2.Construction and Identification of Recombinant Plasmid of p3XFLAG-CMV7-NICD1
Journal of China Medical University 2015;(3):217-220
Objective To construct the eukaryotic expression vector of Notch1intracellular domain,p3XFLAG?CMV7?NICD1,so as to prepare for the further research and exploration of effect of Notch1 on promoting epithelial?mesenchymal transition of human lens epithelial cells. Methods The cDNA fragment was reversely transcribed by RT?PCR from total RNA extracted from the SRA01/04 cells and was encoded with the specific am?plification?targeted NICD1was obtained from the SRA01/04 cells,then the cDNA fragment was inserted into p3XFLAG?CMV7 to transcribe Esche?richia coli DH5α. And the recombinant plasmid was extracted after bacterial screening by LB plating medium and confirmed by the restriction endo?nuclease digestion and DNA sequencing. Results The target gene obtained had the same molecular size as predicted. It was indicated that recom?bined p3XFLAG?CMV7 plasmid contained correct recombinant human Notch1 sequences and p3XFLAG?CMV7?NICD1 was constructed success?fully. The western blotting showed protein NICD1 expressed in SRA01/04 cells transfected with p3XFLAG?CMV7?NICD1. Conclusion The suc?cessful construction of p3XFLAG?CMV7?NICD1 will provide a foundation for a further study studies in on the effect relationship of Notch signaling pathway and in posterior capsular opacification(PCO)after cataract extraction.
3.Comparison of MRI contrast agents gadopentetate dimeglumine, gadodiamide and gadovist: their relaxation rates and effects on imaging
Chinese Journal of Geriatrics 2016;35(9):993-995
Objective To evaluate the relaxation rates and imaging effects of three MRI contrast agents gadopentetate dimeglumine(0.5 mol/L),gadodiamine and gadovist(1.0 mol/L) in the nervous system.Methods Relaxation rate differences between the three contrast agents were assessed using the GE Signa HDx 3.0 T MR scanner and phantom solutions of different albumin concentrations.Twenty leukemia patients whose initial scans had been conducted with the injection of a standard dose(0.5 mol/L)of gadopentetate dimeglumine as the contrast agent were recalled to have a follow-up scan for signs of brain infections with the same imaging protocols,except that a high concentration(1.0 mol/L)gadovist was used this time as the contrast agent.CNR and SNR in the ROI were measured for quantitative analysis.Results Changes in dosage of the three contrast agents produced no difference in intensity of the image signal for each phantom solution of a specific albumin concentration(5.0 g/L:P=0.35,6.5g/L:P =0.27,8.0 g/L:P=0.23).Two sets of scans of the leukemia patients showed that high concentration(1.0 mol/L)gadovist generated higher SNR and CNR in the ROI of the white matter,gray matter and vasculature than standard concentration(0.5 mol/L) gadopentetate dimeglumine(P< 0.05).Conclusions A half dose of high concentration(1.0 mol/L) gadovist generates better imaging enhancement than standard concentration(0.5 mol/L)gadopentetate dimegluminethe.Gadopentetate dimeglumine,gadodiamide and gadovist have no significant difference in relaxation rate.
4.Risk factors of acute organophosphorus pesticide poisoning
Xiao-Wei LIU ; Zhi LIU ;
Chinese Journal of Emergency Medicine 2006;0(05):-
Objective To explore the risk factors of acute organophosphorus pesticide poisoning(AOPP). Method The patients with acute organophosphorus pesticide poisoning,admitted from September 2004 to August 2005,were retrospectively analyzed.The 87 patients were divided into groups according to the presence or absence of combined conditions including hypotension,hypoxemia or metabolic acidosis.Acute physiology and chronic health evaluationⅡ(APACHEⅡ)score was calculated.The chi-square test was used to examine the mortality between those groups.Results The total in-hospital mortality of 87 acute organophosphorus pesticide poisoning patients was 21.8%.The mean APACHEⅡscore was(7.58?5.32)in the 68 survivors and(21.17~9.46)in the 19 dead,there were significant differences between the survivors and the dead(t=9.25,P20 was 65.2%(15/23),andit was 6.3% (4/64)in patients with APACHEII score
5.Determination of Triptolide and Wilforlide A in Biological Samples by LC-MS/MS.
Journal of Forensic Medicine 2015;31(6):445-453
OBJECTIVE:
To determinate triptolide and wilforlide A in biological samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method and to verify the method.
