Objective To study the expression of Ephrin‐B4 receptor (EphB4) in hepatocellular carcinoma (HCC) and its clinical significance ,and to analyze the effect of EphB4 on the proliferation of HCC cells .Methods The expression level of EphB4 in HCC tissues and matched paracancerous liver tissues of 60 cases of HCC patients was assessed by reverse transcriptase‐polymer‐ase chain reaction (RT‐PCR) and immunohistochemistry .The correlation between the expression of EphB4 in HCC tissues and clin‐ical pathologic parameters was analyzed by chi‐square test .Univariate survival analysis was carried out by Kaplan‐Meier Log‐rank test .The effect of EphB4 on the viability of HCC cells was furtherly analyzed by MTS .Results The results of RT‐PCR showed that the mRNA level of EphB4 in HCC tissues (1 .39 ± 0 .80) was significantly higher than in matched paracancerous liver tissues (0 .56 ± 0 .33) ,the difference was statistically significant (P< 0 .01) .Data from immunohistochemistry showed that the positive rates of EphB4 protein in HCC tissues and matched paracancerous liver tissues were 81 .7% and 23 .3% ,respectively .Moreover ,the expression of EphB4 in HCC tissues was relevant to AFP level ,tumor size and TNM stage(P<0 .05) .The three‐year survival rate of HCC patients with positive expression of EphB4 protein was 22 .5% ,and that of HCC patients with negative expression of EphB4 protein was 54 .5% ,the difference was statistically significant (P<0 .05) .Overexpression of EphB4 significantly enhanced the ability of hepatocellular carcinoma cell proliferation (P<0 .05) .Conclusion EphB4 expression was significantly up‐regulated in HCC ,which was associated with HCC progression and prognosis ,and EphB4 could promote the proliferation of HCC cells ,which could be used as a marker of HCC progression .

Objective To investigate the applying comparative values of anticoagulant and thrombolytic Sak-hirudin fusion protein used in cardiopulmonary bypass.Methods From february 2013 to July 2014,selected nitinol sheet into uncoated group (group A),base coated group (chitosan,group B),chitosan/ Sak group (group C),chitosan/hirudin group (group D),chitosan/Sak-hirudin fusion protein group (group E),there were included in the fresh round of blood hemolysis,the blood cell hemolysis rates were calculated.Meanwhile used the healthy newborns umbilical vein endothelial cells were added into various categories nitinol sheet for cell compatibility testing.Results The samples hemolysis rates in the 5 groups were around 1.5% and the RBC,WBC and PLT counts compared in the 5 groups were showed no differences (P <0.05).The ex-tent of proliferation and adhesion in the group E were significantly lower than the other four groups,compared to significant differences (P <0.05).There were no differences to compare group A and group B (P >0.05).Randomized double-blind cell count result shows that the group C,D and E compared to the group A,B were more significant differences (P <0.05), at the same time,the group E compared to the group C,D differences were statistically significant (P <0.05).Conclusion Compared to Staphylokinase and hirudin alone,Sak-hirudin fusion protein used in cardiopulmonary bypass can play stronger anticoagulant and antithrombotic effects,its safety and ensures the effectiveness that interventional treatment of cardiovascu-lar disease.

K. There was no significant difference in the yield among the treatments of N+K, CK, P, N+P, and N, but the results of these five treatments were significantly higher than that of P+K and K. In addition, cichoric acid content did not considerably changed after treatment of various fertilizer combinations. For the second harvest date the yield of N, N+K, P, and N+P were 47.7%, 35.4%, 33.8%, and 12.3% higher respectively than that of CK, the yield of N+P+K, P+K, and K were 7.7%, 10.8%, and 28.5% lower respectively than that of CK. There was significant difference in the yield between the treatment of N and CK, the yield of K was significantly lower than that of CK. Conclusion The results indicate that cichoric acid content is not significantly affected by the treatment of various fertilizer combinations and the yield is strongly influenced by N fertilizer, weakly by P fertilizer, on the contrary the application of potassium chloride results in a decrease in yield.

Objective To design a novel efficient CEA vaccine of fission epitopes with the vehicle of HPV16L1,and construct the recombinant baculovirus in Bac-to-Bac baculovirus expression system.Methods The recombinant baculovirus expression vector pFastBac1-V was constructed.HPV16L1 and CEA-PADRE synthesis primers were inserted into bacmid in E.coli DH10Bac with the help of Tn7 transposition system.Then the recombinant baculovirus was got by infecting insect cell sf9.Results HPV16L1 and CEA-PADRE synthesis primers that were cloned to the baculovirus transfer vector pFastBac1 were assessed by restriction analysis and direct sequencing.The recombinant baculovirus of the novel vaccine had been constructed.After infected sf9,the correct fragment was amplified by PCR.Conclusion The baculovirus of CEA vaccine of fussion epitopes with the vehicle of HPV16L1 was constructed.

