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| 1 | Construction of recombinant plasmid pEGFP-C2-L539fs/47 and its expression in HEK293 cells显示文摘Objective:To reconstruct pEGFP-C2-L539fs/47,a HERG nonsense mutant in eukaryotic expression plasmid,and observe the fusion protein expressed in HEK293 cells(human embryo kidney cells).Methods:After double digestion of pcDNA3-L539fs/47 and pEGFP-C2-HERG with sbf I and Eco91 I,the small product fragment,from pcDNA3-L539fs/47,was subcloned into the big fragment of pEGFP-C2-HERG under T4 ligase.pEGFP-C2-L539fs/47 was identified by agarose gel electrophoresis and sequencing.pcDNA3-L539fs/47 and pEGFP-C2-L539fs/47 were transiently transfected into HEK293 cells by Lipofect,respectively.The expression of fusion protein in HEK293 cells was detected through immunofluorescence,laser confocal imaging scanning in vivo,Western blot and PCR.Results:Mutation region cDNA fragment(about 1 kb) and target vector fragment(about 7.2 kb) were ligated after purification and gel recovery.Agarose gel electrophoresis and sequencing successfully demonstrated eukaryotic expression plasmid pEGFP-C2-L539fs/47,constructed approximately 8.2 kb,sequencing consistent with template gene.The transfection efficiency of recombinant plasmid by fluorescence microscopy was more than60%.Western blot analysis detected pcDNA3-L539fs/47 expression of the protein size 60 KD,the expression of pEGFP-C2 fusion protein size of approximately 90 KD.The L539fs/47 gene expression in HEK293 cells was significant by PCR analysis.Confocal laser imaging showed that pEGFP-C2-L539fs/47 protein was successfully expressed in cytoplasm and cytomembrane of HEK293 cells.Conclusion:pEGFP-C2-L539fs/47 containing the HERG gene mutant was successfully constructed by double digestion method and expressed fusion protein in HEK293 cells,which laid a foundation for the further study on L539fs/47. | Lue Ying Zhang Aifeng Han Wenqi Li Guoliang Zhang Junbo Gao Jie Pan Junqiang Zhang Yong Sun Chaofeng | 2012 | Journal of Medical Colleges of PLA(China)2012，27，3： | 2 |
| 2 | Molecular Characterisation of the Faecal Microbiota in Patients with Type II Diabetes显示文摘 | Xiaokang Wu Chaofeng Ma Lei Han Muhammad Nawaz Fei Gao Xuyan Zhang Pengbo Yu Chang’an Zhao Lianchuan Li Aiping Zhou Juan Wang John E. Moore B. Cherie Millar Jiru Xu | 2010 | Current Microbiology2010，，1： | 2 |
| 3 | Effects of direct aging treatment on microstructure,mechanical properties and residual stress of selective laser melted AlSi10Mg alloy显示文摘Direct aging treatment is an important post-processing procedure,yet little research has been done on how it balances the mechanical properties and the stress removal for selective laser melted(SLMed)AlSi10Mg alloys.Here,we proposed a typical direct aging treatment on SLMed AlSi10Mg alloys,and studied the effects on their microstructure,properties and residual stress evolution.The results indi-cate that the as-built microstructure is mainly composed of fine cellularα-Al and reticulated Si phases,and some pre-existing precipitates and dislocations are found in these cells.The direct aging treatment promotes the precipitation of nano-scaled Si phases and preserves a network-like Si structure.Therefore,the strength of the peak-aged alloy increases while the ductility decreases.As the aging temperature in-creases from 160 to 200℃,aging hardening behavior was accelerated significantly.Aging at 160℃ for 4-9 h removes 32.0%-43.0%of the residual stress,which is attributed to the decomposition of the su-persaturatedα-Al matrix,the precipitation of the nano-Si phase and the exposure of low-angle grain boundaries(LAGBs).Considering the overal alloy performance obtained,over-aging at 160℃ for 4 h is the optimized heat treatment regime.Under this condition,the yield strength(YS),ultimate tensile strength(UTS)and elongation(EL)of the alloy in the transverse and longitudinal direction are 309.5 MPa,464.4 MPa and 8.3%and 286.4 MPa,464.9 MPa and 5.1%,respectively. | Huaping Tang Chaofeng Gao Yong Zhang Nannan Zhang Chuan Lei Yunjie Bi Ping Tang Jeremy Heng Rao | 2023 | Journal of Materials Science & Technology2023，，8： | 1 |
| 4 | Effects of Danqu Capsules on Arterial Blood Pressure,Heart Rate and Myocardial Tension Time Index in Rats显示文摘[Objectives] To explore the effects of Danqu capsules(the prescription of Lei Zhongyi, the master of traditional Chinese medicine for the syndrome of intermingling of phlegm and static blood of coronary heart disease) on arterial blood pressure, heart rate and myocardial tension time index in rats. [Methods] A total of 50 SD rats, weighing(300±20) g, were randomly and evenly divided into Danqu capsule observation groups(low, middle and high-dose groups) and control groups(saline group and captopril group). Under anesthesia, the rats in each group were administered with 0.6, 1.2 and 2.4 g/kg of Danqu capsule suspension, 