1.Construction and Expression of an Eukaryotic Recombinant Plasmid from a Hybrid Gene of Antigen for Plasmodium falciparum
Lang YAO ; Ying-Jie LI ; Ming LI ;
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(02):-
Objective To construct an eukaryotic recombinant plasmid with HGFSP, a hybrid gene encoding the antigen epitopes of MSA1, MSA2, RESA and CSP in different developing stages of Plasmodium falciparum(P.f. ). Methods HGFSP was sub-cloned into an eukaryotic expression plasmid pcDNA3 from a prokaryotic recombinant plasmid pSK-HGFSP to construct the eukaryotic recombinant expression plasmid pc-HGFSP. The identified recombinant was then transfected into HepG2 cells with liposome-mediated method. The G418-selected positive cell clones were tested to identify the immunogenicity of HGFSP-expressing antigen. Results It was evidenced that HGFSP was correctly inserted into pcDNA3 by restriction enzymes map analysis. HGFSP-expressing antigen-specific fluorescent response was observed in pc-HGFSP-transfected HepG2 cells. The results of SDS-PAGE and Western-blotting showed that there was a 23 kD protein band, which can be specifically recognized by anti-sera of HGFSP-expressing protein in pc-HGFSP-transfected HepG2 cell lysis. Conclusion Pc-HGFSP, an eukaryotic recombinant plasmid encoding hybrid antigen epitopes of P.f ., was constructed successfully and the antigenicity of pc-HGFSP-expressing protein was confirmed.
2.A Reusable Chronocoulometric Aptamer Sensor for Detection of Adenosine Triphosphate
Ying LU ; Yan TIAN ; Li WANG ; Yao YANG ; Xiaolin YAO
Chinese Journal of Analytical Chemistry 2017;45(5):721-726
A reusable chronocoulometric adenosine triphosphate (ATP)-aptamer sensor was developed in this work.A short chain of DNA marked as cDNA containing complementary sequence was immobilized on gold electrode based on Au-S self-assembly.The ATP aptamer was hybridized with cDNA.The surface-confined DNA could bind with [Ru(NH3)63+ (RuHex) in the electrolyte via electrostatic interaction.Upon target ATP binding, the aptamer confined onto electrode surface was disassociated from the cDNA oligonucleotides into the solution.Such surface density change of DNA lead to the decrease of chronocoulometric signal for the RuHex which confined on the electrode surface.The chronocoulometric signals showed a linear relationship with logrithm of ATP concentration in the range of 1 nmol/L to 100 μmol/L, and the detection limit of this aptamer sensor could reach 0.5 nmol/L (S/N=3).This aptamer sensor could be regenerated 5 times by simple steps.With this aptamer sensor, the basal level of ATP in the brain cortex micorodialysate was determined to be 19.2±3.7 nmol/L (n=3).
3.Cloning, expression, purification and activity assay of Leucyl-tRNA synthetase from Trypanosoma Brucei
Ying YAO ; Guangwei GAO ; Dawei LI
Chinese Journal of Biochemical Pharmaceutics 2010;31(1):14-18
Purpose To clone and express Trypanosoma Leucyl-tRNA synthetase (LeuRS) gene and to complete the purification and activity assay of LeuRS. Methods We cloned the LeuRS gene from T. Brucei genomic DNA,and cloned it into pBS-T and then into the expression vector pET21a( + ) .The expression of T. Brucei LeuRS was carried out in E. coli BL21 (DE3) RIPL host by optimizing the expression conditions. The products are purified by His-Bind affinity column and verified by Western blot . Enzymatic activity was detected by isotope labeling. Results A DNA fragment at length of 3.2 kb was amplified. Both restriction analysis and sequencing analysis proved that recombinant plasmid pET21a( + )/LeuRS was correctly constructed. The expressed product contained about 20% of total somatic soluble protein and reached a purity of 85% after purification . It was verified by the Western blot. Enzyme unit per millilitre of purified products was about 72. Conclusion Here we report the successful clone, expression and purification of LeuRS from Trypanosoma Brucei ( T. Brucei) in bacterial host. In addition, we tested its enzymatic activity by isotope labeling.This work would be helpful to the design and in vitro screening of Trypanosoma LeuRS inhibitors.
5.The effects of intranasal budesonide on eosinophils in nasal polyps
Shixin ZHENG ; Li YAO ; Ying XIAO
Chinese Journal of Primary Medicine and Pharmacy 2005;0(02):-
Objective To study clinicopathologic significance of eosinophils in human nasal polyps and the effect of corticosteroids.Methods 90 cases of nasal polyps were divided into two group randomly:corticosteriods group(n=30) with topical application of budesonide(BUD,400?g/d) for 14 days before endoscopic polypectomy,and control group(n=60) without BUD before surgery.All cases were stained with HE.Results Eosinophils showed higer in the group of nasal polyps than in the group of norm al inferior turbinate(P
6.Concentrations of Morphine in Colostrum and Plasma During Postcesarean Epidural Anesthesia
Ying WANG ; Xiaomei YAO ; Li LONG
Chinese Journal of Perinatal Medicine 1998;0(03):-
Objective To study the effect of morphine used in epidural anesthesia on puerpera after cesarean section and the safety of morphine to neonate. Method One hundred puerpera undergone cesarean section were randomly divided into test group and control group with continuous 2% lidocaine epidural anesthesia, as soon as the operation were finished, 2 mg morphine was injected into vacum epidurale for test group, nothing for control group. Colostrum and plasma, urine samples of puerpera and neonate were collected, morphine and metabolite level were tested by GC-MS and FPIA. Result Morphine concentrations ranged from 0.05). Conclusion As the epidural analgesia medicine after cesarean section, morphine has no side-effect to neonate and is safe to neonate.
