1.Screening the peptide high-binding VEGF receptor 3 with phage-displayed random peptide library
Journal of Third Military Medical University 1983;0(04):-
Objective To get a high affinity peptide of vascular endothelial growth factor receptor 3 (VEGFR3) as a potent carrier targeted to lymphangiogenesis of ovarian cancer via the technology of phage display. Methods Solid-phase was panned with direct VEGFR3 extracellular protein coating, then the unbound phage was washed away and the eluted phage was amplified. The positive phage clones were identified by ELISA and sequenced, and the affinity and specialty were identified by competent ELISA. Results After four-round bio-panning, the enriched positive phage clones were identified by ELISA. Eight positive phage clones were sequenced and 5 were consensus (WHGSLKQNLWWY). The short peptide displayed on screened positive phage could bind specifically to VEGFR3, and the binding could be inhibited by natural antibody VEGF-D. Conclusion The phage clone (phage-WHGSLKQNLWWY) obtained via bio-panning of peptide library has a high affinity with VEGF receptor 3. The peptide could be a potent carrier targeted to VEGFR3.
2.Research on function of VEGF receptor 3 high-bound peptide
Journal of Third Military Medical University 1988;0(05):-
Objective To assess the function of the peptide (WHGSLKQNLWWY) through measuring the a functional affinity constant between humanized VEGF receptor 3 and peptide, then comparing it to that between VEGF D and peptide. Methods After coating the peptide by BSA binding with glutaral couple, the best concentration, best time of peptide to coat the plate and the coating coefficient were determined. With cognizance of the advantages of solid phase method in time consumption and convenience, we assessed the functional affinity constant of engineered peptide and VEGF-D with non-competitive ELISA method. We plotted the D(410) standard curve of the binding reaction of peptide and VEGFR3/Fc using four grades of concentration of VEGFR3/Fc and a series of consistency of peptide. We got the consistency of peptide at the half of maximal OD value through the standard curve and calculated the affinity constant by law of Mass Action. Results The affinity constant of peptide was between 10 7-10 8 mol/L -1, which was only smaller by 10 times than that of VEGF-D. Conclusion The peptide we got from the peptide liberary can strongly bind to the target (VEGFR3), thus providing a theoretical basis for its pharmacal use.
3.Screening of conjugated peptides of Endoglin from phage display peptide library
Xianmin BI ; Zhiqing LIANG ; Lifeng SHI
Journal of Third Military Medical University 2003;0(07):-
Objective To screen rhEndoglin-binding peptides from phage displayed 12-peptide library. Methods The rhEndoglin was used as target protein for biopanning of phage-displayed 12-peptide library. After three rounds of screening, 16 phage clones were randomly selected and identified by sandwich ELISA. The positive phage clones were sequenced, and the fuse peptides were deduced by the DNA sequence. Further we identified the affinity and speciality by competitive inhibition test. Results Six of 16 phage clones were identified as positive clones by competent ELISA which could bind to rhEndoglin. Five sequences were obtained, and the amino acid sequence in two of these five was AHKHVHHVPVRL. Conclusion The rhEndoglin-binding peptides can be obtained by screening phage random peptide library. It could play an important role in early diagnosis of ovarian cancer.
4.Determination of functional affinity of rhEndoglin conjugated eptide
Xianmin BI ; Zhiqing LIANG ; Lifeng SHI
Journal of Third Military Medical University 2003;0(08):-
Objective To assess functional affinity of rhEndoglin conjugated eptide in order to identify the affinity.Methods We measured the functional affinity of rhEndoglin conjugated eptide with non-competitive ELISA method.After coating the peptide by BSA binding with glutaral couple,affirming the best concentration,best time of peptide to coat the plate and the coating coefficient,and the proper binding time of peptide to Endoglin to reach an equilibrium,we plotted the standard curve of the binding reaction of peptide and Endoglin.Results The affinity constant of peptide and anti-hEndoglin IgG is respectively(2.956?0.749)?106mol/L and(7.403?10.76) ?108mol/L.Conclusion The peptide we got from the peptide liberary can strongly bind to Endoglin,providing a theoretical basis for its clinical use.
