3.Characterization of Bifunctional Chimeric Molecule of PRGDWR Containing Pro-Urokinase
Xin DANG ; Jingxin YANG ; Qiang RU ; Binggen RU
Progress in Biochemistry and Biophysics 2001;28(2):203-209
In order to obtain the bifunctional chimeric molecule of single-chain urokinase-type plasminogen activator (scu-PA) which can inhibit platelet aggregation, PRGDWR peptide was inserted into the site between Gly 118 and Leu119 (called insertion mutant B, InB). The recombinant gene of InB was expressed by Pichia pastoris. The secreted protein was purified by metal chelate affinity and strong cation exchange chromatography. The amidolytic ability of mutant InB is 5 900 IU/mg, the kinetic constants is: KInB m,plg=56.8 μmol*L-1,kInBcat,plg=0.33 s- 1. The kinetic constants of plasminogen activation reaction is: KInB m,plg=0.397 μmol*L-1,kInBcat,plg=0.0164 s-1. Fibrin inhibit the catalytiv ability of InB during plasminogen activatio n, the influence factor is 0.463(means InB remain 46.3% of the catalytic abili ty when fibrin was involved in the reaction system). The mutant not only has alm ost the same catalytic ability as wild type scu-PA, but also has strong ability of anti-platelet aggregation(compared with scu-PA), IC50 of InB is 12.7 μmol*L-1.
4.The Influence of Fibrin on the Reaction of Plasminogen Activation by Mut ant of Pro-Urokinase
Xin DANG ; Jingxin YANG ; Qiang RU ; Hongsheng YUAN ; Binggen RU
Progress in Biochemistry and Biophysics 2001;28(2):210-215
Because the influence of fibrin on the reaction of plasm inogen activation by various plasminogen activators is different, the kinetic co nstant of the reaction of plasminogen activation catalyzed by InB with and witho ut fibrin were detected. The result is: Kfibrinm=4.2 μmol*L -1,greater than the normal Km=0.379 μmol*L-1; kfib rincat=0.107 s-1,greater than the normal kcat=0.0165 s-1. The results suggest that existence of fibrin in the reaction system of plasminogen activation depress the affinity between InB and plasminog en, but accelerates the hydrolysis of plasminogen by InB. The count up effect is inhibition.
5.Quantitative Determination of Saponins in the Root of Panax pseudo-ginseng var.notoginseng by HPLC-ELSD and UV Spectrophotometry
Yingqiao JIANG ; Qiang WANG ; Shiping MA ; Xuedong DANG
Chinese Traditional and Herbal Drugs 2000;31(10):737-739
A reverse phase HPLC-ELSD method for the determination of ginsenoside Rg1 in the rootof Panaa pseudo-ginseng var. notoginseng (Burkill) Hoo et Tseng was reported. Chromatographic condi-tions: Shim-pack CLC-ODS column (6.0 mm×150 mm); acetonitrile-water (30: 70) as the mobilephase; Shimadzu LC-6A with SEDEX-55 ELSD detector. The method was found to be simple and accuratewith recovery rate of 100. 50% and RSD= 1.82 %. The established UV spectrophotometric determinationof total saponins in P. pseudo-ginseng var. notoginseng was also tried and gave an accurate result coinci-dental with that of the HPLC results. The recovery rate was 101.50%, and RSD=1. 44%. It seemed thatboth methods can be used reliably for the quality control of P. pseudo-ginseng var. notoginseng.
6.A death case report of Hamman-Rich syndrome.
Xiu-ying WANG ; Xi-qiang DANG ; Jie-ming LIU
Chinese Journal of Pediatrics 2003;41(5):396-396
Adolescent
;
Cough
;
complications
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Fatal Outcome
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Female
;
Fever
;
complications
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Humans
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Pulmonary Fibrosis
;
complications
;
diagnosis
7.Congenital nephritic syndrome: report of 4 cases.
Chinese Journal of Contemporary Pediatrics 2013;15(1):77-78
Female
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Humans
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Infant
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Male
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Nephrotic Syndrome
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congenital
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pathology
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therapy
10.Biological activity of the cultured rabbit bone marrow-derived mesenchymal stem cells in vitro
Chen ZHANG ; Kunzheng WANG ; Hui QIANG ; Zhibin SHI ; Xiaoqian DANG ; Xianghui HUANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2010;31(2):138-142
Objective To observe the effect of the in vitro isolation, culture and identification of rabbit bone marrow-derived mesenchymal stem cells (BMSCs) and explore the differentiation potentials of BMSCs. Methods BMSCs were isolated and purified by density gradient centrifugation combined with attachment culture method. The biological characteristics of BMSCs were observed under an inverted microscope. The BMSCs were identified with HE staining and CD34/CD44 immunohistochemical staining. The biological activity of BMSCs was detected by osteogenic induction and steatogenic induction. Results BMSCs were isolated successfully in vitro and had good homogenicity. Immunohistochemical staining of CD34/CD44 showed strong positive expression of CD44 and negative expression of CD34. Fortis osteogenic ability and positive staining of ALP could be detected by osteogenic induction, and fortis steatogenic ability and positive staining of axungia could be detected by steatogenic induction. Conclusion BMSCs have the capability of multi-directional differentiation in vitro.