2.Cloning and eukaryotic expression of human CD38
Xinyu WEN ; Cuiling SHU ; Yan LI ; Zhongtian QI ; Beife SHEN
Academic Journal of Second Military Medical University 2001;0(09):-
Objective:To clone and express the full length cDNA of human CD38. Methods:The full length cDNA of the human CD38 antigen was amplified from total RNA of Daudi cell by RT-PCR, and it was inserted into pGEM-T. The validity on the sequences was confirmed by automatic DNA sequencing. Inserting the valid CD38 gene into pcDNA3.1(+) plasmid to obtain recombinant mammalian expression vector pcDNA3.1(+)/CD38Z; Using lipofectin gene transfer technique system, recombinant expression vector containing CD38 gene was transfected into COS7 cells. The expression of CD38 molecules on the surface of COS7 cells was detected by FACS and immunohistochemical technique. Results:DNA sequencing showed that the cloned full length cDNA sequence was identical with reported. The result of FACS and immunohistochemical technique indicated that CD38 molecules were expressed on the surface of COS7 cells. Conclusion:The full length cDNA of human CD38 is obtained, recombinant mammalian expression vector pcDNA3.1(+)/CD38Z is successfully constructed, and the CD38 molecules is expressed on the surface of COS7 cells,this may facilitate studies on the biochemistry and function of CD38 antigen.
3.A novel immunization strategy to induce strong humoral responses against HIV-1 using combined DNA, recombinant vaccinia virus and protein vaccines.
Chang LIU ; Shu-hui WANG ; Li REN ; Yan-ling HAO ; Qi-cheng ZHANG ; Ying LIU
Chinese Journal of Virology 2014;30(6):645-651
To optimize the immunization strategy against HIV-1, a DNA vaccine was combined with a recombinant vaccinia virus (rTV) vaccine and a protein vaccine. Immune responses against HIV-1 were detected in 30 female guinea pigs divided into six groups. Three groups of guinea pigs were primed with HIV-1 DNA vaccine three times, boosted with rTV at week 14, and then boosted with gp140 protein at intervals of 4, 8 or 12 weeks. Simultaneously, the other three groups of animals were primed with rTV vaccine once, and then boosted with gp140 after 4, 8 or 12 weeks. The HIV-1 specific binding antibody and neutralizing antibody, in addition to the relative affinity of these antibodies, were detected at different time points after the final administration of vaccine in each group. The DNA-rTV-gp140 immune regimen induced higher titers and affinity levels of HIV-1 gp120/gp140 antibodies and stronger V1V2-gp70 antibodies than the rTV-gp140 regimen. In the guinea pigs that underwent the DNA-rTV-gp140 regimen, the highest V1V2-gp70 antibody was induced in the 12-week-interval group. However, the avidity of antibodies was improved in the 4-week-interval group. Using the rTV-gp140 immunization strategy, guinea pigs boosted at 8 or 12 weeks after rTV priming elicited stronger humoral responses than those boosted at 4 weeks after priming. In conclusion, this study shows that the immunization strategy of HIV-1 DNA vaccine priming, followed by rTV and protein vaccine boosting, could strengthen the humoral response against HIV-1. Longer intervals were better to induce V1V2-gp70-specific antibodies, while shorter intervals were more beneficial to enhance the avidity of antibodies.
