was decreased (P<0.01).Compared with LIR group,the levels of ET-1,CK,CK-MB in plasma and ET-1,MDA,XOD of myocardial tissue in ADM+LIR group were decreased(P<0.05 or P<0.01),and the content of SOD was increased(P<0.05).The pathological results showed that the myocytes was irregular,the muscle fiber was dissolved,the myofilament was disorder,the marked vascular congestion was observed in LIR group.In ADM+LIR group,the myocardial injury was lightened.The immunohistochemical results showed that the expression of ET-1 was significantly decreased in ADM group compared with LIR group（P<0.01）.Conclusion ADM can protect the myocardium following LIR in rats and the protective effects may be related to anti-oxidation and inhibition of synthesis of  ET-1.

Abstract:Objective
To observe the effects of atorvastatin with different doses on the expression of nuclear factor-kappa B（NF-κB）in spinal cords of rats with experimental autoimmune encephalomyelitis(EAE) and  to explore its protective mechanism.Methods The Wistar  rats were divided into four  groups in the  experiment:control group, EAE control group,high dose  and low dose atorvastatin treatment groups. The rat models of EAE were established by induction with guinea pig spinal cord homogenate.The histological changes were observed under light microscope，then the  changes of incidence rate,latency,clinical symptoms and histology of EAE which were effected by atorvastatin with different doses were compared by method of quantization.And the expression of NF-κB was detected by immunohistochemistry.Results  Compared with  EAE control group,the latency in high dose atorvastatin treatment group was prolonged (P<0.05),the severity of disease in rats was reduced (P<0.05),the  NF-κB expression in the spinal cord was decreased (P<0.05);but   the latency,the clinical symptoms and the expression of NF-κB  in   low-dose atorvastatin treatment group were similar to EAE control group,the differences were not statistically significant (P>0.05).Conclusion High dose atorvastatin plays a protective role in rats with EAE and its mechanism may be involved in some impact on the immunological mechanism of the CNS.

Abstract:Objective
To study the monoclones of SC189 and discuss the role of caspase-8 in glioblastoma resistance to tumor necrosis factor related apoptosis-inducing ligand(TRAIL)-induced apoptosis.Methods The monoclones were obtained from SC189,then the acid phosphate assay was used to detect the cell death rate to TRAIL,the expression of FADD,DR5,DR4 and caspase-8 in monoclones were detected by Western blotting,and then the caspases cleavage was checked after treated with TRAIL.Results The death percentage of cells of the monoclones isolated from SC189 was -5.25%-45.8% after treated with different doses of TRAIL,4 monoclones were resistant cell lines,only one monoclone was sensitive to TRAIL.Western blotting detection showed that the expressrons of FADD,DR5 and DR4 were similar.The expression of caspase-8  in resistant cell lines was decreased,and the cells hadn’t the cleavage reaction,otherwise,the expression of caspase-8 in sensitive cell lines was increased,and the cells had the caspases and DFF-45 cleavage after treated with TRAIL.Conclusion The expression of caspase-8 in SC189 monoclones is directly related to the sensitivity of TRAIL.

Abstract:Objective
 To establish rat model of immune tolerance induced in embryonic stage and to explore the possibility of replacing for human leukocyte antigen （HLA ）matching in hematopoietic stem cell transplantation.Methods The rats were randomly divided into four groups：induced tolerance group,control group,irradiation-transplantation group and irradiation group.Each group had 20 Wistar rats and 20 SD rats.Host female Wistar rats’ peripheral blood mononuclear cells were injected in yolk sac of SD rats during pregnancy to induce immune tolerance.The host female Wister rats were irradiated with lethal dose and accepted bone marrow transplantation from the induced mature male offspring.The survival and the graft- versus- host disease（GVHD） were observed after transplantation.Meanwhile the skin was used to observe the engraftment of solid organ.Results The 30 d　survival rate of the rats accepted the bone marrow of the immune tolerant donor(86.7%)  was significantly longer  than that in control group (0%)(P<0.01),and there was no pathological GVHD occurred. While in control group,the incidence of GVHD was clearly visible.The skin engraftment rate between the host and the immune tolerant donor in induced tolerance group was significantly higher than that of non-related rats (84.6% vs 0%).Conclusion The induction of immune tolerance in embryo can cross the HLA barrier and offer a good donor for hematopoietic stem cell transplantation and solid organ transplantation

Abstract:Objective
To investigate the effects of isoflurane(ISO) preconditioning on the expressions of TNF-α and iNOS mRNA in ischemia reperfusion myocardium of  young rabbits and clarify their mechanisms.Methods 36 young rabbits were randomly divided into three groups,the rabbits in sham operation group were not occluded coronary artery;the rabbits in ischemia-reperfusion (I/R) group were occluded for 30 min and reperfused for 2 h;the rabbits in ISO  preconditioning group were inhaled 1.1% isoflurane for 30 min before ischemia,others were the same to  I/R  group after 15 min.The content of myocardial TNF-α and the expression of iNOS mRNA in myocardioum were determined 30 min after coronary occlusion and 2 h after reperfusion. Results Compared with sham operation group,the contents of myocardial TNF-α in I/R and ISO groups were significantly increased(P<0.05);the content of myocardial TNF-α in ISO group was significantly lower than that in I/R group(P<0.05).The expressions of iNOS mRNA in I/R and ISO groups were significantly higher than that in sham operation group(P<0.05),the expression of iNOS mRNA in ISO group was significantly lower than that in I/R group(P<0.05).Conclusion ISO could reduce the TNF-α content and the expression of iNOS mRNA during I/R injury,which may be one of molecular mechanisms of  ISO  preconditioning in myocardial protection.

