Abstract:Objective To investigate the effect of recombinant plasmid pEgr-hp53 transfected stably in combination with X-ray irradiation on the cell cycle progression and the proliferation in human SKOV-3 tumor cells.Methods pEgr-hp53 and pcDNA3.1 packaged with liposome were stably transfected into SKOV-3 cells in vitro.SKOV-3-hp53 and SKOV-3-vect were irradiated with 0,0.5, 2.0 and 5.0 Gy X-rays, respectively, i.e.8 experimental groups.The SKOV-3 cell proliferation and the cell cycle progression were measured with flow cytometry and cell growth curve, respectively.Results Compared with 0 Gy group, the cell counts in SKOV-3-hp53 plus different doses of irradiation groups 2 d after irradiation decreased significantly (P<0.05 or P<0.01), and the decrease of the cell counts was more significant with the increasing of dose and the prolongation of time after irradiation; the percentage of G₀/G₁ cells increased significantly (P<0.001), while that of G₂/M cells decreased in varying degrees.The cell counts in SKOV-3-hp53 plus irradiation group were significantly lower than those in corresponding SKOV-3-vect plus irradiation group, the cell counts 4-8 d after irradiation with 0.5 Gy, 2 d after 2.0 Gy irradiation and 6 d after 5.0 Gy irradiation decreased significantly (P<0.05 or P<0.01); the percentage of G₀/G₁ cells increased significantly (P<0.01), while that of G₂/M cells decreased significantly (P<0.01).Conclusion pEgr-hp53　 transfected stably in combination with X-ray irradiation leads to p53-induced G₁ arrest in SKOV-3 cells and inhibits the cell proliferation.Ionizing radiation can activate early growth response-1 (Egr-1) gene promoter and increase the expression of p53 gene, and enhance the inhibition of tumor cell growth.

Abstract:Objective To study the effect of small heterocyclic compound 8-oxo-3-thiomorpholino-8H-acenaphtho ［1,2-b］ pyrrole-9-carbonitrile (S1) on apoptosis of human androgen independent prostatic carcinoma cells line(PC3) and its mechanism.Methods PC3 cells were cultivated, and divided into different groups: 10.00,5.00,1.00,0.50,0.10,0.05 and 0.01 μmol•L^-1 S1 groups, meanwhile, PC3 control group and cyclophosphamide group were set up. MTT was used to detect the inhibitory rate of PC3 cell proliferation.Flow cytometry was used to detect the inducing effect of S1 on apoptosis of PC3 cells.Caspase 3, 8, 9 kits were used to detect apoptosis route.Results The inhibitory rates of PC3 cells induced by 0.10-10.00 μmol•L^-1 S1 were significantly higher than that in cyclophosphamide group （P<0.01）.The apoptotic rates induced by 5.00 and 10.00 μmol•L^-1S1 were significantly higher than that in PC3 control group（P<0.01）.When PC3 cells were treated with 10.00 μmol•L^-1 S1 for 6 h, the activity of Caspase3 was significantly higher than that in PC3 control group（P<0.01）.When PC3 cells were treated with 10.00 μmol•L^-1 S1 for 4-6 h, the activities of Caspase 9 were significantly higher than that in control group（P<0.01）, but the activity of Caspase 8 did not change significantly.Conclusion S1 compound can induce apoptosis of PC3 cell line and death,and its mechanism may be related to mitochondria.

Abstract:Objective To construct a recombinant adenoviral shuttle vector pshuttle-Egr1-hTRAIL containing radiation-sensitive Egr-1 promoter and TNF-related apoptosis-inducing ligand (TRAIL). Methods The TRAIL gene fragment was acquired from the plasmid pACCMV-hTRAIL by RT-PCR.Then the TRAIL gene was ligated to pMD19T vector and sequenced.With the gene recombinant technique, the recombinant plasmid pshuttle-Egr1-hTRAIL with radiation-inducible promoter Egr-1 was constructed. Results A fragment about 820 bp was amplified by PCR, and the sequence of acquired hTRAIL gene was totally in concordance with that published in GenBank（NM_003810）.Moreover, the recombinant plasmid pshuttle-Egr1-hTRAIL was digested by EcoRⅠand KpnⅠdouble-enzyme and BamHⅠsingly both into two fragments, with the length of 3 540 and 4 299 bp, 3 304 and 4 535 bp,　 respectively.The SmaⅠenzyme could digest it into three fragments with lengths of 1 517, 2 282 and 4 040 bp.The results of enzyme identification were all in concordance with that expected. Conclusion The hTRAIL gene is cloned and the recombinant plasmid pshuttle-Egr1-hTRAIL is constructed successfully.

Abstract:Objective To clone the rat second mitochondria-derived activator of caspase（rSmac）, construct its eukaryotic expression vector, and express it in cardiocytes. Methods The rSmac CDs gene was amplified from the rat kidney tissue by reverse transcriptase-polymerase chain reaction（RT-PCR）.It was cloned into pcDNA3.1⁺ vector to construct the recombinant plasmid pcDNA3.1⁺-rSmac.Then the recombinant plasmid pcDNA3.1⁺-rSmac, pcDNA3.1⁺, and pcDNA3.1⁺-EGFP were transfected into the rat myocardial cell line, H9C2 mediated by lipofectamine 2000.Among them, pcDNA3.1⁺-EGFP was used to detect the transfection efficacy indirectly through fluorescence microscoy（FM）and flow cytometry（FCM）.After transfection, RT-PCR method was used to detect the expression of exogenous Smac gene.Results The sequencing result indicated that the acquired sequence was in concordance with that published on GenBank.The recombinant plasmid pcDNA3.1⁺-rSmac was idenfied by PCR and enzyme digesting, and the results were coincident with anticipation.The results detected by FM and FCM showed that the transfection was effective, and the efficacy was up to 38.36%.The mRNA level was increased after transfection with the recombinant plasmid pcDNA3.1⁺-rSmac for 48 h, and the ratios of Smac/β-actin in control, vacant vector, and recombinant plasmid pcDNA3.1⁺-rSmac group were 1.19, 1.31, 1.67, respectively（P>0.05, P<0.01 vs control group）.Conclusion The rSmac gene is cloned and the recombinant plasmid pcDNA3.1⁺-rSmac is constructed successfully.Moreover, it could be effectively expressed in transfectant H9C2, which establish fundament and benefit the further study on the mechanism of Smac in apoptosis of cardiocytes.