1 mL/100 g of saline and 10 mg/kg of captopril, respectively through duodena, and their systolic blood pressure, diastolic blood pressure, mean arterial pressure, pulse pressure difference and heart rate were measured and recorded by RM6240 multi-channel physiological signal acquisition and processing system before administration, 10, 20, 30, 40, 50, 60, 90 and 120 min after the administration, respectively, and the myocardial tension time index was calculated. [Results] Compared with the saline group, the systolic, diastolic and mean pressure reduced 10, 20, 30, 40, 50, 60, 90 and 120 min after the administration, and the pulse pressure difference did not change significantly in the Danqu capsule groups(low, middle and high-dose groups), and the heart rate slowed down and the myocardial tension time index reduced in the high-dose Danqu capsule group. Compared with the captopril group, the Danqu capsule groups showed similar blood pressure-reducing effect. [Conclusions]Danqu capsules have obvious effect of lowering blood pressure and myocardial tension time index. High-dose Danqu capsules has the effect of slowing down the heart rate. The blood pressure-lowering effect of Danqu capsules shows a dose-dependent manner. Clinically, they can be used in patients with coronary heart diseases and hypertension, suggesting that they should be used with caution in patients with hypotension. | Jinfeng CHEN Li JING Cong GAO Renming XIE Zhongyi LEI Chaofeng LIU Hong FAN | 2019 | Medicinal Plant2019，10，4： | 1 |
| 5 | One-stage surgical treatment for upper thoracic spinal tuberculosis by internal fixation, debridement, and combined interbody and posterior fusion via posterior-only approach显示文摘 | Hongqi Zhang Bin Sheng Mingxing Tang Chaofeng Guo Shaohua Liu Shu Huang Qile Gao Jinyang Liu Jianhuang Wu | 2013 | European Spine Journal2013，，3： | 1 |
| 6 | Numerical simulation and damage analysis of fissure field evolution law in a single coal seam mining显示文摘Based on the different deformation and failure laws of coal and rock materials under tensile or compressive loads,the damage variable was defined to divide three phases from the damage and rupture point of view in order to reveal the fissured field evolution characteristics and spatial distribution of coal and rock in the single coal seam mining with low permeability and rich methane.According to the corresponding damage constitutive equations,the secondary development of the finite element program was completed.The fissures field evolution law of a coal mine with single coal seam mining was calculated and analyzed by this new program and the distribution areas and failure degree of the surrounding coal and rock structures with damage,fissure or rupture are given on the condition of mining.This paper provides a scientific basis for quantitative research and evaluation of the safe simultaneous production of coal and gas. | Feng Gao Xingguang Liu Chaofeng Ge Hongmei Cheng | 2012 | International Journal of Mining Science and Technology2012，22，6： | 1 |
| 7 | Characterization of four diol dehydrogenases for enantioselective synthesis of chiral vicinal diols显示文摘Enantiopure vicinal diols are important building blocks used in the synthesis of fine chemicals and pharmaceutical compounds. Diol dehydrogenase(DDH) mediated stereoselective oxidation of racemic vicinal is an efficient way to prepare enantiopure vicinal diols. In this study, four new bacterial DDHs(AnDDH from Anoxybacillus sp. P3 H1 B, HcDDH from Hazenella coriacea, GzDDH from Geobacillus zalihae and LwDDH from Leptotrichia wadei) were mined from the GenBank database and expressed in E. coli T7.The four DDHs were purified and biochemically characterized for oxidation activity toward(R)-1-phenyl-1,2-ethanediol, with the optimal reaction condition of pH9.0(AnDDH), 10.0(HcDDH) and 11.0(GzDDH and LwDDH) and the temperatures at 40 ℃(AnDDH), 50 ℃(HcDDH) and 60 ℃(GzDDH and LwDDH), respectively. The four enzymes were stable at the pH from 7.0 to 9.0 and below 40 ℃.Kinetic parameters of four DDHs showed that the HcDDH from Hazenella coriacea had high activity toward a broad range of vicinal diols. A series of racemic vicinal diols were successfully resolved by recombinant E. coli(HcDDH-NOX) resting cells co-expression of an NADH oxidase(NOX), affording(S)-diols and(1 S, 2 S)-trans-diols in ≥99% ee. The synthetic potential of HcDDH was proved by E. coli(HcDDH-NOX) via kinetic resolution of racemic trans-1,2-indandiol on a 100 ml scale reaction,(S, S)-trans-1,2-indandiol was prepared in 46.7% yield and >99% ee. In addition, asymmetric reduction of four α-Hydroxy ketones(10–300 mmol·L^(-1)) by E. coli(HcDDH-GDH) resting cells resulted in >99% ee and69–98% yields of(R)-vicinal diols. The current research expands the toolbox of DDHs to synthesize chiral vicinal diols and demonstrated that the mined Hc DDH is a potential enzyme in the synthesis of a broad range of chiral vicinal diols. | Jiandong Zhang Rui Dong Xiaoxiao Yang Lili Gao Chaofeng Zhang Fan Ren Jing Li Honghong Chang | 2022 | Chinese Journal of Chemical Engineering2022，35，7： | 0 |
