Objective To construct expression vectors that Renilla reniformis (Rluc) fused with neurotensin type 1 receptor (NTSR1),and to investigate the interaction between NTSR1 and other receptors,as well as intercellular signal transduction mechanism mediated by neurotensinl-R.Methods The human NTSR1 gene was amplified by PCR using the plasmid pcDNA3.1-hNTSR1 as template.The PCR product was digested,ligased with the plasmid pRluc and then be transformed into the competent cell Top10.The construct was identified by DNA sequencing.The recombinant plasmid was transiently transfected into human embryonic kidney 293 ( HEK293 )cells,and the expression of pRluc-hNTSR1-pcDNA3.1 was detected by confocal microscopy and Western blot.Results The fragment of 1257 bp was amplified by PCR,and the DNA sequences were identical with the gene in GenBank ( NM_002531 ).Western blot showed a band about 90kDa.Confocal microscopy showed that NTSR1 was expressed on the plasma membrane.Conclusion The pRluc-hNTSR1-pcDNA3.1 eukaryotic expression vector is successfully constructed,and the expression vector can be used to investigate the interaction between NTSR1 and other receptors,as well as intercellular signal transduction mechanism mediated by neurotensinl-R,which will provide new target for drug development.

Objective: To evaluate the mechanical properties of Rene95 alloy component made by laser engineered net shaping (LENS). Methods: The component was fabricated from Rene95 alloy powder by LENS, then the mechanical properties of the component, Rene95 alloy casting and dental Ni-Cr alloy casting were studied. Results: The tensile strength(MPa), yield strength(MPa), elongations(%) and the microhardness(Hv) of Rene 95 alloy component made by LENS were 1 354.27?63.44, 1 175.29?34.32, 16.53?5.85, and 464.33?10.95, those of Rene95 alloy casting 1 430.28?55.73, 1 145.52?29.78, 10.31?4.25 and 457.56? 11.38, those of dental Ni-Cr alloy casting 612.05?18.46, 426.06?12.70, 6.21?1.21 and 260.14? 6.08, respectively.Conclusion: The mechanical properties of Rene95 alloy component made by LENS were same as those of Rene95 alloy casting, better than dental Ni-Cr alloy casting.

Objective:To evaluate the anti-corrosion property of Ti-Zr alloy fabricated by laser solid forming technic in artificial saliva.Method:The corrosive behavior of Ti-Zr alloy fabricated by laser solid forming was studied by potentiadynamic polarization technique in artificial saliva at 37 ℃(pH=6.8).Ti-Zr alloy formed by casting was applied as the control.With the polarization curves given by the electrochemical corrosive measure system and the picture of corrosive plane given by SEM, the corrosive property of the Ti-Zr alloy were analyzed.Results:Ti-Zr alloy fabricated by laser solid forming showed more relatively positive corrosion potential, smaller corrosion current density and lower passive potential, and did not show any evidence of corrosive phenomena.Conclusion:Ti-Zr alloy fabricated by laser solid forming has good corrosive resistance in oral circumstances.

OBJECTIVE: To analyze the use situation and trend of cardiovascular and cerebrovascular drugs during 2007～2009 in a military sanatorium, and provide reference for the rational use of cardiovascular and cerebrovascular drugs and its essential medicines. METHODS: Cardiovascular and cerebrovascular drugs in the rehabilitation center during 2007～2009 were analyzed statistically in the respect of category, consumption sum and DDDs. RESULTS: The consumption sum and DDDs of cardiovascular and cerebrovascular drugs were increasing in the past three years. And its essential drugs account for 94.8%.The most widely used drugs were cerebrovascular drug, brain metabolism drug, nootropics and antihypertensive drugs. Oxiracetam and Xueshuantong were widely used cerebrovascular drug, brain metabolism drug and nootropics. Amlodipine were widely used antihypertensive drugs in the clinic. CONCLUSION: Cardiovascular and cerebrovascular drugs and its essential drugs were characterized with complex classification, numerous categories, extensive application and potential market demand, which require rational use of drug should be strengthened.

objective: To study the effects of laser engineered net shaping(LENS) in the manufacture of dental prosthesis. Methods: An experiment LENS system, including a 5 kW continuous wave CO 2 laser, a four-axes numerical control working table and powder feeding system, was built up, by which, a rectangular LENS component was made from Rene95 alloy powder. The microstructure of the component was observed with scanning electron microscope and ESA analysis. Results: The size of the component was 120 mm?15 mm?15 mm , and the wall of the component was perpendicular to the base. The microstructure of the component was made up of fine columnar dendrite with the length of 10～20 ?m, 10 times less than that of the cast made from the same material. The main elements of the powder distributed homogeneously,the negative effect of segregation could be limited and the properties of the component could be improved.Conclusion: The microstructure of components made by LENS is much better than that of those made by casting from the same material.

Aim To investigate the roles of intracellu-lar reactive oxygen species ( ROS ) and Nrf2 pathway in shikonin-induced A549 cell apoptosis. Methods The cytotoxicity was analyzed by MTT assay. The ap-optosis of A549 cells was analyzed by both cellular morphological and biochemical methods. The relative changes of the redox marks ( ROS/GSH) were studied by fluorescence assay in the shikonin-treated A549 cells in accompany with the changes of the intracellular redox homeostasis by GSH/GSSG ratio. ROS inhibitor was also employed in the treatment to find the role of ROS in shikonin-induced A549 cell apoptosis. Real-time PCR analysis and ELISA assay were performed as well to determine the role of Nrf2 pathway in the shiko-nin-induced A549 cell apoptosis. Results The IC50 of shikonin on A549 cells was 3. 2 mg·L-1 . The cellu-lar redox homeostasis shifted toward oxidation signifi-cantly in shikonin treatment in a time-dependent man-ner. The expression of the Nrf2 pathway related genes was up-regulated by shikonin ( 3 . 2 mg · L-1 , 8 h ) . The expression of the anti-apoptotic genes was down-regulated , and proapoptotic genes were up-regulated by shikonin (3. 2 mg·L-1, 24h). Futhermore, the inhi-bition of intracellular ROS alleviated the cytotoxicity of shikonin in A549 cells. Conclusion The critical role of shikonin-induced redox imblance in A549 cell, coped with the secondary produced ROS and Nrf2 path-way antioxidants, result in A549 cell apoptosis.