AIDS Vaccines
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administration & dosage
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genetics
;
immunology
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Animals
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DNA, Viral
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administration & dosage
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genetics
;
immunology
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Female
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Guinea Pigs
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HIV Infections
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immunology
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prevention & control
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virology
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HIV-1
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genetics
;
immunology
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Humans
;
Immunization
;
methods
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Vaccines, DNA
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administration & dosage
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genetics
;
immunology
;
Vaccinia virus
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genetics
;
immunology
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env Gene Products, Human Immunodeficiency Virus
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administration & dosage
;
genetics
;
immunology
4.Establishment of rat model of syndrome of stagnation of liver qi and primary study using metabonomics
Shu XU ; Hebing CHEN ; Hong LI ; Qi ZHANG ; Hongbing CAI ; Xianzhong YAN ; Zhiping LV
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(06):-
Objective:To establish and validate the rat model of syndrome of stagnation of liver qi,followed by a primary study on this model with 1H NMR based on metabonomics to explore the essence of syndrome of stagnation of liver qi. Methods:Twelve Wistar male rats were randomly divided into two groups(model group and control group).The rats of model group were restrained by special equipment for 21 days to get into stagnation of liver qi.The behavior,fluid consumption test and plasma CORT of rats were recorded.At 22th day,animals were sacrificed and biopsies of gastric mucosa and adrenal gland were collected for pathological check,and serum samples for 1H NMR spectroscopy.The NMR data were analyzed using principal component analysis.Results:There were abnormal behaviors,such as decrease of elusion,slackness,looser stools,and matte fur were observed among model group rats.After one week the body weight of model group was significantly lower than that of control group(P
5.Investigation and analysis of trace element levels in children with osteomalacia malformation from endemic fluorosis area in Shuicheng, Guizhou
Fu-cheng, LI ; Liang, QI ; Xu-zi, CONG ; Ying, YANG ; Shu-lin, YAN
Chinese Journal of Endemiology 2011;30(2):197-201
Objective To investigate the levels of trace elements such as fluorine(F), and aluminium (Al)etc. of osteomalacia malformation children and to make etiological diagnosis in reference with clinical manifestations.Methods Urine and occipitalia hairs of 14 diseased children(patient group) from endemic fluorosis area and 13 healthy children(control group) from non-endemic area were included in the study on November, 2008, and contents of 10 elements of fluorine(F), aluminum(Al), chromium(Cr), manganese(Mn), ferrum(Fe), cuprum(Cu), zinc(Zn), arsenic (As), selenium(Se), strontium(Sr), and barium(Ba) were tested. The data were analyzed with medical soft package PEMS 3.1. Results Urinary contents of F, Al, Mn, Cu, Sr, and Se(1.18 mg/L, 112.6 μg/L,6.62,29.86 mg/L, 177.5,4.23 ng/L) in patient group were significantly different from those in control group (0.48,47.1,2.04,16.61 mg/L, 55.17,15.52 ng/L, t = 4.592,2.486,4.850,2.210 2.078,2.912, all P< 0.05); Hair contents of Al, Mn, As, Sr, Ba, Fe, and Se in patient group(59.27,5.26,0.96,1.50,1.29,297.13,0.45 mg/kg)were significantly different from those of control group( 18.69,0.72,1.09,0.62,0.68,69.02,1.323 mg/kg, t = 4.583,6.318,3.309,2.704,5.606,6.294, all P < 0.05); in patient group, the correlation coefficients of urinary Fe to Al,Zn, As, and Se were all bigger tan 0.662(all P< 0.05), those of urinary Se to Mn, Ba, Cu, Zn, Sr, and As were all bigger than 0.694(all P< 0.05), those among urinary Mn, Sr, As, and Ba were bigger than 0.550(all P<0.05), those of hair Al to Mn, Cr, Fe, and Cu were bigger than 0.732(all P< 0.05), those of hair Ba to Mn,Cr, Fe, and Sr, and of hair Mn to Cr and Fe, and those between Cr and As, between Cu and Sr were all bigger than 0.686 (all P < 0.05). In control group, the correlation coefficients of urinary Cu to Zn, Se, and Ba, those of Zn to Se and Ba, and those of Cr to Mn and Ba were all bigger than 0.516(all P < 0.05), those of hair Al to Mn,Fe, Cu, As, and Se, and those of hair Se to Fe, Cu, and As, those of hair Fe to Mn, Cu, and As, those of hair Cu to Zn and As, and that between Zn and As were bigger than 0.739(all P < 0.05). The correlation coefficient of urinary F to Se in patient group(0.762) was significantly different from that in control group( - 0.469, u = 2.079,P < 0.05). Conclusions The burden of F and Al of osteomalacia malformation children in endemic fluorosis area of Shuicheng county is too high. The contents of multi-elements in urine and hairs and their correlation are coincident with high levels of Al and F and they cause network increase of multi-element content changes and their correlation. According to bone X-ray features combining with the living environment, the diagnosis of endemic Al-F fluorosis can be made. The biological significance of reducing urinary and hair Se levels and the correlations of F and Al need to be further studied.
