1.Construction of pRluc-hNTSR1-pcDNA3.1 eukaryotic expression vectors and its expression in isolated cells
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(12):1140-1142
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.
2.Basic study of fabricating dental prosthesis with laser engineered net shaping (LENS) method——The microstructure of LENS components made from Rene95 alloy
Bo GAO ; Yanmin LI ; Jing CHEN
Journal of Practical Stomatology 2000;0(06):-
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.
3.Basic study of fabricating dental prosthesis with laser engineered net shaping (LENS) method-The mechanical properties of LENS components made from Rene95 alloy
Bo GAO ; Yanmin LI ; Jing CHEN
Journal of Practical Stomatology 2001;0(03):-
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.
4.Corrosion behavior of Ti-Zr alloy formed by laser solid forming technology
Bo GAO ; Xiaodong YAN ; Jing CHEN
Journal of Practical Stomatology 2001;0(03):-
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.
5.Analysis on the Utilization of Cardiovascular and Cerebrovascular Drugs in a Military Sanatorium During 2007~2009
Jing LING ; Fangfang LU ; Bo CHEN
China Pharmacy 2001;0(12):-
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.
6.Up-regulation of Nrf2 pathway and intervention of intracellular redox homeostasis in shikonin-induced A549 cell apoptosis
Chen XIE ; Hanying CHEN ; Jing ZHONG ; Xiaoqin WANG ; Bo ZHANG
Chinese Pharmacological Bulletin 2014;(10):1445-1451
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.
7.Expression of MIP-3? gene regulated by irradiation via Egr-1 promotor in Lewis lung cancer
Lan ZOU ; Bo ZHU ; Jing CHEN ; Jianping DU ; Zhengtan CHEN
Journal of Third Military Medical University 2003;0(24):-
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.
8.Engineering of the xylose metabolic pathway for microbial production of bio-based chemicals.
Weixi LIU ; Jing FU ; Bo ZHANG ; Tao CHEN
Chinese Journal of Biotechnology 2013;29(8):1161-1172
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
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genetics
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metabolism
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Escherichia coli
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genetics
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metabolism
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Ethanol
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metabolism
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Fermentation
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Fungi
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genetics
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metabolism
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Industrial Microbiology
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methods
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Metabolic Engineering
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Metabolic Networks and Pathways
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genetics
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Saccharomyces cerevisiae
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genetics
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metabolism
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Xylose
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metabolism
9.Role of G-protein-coupled receptor kinases in cell migration and signal transduction
Xin CAI ; Jing CHEN ; Jingmei ZHANG ; Bo BAI
Chinese Journal of Pathophysiology 2011;27(7):1438-1444
G-protein-coupled receptor kinases (GRKs) are a family of serine/threonine protein kinases. The investigators pay much attention to the roles of GRKs in the signal transduction through G-protein-coupled receptors (GPCRs) with arrestin ever since a long time ago. Due to the physiological and pathological observations with the methods of deletion or overexpression, GRKs are considered as new drug targets. The kinases play a role in the pathogenesis of hypertension and cell migration through GPCRs and Hedgehog signaling pathways. As the development of research techniques, especially bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET), the special mechanism of GRKs for GPCRs is more evident. In this review, we discuss the recent achievement in the roles of GRKs signaling and the related newest research techniques.
10.Multivariate analysis of elderly patients with thoracic malignancies caused by radioactive radiation pneumonitis
Jing WU ; Bo LI ; Fei CHEN ; Xiaolu REN
Journal of Chinese Physician 2016;18(6):809-812
Objective To investigate elderly patients with thoracic radiotherapy nausea result after risk factors associated with radiation pneumonitis.Methods The clinical data of a total of 440 cases of cancer patients with chest radiation therapy during January 2010-January 2014 were collected retrospectively.Of them,76 cases of radiation pneumonitis after radiotherapy were compared with other patients.The unconditional Logistic regression analysis was used to analyze the relationship of radiation pneumonitis and different factors including smoking,pulmonary dysfunction,combination with chemotherapy,radiation dose,and radiation sites.Results Elderly incidence of radiation pneumonitis was 17.27%.Multivariate Logistic regression analysis revealed the chi-square value of smoking,pulmonary dysfunction,combined with chemotherapy,radiation dose,and radiation sites was significant correlation (x2 =16.936,19.633,11.531,17.133,10.178,P <0.05),and the correlation degree was gradually decreased from pulmonary dysfunction,radiation dose,smoking,combined chemotherapy,to radiation site.Conclusions Elderly patients with thoracic malignancies after radiotherapy had more radiation pneumonitis,which was related to smoking,previous chemotherapy PO2 < 80%,combined with chemotherapy,radiation dose ≥ 55 Gy,and low-lung radiation.The correlation degree was gradually decreased from chemotherapy before PO2 < 80%,the radiation dose ≥55 Gy,smoking,combined chemotherapy,to low-lung radiation.