Abstract:ObjectiveTo observe the expression of  soluble resistance-related calcium binding protein(sorcin) in the model of Parkinson’s disease (PD) induced by 1-methyl-4-phenyl-pyridinium ion (MPP+) in SH-SY5Y cells,in order to identify proteins related to degeneration of dopaminergic neuron for revealing the potential pathogenesis of PD.Methods The undifferentiated SH-SY5Y cells were treated with 1.0 mmol·L^-1 MPP+ for 24 h to set up in vitro PD cell models.Then,quantitative two-dimensional difference in-gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was performed to identify the differentially expressed proteins in PD models.Results Proteomics analysis revealed that 22 proteins were differentially altered in MPP+-treated SH-SY5Y cells,of which one protein spot was nambiguously identified as sorcin.Sorcin showed a significant increase in level in this PD model,and the fold changes in protein expression between MPP+-treated SH-SY5Y cells and controls was 1.97 (P<0.01).Its coverage,expectation and theoretical Mr and pI was 25.7,0.006,21 940 and 5.3,respectively.Conclusion
The function role of sorcin is correlated with calcium homeostasis and apoptosis,which may be pertinent in the toxicity induced by MPP+ in SH-SY5Y cells and  underlying pathogenesis of PD.

Abstract:Objective To clone the GRA1 gene and construct the prokaryotic expression vector PET-28a-GRA1,and to transform the expression vector  into E.coli BL21,then  induce and identiy,and to lay a foundation for the study on the biological characteristics and immune protection of GRA1.Methods A pair of primers were designed according to the sequence of GRA1 from GenBank.The GRA1 gene was amplified by PCR.The amplified GRA1 gene was inserted into prokaryotic expression vector PET-28a,and the constructed recombinant plasmid PET-28a-GRA1 was transformed to E.coli BL21 for expression under induction of IPTG.The expressed product was identified by SDS-PAGE and Western blotting.Results:The sequencing result showed the homology of 100% of the cloned GRA1 gene to that reported in GenBank.The positive recombinant plasmids PET-28a-GRA1 were identified by restriction endonuclease digestion and PCR,the objective gene fragment with the size of 573 bp was acquired,in accordance with the expected results,and the recombinant plasmid PET-28a-GRA1 was constructed successfully.The results of SDS-PAGE  revealed that the molecular  weight of recombinant protein PET-28a-GRA1 was 24 000 and Western blotting proved that the recombinant protein was recognized by murine antisera against Toxoplasma gondii.Conclusion The GRA1 gene has been successfully cloned,the recombinant
 plasmid PET-28a-GRA1 is generated and expressed highly in prokaryote.

Abstract:Objective To investigate the effects of signal transducer and activator of transcription 3 (STAT3) on the invasiveness of pancreatic cancer cell line SW1990 and its mechanism.Methods  The SW1990 cells were divided into control group,negative group and siRNA group.The cells in siRNA group  were transfected with STAT3 siRNA,the expressions of STAT3 mRNA and protein were assayed by RT-PCR and Western blotting;the invasive ability of cancer cells was detected by Transwell assay.The expressions of MMP-2 and MMP-9 of pancreatic cancer cells were detected by RT-PCR and Western blotting.Results After transfected with STAT3 siRNA for 72h,the expressions of STAT3 mRNA and protein were significantly decreased,and there were significant differences compared with control group and negative group (P<0.05).The number of invaded cells in siRNA group was decreased significantly compared with  control group and negative group (P<0.05).At mRNA and protein levels,the expressions of MMP-2 and MMP-9 were significantly decreased in siRNA group,and there were significant differences compared with control group and negative group (P<0.05).Conclusion STAT3 siRNA could specifically block the activaty of STAT3 signal on SW1990 and  also inhibit the invasiveness of pancreatic cancer cells by down-regulating the MMP-2 and MMP-9 expressions.

Abstract:Objective To study the anti-tumor effects of cadmium chloride (CdCl₂) and the influence in proliferating cell nuclear antigen (PCNA) and metallothionein(MT) expressions,and to  provide theoretical basis for development of CdCl₂ as anti-tumor drug.Methods Five kinds of carcinoma cell lines,SMMC-7721,BEL-7402,Hela,A549 and MCF7 were treated with 1,5,10,50 and 100  μmol·L-1 CdCl₂ for 24 h,MTT assay was used to measure the inhibitory rate. SMMC-7721 cells were treated with 5,10,20,40 and 80 μmol·L-1CdCl₂ for 24,48 and 72 h,MTT assay was used to detect the time-dose-effect relationship.SCGE was applied to observe DNA damage.Immunocytochemistry assay was performed to detect positive expressions of PCNA and MT.Results The malignant hepatocarcinoma SMMC-7721 cell line showed a significant sensitivity to CaCl₂ with a time-dose-effect relationship (IC₅₀=12.43 m L^-1).Compared with control group,the inhibitory rates in 40 and 80  μmol·L^-1 groups were significantly decreased(P<0.01).Compared with  control  group,after treated with CdCl2 for 12 and 24 h,the DNA damage rates in CdCl2 groups were  increased(P<0.05 or P<0.01) with the increase of CdCl2 except for 80 μmol·L^-1 group.Compared with control
 group,the expressions of PCNA in CdCl2 groups were  decreased (P<0.05 or P<0.01) with the increasing of CdCl2. SMMC-7721 could hardly express MT, but the expression could be induced by CdCl2 with a dose-effect relationship.Conclusion CdCl2 shows a significant anti-tumor effects in vitro,and  cause proliferation depression through inducing DNA damage and inhibiting PCNA protein expression,and its
 side effects may be alleviated by inducing MT expression.