Abstract:Objective To study the characters of degradation of poly-L-lactic acid (PLLA) internal fracture-fixing system.Methods The mandible fracture of dogs was fixed with PLLA internal fracture-fixing system.In 1,3,6,12 months after operation , the materials were taken out from dogs, and the changes of ultrastructures of miniplates were observed using the scanning electron microscope after macroscopic observation.Results Under macroscopic observation,the miniplate was elastic,hard and semitransparent,there was no obvious changes on the surface one month later.The surface of PLLA miniplate became rough, soft,cretaceous, opaque and hard to break three months later.But the miniplate became even softer,still opaque and easy to break with some gabs on the surface six months later.The miniplate became into something like cheese,softer and exquisite twelve months later.Observed by scanning electron microscope, the surface of the miniplate was smooth,there was no fissuaring and hole on it before implantation.One month later,the surface became rough.Three months later there were many gabs on it.Six months later,the gabs entered deeply into the materials.Twelve months later the miniplate had degradated and broken completely.And much of micropore could be seen on the cross-section of miniplate.Conclusion PLLA miniplate degrads on the surface at the beginning and then at 6 months after the operation,the material begins to degrade deeply.The decrease of the mechanic strength speed of degradation is moderate,it matches with the speed of fracture healing.It will help to avoid stress shielding and accelerate the speed of fracture healing.

Abstract:Objective To detect the optimum reaction temperature ,pH, Km of N-V proteinase and determine its enzymological classification.Methods The fibrin plate method was used to determine the optimum reaction temperature and pH of N-V proteinase.The Km of N-V proteinase was detected by using Leu-pNA as substrate.E64, PMSF, zymofren and 1,10-phenanthroline were used as inhibitors to determine proteinase classification of N-V proteinase.Results The optimum reaction temperature was 50℃, pH was 8-9,Km was 1.9710^-3mol•L^-1.PSMF inhibited the fibrinolytic activity of N-V proteinase （P<0.01）, whereas E64, zymofren and 1,10-phenanthroline had not significantly inhibitory effects on its activity（P>0.05）.Conclusion N-V proteinase belongs to serine proteinase.

Abstract:Objective To explore the changes of the expression of connexin 43 in rat glioma treated by antigliomatin and research multi-originated regulatory mechanism of antigliomatin’〖KG-*3〗s suppression on glioma.Methods C6 glioma cells were cultivated in vitro,which were used to make model of rat glioma.These cells were divided into control group and experimental groups treated with different concentrations of antigliomatin（8,16,32 and 64 μg•kg^-1）.Antigliomatin was given by peritoneal injection in experimental groups.The effects of antigliomatin with different concentrations on the expression of CX43-immunoreaction positive cells were determined by immunohistochemical staining method and Western blotting.Results The number of CX43-immunoreaction positive cells in control group and 8,16,32 and 64 μg•kg^-1 antigliomatin groups were 4.31±1.24,5.23±2.01,11.58±3.21,20.13 ±3.04 and 23.58±4.32 in each visual field,there were significant differences between control group and experimental groups（P<0.05 or P<0.01）,and there also were significant differences between experimental groups （P<0.01）.The amounts of CX43-expression（cross product of albumen-strap’〖KG-*3〗s area and average gray scale） in control group and 8,16,32 and 64 μg•kg^-1 antigliomatin groups were 12 241, 12 377,16 183,17 138 and 28 208,there were significant differences between control group and experimental groups（P<0.05 or P<0.01）,and there also were significant differences between experimental groups （P<0.01）.Conclusion The inhibitory effects of antigliomatin on glioma may be related to influence of connexin function.

Abstract:Objective To study large-scale fermentation of recombinant human HSP110(rhHSP110) in Pichia pastoris, and the purification of expressed target protein in Pichia pastoris.Methods rhHSP110(2 L) was prepared in shake flask.Using fed-batch fermentation,the high-density fermentation of genetically engineered Pichia pastoris was processed in a 80 L bioreactor.After induction for 72 h, the target protein was purified by SP Sepharose XL.Results The fermentation temperature was set at 30℃,the pH value was 4.2, and the DO was kept over 20%.When the wet weight of the cells reached 200 g•L^-1, the methanol-induced phase was initiated.By cation exchange chromatography it was possible to purify the rhHSP110 produced in Pichia pastoris, and a yield of about 500 mg•L^-1 was reached.Conclusion The success of large-scale fermentation lays a foundation for mass production and clinical applications of rhHSP110.

Abstract:Objective To study the designing principles and screening method of immunoregulatory oligodeoxynucleotides (ODN) and provide information for the research and development of novel immunoregulatory ODN.Methods The human peripheral blood monouclear cells (hPBMC) were stimulated by CpG ODN and/or immunoregulatory ODN.The CpG ODN-induced cell proliferation was examined by thymidine incorporation to evaluate the immunoregulatory activity of self-designed ODNs.Results ODN 667B with relatively high immunosuppressive activity was obtained by designing and screening of two groups of immunoregulatory ODNs.Compared with CpG BW006 group, the inhibitory rate of 667B reached 66.3% (P<0.01) and the suppression was concentration-dependent. Conclusion The designing principle of immunoregulatory ODN is summaried and the screening method is proved to be available.The results set the foundation for the research and development of novel immunoregulatory ODN.

Abstract:Objective To investigate the expression pattern of activin and type ⅡA receptor of activin (ActRⅡA) and its possible effect on liver injury of Con A-induced model mice.Methods 24 mice were injected with Con A (10 mg•kg^-1) in the tail vein weekly for 4 consecutive weeks to cause liver injury and normal control mice (n=24) were used as control.Then the levels of serum ALT and AST and severity of liver injury were examined, as well as the expression levels of activin and ActR ⅡA mRNA in liver were detected by using fluorescent quantitive PCR. Results The liver injury was most obvious at 72 h after the last injection, such as structure of liver lobules disordered, hepatic cord disappeared, balloon-like hepatocytes swelled, and then gradually repaired from 120 h to 168 h.The activin A protein level and its mRNA expression paralleled with ActRⅡA mRNA expression, and also reached the peak at 72 h after the last injection, which had significant difference compared with control mice (P<0.05).Conclusion The changes of activin A and ActR ⅡA are positively correlated with the severity of liver pathological changes, which indicate that activin may take part in Con A-induced mouse liver injury.