Objective Construction of human IL‐4 recombinant expression vector and then conduct the expression ,purification and identification of human recombinant IL‐4 .Methods the open reading frame of IL‐4 was amplified by nest PCR with total RNA from PBMC of healthy volunteer .And then the amplified IL‐4 was inserted into pET101/D‐TOPO ,transformed into BL21 ,ex‐pressed ,purified and indentified .Results The size of amplified open reading frame of IL‐4 was about 460 bp and the sequence was correct .After transformed into BL21 ,the IL‐4 clone with higher expression level was selected by selection of different clones insert‐ed with IL‐4 and the size of expressed ,purified IL‐4 was about 28 × 103 .Western blot results showed that the size of single band was identical with the expected protein .Conclusion Human IL‐4 recombinant protein was got successfully .

Objective To investigate the advantage of separate bolus injection technique in CT urography (CTU) improving the display of the whole urinary tract.Methods Sixty cases of CTU examination,were divided into observation group and control group by random digits table with 30 cases each,observation group used separate bolus injection technique and control group used single bolus injection technique.The scanning included routine scanning,cortical phase,medullary phase and lag phase.Reconstruction of lag phase displayed the whole urinary tract.Then the image quality was compared between two groups.Results The whole urinary tract showed excellent in 12 cases (40.0％,12/30),good in 17 cases (56.7％,17/30),normal in 1 case (3.3％,1/30) in control group,which showed excellent in 23 cases (76.7％,23/30),good in 7 cases (23.3％,7/30) in observation group,there was significant difference in excellent rate between two groups (P ＜ 0.01).The CT value of starting of ureter was (238.6 ± 82.5) HU,middle-lower ureter was (245.9 ± 112.3) HU in control group and (239.0 ± 93.8),(235.3 ± 74.6) HU in observation group,there was no significant difference between two groups (P ＞0.05).There was no difference in developing of urolithiasis between two bolus injection techniques.Conclusion The application of separate bolus injection technique in CTU examination can reduce the dose of the first contrast material and contrast material reaction,and receive high-quality image of the whole urinary tract.

AIM: In order to search carrier material and better tissue culture method, the morphology and structure of the cultured cat corneal endothelium was observed. METHODS: The cat corneal endothelial cells were seeded on the Descemet's membrane of the dehydrated swine corneal stroma, and then cultured in the medium with epidermal growth factor and laminin for 7 days. The morphology and structure of reconstructed tissue was tested by the inverted microscope and the scanning electron microscope. RESULTS: As shown by the morphological observations, the cultured cat corneal endothelial cells formed an integrated membrane, and attached to the Descemet's membrane, which was similar to the nature tissue. The cells connected tightly to each other, and some of them arranged in hexagon approximately. CONCLUSION: The cat corneal endothelial cells could be rebuilt on the carrier of the dehydrated swine corneal stroma successfully, which is similar to the nature tissue.

Objective To investigate the feasibility, safety and short-term efficacy of endoscopic implantation of 5-FU Slow-release Particles for advanced gastroenteric tumor. Methods During the endoscopy procedure,slow-releasing 5-FU agents were implanted densely into the tumors and infiltrated area. Forty-five to sixty pieces of agents ( each piece equivalent to 1.67 mg 5-FU)were injected, which containing an average dose of 100 mg 5-FU. Results A total of 13 advanced gastric cancer patients were enrolled into this study. Significant effects were observed in 3 patients and good effects in 8 patients, but 2 cases with no effects. The total effective rate was 84.62%. The endoscopy examination showed that the size of tumors reduced in various degree at 1 - 2months after the implantation. No hemorrhage or perforation was observed. Parameters of hepatorenal function and routine blood test were stable after implantation. Conclusion Endoscopic implantation of 5-FU Slow-release Particles can relieve the symptoms of patients and limit tumor growth in advanced gastroenteric tumor with no marrow and hepatorenal functional impair.

Objective To study the protective effect on CCl4-injured L-02 hepatocytes of polysaccharides from Hyriopsis cumingii(HCP) and try to explain the mechanism.Methods The L-02 hepatocytes were incubated and then injured by CCl4.The levels of AST,ALT,MDA and SOD in cultural supernatant were detected by general methods.Cell viability was assayed by MTT method.Results The polysaccharides(25,250 and 1000?g?L-1)could reduce the levels of AST,ALT,MDA in cultural supernatant which increased by CCl4.It also could boost the hepatocytes viability and elevate the level of SOD which reduced by CCl4.Conclusion The results suggest that polysaccharides from Hyriopsis cumingii have direct protective activity on hepatocytes injured in vitro.It might be associated with the anti-oxidative activity of polysaccharides from Hyriopsis cumingii.

Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.