Abstract:Objective  To study the antagonism of zinc sulfate on damage effect of ultraviolet  B (UVB) on HaCaT cells,and to provide theoretical basis for development of drug to prevent UVB damage. Methods The HaCaT cells were cultivated in RPMI 1640 and randomly divided into control group,Zn group,UVB group and Zn+UVB group.Nothing was administrated to the  cells in control group,the cells in UVB group  were radiated by UVB,the cells in Zn group  were administrated with 50 μmol·L^-1 ZnSO4 and the  cells in  Zn+UVB group were administrated with 50 μmol·L^-1 ZnSO4 24 h before UVB radiation.The  UVB radiation time was 6 min,the  cells in every group were collected 24 h after UVB radiation.The cell viability was measured by trypan blue exclusion assay,the  MDA level was measured by TBA method,the DNA damage in HaCaT cells was me
asured by single cell gel electrophoresis(SCGE).Results  Compared with control group,24 h after radiation,the cell viabilities in UVB group and Zn+UVB group were significantly decreased (P<0.05),the MDA levels were increased (P<0.05).Compared with UVB group,the cell viability in Zn+UVB group was significantly increased (P<0.05),the MDA level was decreased (P<0.05).The SCGE results revealed that  the DNA fragmentation(comet) of the cells in UVB group and Zn+UVB group were induced by UVB. The tail extent moment and Olive tail moment of the cells in UVB group were significantly higher than those in Zn+UVB group (P<0.05).
Conclusion UVB can lead the decreasing of the viability of HaCaT cells,the increasing of MDA level and DNA damage,while zinc sulfuric can antagonize the damage effect of UVB on HaCaT cells.

Abstract:Objective To investigate the cytotoxicity of gold nano-flower particle which has the absorption ability in near-infrared (NIR) region to human fibroblasts(HF) in vitro，and to provide basis for its  application
 in biomedical field.Methods HF were cultured with medium contained different doses of gold nano-flower particles （1.875,3.750,7.500,15.000,30.000 mg·L^-1） for 24 h,the HF cultured with medium free of nanoparticles as control group,the cytotoxicity effect was tested by MTT method，the morphology of HF was observed under inverted microscope.Results The relative growth rates（RGR）were 92.245%,88.980%,86.939%,75.782% and 71.756% respectively in different doses of gold nano-flower particle groups,the RGR in 30.000 mg·L^-1 group had significant difference compared with control group（P<0.05）,while all groups were the first cytoxicity grade.The HF had normal morphology and well growth under inver
ted microscope observation in low dose gold nano-flower particle group.Conclusion The cytotoxicity of the gold nano-flower particles has does-effect response,and low dose of gold nano-flower particles have well biocompatibility with HF.

Abstract:Objective To construct the  recombinant expression vector pET32a-CP6 and  transform  into the Escherichia colic （E.coli） BL21 (DE3) and express the recombinant protein at a high level,in order to provide reliable basis for obtaining a large number of soluble collagen. Methods The recombinant plasmid pET32a-CP6 was transformed into E.coli BL21 and induced by IPTG.The expression conditions of the recombinant protein were optimized in this test,which included temperature,time for induction,concentration of ITPG,inoculation volume,adding time of IPTG.The recombinant E.coli was fermented under the optimal condition,and the lysate of bacteria was purified by nickel ion affinity.The purified target protein was determined for purity by SDS-PAGE.Results The optimal temperature and time for induction of recombinant E.coli were 37℃ and 5 h respectively,while the optimal concentration of IPTG as an inducer was 0.5 mmol·L^-1.The expression level of target protein reached 31.52 mg·L^-1 under the optimal condition.The Western blotting results showed that the recombinant protein had specific reaction with anti-human COL6A2 antibody.Conclusion The optimized recombinant E.coli　can efficiently express the recombinant human collagen,the recombinant protein is purified successfully.

Abstract:Objective To construct prokaryotic expression vector harboring human papillomavirus-16 E7(HPV-16) gene and   observe the E7 protein expression.Methods Synthetic E7 gene was amplified using polymerase chain reaction and subcloned into prokaryotic expressing vector pGEX-4T1.The recombinant plasmids,pGEX-4T1-HP
V16-E7,were transferred into engineering bacteria Rosetta,and the fusion proteins E7-GST were analyzed on SDS-PAGE and purified by　Ni2+-NTA affinity chromatography under induction with IPTG.Results The target gene fragment at a length of 300 bp was observed on the agarose gel electrophoresis pattern and PCR.SDS-PAGE analysis showed that the expressed recombinant protein,with a relative molecular mass of about 36 000,could be found.Conclusion The recombinant protein,E7-GST,is effectively expressed in prokaryotic expression system.

Abstract:Objective To study the effects of oviductus ranae on the immune system and prepare its microspheres.
Methods Fifty ICR mices were randomly divided into control group，model group，low dose oviductus  ranae group（0.1 g·kg^-1），middle dose oviductus ranae group（0.4 g·kg^-1）and high dose oviductus ranae group（1.0 g·kg^-1），the contents of spleen T lymphocyte subpopulations in mice were determined by flow cytometry，and the content of IL-2 in spleen T lymphocytes was detected by ELISA.The microspheres of oviductus ranae were prepared by spray drying, orthogonal experiment was used to investigate  the material ratio，inlet temperature，outlet temperature，feed flow rate with average particle size，range of particle size，drug loading and the degree of encapsulated efficiency，the optimum preparation of oviductus ranae microspheres was determined.Results Compared with  control group， the contents of spleen T lymphocyte subpopulations  CD3+，CD4+ levels and CD4+/CD8+ ratio in the mice in model group  were