Abstract:Objective To study activation effect of a natural compound baicalein on cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel.Methods A cell-based fluorescence assay was used to determine CFTR-mediate iodide influx rate［d［I^-］/dt（mmol•L^-1•s^-1）］ activated by baicalein（the concentrations were 0.18,0.55,1.65,5,15,44,133 and 400 μmol•L^-1）.Results The Ka of flavonoid baicalein stimulating CFTR was about 16 μmol•L^-1.The half of maximal activity was reached in ten minutes and the activation disappeared in 20 min after baicalein was washed out.The activation of baicalein was not affected obviously under different concentrations of Forsklin （0,20,50 and 100 nmol•L^-1）and the activation could be totally inhibited by CFTRinh-172.Conclusion Baicalein can stimulate CFTR-mediated iodide influx in a dose-dependent way and its activity manifests a rapid and reversible characteristic.It might work in both elevating CFTR protein phosphorylation and direct binding way.

Abstract:Objective To study the inhibitory effects of total flavonoids of scutellaria baicalensis georgi (TFSB) on S₁₈₀, Hep-A-22 and Bcap-37 tumor cell proliferation in vitro and on S₁₈₀, Hep-A-22 in mice bearing tumor in vivo.Methods In vitro, S₁₈₀, Hep-A-22 and Bcap-37 cells were divided into control group and TFSB groups (12.5，25.0，50.0，100.0 mg•L^-1).The inhibitory effects of TFSB on proliferation of S₁₈₀ and Hep-A-22 were measured by XTT colorimetric assay, and Bcap-37 cells were measured by MTT colorimetric assay.In vivo, the mice bearing tumor were divided into control group, CTX group (30 mg•kg^-1)，high, middle, low doses TFSB groups (200, 100，50 mg•kg^-1).After the mice bearing S₁₈₀ and Hep-A-22 tumor cells were treated with TFSB for 15 d, the tumor weights were measured, the inhibitory rates of S₁₈₀ and Hep-A-22 were calculated and survival of Hep-A-22 was measured after administration of TFSB for 10 d.Results TFSB inhibited the proliferation of S₁₈₀, Hep-A-22 and Bcap-37 cells, IC₅₀ values were 16.04, 17.74 and 9.05 mg•L^-1, respectively.The tumor weight of mice bearing S₁₈₀ and Hep-A-22 cells in TFSB groups(200, 100, 50mg•kg^-1) were lowered than that in control( P<0.01 or P<0.05). The tumor inhibitory rates were increased in dose-dependent manner.The survival of mice bearing Hep-A-22 in TFSB groups were higher than that in control（P<0.01 ).Conclusion TFSB may possess significantly inhibitory effect on proliferation of S₁₈₀, Hep-A-22 and Bcap-37 cells in vitro and the growth of tumor in mice bearing S₁₈₀ and Hep-A-22 in vivo.

Abstract:Objective To study the preparation method of compound tissue-engineering scaffolds of the PLA/silk fibroin and evaluate its biological features.Methods The PLA scaffolds matrix were dipped into the silk fibroin solution,then dried,and PLA/silk fibroin scaffolds were prepared. There were two groups in the experiment,one group was PLA group,and the other one was compound scaffolds group.According to ISO-10993 standard, hematolysis test,dynamic coagulation time test, cell toxicity test，stimulation test and pyrogen test were performed in two groups,and the results were compared betwen two groups. Results In the stimulation test，the two kinds of materials had equally not aroused the obvious animal skin stimulation, it showed that the experiment was in accordance with the standard. In the pyrogen test, the two scaffolds material aroused the animal temperature rising without exception under 0.2℃ and the total number of degree was under 1.0℃,therefore there was no obvious difference between two groups. In the hematolysis test, the hemolysis rates of the two scaffolds samples were smaller than 5% equally(P=0.000),which indicated that the hemolysis of the compound scaffolds was better than that of the PLA scaffolds.In the dynamic coagulation time test,the coagulation time of the compound scaffolds(37 min) was longer than that of the PLA scaffolds(26 min). The anti-coagulation ability of the compound scaffolds was better than that of the PLA scaffdds.In the cell toxicity test,the cell growth situation of the compound scaffolds group was obviously better than that of the PLA group, and at the meantime the cell toxicity of the compound scaffolds was obviously smaller than that of the PLA scaffolds. Conclusion The material of PLA/silk fibroin compound scaffolds has the advanced biological consistent compared with the simplex scaffolds.Accordingly,the PLA/silk fibroin can be used as a scaffolds matrix to be transplanted into the body.

Abstract:Objective To construct the specific high efficiency small interfering RNA (siRNA) expression vector that can silence PTTG gene. Methods Using vector based RNA interference technique, vectors were constructed to transcribe functional siRNA specially targeting PTTG. The vectors were used to transfect U251 cells by lipofectmine2000 reagent. And the cells were dividedinto five groups: normal control, HK negative group and three siRNA interfering groups (pGenesil2-PTTG siRNA1, pGenesil2-PTTG siRNA2 and pGenesil2-PTTG siRNA3). The expression levels of mRNA and protein of PTTG were analyzed by RT-PCR and flow cytometry methods. Results By restriction endonuclease and DNA sequencing analyzing, eukyaryotic expression plasmid of PTTG was successfully constructed. PTTG mRNA and its protein level in transfected U251 cells with PTTG RNAi plasmid decreased significantly compared with the normal control group (P<0.01). Conclusion pGenesil2-PTTG siRNA vectors were successfully constructed, and they can silence the PTTG gene in U251 cells with high efficiency.

Abstract:Objective To clone the human TNF-like molecular 1A (TL1A) gene, and construct prokaryotic plasmid of TL1A and express it in E.coli, further more, to obtain high pure HCA661 protein.Methods The gene encoding TL1A was amplified using the total RNA of human umbilical vein epithelial cells (HUVECs) as template by RT-PCR, and inserted into pTA2 vector, then identified by restriction enzyme and sequencing.The recombinant expression plasmid PQE-TL1A was constructed and transferred into E.coli M15.The recombinant protein was expressed under the induction of IPTG, identified by Western blotting, purified by Ni-NTA affinity chromatography column.Results The identification of target gene by restriction enzyme was the same as the expectation.The sequence of target gene was identical with that registered in GenBank.With induction of IPTG, a new fusion protein with relative molecular mass of 22 000 was expressed and mainly located in inclusion bodies; the expressed 6×his-rhTL1A fusion proteins were identified by Western blotting with anti-His monoclonal antibody.Conclusion The　 recombinant prokaryotic expression plasmid pQE-Tl1A is constructed successfully, and recombinant TL1A protein with high purity coefficient is gained.