5.Analysis of cognition and behavior in postpartum rehabilitation of pelvic floor
Zhengcheng ZHANG ; Caiyun LI ; Jingye MAO ; Lifeng SHI
Chinese Journal of General Practitioners 2015;14(7):548-551
From January 2014 to June 2014,a total of 178 primiparas and biparas scheduled for a 42-day postpartum follow-up visit at our hospital were examined.And their pelvic floor dysfunctions were evaluated according to the surveys of incontinence impact questionnaire,short form (IIQ-7),pelvic floor impact questionnaire-7 (PFIQ-7) and pelvic organ prolapse/urinary incontinence sexual questionnaire (PlSQ.12).And 130 primiparas and 48 biparas lacked an understanding of performance,prevention and treatment for postpartum pelvic floor dysfunction.And two groups showed no significant difference.The selfdiscovery and detection rates of decreased pelvic myodynamic were 9.2% and 14.6% in primiparas versus 10.4% and 16.7% in biparas.The self-discovery and detection rates of pelvic organ prolapse were 0.8% and 2.3% in primiparas versus 0.0% and 8.3% in biparas.And the difference was statistically significant.Most related knowledge came from parents and colleagues in 53.1% (95/178) and doctors and nurses in 15.5% (27/178).Only about 44.4% (12/27) women with prominent symptoms of pelvic floor dysfunction requested treatments.Therefore the related knowledge of postpartum pelvic floor dysfunction should be strengthened.
6.Therapeutic effects of erythropoietin and different doses of vitamin E on the prevention of anemia in premature children
Lifeng SHI ; Ping WANG ; Delong FU ; Yunling ZHANG ; Bingping QIU
International Journal of Pediatrics 2013;(2):204-207
Objective To research the therapeutic effect of vitamin E in prevention and treatment of anemia in premature children.Methods We investigated 90 cases of premature children admitted to the neonatal ward of our hospital from December 2010 to December 2011.According to hospital successively order these premature were randomly divided into control group (Group A),high-dose group (Group B),low-dose group (Group A).Group A including 30 cases,subcutaneous injection of erythropoietin(EPO) from the 7th days after admission,750IU/kg weekly for 3 times,for 4 weeks.At the same time,plus iron 6mg/(kg·d).On the basis of the above-mentioned control treatment,oral administration of different doses of vitamin E was in the treatment group,for 4 weeks.Group B including 30 cases was given vitamin E 15mg/(kg·d).Group C including 30 cases was given vitamin E 2.5mg/(kg· d).The changes of each group during the treatment in hematocrit (Hct),hemoglobin(Hb),red blood cell(RBC),reticulocyte(Ret) and the changes of serum concentrations of vitamin E before and after treatment were observed.Results Hct,Hb,RBC,Ret in group B,group C patients were both improved.The improvement of the indicators was better than that of group A (P < 0.05).There was no significant difference between the improvement of each index of Group B and Group C(P > 0.05).Serum concentrations of vitamin E after treatment in group B and group C patients were significantly higher and the difference was statistically significant(P < 0.05).Serum concentration of vitamin E in group A didn't increase obviously.Conclusion Additional small doses of VitE should be given during EPO prevention of anemia in premature children.