significantly increased (P<0.001)；Compared with model group，the contents of spleen T lymphocyte subpopulations   CD3+，CD4+ levels in the mice in  low dose oviductus ranae group were decreased significantly（P<0.001），CD4+/CD8+ ratio was also decreased significantly（P<0.05）.Compared with  control group，the content of IL-2 in  spleen T lymphocytes   in the mice  in model group was decreased
significantly （P<0.001）；Compared with  model group，the contents of IL-2 in spleen T lymphocytes
in the mice in  low，middle and high dose  oviductus ranae groups   were significantly increased（P<0.001）；Spray drying was used to prepare microspheres，the optimum preparation was  A3B1C1D1，the material ratio was 1∶3，the inlet temperature was 130℃，the outlet temperature was 100℃，and the feed flow rate was 30 mL·min^-1.Conclusion Oviductus ranae has immune regulation,they can be  prepared into microspheres,  the technology is stable and suitable for industrial production.

Abstract:Objective
To construct the rat NKX2.5-pEGFP fusion eukaryotic expression plasmid and screen bone marrow mesenchymal stem cells（MSCs） xpressing rat fusion protein NKX2.5-pEGFP and provide theoretical foundation for the research of cell and gene therapy of myocardial infarction. Methods The total RNAs were extracted from myocardial cells of embryo rats,RT-PCR amplification was performed to produce rat gene of NKX2.5.The recombinant plasmid NKX2.5-pEGFP was obtained
 by inserting the aim gene into pEGFP-N3.The MSCs derived from bone arrow mononuclear cells were obtained by gradient centrifugation and adherent culture.The recombinant plasmids NKX2.5-pEGFP were ransfected into rat MSCs by Lipo2000 and screened with G418 for 2 eeks.The expression of recombinant plasmid in MSCs was detected by Western blotting and fluoroscope analysis.Results The length of NKX2.5 gene obtained by RT-PCR was 981 bp which was consistent with theoretical  numerus.The recombinant plasmid NKX2.5- pEGFP was identified by endonuclease digestion and PCR,which could be digested into two fragments with  BamHⅠ and SalⅠ,about 981  and  4 700 bp,the results were coincident with the anticipated result.The protein with molecular weight of 65 000  was found in MSCs transfected with the recombinant plasmid NKX2.5- pEGFP by Western blotting which was coincident with theoretical numerus. Green fluorescence was positive expression in MSCs transfected with the recombinant plasmid NKX2.5-pEGFP by fluorescence detection.
Conclusion The rat MSCs expressing NKX2.5-pEGFP fusion protein is successfully constructed.

Abstract:Objective
To  compare  the contents of baicalein and galangin in propolis of different origins.Methods The contents of baicalein and galangin in 13 kinds of prololis of different origins were determinated by high performance liquid chromatography (HPLC).The　samples were separeted by C18 cloumn with the mobile phase of methanol-0.15% phosphoric (60∶40) and methanol-0.15% phosphoric (65∶35),the wavelength of detection was  268 nm.Results  Five samples were collected in each  area.The contents of baicalein and galangin in propolis of Jilin Jiaohe and Henan Changge were higher,all  over　2.0% and 1.8%,and the former was higher than the later(P<0.05);compared with the other 11 areas,there were significant differences（P<0.05）.Conclusion The contents of baicalein and Galangin in propolis of Jilin Jiaohe and Henan Changge are higher.

Abstract:Objective To evaluate the association of 5-hydroxytryptamine 1A(5-HT1A) receptor gene polymorphism and schizophrenia with Meta-analysis and  provide basis for evidence-based medicine for the genetic background of schizophrenia.Methods PubMed,Cochorane,CNKI and Wanfang Database were used to search all the relevant case-control trials of full text about the relationship between 5-HT1A receptor gene polymorphism and schizophrenia. The values of OR and 95%CI were used as effect indicators. It was based on reviewing full text to search,evaluate and adopt data. Statistical analysis was performed with RevMan5.0 including the heterogeneity inspection,effect consoliating and evaluatation on publication bias. Results A total of 5 articles were   included after the analysis. Two of the studies were about the association between  schizophrenia and  5-HT1A receptor gene rs6294,and the other 3 about 5-HT1A receptor gene rs6295. The Meta-anaylsis results of 5-HT1A receptor gene rs6294 genotype showed OR=0.35,95%CI=0.12-1.08，P=0.07;and the allele results were OR=0.34,95%CI=0.11-1.03，P=0.06;the  results of 5-HT1A receptor gene rs6295 genotype  were OR=1.78,95%CI=0.69-4.59，P=0.23;and the allele results were OR=1.80,95%CI=1.03-3.16，P=0.04. Conclusion  5-HT1A receptor gene polymorphism is associated with schizophrenia.