Abstract:Objective To clone different lengths cell division autoantigen1( CDA1) promoters(CDA1P) and measure their activities in order to provide basis for fumctional study of CDA1P. Methods Different lengths CDA1P were amplified by PCR using mouse liver genomic as template and cloned into BglⅡ digested pGL3-basic plasmid to produce recombinant plasmid pGL3-basic-mCDA1P, then they were transfected into Lewis lung cell line (LLC) and RAW 264.7 cells differently, these cells were collected after transfected for 48 h, finally luciferase activities of pGL3-basic-mCDA1P were measured. Results ①Different lengths CDA1P were cloned and characterized by restriction enzymes. ②Different lengths CDA1P were ligated into pGL3-basic vector,and were proved with PCR and DNA sequencing .③Activities of pGL3-basic-mCDA1A were different in the two kinds of Leuwis and mononuclear macrophage cells (RAW264.7 cells). Conclusion The different activities of CDA1P may be related with different cellular types.

Abstract:Objective To determine the role of epidermal growth factor receptor (EGFR) signaling in bone development in mice.Methods Long bone from EGFR wild type (EGFR+/+) and knockout mice (EGFR-/-) were collected and analyzed by Trichrome Masson staining, in situ hybridization was used to observe trabecular bone formation and recruitment of osteoblasts.Results EGFR deficiency can cause delayed trabecular bone formation in EGFR-/- mice and delayed osteoblast recruitment compared with EGFR+/+ mice.There were different expressions of core binding factor A (Cbfa), osteocalcin (OC) and collagen I (Col I) between EGFR+/+ and EGFR-/-.Conclusion EGFR signaling play an important role in trabecular bone formation, the partial mechanisms for which might be related to the regulation of osteoblast recruitment.

Abstract:Objective To investigate the neuroprotection of antidepressants venlafaxine and fluoxetine on the hippocampus neurons injured by glutamate.Methods Primary cultured hippocampus neurons were injured with glutamate to mimic the pathological changes in depression.Four groups were divided as control, glutamate, venlafaxine and fluoxetine administration group.The survival rate of neurons was detected with MTT method.LDH release and SOD activity were detected by spectrophometry.Results The survival rate of neurons injured with the excitatory neurotoxicity of glutamate decreased to 50% of control (P<0.05).The survival rates of neurons injured with glutamate increased dramatically after administration with venlafaxine as compared with those in glutamate group (P<0.05 or P<0.01).However, the survival rates of neurons were not increased after administration with fluoxetine.The LDH release of neurons injured with glutamate increased significantly, which was 7.4-fold of control group.However, after administration with two kinds of antidepressants LDH release from neurons injured with glutamate decreased in varying degrees as compared with that in glutamate group.The SOD activity of neurons in hippocampus injured with glutamate decreased significantly（P<0.01）, nevertheless, the administration of venlafaxine can reverse the parameter（P<0.001）, but the activities of SOD in fluoxetine groups (100 and 500 μmol•L^-1)decreased significantly (P<0.05).Conclusion Antidepressant venlafaxine and fluoxetine both have certain neuroprotection, however, venlafaxine has greater neuroprotection compared with fluoxetine.

Abstract:Objective To investigate the effects of 15-deoxy-Δ12,14-prostaglandin J₂(15d-PGJ₂) and DDP on the growth of human pulmonary carcinoma PLA-801D cells and the mechanisms of apoptosis.Methods The human pulmonary carcinoma PLA-801D cells were selected and added to each well of 96-well place and cultivated for 24 h.Then the cells were treated with different concentrations of 15d-PGJ₂(0,5,10,20,40 and 80 μg•L^-1) or 15d-PGJ₂ combined with DDP(3 mg•L) for 24 h.0 μg•L^-1 15d-PGJ₂ group was control group.The morphological changes of cells were observed under inverted microscope.Microculture tetrazolium (MTT)dye was applied to detect the proliferation of the human pulmonary carcinoma PLA-801D cells treated with 15d-PGJ₂ and DDP.Diphenylamine assay (DPA) was used to evaluate the activation.Flow cytometry assay( FCM) was used to detect the apoptosis proportion and the changes of cell cycle.Results When the human pulmonary carcinoma PLA-801D cells were treated with low- concentration 15d-PGJ₂ alone( 5,10 and 20 μg•L^-1), no significant difference was observed in the inhibitory rate of cell growth and the apoptotic indexes such as the apoptosis proportion, the percent of DNA fragmentation and the activity of caspase-3 compared with control roup（P<0.05）；when 15d-PGJ₂ concentration was 40 μg•L^-1 ,the inhibitory rate of cell growth the apoptosis proportion and the percent of DNA fragmentation were higher than those in control group （P<0.01）.While the cells were treated with 15d-PGJ₂ (10 μg•L^-1)combinated with DDP(3 mg•L^-1), the inhibitory rate of cell growth, the apoptosis proportion, the percent of DNA fragmentation and the activity of caspase-3 were significantly higher compared with 15d-PGJ₂ alone（P<0.01）.when the human pulmonary carcinoma PLA-801D cells were treaed with 15d-PGJ₂ alone for 24 h, and the 15d-PGJ₂ concentration was 40 μg•L^-1, the cell proportion in G₀/G₁ phase was significantly higher than that in control group,and the cell proportion in S and G₂/M phase was greatly lower than that in control group（P<0.01）.Conclusion High- concentration 15d -PGJ₂ can induce the apoptosis of human pulmonary carcinoma PLA-801D cells , but the effects inducing apoptosis of 15d -PGJ₂ alone are weaker than those of 15d -PGJ₂ low- concentration combined with DDP.

Abstract:Objective To explore the effects of oxymatrine(OMT) on injury induced by focal cerebral ischemia in rats and its mechanism. Methods Rats were randomly divided into sham(n=8),cerebral ischemia model(n=8),Lulu Tong(LLT,31.25 mg•kg^-1,n=8) and OMT 35,70 and 105 mg•kg^-1 groups(n=8). The focal cerebral ischemia was induced by middle cerebral artery occlusion in rats. The drugs were administrated by intraperitoneal injection. The neurological score and infarct volume were used to evaluate the protective effects of OMT in focal cerebral ischemia rats. The NO content in serum and myeloperoxidase(MPO) content in brain tissue were determined to study the its mechanisms. Results Compared with model group,the infarct volumes were decreased in OMT 35,70 and 105 mg•kg^-1 groups(P<0.05，P<0.01，P<0.001) and the neurological scores were decreased in OMT 70 and 105 mg•kg^-1 groups(P<0.01); The NO content in serum and MPO content in brain tissue in OMT groups were decreased (P<0.05，P<0.01). Conclusion OMT has protection against injury induced by cerebral ischemia in rats. The anti-inflammation effects may be one of the mechanisms.