6.Immigration and proliferation of Schwann cells in chemical acellular xenogeneousnerve grafts in rats
Bing-Yao CHEN ; Shu-Xun HOU ; Min ZHAO ; Yan-Xia QI ; Hong-Bin ZHONG ;
Chinese Journal of Microsurgery 2006;0(05):-
Objective To observed the immigration and proliferation of Schwann cells in acellular xe- no-nerve graft in rats.Methods The sciatic nerves on the right side of the rats were exposc.d and 0.8cm long segments of the nerves were removed from the mid-thigh level and replaced by 1.0cm long rabbit nerves made acellular through chemical extraction.After 4 months,the immigration and proliferation of Schwann cells in the graft were revealed by HE staining,S-100 immunohistochemieal staining and transmission electromicro- scope.Results In the rats repaired by acellular nerves,regenerated axons upgrow into the graft,and a- round regenerated axons there were abundant cells aligned,the cytoplasm of which were S-100 immunoreac- tive.Electromicroscope observing showed that regenerated axons were surrounded by myelin formed by the mi- grated cells reoccupied the acellular segments.Conclusion The host Schwann cells can immigrate into rab- bit nerve grafts made acellular through chemical extraction and form myelin enwrapping regenerated axons in rats.
7.Heat shock cognate 71 (HSC71) regulates cellular antiviral response by impairing formation of VISA aggregates.
Zhigang LIU ; Shu-Wen WU ; Cao-Qi LEI ; Qian ZHOU ; Shu LI ; Hong-Bing SHU ; Yan-Yi WANG
Protein & Cell 2013;4(5):373-382
In response to viral infection, RIG-I-like RNA helicases detect viral RNA and signal through the mitochondrial adapter protein VISA. VISA activation leads to rapid activation of transcription factors IRF3 and NF-κB, which collaborate to induce transcription of type I interferon (IFN) genes and cellular antiviral response. It has been demonstrated that VISA is activated by forming prion-like aggregates. However, how this process is regulated remains unknown. Here we show that overexpression of HSC71 resulted in potent inhibition of virus-triggered transcription of IFNB1 gene and cellular antiviral response. Consistently, knockdown of HSC71 had opposite effects. HSC71 interacted with VISA, and negatively regulated virus-triggered VISA aggregation. These findings suggest that HSC71 functions as a check against VISA-mediated antiviral response.
Adaptor Proteins, Signal Transducing
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biosynthesis
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chemistry
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genetics
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metabolism
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Cell Aggregation
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genetics
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GPI-Linked Proteins
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metabolism
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Gene Knockdown Techniques
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HEK293 Cells
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HSC70 Heat-Shock Proteins
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genetics
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metabolism
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Heat-Shock Response
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genetics
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Humans
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Interferon Regulatory Factor-3
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genetics
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metabolism
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Interferon-beta
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genetics
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NF-kappa B
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genetics
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Prions
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metabolism
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Receptors, Retinoic Acid
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metabolism
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Viruses
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drug effects
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metabolism
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pathogenicity
8.Brief introduction of research methods of commodity specification and grade of Chinese medicinal materials.
Hua-ye ZHAO ; Pei-pei YAN ; Wen-hua YANG ; Chao-feng LI ; Shu-ya QI ; Yan-qi LI ; Jun-ling CAO
China Journal of Chinese Materia Medica 2015;40(4):765-769
The commodity specification and grade of Chinese medicinal materials is a measure of the quality of traditional Chinese medicines (TCMs), which directly impacts on the safety and effectiveness of clinical medicines. It is an urgent problem to establish a set of standards which can both interpret the scientific connotation of the commodity specification and grade of Chinese medicinal materials and play a significant role on clinical medicines as well as markets. This paper reviews the research methods of the commodity specification and grade of Chinese medicinal materials such as sensory evaluation, chemical assessment, biological evaluation, and cited the applications of various methods for the classification of TCMs. It provides technical support for establishing standards of the commodity specification and grade of Chinese medicinal materials, and also constructs scientific basis for clinical rational drug use.