Objective To construct the expression vector of early growth response-1 promoter-macrophage inflammatory protein 3? (pEgr-1-MIP-3?, pEM) to study MIP-3? expression in transfected Lewis lung cancer cells. Methods MIP-3? cDNA was inserted into plasmid with Egr-1 promoter by directional cloning. The vectors were transfected into Lewis lung cancer cells with liposomes. The expression of MIP-3? in transfected Lewis lung cancer cells induced by electron ray irradiation at different doses were confirmed by RT-PCR and Western blotting. Results The recombinant vector was sequenced, and the result indicated the correct insertion in the expression vector. After electron ray irradiation at different doses, MIP-3? expression in the transfected Lewis lung cancer cells increased significantly. Conclusion Electron ray can enhance the expression of MIP-3? in Lewis lung cancer cells. This has laid the experimental foundation for the studies of pEM for radio-genetic therapy in vivo.

As the rapid development of economy necessitates a large number of oil, the contradiction between energy supply and demand is further exacerbated by the dwindling reserves of petroleum resource. Therefore, the research of the renewable cellulosic biomass resources is gaining unprecedented momentum. Because xylose is the second most abundant monosaccharide after glucose in lignocellulose hydrolyzes, high-efficiency bioconversion of xylose becomes one of the vital factors that affect the industrial prospects of lignocellulose application. According to the research progresses in recent years, this review summarized the advances in bioconversion of xylose, which included identification and redesign of the xylose metabolic pathway, engineering the xylose transport pathway and bio-based chemicals production. In order to solve the energy crisis and environmental pollution issues, the development of advanced bio-fuel technology, especially engineering the microbe able to metabolize xylose and produce ethanol by synthetic biology, is environmentally benign and sustainable.
Bacteria ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Ethanol ; metabolism ; Fermentation ; Fungi ; genetics ; metabolism ; Industrial Microbiology ; methods ; Metabolic Engineering ; Metabolic Networks and Pathways ; genetics ; Saccharomyces cerevisiae ; genetics ; metabolism ; Xylose ; metabolism

BACKGROUND: Apelin/APJ system has a wide range of physiological functions,but its intracellular signal transduction,in particular,apelin receptor desensitization,internalization,resensitization degradation,have still no consistent opinion.OBJECTIVE: To construct eukaryotic expression vector expressing human apelin receptor(APJ)tagged to red fluorescent protein(pDsRED-express-C1),and to determine the expression in human embryo kidney 293 cells.METHODS: The plasmid pcDNA3.1-hAPJ was used as a template for PCR amplification of human APJ.Following PCR amplification the PCR product were removed and enzymatic digestion with EcoR I and BamH I.Same enzymes were used to cut vector pDsRED-express-C1.The digestive product was ligated by conventional methods of connection,then transfected into Competent E.coli TOP10.Single clones were picked plasmid extraction,followed by restriction enzyme digestion and finally DNA sequencing.The recombinant plasmid with correct sequencing was transfected into human embryonic kidney cells,PI staining,followed by the observation under a confocal microscope.RESULTS AND CONCLUSION: PCR amplified a 1.2-kb fragment,which was consistent with the expected size of the human APJ.The pDsRed-hAPJ recombinant plasmid was cut into two fragments,one corresponded to the pDsRED-express-C1 vector size,and the other fragment corresponded to APJ target fragment.Confocal microscopy analysis showed that,APJ was expressed mainly in the membrane of human embryo kidney 293 cells.The pDeRed-hAPJ eukaryotic plasmid expression vector was successfully constructed and effective expression of this fusion protein is achieved,which might be instrumental in the study of displacement and intracellular localization of human APJ.

AIM:To investigate the clinical effect of panretinal photocoagulation (PRP) combined with calcium dobesilate in treatment of diabetic retinopathy (DR).METHODS:Selected 120 cases (240 eyes) of DR diagnosed in our hospital from January 2011 to January 2016 were retrospectively analyzed.According to whether calcium dobesilate was used, the treatment group was divided into two groups.Sixty cases were treated with PRP combined with calcium dobesilate, and 60 cases in the control group were treated with PRP only.The BCVA, CMT and clinical efficacy of the two groups were compared.RESULTS:Before treatment, there was no significant difference on BCVA between combined group and control group (P>0.05).After treatment, BCVA of combined group was higher than that of the control group (P<0.01).Before the treatment, no statistically significant difference between combined group and control group on CMT, neovascularization and fluorescein leakage area were found (P>0.05);after treatment, the combination group on CMT, neovascularization and fluorescein leakage area value were less than the control group (P<0.01).After treatment, combined group was appearance effect in 65.0%, effective in 30.0%, invalid in 5.0%, the control group was 50.8%, effective 36.7%, invalid 12.5%, all statistically significant differences between the two groups (P<0.05).CONCLUSION:PRP combined with calcium dobesilate in treatment of DR has a more significant clinical effect than PRP alone.