Abstract:Objective To study the inhibitory effects of siRNA on MDM2 expression in osteosarcoma cell line U20S and cell proliferation. Methods siRNA targeting MDM2 (PGCsilencer^TM-MDM2-siRNA) was constructed and transfercted into U20S cells.Meanwhile,control siRNA was transfected and the transfection agent was used as the comparison.MDM2 gene and protein expressions were measured using RT-PCR and Western blotting.Cell proliferation was measured with MTT assay.Results RT-PCR and Western blotting results showed that MDM2 mRNA expression levels were reduced to 32.61% and 39.06% after being transfected with siMDM2-1 and siMDM2-２; MDM2 protein expression levels decreased to 35.76% and 42.20%,respectively.The difference betweencontrol siRNA and liposome group was not obvious（P>0.05）.MTT result showed that the cell growth was inhibited by being transfected with siMDM2,the difference was obvious compared with liposome group（P<0.05）.The difference was not significant between control siRNA and liposome group (P>0.05).Conclusion siRNA　 can effectively down-regulate MDM2 expression in U20S cell line and inhibit the cell proliferation.

Abstract:Objective To study the effect of candesartan on the ERK1/2 protein expression in myocardium of epilepsy rats induced by kainic acid(KA) and its mechanism.Methods 105 male Wistar rats were divided into: control g₁₈₀roup（A）;epilepsy groups(B_1-5);candesartan groups(C);1-5 meaned 0，0.5，2，4，6 h after epilepsy,respectively.Epilepsy rat models were made by injecting KA into amygdale under stereotactic instrument.The ERK1/2 protein expression in various groups were tested with immunohistological method.Results Compared with control group,the protein expressions of ERK1/2 increased significantly in the 0.5 h groups of B and C （P<0.05）,and attained their peaks respectively in 2 h groups of B and C（P<0.01）,then decreased gradually in 6 h groups of B and C（P>0.05）.The differences in the level of ERK1/2 protein expression between B and C groups were significant（P<0.05）.Conclusion The protein expression of ERK1/2 in rat myocardium increases after epilepsy in a short time;candesartan can decrease the protein expression of ERK1/2 in the myocardium of epilepsy rats.

Abstract:Objective To investigate the alteration of aquaporin 1 (AQP1) expression in lung tissues in hemorrhage-lipopolysaccharide(LPS) two-hits rats and the effects of panaxadiols(PDS)and dexamthasone(Dex) on it.Methods The rat model of acute lung injury was built with hemorrhage-LPS two hits.The experiment was divided into control group(S),two-hits model group(HL),DEX group(HLD),and PDS group(HLP).The pathological changes of lung tissue were examined by HE staining.The expression of AQP1 was analyzed by RT-PCR,Western blotting and immunohistochemical staining. Results ① Significant inflammatory changes in pulmonary interstitial of rats in HL group were observed.However,in HLD group and HLP group,the pulmonary pathologic changes were much slighter.② AQP1 mRNA and protein expressions in lung tissues in HL group were significantly decreased compared with others groups(P<0.05),immunohistochemistry analysis got the similar result.Positive AQP1 expression in the endothelium of capillary vessels on the wall of alveolus was merely very weak expression in HL group.AQP1 mRNA and protein expression levels in lung tissues in HLD group and HLP group were significantly higher than those in HL group(P<0.05).Conclusion The hemorrhage-LPS two hits can inhibit the expression of AQP1,while PDS could increase AQP1 expression in lung tissue,lighten the edema,and maintain the intact structure of lung tissue.

Abstract:Objective To investigate anti-inflammatory effects and analgesic effects of Qianlie Shutong Tablets(QLS) and provide medical basis for its clinical use.Methods A total of 40 rats or mice were randomly assigned into control group(n=8),drug control (Aspirin,n=8) group,low -dose QLS (mice: 25 mg•kg^-1•d^-1,n=8;rats: 17.5 mg•kg^-1•d^-1,n=8) group,mid-dose QLS (mice: 50 mg•kg^-1•d^-1,n=8;rats: 35.0 mg•kg^-1•d^-1,n=8) group and high -dose QLS (mice: 100 mg•kg^-1•d^-1;rats: 70.0 mg•kg^-1•d^-1) group.Anti-inflammatory effects were tested by xylene-induced ear edema in mice,ovoalbumin-induced feet edema and cotton pellet-induced granuloma in rats.Analgesic effects were tested by hot-plate method and 0.6 % acetic acid solution injected into the mice abdominal cavity to produce pain in order to cause the body bend.Different doses of QLS were given and their actions were observed.Results QLS had obvious anti-inflammatory effectts on the xylene-induced ear edema.Compared with control group,ear edema was significantly decreased by QLS with the doses of 50 and 100 mg•kg^-1•d^-1 (P<0.05 or P<0.01),the ear edema inhibitory rates were 53.97 % and 60.09 % in mice,respectively.Ovoalbumin-induced rat feet edema was significantly decreased by QLS with the doses of 35 and 70.0 mg•kg^-1•d^-1 (P<0.05).Cotton pellet-induced rat granuloma was significantly decreased by QLS with the doses of 35.0 and 70.0 mg•kg^-1•d^-1 (P<0.05 or P<0.01 ) and the granuloma inhibitory rates were 32.40 % and 35.65 %,respectively.Pain thresholds 90,90 and 30min after administration were markedly elevated (P<0.05 or P<0.01) and the licking-paw latencies in hot plate method were extended by QLS with the doses of 25,50 and 100 mg•kg^-1•d^-1 (P<0.05),the inhibitory rates of pain threshold were 36.17%,44.13%,and 46.64%,respectively.Conclusion QLS has significant anti-inflammatory and analgesic effects.