METHODS:
After 0.4 mL blood, urine or 0.4 g hepatic tissues with internal standard were extracted by ethyl acetate, they were separated on a Allure PFP Propyl (100 mm x 2.1 mm, 5 µm) with a mobile phase of methanol-20 mmol/L ammonium acetate using gradient elution. For mass spectrometric detection, electrospray ionization (ESI⁺) in positive mode was elected and the data was collected using multiple-reaction monitoring (MRM).
RESULTS:
The linearity was good (r > 0.995 0) and the limit of detection was 2 ng/mL or 2 ng/g for triptolide and wilforlide A. The recovery was 61.08%-102.98%. The intra-day and inter-day precision was less than 12.58% for each biological sample, and the accuracy was 90.61%-105.80%.
CONCLUSION
This method is simple, convenient and good selective, and could be applied to analysis of triptolide and wilforlide A in different biological samples. And the method may provide technical support for forensic medicine identification, clinical diagnosis and treatment of tripterygium wilfordii Hook. f. poisoning.
Chromatography, High Pressure Liquid
;
Diterpenes/urine*
;
Epoxy Compounds/urine*
;
Humans
;
Oleanolic Acid/urine*
;
Phenanthrenes/urine*
;
Sensitivity and Specificity
;
Tandem Mass Spectrometry
7.Determination of 6-Methyl Coumarin in Cosmetics by High Performance Liquid Chromatography
Wei HUANG ; Xiangping LIU ; Shangjia XIAO
Journal of Environment and Health 1992;0(02):-
Objective To establish a high performance liquid chromatographic method for determination of 6-methyl coumarin in cosmetics. Methods Samples were extracted with ethanol, On C18 column CH3CN-0.02 mol/L NaH2PO4(pH=3.5, 35∶65) was applied as mobile phase, 6-Methyl Coumarin in cosmetics was determined by photo-diode-array(PDA) detector. Results The linear range of the method was 1.0-25.0 mg/L. The lowest detected limit of 6-Methyl Coumarin in 1.0 g samples was 0.000 1%. The recovery rates were 96.6%-99.2%, and the relative standard deviation were 2.0%-4.4%. Conclusion The method was simple, rapid and accurate, which was suitable for determination of 6-methyl coumarin.
8.Relationship Between the Invasion of Pituitary Adenomas and Expressions of MMP-9,Ki-67 and nm23
Aihua LIU ; Wei HUANG ; Shaowen XIAO
Journal of Chinese Physician 2001;0(04):-
Objective To study the relationship between the invasion of pituitary adenomas and expressions of MMP-9,Ki-67 and nm23. Methods The expressions of MMP-9,Ki-67 and nm23 were examined by immunohistochemical Elivision methods in 78 cases of pituitary adenomas. Of them,40 cases were invasive pituitary adenomas and, 38 cases were noninvasive pituitary adenomas. Results The expression levels of MMP-9 and Ki-67 in the invasive pituitary adenomas were significantly higher than those in the noninvasive ones (P
9.Analysis of the mutation of rhodopsin gene in an inbreeding family with autosomal recessive retinitis pigmentosa
Jing LIU ; Lin XIAO ; Wei WANG
Chinese Journal of Ocular Fundus Diseases 2001;0(03):-
Objective To detect characteristics and the pathogenesis of rhodopsin (RHO) gene mutation in an inbreeding family with autosomal recessive retinitis pigmentosa (ARRP). Methods Peripheral venous blood 5-8 ml was abstracted from 8 members in the inbreeding ARRP family and 10 control individuals. DNA gene group was picked. Extron 1-5 of RHO gene was amplified by polymerase chain reaction (PCR),and the mutation of RHO gene was screened by direct DNA sequence measurement. Results The Gln-344-Arg mutation in the RHO gene was detected in 3 patients with ARRP and homozygotes of the mutation in 3 patients were found. Heterozygous of the mutation was detected in the parent of patients and 1 healthy family member. No mutation of RHO gene was found in 2 healthy family members and 10 control individuals. Conclusions The Gln-344-Arg mutation in the RHO gene may be the pathogenic factor of the ARRP family; the frequency of the mutation of RHO gene may increase in the inbreeding ARRP family.
10.METHODS OF THE DIRECTED MOLECULAR EVOLUTION OF ENZYNE IN VITRO
Microbiology 1992;0(02):-
The directed molecular evolution of enzyme in vitro can not only improve the efficiency of evolution, but also evolve enzyme according to the investigator's desire. This review summed up the feasible methods of this novel technique.