7.Practice of concept maps in nursing teaching
Ying LI ; Jing ZHANG ; Li GUI ; Yao CHEN ; Lingjun ZHOU
Chinese Journal of Medical Education Research 2011;10(5):598-600
This article is to explore the practical application of concept maps in nursing teaching practice to make it as a learning tool to promote undergraduates to make a meaningful study. Besides, the results is applied in research on improving the teaching method so as to provide an effective teaching policy and evaluation tools to promote the scientific research and clinical practice in nursing care.
8.A respective analysis on 462 cases of leukemia typing
Wen LIU ; Jun′an LI ; Bin GUO ; Ying LI ; Lihua YAO ;
International Journal of Laboratory Medicine 2015;(12):1708-1709,1712
Objective To study the relationship between FAB‐classifying standard and immunophenotype of leukemia in clinical diagnostic and treatment .Methods There were 462 cases of leukemia according to FAB‐classifying standard and 34 cases by immu‐nophenotype in our hospital from 2011 to 2013 .The results of FAB‐classifying standard and immunophenotype were statisticed and compared .Results Among the 462 cases ,there were 171 acute myeloid leukemia (AML) ,50 acute lymphoblastic leukemia (ALL) , 76 chronic leukemia ,3 rare types ,56 unclear types and 27 mismatched according the FAB‐classifying standard .The diagnostic rate was 82 .03% .34 cases of immunophenotype included 29 single phenotype ,4 double phenotypes and 1 unknown .The diagnosis rate was 97 .06% .CD13 and CD33 have high positive express in AML ;CD34 and HLA‐DR do not express in M3 ;CD7 which was known as a lymphatic antigen also was founded in AML .Conclusion The combing use of FAB‐classifying standard and immunophenotype can improve diagnostic rate ,and immunophenotype has an important role in differentiating different subtypes of leukemia .
9.Effects of Shenmai injection on hemodynamics of isolated heart in rats of myocardial infarction
Hongying CHEN ; Wenjie WANG ; Zhongjun YAO ; Ying LI ; Xianyu LI
International Journal of Traditional Chinese Medicine 2015;(1):61-64
Objective To observe the effects of Shenmai injection on cardiac hemodynamics of isolated heart in rats of myocardial infarction model. Methods 30 myocardial infarction rat models created by ligation of left anterior descending coronary artery were randomly divided into a model group, a captopril group and a Shenmai group. 24 hours after successfully setup the models, all rat models were executed and their hearts were taken. Langendorff isolated heart perfusion method was adopted, and each group was perfused with 0.05mol/L amount of corresponding medicine, the model group was perfused with KH nutritive medium. left ventricular peak pressure(LVSP)and left ventricular ejection fraction(LVEF), left ventricular pressure increase/decrease rate(dp/dtmax±LV)(mmHg/s)and left ventricular end systolic diameter(LVESD)(%)and left ventricular end diastolic diameter(LVEDD)(%) was observed 5 min before the perfusion and 0.5 h, 1 h, 2 h after the perfusion. Results LVSP(9.47%± 0.15%vs. 3.34%± 0.05%),(11.25%± 1.31%vs. 3.79%± 0.04%) and LVEF reducing rate (7.44%± 0.10%vs. 4.94%± 0.04%), (10.24%± 0.31%vs. 5.34%± 0.05%) were elevated, and ±dp/dtmax(5 011 ± 253 mmHg/s vs. 5 827 ± 227 mmHg/s), (4 732 ± 212 mmHg/s vs. 5 837 ± 254 mmHg/s);(3 139 ± 127 mmHg/s vs. 4 722 ± 231 mmHg/s), (2 997 ± 125 mmHg/s vs. 4 793 ± 241 mmHg/s) was reduced in the Shenmai group 1 and 2 hours after the administration, which showed statistical difference compared to the model group (P<0.05). Conclusions Shenmai injection reduce myocardial contractility and intraventricular pressure of isolated heart o myocardial infarction rat models.
10.Analyze compensation patterns and compensation effects of catastrophic disease insurance in L City
Li XIANG ; Huiqiu LUO ; Yao PAN ; Cong LI ; Ying ZHANG
Chinese Journal of Health Policy 2015;8(3):29-33
Objective:This paper designs to analyze compensation patterns and compensation effects of cata-strophic disease insurance in L City, and put forward feasible suggestions to improve the compensation patterns of cat-astrophic disease insurance. Methods:We combined the relevant policy documents to analyze compensation patterns, and used benefit rate, OOP and The effective reimbursement rate to analyze compensation effects of catastrophic dis-ease insurance. Results:Catastrophic disease insurance benefit rate in L city in 2013 was 3. 2%; Rates of NCMS fund unilization was 92%, which diversed from county to county. Patients' OOP decreased significantly after reim-bursement of catastrophic disease insurance;Catastrophic disease insurance and NCMS total effective reimbursement rate reached 84.8%;The NCMS compensation rate reached 68. 9%, while fund incurred a financial deficit at the same time. Conclusion:Set deductibles, compensation rate and compensation range scientifically, and cancel ceiling level,improve the program of catastrophic disease insurance. Take measures to reduce the unfair between the districts and counties at city level. Establish effective link-up between catastrophic disease insurance and NCMS.