Objective To explore the relationship between Ghrelin and growth hormone（GH）-insulin-like growth factor（IGF）axis and the effect on the growth and development in 2-7 years old children with small for gestational age（SGA）at birth. Methods Thirty 2-7 years old children with SGA at birth were used as SGA group,and according catching up growth（CUG）or not they were divided into SGA-CUG group and  SGA-no-CUG group.Twenty short stature children with appropriate for gestational age（AGA）at birth were used as AGA short stature group,and nineteen normal height children with AGA at birth were used as control group.Height and
 weight were measured and the standard deviation score（SDS）were calculated.The levels of Ghrelin,IGF-1,insulin-like growth factor binding protein-3（IGFBP-3）and blood sugar were  measured and compared and the correlation was analyzed in all the subjects.Results The levels of plasma Ghrelin（mg·L^-1）in SGA-CUG group and SGA-no-CUG group (0.371±0.093 and 0.446±0.076）were much higher than those in AGA short stature group and control group(0.263±0.174 and 0.245±0.080) （P<0.05）and highest in SGA-no-CUG group.There were no significant differences between SGA-CUG group and SGA-no-CUG group as well as AGA short stature group and control group（P>0.05）.The levels of serum IGF-1（mg·L^-1）and IGFBP-3（mg·L^-1）in SGA-no-CUG group（123.882±46.275 and 33.310±8.836） and AGA short stature group（171.83±65.77 and 42.150±13.515） were significantly lower than those in SGA-CUG group（380.037±137.548 and 147.389±46.430）and control group（354.121±95.023 and 145.058±30.706）（P<0.05）. The correlation analysis in SGA-CUG group and control group showed negative correlation between Ghrelin and calory,weight SDS,IGF-1,IGFBP-3(SGA-CUG group:r=－0.628，P<0.01；r=－0.658，P<0.05；r=－0.854，P<0.000 1；r=－0.685，P<0.001； control group:r=－0.808，P<0.000 1；r=－0.664，P<0.001；r=－0.771，P<0.000 1；r=－0.502，P<0.05).There were no correlation between Ghrelin and year old,height SDS and blood sugar（SGA-CUG group:r=0.255，－0.218，－0.135，all P>0.05； control group:r=0.203，－0.328，－0.228， all P＞0.05）.Conclusion  There is interaction between Ghrelin and GH-IGF axis，and they regulate the growth and development together. There is resistance to Ghrelin and aggression to GH-IGF axis and it will interfere the growth and development untill prepubescence.

Objective To detect the expressions of neurofilament and myelin basic protein in acoustic tumor tissues and explore the growth features of cells in acoustic tumor tissues. Methods immunofluorescence labeling and immunoblotting analysis were performed to detect the expressions of neurofilament and myelin basic
protein in acoustic tumor tissues in 15 patients with acoustic tumor(acoustic tumor group) and  5 cases of normal peripheral nerves (control group). Results  The expression of myelin basic protein  indicating the be
ginning of myelination in acoustic tumor tissues was negative. The expression ofaxon marker neurofilament
 was also negative. But the expressions of neurofilament and myelin basic protein in control group were positive. Conclusion The axon or myelin are compact abscent in acoustic tumor tissues,it indicates that the cells of acoustic tumor is in a premyelinating state,and lose the ability to form compact mylin or remyelination.

Abstract:Objective To evaluate the correlation of thyroglobulin(Tg) with 131-iodine whole body scan(^131I-WBS) in the differentiated thyroid carcinoma(DTC) patients after thyroid ablation and  provide reference index for judging  recurrence and metastasis of DTC. Methods  596 patients with DTC were cut off partial thyroid and then undergone remanent thyroid ablation with ^131I and took thyroid hormone regularly. The serum Tg levels were measured and ¹³¹I-WBS was offered after administration of 185 MBq radioactive iodine for 24，48 and 72 h. Semiquantitative visual analysis was used to grade the extent of metastasis showed in131I-WBS．The correlation of Tg levels with ^131I-WBS scores was analyzed. Results In 596  DTC patients,64 cases were confirmed with recurrence and metastasis. In these patients,the results of Tg were positive in 63 cases and negative in 1 case;the results of ^131I-WBS were positive in 56 cases and negative in 8 cases. In 63 cases with positive Tg,^131I -WBS were also positive in 55 cases. In 56 cases of patients with positive^131I-WBS,the levels of Tg in 55 cases were also positive. Nobody was both negative in two checks. The serum Tg levels had positive correlation with ^131I-WBS scores (r=0.718-0.812，P<0.01)Conclusion The serum levels of Tg are correlated with ^131I-WBS after thyroid ablation，the raising of serum Tg level indicates massive metastasis in DTC patients，a close follow-up and proper treatment are suggested．

Objective  To guide the clinical treatment and evaluate prognosis through screening several biological indicators related with nasal and sinusal salivary gland carcinoma and analyzing the clinical data. Methods The Logistic regression analysis was employed to analyze the data of the expressions of several biological indicators in nasal and sinusal salivary gland carcinoma,para-cancer tissues and nasal polys and cyst detected with immunohistochemical technique. Results There were  significant differences in the expressions of laminin(LN) and type Ⅳcollagen(ColⅣ) and micrangium in the occurrence of the nasaland sinusal salivary gland carcinoma (P<0.05),and there was a poorly prognosis of nasal and sinusal salivary gland carcinoma if the levels of LN and ColⅣ reduced. Conclusion The LN and ColⅣare  protective factors and the micrangium is a risk factor in the occurrence of the nasal and sinusal salivary gland carcinoma.The decreasing in LN and ColⅣexpressions have relationship with recurrence and metastasis of nasal and sinusal salivary gland carcinoma.The combined detection of LN and Col Ⅳ in nasal and sinusal salivary gland carcinoma can guide its clinical treatment and prognostic evaluation.