7.Imaging and biodistribution of ~(99)Tc~m labeled peptide high-binding VEGF receptor 3 in nude mice with ovarian cancer
Ling WANG ; Zhiqing LIANG ; Lifeng SHI ; Mingfu YANG ; Guangyua LIU
Journal of Third Military Medical University 2003;0(21):-
Objective To investigate the imaging and biodistribution of 99Tcm labeled peptide ( phage-SHSWHWLPNLRHYAS) high-binding VEGF receptor 3 in nude mice with ovarian cancer. Methods We used NHS-MAG3 as the bifunctional chelating agent and synthesized the peptide above. It was completed with pre-stannous direct labeling by 99Tcm and then the labeling efficiency was determined by paper chromatography. The tumor bearing mice were injected via tail veins with radiolabeled peptide. The tumors were imaged with single photon emission computed tomography ( SPECT) . We measured and calculated the biodistribution of the radiolabeled peptide. Results The labeled rate was 95. 27% and the radiochemical purity was 96% . Under a SPECT apparatus,we observed tumor location image at 1 h post injection and the tumor images was the clearest at 3 h post injection,with the injected dose per gram of tissue ( % ID/g) of ( 30. 20 ? 6. 89) at the tumor sites. The tumor/muscle ratio was 13. 13. Conclusion Our peptide can accumulate in the tumor masses. It is hopeful to be used as an ovarian cancer targeting vector for diagnosis and therapy of the cancer.
8.Drug Quality Characterization Consideration of Preparations of Oil Tea Camellia Defatted Cake based on Antithrombotic Effect
Yali WANG ; Lifeng SHEN ; Linfu LI ; Qiuhong WU ; Dongbo LU ; Wenxia LI ; Renbing SHI
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(3):516-521
This study was aimed to select the best preparation form of medicinal in camellia cake,based on the optimal antithrombotic agents in oil tea camellia defatted cake to establish the foundation of research.The content of effective index component of the oil tea camellia defatted cake preparation was detected by HPLC-PDA.The quality of medicinal preparation from oil tea camellia defatted cake was characterized based on the content,relative quantity,relative ratio of content,yield rate and antithrombotic effect.The results showed that the effect of antithrombotic of three medicinal preparations from oil tea camellia defatted cake was listed in the following descending order of enrichment,ethanol extract,and water extract.The enrichment was the best preparation form of medicinal in oil tea camellia defatted cake preparation with the highest content of effective index components,the lowest extract rate and the lowest dosage.But it had the best effect of antithrombotic.It was concluded that the significant improvement of efficacy was due to the increase of the content of the components in the preparation and the rationality of the proportion.At the same time,analysis of component quality based on basic composition of drug system through the consideration of effective components of type and quantity,composition and other quality factors can further guide the preparation of drugs.
9.Research on dendritic cells to soak gallbladder carcinoma tissue
Lifeng AN ; Li SHENG ; Ping HE ; Lin SHI ; Guixiang FAN ; Yukang YUAN ; Min LI
Chinese Journal of Immunology 1985;0(03):-
Objective:Explore the dendritic cells to soak gallbladder carcinoma tissue,to explain the relation dendritic cells and tumor immunity.Methods:Adopt immune histochmistry S-P method.Results:The extent of dendritic cells to soak gallbladder carcinoma tissue is not relate to age,gender and types of pathhistology but negative contact with degree of pathological differentiation (P
10.Effect of cilostazol and aspirin on function of platelet assembly rate and change of protein kinase B activity in elderly patients with acute coronary syndrome
Peiliang LIU ; Tao JING ; Zhuo ZHOU ; Yao CHEN ; Xuan LI ; Chunlai SHI ; Lifeng PEI
Chinese Journal of Postgraduates of Medicine 2006;0(25):-
Objective To examine the effect function of platelet(Pt)assemble rate(PLTAR) and the change of protein kinase B(PKB) active by cilostazol (CS)and aspirin (AS)on elderly patients with acute coronary sydrome(ACS). Methods Forty-eight elderly patients with ACS were divided randomly into two groups:CS group (100 mg,n=26),AS group (300 mg,n=22).Twenty-six healthy elderly were into the group of normal control(NC group) . The CS group and AS group were treated by routine anticoagulation and antiplatelet.PLTAR and PKB activity were measured at 10 minutes before treatment and at 7 days after treatment 3.5,6.0,24.0 hours. Results The maximum PLTAR in elderly CS group and AS group was elevated significantly compared with NC group(P