Abstract:Objective To analyze the stress distribution of abutment and alveolar ridge between two kinds of distal-extention absence dentures,and provide evidence for the application of different dentures in clinic. Methods Attachment　 denture and cantilever fixed bridge were used to restore the distal-extention absence respectively,the stress distribution of abutment and alveolar ridge was compared by three-dimensioal finit element method.Results The stress peak values of abutmen root,periodontal membrance,alveolar ridge were (1.42E+5 )MPa,(1.33E+4)MPa and (3.49E+5)MPa, respectively, in attachment denture;while in cantilever fixed bridge,the stress peak values of them were (1.45E+7)MPa and (2.25E+6) MPa, (1.45E+3)MPa, respectively.And the stress peak values of these two kinds of dentures were all located in the mid-upper 1/3 and the cervical of abutment.Conclusion Compared with cantilever fixed bridge, the attachment denture could decrease the stress of abutment and be helpful to its protection,the hight of residual alveolar ridge should be taken into account when the two kinds of dentures are used for distal-extention absence.

Abstract:Objective To investigate the effect of dexamethasone on proliferation of human periodontal ligament fibroblasts(hPDLF) cultivated in vitro,and search for optimal culture condition for hPDLF in vitro， and provide basis for further study on regeneration of periodontinum.Methods The hPDLF cultivated in vitro were divided into 5 groups and cultivated in 5 different culture media separately which all included 10% fetal bovine serium: ①control group(no DEX）；②5 mg•L^-1DEX；③10 mg•L^-1 DEX；④20 mg•L^-1 DEX；⑤50 mg•L^-1 DEX. The proliferation of hPDLF was assyed by MTT method.The phase contrast microscope was used to observe the morphological changes of the cells.Results The PDLF appeared aggregation and cells on bottom of culture bottle showed squamae-shape after inoculated on plate with 24 hole for 3 d.There were more layers of smaller hPDLF with blunt prominency came forth after cultivated in DEX culture media.MTT method showed that DEX promoted the proliferation of hPDLF obviously compared with control group(P<0.05).The different concentrations of DEX had different effects on proliferation of hPDLF(P<0.05),the proliferation ability was enhanced with the increasing of DEX doses.Conclusion DEX could improve the proliferation ability of hPDLF cultivated in vitro.

Abstract:Objective To discuss the role of matrix metalloproteinase-8(MMP-8) mRNA in apical granuloma and periapical cyst by detecting the its expressions in the periapical granuloma and periapical cyst. Methods Reverse transcription polymerase chain reaction(RT-PCR) technique was used to measure the expressions of MMP-8 mRNA in 22 cases of periapical granuloma,11 cases of periapical cyst and 10 cases of normal periapical tissues. Results The positive rates and the levels of MMP-8 mRNA expression in apical granulomas and periapical cysts were significantly higher than those in normal tissues(P<0.01). However, the expression of MMP-8 mRNA was not obviously different between apical granuloma and periapical cyst(P>0.05). Conclusion The expression of MMP-8 may play an important role in the pathogenesy of apical granuloma and periapical cyst,and it may be involved in the process of bone destruction.

Abstract:Objective To investigate the effect of 2-methoxyestradiol(2-ME) on U937 myeloid leukemia cell line and its mechanism. Methods The experiment was divided into control group (myeloid leukemia U937 cell in RPMI 1640 culture medium with equal DMSO), 2-ME-treated group, NAC-treated group, and 2-ME+NAC-treated group. The cytotoxicity was analyzed by MTT assay. Apoptosis and cellular nitric oxide(NO) were detected by flow cytometry using annexin V and NO sensor dye. Superoxide anion was measured with a fluorescent plate reader by DHE. Results Viabilities of U937 cells treated with 2-ME (0.25, 0.50, 1.00, and 2.00 mol•L^-1 ) for 48 h were gradually reduced to 0.68±0.05, 0.28±0.07, 0.18±0.07, and 0.11±0.04，respectively. The differences were significant compared with control group (1.00±0.05) (P<0.05). 2-ME resulted in viability decrease in a dose-dependent manner. The apoptotic rates in 2-ME (2.00 μmol•L^-1 )-treated group were 4.64％±0.21％, 9.86％±0.9％, 14.62％±0.67％, and 19.49％±0.90％ at 8, 12, 18, and 24 h time points，respectively, and significantly higher than that in control group (1.74％±0.08％) (P<0.05). There were no significant differences of apoptotic rate between 2-ME-treated group and control group at 1, 3, and 6 h time points（P>0.05）. 2.00 μmol•L^-1 2-ME also significantly increased the mean fluorescence of NO sensor dye and superoxide anions in U937 cells compared with control group (P<0.05). The percentage of NO positive cells increased from 0.52％±0.21％ to 46.74％±0.15％ after treatment for 1 h with 2.00 μmol•L^-1 2-ME (P< 0.05), whereas, there was no significant difference of apoptotic cells at this point between 2-ME group and control group (1.28％±0.07％ vs 1.59％±0.12％, P>0.05). An markedly increase in apoptotic cells was detected after 2-ME treatment for 3 h(6.78％±1.01％ vs 1.59％±0.12％, P<0.05).These results indicated that the generation of NO was earlier than apoptosis underwent. Furthermore, quenching of ROS with NAC protected leukemia cells from the cytotoxicity of 2-ME and prevented apoptosis induced by 2-ME. The viabilities and apoptotic rate of 2.00 μmol•L^-1 2-ME-treated group were 0.47±0.02 and 13.87％±0.69％, respectively. The differences were significant compared with control group or 2-ME＋NAC group( 0.82±0.08 and 2.98％±0.19％, respectively) (P<0.05). Conclusion 2-ME can induce apoptosis in U937 cells through generation of ROS. It is possible to use ROS-generation agents to enhance the antileukemic effect.

Abstract:Objective To investigate the stress in the periodontal tissues using three-dimansional finite element method when Richtts Arch was used. Methods Three-dimensional finite element models of the lower left central incisor and first molar were set up by means of CT.Stress distribution in root,PDL and alveolar bone,and the tendency of the tooth movement were obtained by calculation under Richtts Arch.Results The value of the force at the left lower incisor was －0.34 Newton and the value of the force at the left lower molar was 0.65 Newton.The molar model revealed that the tensile stress concentration was at the mesial root cervix and the compressive stress was at the distal root cervix.The incisor model showed that the tensile stress was concentrated at the lingual cervix and the compressive stress was concentrated at the apical tip.The arch wire exerted intruding force,lingual force and the incisor tended to lingual movement and intruding movement.The molar had the tendency of lingual movement and distal tipping movement.Conclusion The anchorage molar will incline mesially when Richtts Arch is used in treatment of deep overbite,the additional torque must be used in the anchorage molar.