Abstract:Objective To investigate the potential role for Hath1 in pathogenesis of  colon adenocarcinoma. Methods The  sections of paraffin-embedded tissues were used to investigate the goblet cell population of normal colon mucosa,mucosa adjacent colon cancer and colon cancer tissues obtained  from 48 patients with non-mucinous colon adenocarcinoma. ThemRNA and protein  expressions of hath1 and Muc2   in these samples were tested by quantitative real-time reverse transcription -PCR,immunohistochemistry andWestern blotting. The Hath1 gene was transfected into  colon cancer cells HT29,then three clones were selected randomly to test the expressions of mRNA and protein of Hath1 and Muc2 by quantitative real-time reverse transcription-PCR and Western blotting.The protein expressions of cyclin D1 and p27 were also tested by Western blotting. Moreover,the proliferativeability of HT29 cells transfected with Hath1 was observed by means of colony formation assay and xenograft. Results There were not any goblet cells to be found in colon cancr. The expressions of mRNA and protein of Hath1 and Muc2 were significantly down-regulated in colon cancer（P<0.05）. Hath1 could decrease the expression of cyclin D1 protein（P<0.05）,increase the expressions of p27 protein（P<0.05） and Muc2 protein（P<0.01） in HT29 cells,inhibit these cells to form colonies（P<0.01） and decrease the size of xenografts（P<0.01）. Conclusion Down-regulation of Hath1 may concern with non-mucinous colon adenocarcinoma,Hath1 may be an anti-oncogene in pathogenesis of non-mucinous colon adenocarcinoma.

Objective To study the relevance between single-nucleotide polymorphism（SNP）of dihydrogenpyrimidine dehydrogenase gene （DYPD） and 5-fluorouracil（5-FU） toxicity in colorectal cancer patients and provide the evidence for individualized treatment in colorectal cancer patients.Methods  The genomic DNA was extracted from EDTA anticoagulation peripheral blood in a total of 60 patients with colorectal cancer and then real-time quantitative polymerase chain reaction (PCR) was performed.The SNPs of 14G1A,G2194A,T85C,G1156T and T464A sites of DPYD were detected,the side effects of chemotherapy with 5-FU in 60 patients were recorded,the relevance between the SNP of DPYD and side effects of 5-FU chemotherapy was analyzed. Results ①14G1A and G1156T polymorphism loci transformation were not found in 60 patients. G2194A heterozygous type was found in 6 cases (10%),wild type in 54 cases (90.0%);T464A heterozygous type in 2 cases (3.3%),wild type in 58 cases (96.7%);T85C mutations type in 2 cases (3.3%),heterozygous type in 8 cases (13.3%),wild type in 50 patients(83.4%).②The incidence of gastrointestinal tract toxicity in patients with T85C polymorphism sites without SNP mutation was 14% (7/50),the incidence of hematological toxicity was 4% (2/50);the incidence of gastrointestinal tract toxicity in patients with T85C polymorphism sites with SNP mutation was 60% (6/10),the incidence of hematological toxicity was 70% (7/10),the differences were statistically significant (P<0.05);the incidences of gastrointestinal reaction andbone marrow restrain in patients with T464A SNP mutation were 50% (1/2) and 100% (2/2),in patients without SNP mutation were 21% (12/58) and 12% (7/58),the differences were statistically significant (P<0.05). ③ The incidences of hematological toxicity and bone marrow restrain in patients with G2194A SNP mutation were33% (2/6) and 50% (3/6), in the patients without SNP mutation were 20% (11/54) and 11 (6/54),the differences were statistically significant (P<0.05). Conclusion The SNP of DPYD has correlation with the adverse reactions of 5-FU chemotherapy.The detection of SNP of DPYD may predict the serious adverse  reactions of 5-FU chemotherapy.

Abstract:Objective To study the expressions of cyclooxegenase-2（COX-2）,matrix metalloproteinases-9（MMP-9）,vascular endothelial growth factor（VEGF）,transfection rearrangerment gene（RET） in papillary thyroid carcinoma and significances in primary diagnosis.  Methods The expressions of COX-2,MMP-9,VEGF and RET were detected in papillary thyroid carcinoma,thyroid adenoma,nodular goiter and normal thyroid tissues by immunohistochemical method.The diagnostic sensitivities and specificities of these genes in thyroid papillary carcinoma were discussed as well as the relativities betweenthem. Finally, the most sensitive index of papillary thyroid carcinoma’s  primary diagnosis and the assessment of lymph node metastasis was selected.  Results The expressions of COX-2,MMP-9,VEGF and RET in papillary thyroid carcinoma group were obviously higher than those in  thyroid benign tumors,which presented  an intimate relationship with age,tumor size,mem
brane integrality and clinical stage (P<0.05),the expressions of COX-2 and  MMP-9 in thyroid adenoma were higher than those in  nodular goiter(P<0.05),the expression of COX-2 was correlated to lymph node metastasis(P<0.05)，but all of them were negatively correlated to patient gender (P> 0.05). MMP-9 was regarded as a sensitive marker of  diagnosis  for early thyroid carcinoma by gene filtration,the sensitivity was 90.9% and the specificity was 90.0%.COX-2 could  work as a marker to evaluate lymph node metastasis，the sensitivity was 77.8% and the specificity was 66.7%.Conclusion Combined detection of COX-2 and MMP-9 can be used as the most sensitive biological index in preoperative diagnosisof papillary thyroid carcinoma and assessment of lymph node metastasis.

Objective To investigate the polymorphism of killer cell immunoglobulin-like receptor KIRgene expression in Han population of Jilin province and lay a theoretical basis for the study of thecorrelation between KIR genes and disease. Methods The DNA genome samples were extracted from the 98 health people who were received physical examination in Han population of Jilin province separately. Then the genome samples were genotyped using low resolution polymerase chain reaction with sequence-specific primers(PCR-SSP) typing method. The gene loci of KIR2DL1—KIR2DL5，KIR2DS1—KIR2DS5，KIR3DL1—KIR3DL3，KIR3DS1 and KIR2DP1 were examined,the genotypes and haplotypes were assigned according to standard model of Hsu. Results 15 kinds of KIR genes were observed in all samples,among them 3 kinds of framework genes,ie KIR2DL4，KIR3DL2，KIR3DL3 were all expressed in all individuals. 2DL1，2DL3，2DL4，3DL1，3DL2，3DL3，2DP1，2DS4*001 genes were relatively common. A total of 15 different kinds of KIR haplotypes were identified. The haplotype 2 was predominant with a frequency of 53.1％,and the second kind was haplotype 1 with a frequency of 14.8％,both haplotype 1 and 2 all belonged to haplotype A. The frequency of haplotype A was 67.9％.Conclusion The KIR gene distribution in Han population of Jilin province is different from White people，but it has some similarity with the Chinese Han population in other araes,meanwhile，there are distinctive frequencies of KIR gene，haplotype as well as genotype，in Jilin Han population.