| 8 | Construction of a HERG mutant L539fs/47-*558W pEGFP vector and the expression of the fusion protein in HEK293 cells显示文摘Objective:To construct a human ether-a-go-go-related gene(HERG)nonsense mutant L539fs/47-*558W into the autonomously fluorescent,eukaryotic expression vector pEGFP-C2,and to verify expression of the reconstruct in human embryonic kidney-293(HEK293)cells.Methods:The mutational fragment was subcloned into pEGFP-C2-HERG by double digestion of SbfⅠ,Eco91Ⅰand rejoining of T4 ligase.After verification,the recombinant pEGFP-C2-L539fs/47-*558W and pEGFP-C2-HERG were respectively transfected into HEK293 cells for 48 h by the Lipofect method to observe the expression location of the fusion protein by laser confocal imaging scanning in vivo.pcDNA3-L539fs/47-*558W and pcDNA3-HERG were transfected to observe the expression location of the HERG protein by immunofluoresceoce.The mutant protein size was determined by Western blotting.Results:The about 1 kb-sized mutation region cDNA fragment from pcDNA3-L539fs/47-*558W and the about 7.2 kb-sized target vector fragment from pcDNA3-HERG were ligated after purification and gel recovery.pEGFP-C2-L539fs/47*-558W,approximately 8.2 kb,was demonstrated successfully been constructed under agarose gel electrophoresis and further sequencing.Laser confocal imaging showed that pEGFP-C2-HERG was mainly expressed in the membrane,whereas truncated mutant-type HERG in the pEGFP-C2 vector was partially located in the cytoplasm,the others were transported to the cell membrane in living HEK293 cells.The same as the immunofluoresceoce results after transfection of pcDNA3-HERG and pcDNA3-L539fs/47-558W.Wild-type HERG-GFP fusion protein expressed 160 and 180 kDa bands.The mutant and mutant-GFP fusion proteins were 70 and 100 kDa,respectively.Conclusion:pEGFP-C2-L539fs/47-*558W was successfully constructed by double digestion method GFP had no effect on its protein expression and trafficking in HEK293 cells,which laid a foundation for the further study on L539fs/47-*558W. | ZHANG Junbo Lü Ying ZHANG Aifeng SUN Chaofeng HAN Wenqi LI Guoliang GAO Jie HUO Jianhua PAN Junqiang ZHOU Xin NIU Xiaolin | 2013 | Journal of Medical Colleges of PLA(China)2013，28，4： | 0 |
| 9 | Spatiotemporal evolution of sustainable development of China’s provinces:a modelling approach显示文摘Three sustainable development sub-indicators were established along with 49 indicators of sustainable development at the provincial level.We collected data for the key years of 2010,2015,and 2018 and evaluated the sustainable development level of 31 provinces in China by using the best and worst value improved equal-weight TOPSIS method.With spatial autocorrelation and hotspot analysis,the spatial characteristics of the sustainable development level were studied.Grey correlation analysis and the offset degree index were also used to find out which indicators should be of most concern to improve the level of sustainable development.The results showed the following.(1)The level of sustainable development of China’s provinces is increasing,but there is a decrease from east to west.(2)In terms of sustainable development,the gap between the four major economic regions is narrowing.(3)The overall sustainable development level is highly consistent with the mainstream green development and the human development index,indicating that exploiting the driving force behind developing and improving the quality of development is the key to China’s sustainable development strategy.(4)Different measures should be taken to promote the realization of Sustainable Development Goals in different areas. | Junli Gao Chaofeng Shao Sihan Chen Xiaotong Zhang | 2021 | Ecosystem Health and Sustainability2021，7，1： | 0 |
| 10 | Doctors' faith显示文摘We make some efforts to find some connections between religion and medical science. Medical science can be regarded as sacred and holy as a religion for doctors to believe. The common points between them are the basis to successfully integrate them into doctors' faith. Furthermore, we explain that doctors need a faith to believe so as to meet the demand of humanitarian traits we should have and to do better in the scientific work and win in the battles with the diseases. | ZHANG Junbo Lü Ying ZHANG Aifeng SUN Chaofeng HAN Wenqi LI Guoliang GAO Jie HUO Jianhua PAN Junqiang ZHOU Xin NIU Xiaolin | 2013 | Journal of Medical Colleges of PLA(China)2013，28，1： | 0 |
| 11 | The exploration of the innovation mode of school enterprise cooperation in game analysis显示文摘 | GAO Chaofeng YANG Ting DUAN Longlong | 2015 | International English Education Research2015，，12： | 0 |
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