Abstract:Objective To investigate the inhibitory effect of silencing survivin gene with siRNA on the growth of gastric cancer cells and its mechanism,and provide evidence in treatment for gastric cancer.Methods DNA template coding survivin-specific siRNA was designed and synthesized.Two recombinant plasmids (pGCsilencerU6/GFP/survivin-siRNA-１ and -2) were constructed. The gastric carcinoma cel1 line SGC-7901 were divided into three groups: liposome-treated control group,empty plasmid-transfected control group and survivin-siRNA-1 transfected group. In order to observe the effect of survivin-siRNA, the expressions of survivinmRNA and protein were detected by RT-PCR and Western blotting,respectively.Methyl thiazolyl tetrazolium (MTT) assay was applied to determine the cell growth status.Apoptotic rates were evaluated by flow cytometry (FCM).Results The results of Western blotting and RT-PCR indicated that the inhibitory rates of protein and mRNA in pGCsilenerU6/GFP/survivin-siRNA-1 transfected group （78.25% and 88.75%) were higher than those in liposome-treated control group（5% and 2%） and empty plasmid-transfected control group （1% and 6%）（P<0.01）.The inhibitory rates of protein and mRNA in pGCsilenerU6/GFP/survivin-siRNA-2 transfected group （42% and 77%）were higher than those in liposome-treated control group and empty plasmid-transfected control group also (P<0.01).The MTT assay results showed that the inhibitory rate of cell proliferation in survivin-siRNA-1 transfected group （64%）was higher than that in empty plasmid-transfected control group （11%）（P<0.01）；The FCM results showed that the apoptotic rate in pGCsilenerU6/GFP/survivin-siRNA-1 transfected group（21.20±3.21） was higher than those in liposome-treated control group（0.830±0.001） and empty plasmid-transfected control group（1.98±0.67）（P<0.01）.Conclusion pGCsilencerU6/GFP/survivin-siRNA can not only suppress the expression of Survin in gastric cancer cells and proliferation of SGC-7901 cells,but also induce apoptosis of gastric cancer cells.

Abstract:Objective To study the security of low dose and half-body irradiation by ⁶⁰Co γ-rays as a new method of clinical radiotherapy．Methods The simulated manikin was used to simulate human body and two radiation modalities of facing and backing on radioactive source were adopted．Half-body irradiation was done by ⁶⁰Co γ-rays with doses of 9，10 and 11 cGy．The exposure dose of every layer and important target organs in the simulated manikin were detected,and the security of low dose and half-body irradiation as a therapeutic method was evaluated．Results The exposure dose of every layer and sensitive organs were all within safety margin,when simulated manikin facing or backing on the radioactive source was irradiated by ⁶⁰Co γ-rays with doses of 9,10 and 11 cGy．Further，the exposure dose of sensitive organs in the simulated manikin backing on the radioactive source was lower than that in those facing the radioacive source．Conclusion The method of low dose and half-body irradiation as a radiotherapeutic method is safe and feasible and the radiation modality of backing on the radioactive source is more safe．

Abstract:Objective To observe the serum level of C-reactive protein(CRP) in transient ischemic attack(TIA) patients,and to assess the correlation between CRP and other dangerous factors of cerebral infarction.Methods 102　 TIA patients were divided into two groups: TIA deteriorated into cerebral infarction in two weeks (group A,n=40) and TIA could be relieved in two weeks (group B,n=62).Many factors were measured within 24 h,such as CRP,BP,BS,FIB,TC,TG,HDL-L, and LDL-L.67 healthy subjects were usded as control group.Relative analysis was performed between CRP and 8 parameters mentioned above.Results ①The serum CRP level of TIA patients was higher than that of healthy controls (P<0.05).It was also higher in group A than that in group B (P<0.05).②The correlative analysis demonstrated that the serum CRP level had obviously positive relationships with BP,BS,FIB and TG(P<0.01),the　γ　 values were 0.517,0.439,0.533,and 0.406, respectively.Conclusion CRP is a non-differential,but very important new serum symbol to evaluate whether TIA evolves to cerebral infarction in a short period of time.The higher serum CRP level is,the more probably TIA evolves to complete stroke.

Abstract:Objective To determine the plasma DNA level of patients with ovarian can cer and evaluate its potential clinical value.Methods Blood samples were collected from 30 ovarian cancer patients before and after surgery,and from 20 patients with ovarian benign tumor and 20 healthy women.The plasma DNA was extracted by commercial kit and detected by fluorescntmeter.Results The mean plasma DNA level in the cancer patients (25.33 μg•L^-1±17.69 μg•L^-1) was significantly higher than those in the benign tumor patients (10.28 g•L^-1±4.80 μg•L^-1) and normal control(7.60 μg•L^-1±3.87 μg•L^-1) (P<0.001).There was no significant difference between the benign and normal groups.The plasma DNA level of the patients in stage Ⅰ-Ⅱ (15.83 μg•L^-1±5.30 μg•L^-1) was significantly higher than that in the normal control(P<0.01) and was significantly lower than that in stage Ⅲ-Ⅳ(30.83 μg•L^-1±20.04 μg•L^-1) (p=0.005). The plasma DNA level of patients with metastasis(35.43 μg•L^-1±21.08 μg•L^-1) was significantly higher than that of patients with no metastasis(16.49 μg•L^-1±6.44 μg•L^-1) (P=0.006). The DNA level in the cancer patients one month after surgery (10.30±2.45 μg•L^-1) was much lower than before surgery(P<0.001),near that in the normal control.The overall survival of the patients with high plasma DNA level was much shorter than that of the patients with low DNA level(P=0.027).When the cut-off for diagnosis of ovarian cancer was 15.70 μg•L^-1,the sensitivity and specificity were 63.30% and 90.00%,respectively. Conclusion The plasma DNA level may have a potential use in early diagnosis and serve as a predict marker for metastasis potential and prognosis of ovarian cancer.