Objective To investigate the curative effect of meglumine cyclic adenylate(MCA) combined with recombinant human brain natriuretic peptide(rhBNP) in treatment of coronary artery disease patients with heart failure,and the changes of hemodynamics and the  BNP concentration. Methods Fifty-four patients with coronary atherosclerotic heart disease whose heart function was Ⅲ-Ⅳ grade were divided into three groups(n=18):groupⅠ,conventional therapy;group Ⅱ:conventional therapy plus MCA;group Ⅲ,conventional therapy plus MCA combined with rhBNP. Results ① Clinical curative effect:the total effective rates of group Ⅱ(77.8%) and group Ⅲ(88.9%) were higher than that of group Ⅰ(61.1%), but the total effectively rate of group Ⅲ was obviously higher than that of groupⅠ(P<0.05). ② The changes of hemodynamics:after treatment,the stroke volume (SV),cardiac output (CO),cardiac index (CI) and left ventricle ejection fraction (LVEF) were increased in groupⅠ,Ⅱ,Ⅲ;they were significanthy increased in groupⅡ and group Ⅲ(P<0.01);compared with group Ⅱ,they were significanthy increased in group Ⅲ(P<0.05,P<0.01). The ratios of the peak flow rate of bicuspid blood current diastolic phase to atrial systole period (E/A)  in group Ⅱand group Ⅲ were increased(P<0.01);compared with group Ⅱ,the E/A in group Ⅲ was significanthy increased (P<0.05). ③ The changes of BNP concentration:after treatment,the BNP concentrations  were decreased in three groups,especially in group Ⅱ(P<0.01) and group Ⅲ(P<0.001);compared with groupⅡ,it was significant decreased in group Ⅲ(P<0.01). Conclusion MCAcombined with rhBNP has utility,safety and synergistic effect in treatment of  coronary artery disease patients with heart failure. 

Objective To compare the anesthetic effects between sufentanil based total intravenous anesthesia and epidural anesthesia with ropivacaine in conserving operation for breast cancer. Methods Thirty-eight ambulatory adult female patients in ASA ⅠorⅡphysical statuses scheduled for breast operation were randomly divided into two groups. Nineteen cases adopted sufentanil based total intravenous anesthesia (sufontanil group)and others were treated with ropivacaine based  epidural anesthesia(ropivacaine group). The systolic blood pressure (SBP),diastolic blood pressure (DBP),heart rate (HR ),respiratory rate (RR),tidal volume (TV),oxyhemoglobin saturation (SpO2),pH,partial pressure of arterial carbon dioxide (PaCO2) and arterial oxygen pressure (PaO2)were detected  before anesthesia (t0),15 min after anesthesia (t1),the beginning of operation (t2), 1 h after anesthesia (t3),the end point of operation (t4).The application of ephedrine and atropine and the revive time were recorded. Results① Comparison in each group:compared with t0,SBP  at t1,t2,t3 and t4 points were decreased significantly(P<0.05);and DBP  at t1,t2 and t3 time points were decreased(P<0.05） in two groups. In ropivacaine group,HR showed a decline trend  at t1,t2 and t3 time pints compared with  t0 (P<0.05);and in sufentanil group,HR showed a decline trend at t1 and t3 compared with  t0 (P<0.05). Both in two groups,compared with t0,SpO2  showed no significant difference at four time points. At t3 and t4 time points,PaO2 was increased significantly(P<0.05),while pH was decreased(P<0.05). At t2 and t2 time points,PaO2 were increased significantly in ropivacaine group(P<0.05),and PaCO2 were increased significantly in two groups at t1,t2,t3 and t4 time points(P<0.05). Compared with t0,TV were increased at t2,t3 and  t4 in two groups,and TV was also increased in sufentanil group at t1,while RR were decreased significantly at t1,t2,t3 and t4 time points in two groups(P<0.05). ②Compared with  ropivacaine group: higher SBP and PaO2 were found in sufentanil group at t2 time point (P<0.05),and HR were also increased in sufentanil group at t2 and t3 time points. The rate of HuoNa’s levy in ropivacaine group was higher than that in sufentanil group. Other indexes were not significantly different. Conclusion  The two anesthesia methods have a similar blood gas analysis results;compared with epidural anesthesia with ropivacaine,sufentanil based total intravenous anesthesia shows a lower rate of complication and tiny homodynamic changes. Sufentanil can be used in breast conserving operation for breast cancer safely.

Abstract:Objective To evaluate the clinical effects of Ankylos implant system to restore the loss of single-tooth.Methods 90 cases with loss of single-tooth were selected and treated with  routinely two-stage surgery.When the patients presented with deficient alveolar ridge, guided bone regeneration (GBR), osteotome sinus floor elevation, lateral antrostomy surgery with simultaneous placement of implant were applied.They were restored with platinum ceramic crown.All the implants were followed up, and the records were kept about stability of the implant and abutment, the status of  surrounding soft tissue, sealability of implant abutment junction and the marginal bone lossing through X-ray checking, and satisfaction of the patients  to mastication and aspect of the restorations.The follow- up time was 1-2.5 years. Results Among the cases, one case had peri-implant inflammation, and one case had porcelain dropped.No loosening occurred in the other implants and abuments.Implant abutment junction was sealed well.The marginal bone loss 1 year after final restoration was less than 1 mm.Soft tissue surrounding implants was healthy.The satisfaction rate was 98.9%（89/90）.According to standard of implant success,88 cases were successful,the 2.5-year cumulative success rate was 97.8%, 2 cases failed, and the failure rate was 2.2 %. Conclusion A satisfactory treatment effects could be gotten by using Ankylos implant system to restore the loss of single-tooth.