Drugs, Chinese Herbal
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chemistry
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economics
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Humans
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Medicine, Chinese Traditional
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economics
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standards
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Plants, Medicinal
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chemistry
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Quality Control
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Research Design
9.Radiofrequency catheter ablation of the premature ventricular contractions originating from His bundle region.
Yu-bin WANG ; Jian-min CHU ; Jing WANG ; Shu-kai SONG ; Xiao-yan LIU ; Qi GUO ; Jie-lin PU ; Shu ZHANG
Chinese Journal of Cardiology 2013;41(5):382-386
OBJECTIVETo explore the electrocardiogram and 3-dimensional electroanatomic mapping features and radiofrequency catheter ablation efficacy of patients with premature ventricular contractions (PVCs ) originating from His bundle region.
METHODSBetween February 2009 and February 2011, 10 consecutive patients ( 4 male, aged from 19 to 59 years) who underwent ablation for frequent PVCs originating close to His bundle region in our department were included. Electroanatomic mapping of RVOT and ASC, ablation was performed with the 7F 4-mm-tip ablation catheter.
RESULTSAmong these 10 patients with PVCs originating from His bundle region, 6 originated from the RVOT, 1 from NCC and 3 from RCC. Eight patients showed LBBB morphology,1 patient with PVCs originated from RCC and 1 patient with PVCs originated from NCC showed RBBB morphology. At the successful ablation sites, local ventricular activation v wave was detected 22-52 (32.6 ± 10.2) ms earlier than the QRS wave in the surface electrocardiogram. The distance between target and His bundle was 5.0-8.4(7.0 ± 1.1)mm. Ablation was successful in all 10 patients without complications (PVCs < 500 beats/24 h post ablation).
CONCLUSIONPVCs originating near the His bundle have similar electrocardiographic and electrophysiological characteristics for PVSc originated from the RVOT or ASC. Because of the close anatomical relationship between RVOT and ASC, it is necessary to mapping both RVOT and ASC to accurately identify the site of PVCs origin and to guild successful ablation.
Adult ; Bundle of His ; surgery ; Catheter Ablation ; methods ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Ventricular Premature Complexes ; surgery ; Young Adult
10.Research on resources chemistry of Chinese medicinal materials and resources recycling utilization ways and goals and tasks.
Jin-ao DUAN ; Shu-lan SU ; Sheng GUO ; Shu JIANG ; Pei LIU ; Hui YAN ; Da-wei QIAN ; Hua-xu ZHU ; Yu-ping TANG ; Qi-nan WU
China Journal of Chinese Materia Medica 2015;40(17):3395-3401
The objects of research on the resources chemistry of Chinese medicinal materials (RCCMM) are promotion of efficient production, rational utilization and improving quality of CMM and natural products. The development of TCM cause depends on the efficient utilization and sustainable development of CMM, hinges on the technologies and methods for using and discovering medicinal biological resources, stand or fall on the extension of industy chains, detailed utilizaion of resource chemical components by multi-way, multi-level. All of these may help to the recycling utilization and sound development of RCMM. In this article, five respects were discussed to the RCCMM researches and resources recycling utilization ways and goals and tasks. First, based on the principle of resource scarcity, discovering or replacing CMM resources, protecting the rare or endangered species or resources. Second, based on the multifunctionality of CMM, realizing the value-added and value compensation, and promoting the utilization efficiency through systermatic and detailed exploitation and utilization. Third, based on the resource conservation and environment-friendly, reducing raw material consumption, lowering cost, promoting recycling utilization and elevating utilization efficiency. Fourth, based on the stratege of turning harm into good, using the invasive alien biological resources by multi-ways and enriching the medicial resources. Fifth, based on the method of structure modification of chemical components, exploring and enhancing the utility value of resouces chemical substances. These data should provide references and attention for improving the utilization efficiency, promoting the development of recycling economy, and changing the mode of economic growth of agriculture and industry of CMM fundamentally.
Agriculture
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economics
;
trends
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China
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Conservation of Natural Resources
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economics
;
trends
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Drugs, Chinese Herbal
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chemistry
;
economics
;
Materia Medica
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chemistry
;
economics
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Medicine, Chinese Traditional
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economics
;
trends
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Plants, Medicinal
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chemistry
;
growth & development