To explore the role and possible mechanism of survivin gene during the progress of originating and developing of human hepatocellular carcinoma(HCC). Methods Reverse transcriptase-polymerase chain reaction(RT-PCR) was used to detect mRNA expressions of survivin and related genes included STAT3，c-myc，p53 and VEGF in HCC tissues，tissues surrounding tumor and normal tissues of fifteen HCC cases.The protein expression of survivin gene was detected by Western blotting and immunohistochemical method.Results The mRNA expression levels of survivin gene in HCC tissues and tissues surrounding tumor were significant higher than that in normal liver tissues(P<0.01),and there was also significant difference between HCC tissues and tissues surrounding tumor (P<0.01).The mRNA expressions of c-myc，STAT3 and VEGF genes were up-regulated while the mRNA expression of p53 was down-regulated in HCC tissues and tissues surrounding tumor compared with normal tissues (P<0.01), there was also significant difference between HCC tissues and tissues surrounding tumor (P<0.01).The expression levels of survivin protein in HCC tissues and tissues surrounding tumor were higher than that in normal liver tissues (P<0.01), there was also significant difference between HCC tissues and tissues surrounding tumor (P<0.01).Conclusion The persistent activation of survivin gene plays a promotion role in the carcinogenesis of HCC，and survivin gene may act as a new target in therapy of HCC.

To detect the polymorphism of CYP1A1-MspI gene in patients with ovarian cancer.and discuss the relationship between the polymorphism of CYP1A1-MspI gene and correspond cases’ general materials and clinical materials. Methods The free peripheral blood samples of 81 cases confirmed to be ovarian cancer by postoperative pathology were collected preoperatively and the polymorphism of CYP1A1-MspI gene was detected.The clinical materials of the 81 cases with different genotypes were compared.The relationship between the polymorphism and clinical materials was analyzed. Results Among the 81 cases of ovarian cancer, there were 47 cases of wild type-genotype A(T/T)(58%),25 cases of mutation heterozygosis-genotype B(T/C) (31%), and 9 cases of mutation homozygosis-genotype C (C/C)(11%). The genotypic frequency distribution in patients aged from 12 to 29 was one case of genotype A (2.1%), 5 cases of genotype B(20.0%), and no case of genotype C. The genotypic frequency distribution in patients aged from 30 to 49 was 12 case of genotype A (25.5%),8 cases of genotype B(32.0%),and 3 cases of genotype C(33.3%) . The genotypic frequency distribution in patients aged from 50 to 69 was 31 case of genotype A (66.0%), 8 cases of genotype B(32.0%) and 4 cases of genotype C(44.4%) . The genotypic frequency distribution in patients aged more than 70 years was 3 case were of genotype A (6.4%), 4 cases of genotype B(16.0%), 2 case of genotype C (22.2%). There were significant differences of the ages of onset between patients with different CYP1A1-MspI genotypes (P<0.05).There were 40 cases of genotype A (85.1%),15 cases of genotype B(60.0%), and 9 cases of genotype C (100%) in epithelial cancer patients. There significant differences of the constituent ratio of the ovarian cancer pathological types between patients with different genotypes(P<0.05) . Conclusion Polymorphism of CYP1A1-MspI gene exisits in patients with ovarian cancer. The age of onset of patients with ovarian cancer is related with CYP1A1-MSPI genotypes; The incidence of epithelium tumor in patients with genotype A is higher than that of other ones. The ratio of non-epithelial tumor in the patients with mutation allele gene C has an increasing tendency.

To observe the effect of acute hypervolemic hemodilution (AHH) combined with nicardipine controlled hypotension on hemodynamics changes in revision operation of total hip replacement. Methods Forty patients were divided into two groups according to the sequence of the surgery (odd number came into group A,even number came into group B,20 in each group).In group A,AHH was carried out with nicardipine controlled hypotension;group B was control group.AHH was carried out in both groups before anesthesia induction.6%voluven was transfused in group A at 15 mL•kg^-1 and 30 mL•min^-1 and nicardipine was transfused at 0.3-0.5 mL•kg^-1•h^-1 with MAP altering between 60 and 65 mmHg;Same quantity of physiological saline was transfused in group B.HR,MAP,CVP,Hb,Hct and Plt were recorded and observed at preoperation,1 h after AHH and 24 h after the operation. Results In group A after AHH with nicardipine controlled hypotension,MAP was obviously lower than pre-operation (P<0.01) and group B (P<0.01) and higher than group B when controlled hypotension ceased. There was no much variation in HR.There was an evident rise of CVP in both groups (P<0.01),it was lower in group A than that in group B (P<0.05).Hb,Hct and Plt decreased evidently after AHH (P<0.01,P<0.05) and went up 24 h post-operation,and there was no statistical significance when they went lower than preoperation in group B,while Hb in group A was still lower than that pre-operation (P<0.05).Conclusion AHH combined with nicardipine co ntrolled hypotension can maintain stable hemodynamics in revision operation of total hip replacement.

To study the relationships between the expression level of PTEN gene, gene mutation and occurrence and development of glioma.Methods Fifteen glioma tissues were selected,and the normal 4 non-glioma tissues were used as control.The expression level of PTEN mRNA was measured by RT-PCR method,and the mutation of PTEN gene was screened by SSCP analysis in various histopathological grades of glioma. Results Compared　 with control group(0.126±0.015),the level of PTEN mRNA in glioma tissues(0.063±0.006) was decreased（P<0.05）.SSCP analysis showed that there was not mutation of the bands of exon 8 and 9 of PTEN gene .The band shift of exon 5 of PTEN gene of one specimen was found on ployacrylamide gel electrophoresis. Conclusion The low expression and mutation of PTEN gene probably promote the occurrence and development of glioma.

To explore the rehabilitation effect of constraint-induced movement therapy(CIMT) on upper extremity motor function of stroke patients with hemiparasis. Methods 20 chronic stroke patients were divided into 2 groups: CI group(10 cases),the patients were treated with CIMT for 3 weeks,3 h per day;control group(10 cases),the patients were treated with repetitive occupational therapy for 3 weeks,3　h per day.MAL score and WMFT score were measured in two groups before treatment and 1 week,6 months and 12 months after treatment. Results There were improvement of MAL score in both CI group and control group 1 week after treatment compared with pre-treatment(P<0.01),and the scores of MAL on post-treatment 6 months and 12 months　 were higher than those of pre-treatment in CI group(P<0.05,P<0.01),but no significant difference in MAL score between post-treatment 6 months and 12 months and pre-treatment in control group;there was no significant difference in WMFT between pre-treatment and post-treatment 1 week,12 months in CI group and control group(P>0.05). Conclusion CIMT can significantly increase the use ability on the upper extremity of hemiplegic patients after stroke,but can not improve the motor function of upper extremity in stroke patients.