Abstract:Objective Using proton magnetic resonance spectroscopy (1H-MRS) method ，to evaluate the difference of radiation brain injury between volumetric modulated arc therapy (VMAT) and three-dimensional conformal radiation therapy (3DCRT) in patients with postoperative glioma after radiation therapy.Methods 24 patients with malignant glioma (WHOⅡ-Ⅳ grade glioma) confirmed with clinical surgery were selected,among them 12 patients were treated with VMAT technique,and another 12 patients with 3DCRT technique,all received DT60-66GY/30-33F dose prescriptions. 1H-MRS examination was performed to analyze the change of metabolites in the brain tissues of region of interest (ROI) before and after radiotherapy,and the ratios of NAA/ Cr,Cho / Cr,NAA / Cho were computed.Results The dose distribution of VMAT group was superior to 3DCRT group,the NAA/Cr in two groups after radiation were decreased compared with before radiation,there was a statistically difference in NAA/Cr after radiation between two groups (P<0.01).The  Cho / Cr and  NAA / Cho in two groups were increased compared with before radiation;after radiation,only NAA/Cho  had a statistical difference between  two groups (P<0.01).Conclusion VMAT technique is superior to  3DCTR to reduce radiation brain injury in patients with postoperative glioma.

Abstract:Objective  To compare the effects between 18F-fluorodeoxyglucose(18F-FDG)PET-CT and MRI  in diagnosis of breast cancer．Methods 29 patients with suspicious masses in breast were selected,18F-FDG PET/CT and MRI scanning were performed and their pathological results were gotten within 10 d．The sensitivities,specificities and accuracies of PET/CT and MRI in detecting breast cancer were calculated by using surgery or biopsy results as diagnostic golden standard．Results 20 malignant and 9 benign breast lesions of the 29 patients were proven histologically after surgery or biopsy．The sensitivity，specificity and accuracy value obtained by MRI were 85.0％,66.7％ and 79.3％ respectively,and they  were 90.0％,88.9％ and 89.6％ for PET/CT．The accuracy of PET/CT was a little higher than that of MRI,but there was no significant difference between them (χ2=1.17，P>0.05). PET/CT found 3 patients with left infraclavicular lymph node metastasis and bone metastasis. Conclusion There is no significant difference between PET/CT and MRI in detecting breast cancer. But PET/CT is superior to MRI in evaluating distant metastasis and accurate staging to guide therapy.

Abstract:Objective To study the mechano-electric delay phenomenon,an unrecognized phenomenon and its mechanism in cardic cycle with M-mode echocardiography and tissue Doppler imaging (TDI). Methods The mitral valve closure point to the onset of QRS complex by M-mode and TDI（C-Q interval,C’-Q interval）and the onset of isovolumic contraction wave S1 to the onset of QRS complex（S1-Q interval）by TDI of 26 patients with first-degree AV block were obtained,as well as the displacements and strains of myocardium in C’-Q interval and  S1-Q interval. Results The  onset of S1 of 26 patients were ahead of the onset of QRS complex,furthermore,the mitral valve closure point of 23 patients were ahead of QRS complex too. The obvious displacements and strains of myocardium in C’-Q interval and S1-Q interval could be seen. Conclusion Because of  passive contraction not controlled by electrical signals,another contraction fashion except active contraction,there should be exist a mechano-electric delay phenomenon in cardic cycle.

Abstract:Objective To estimate the number of people who lives with HIV and  AIDS （PLWHA）in 2009 and the HIV epidemic trend in next six years (2010-2015) in Jilin province and provide evidence for HIV prevention and control. Methods The data from sentinel surveillance,case report,epidemiological survey and scientific lectures were used in Workbook method,Estimation and Projection Package and Asia Epidemic Model to estimate the number of HIV/AIDS and to project the HIV epidemic trend in Jilin province. Results By the end of 2009,it was estimated that among high-risk population the number of cumulative PLWHA was 4 388,the total prevalence of HIV was 0.024% in Jilin province and over 50% of PLWHA were men who had sex with men. HIV epidemic trend will be rising from 2010 to 2015 in Jilin province,where the trend among MSM will go upward markedly,the rising trend among IDU will be slowed down and the rising trend among FSW and their clients will be relatively slow. Conclusion The HIV epidemic in Jilin province was still at a lower level in 2009 and the sexual transmission was the main transmission way. MSM and clients of FSW are the focus population who must be given more attention in the future.

Abstract:Objective To observe the rapid detection effccts of resistance to isoniazide(INH),rifampcin (RFP)and streptomycin(SM) in mycobacterium tuberculosis (MTB) by  membrance reverse dot blot hybridizd technique. Methods The probes of  ropB,katG,inhA and rpsL resistant genes were mixed with PCR products of MTB .The mixture were detected by membrance reverse dot blot hybridization technigue.Then the results were compared with conventional drug susceptibility tests. Results The sensitivities of INH,RFP and SM were 84.88%,87.10% and 79.17%，and the coincident rates were 97.33%,94.74% and 88.37%. Conclusion The membrane-reverse dot blot hybridization   can be used to detect resistance of partial MTB  clinical strains rapidly and simply.