1.Targets of molecularimaging for detection of atherosclerosis vulnerable plaques
Chunbao LIU ; Xiaoli LAN ; Yongxue ZHANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2016;36(6):560-564
Early detection and accurate evaluation of vulnerable plaques is important to clinical prevention and in time intervention of atherosclerosis plaque rupture,which is the main reason of cardiovascular and cerebrovascular emergency events.Molecular imaging reveals the formation and progression mechanisms of atherosclerosis at the molecular level,and thus has obvious superiority in early detection and evaluation of vulnerable plaques.Suitable targets are the major contents of molecular probe research.Probes of different imaging modalities have been used to detect vulnerable plaques.The targets including low density lipoprotein,macrophage,adhesion molecule,micro calcification,activated protease,apoptosis,proliferation gene,integrin and thrombus.The mechanism of detecting different targets is different,and the effectiveness varies as well.This review summarizes the development of imaging probes for molecular detection of atherosclerosis vulnerable plaques.
2.Screening scFv Specific to Vcam-1 by Phage Display Library and Its Activity Evaluation
Chunbao LIU ; Yiling SONG ; Yongxue ZHANG
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2015;(4):390-394
Objective To screen out single chain variable fragment antibody (scFv)specific to vascular cell adhesion mole‐cule 1(Vcam‐1)from phage recombinant antibody library ,and to evaluate its activity and compare its activity with full‐length monoclonal antibody.Methods Amplification of Vcam‐1 was performed by PCR and Vcam‐1 gene plasmid was transferred into eukaryotic cells to express Vcam‐1 antigen protein.Immune cuvette was coated with purified Vcam‐1 antigen ,and the positive clones were screened out by 4 rounds of “adhesion‐elution‐proliferation” process with gradually increasing pressure.The posi‐tive clones were tested by ELISA method and high titer clones were chosen for gene sequencing.Then the high‐titer clones were transferred into E.coli ,and the clone with the highest expression was regarded as the final requisite one.Competent cells were infected by the final requisite clone and scFv was expressed.After purification ,the activity of scFv was tested by ELISA and its affinity was evaluated.Results Molecular weight of Vcam‐1 antigen protein was 85-90 kD.Positive clones were screened out by taking Vcam‐1 protein as the antigen ,and 9 high titer clones were obtained by single phage ELISA.Gene sequencing of these clones was carried out and 3 sequences were obtained ,1 of which got the highest expression.Molecular weight of the expressed scFv was about 30 kD.The scFv got high affinity to Vcam‐1 antigen according to ELISA ,in spite of its lower activity than full‐length monoclonal antibody.Conclusion scFv antibody specific to Vcam‐1 was successfully obtained from phage display librar‐y ,which laid the foundation of subsequent in vivo diagnosis and therapy.
3.Effect of Oral Ambroxol on Pharmacokinetics of Aminophyllin
Chunbao ZHANG ; Ning LIU ; Chaobo CUI ; Wenli LIU
China Pharmacy 1991;0(02):-
OBJECTIVE:To study whether there is interaction between ambroxol hydrochloride and amino?phylline.METHODS:16volunteer were randomly divided into trial group and control group.The blood concentration of amin_ ophylline was determinated with HPLC method,then the main parameters of pharmacokinetics of aminophylline were calculated with3p97software.RESULTS:The main parameters of pharmacokinetics of aminophylline in trial and control group were:K e (0.062?0.023)/h and(0.0905?0.013)/h;V(22.83?7.85)and(23.27?3.78)L;T 1/2 (12.51?4.2)and(7.77?0.89)h;AUC 0~21 (426.59?186.92)and(245.74?48.6)(mg?h)/L;CL(s)(23.17?9.83)and(34.84?6.00)ml/min respective?ly.CONCLUSION:Although ambroxol hydrochloride did not influence the distribution of aminophylline,but it could prolong the half time of aminophylline and increase the stagnation time of aminophylline in vivo.
4.A technique of screw retained implant denture for edentulous patients
Zaixi DENG ; Chunbao ZHNAG ; Wei MA ; Chao XIE ; Yan LIU
Journal of Practical Stomatology 2015;(1):136-138
Treatment of edentulous jaws with fixed implant supported denture is a complex procedure with high technical difficulty.In the present paper,the characteristics of screw retained implant denture and key points of manufacture are introduced in details.
5.Thoughts and practice in the emphatic course construction of medical immunology
Xiaojuan WANG ; Bo LI ; Aiping LIU ; Chunbao GAN ; Xueqin WAN
Chinese Journal of Medical Education Research 2002;0(01):-
Course construction is the central part in the teaching innovation and an important indicator of the quality of a school teaching. Based on the emphatic course construction ,the department of immunology has carried out practices from the educational philosophy,teaching content,teaching methods,teaching staff and other aspects of the courses in the construction of immunology,and has got better results.
6.Imaging atherosclerosis model rabbits with 99Tcm-single chain antibody fragment against vascular cell adhesion molecule-1
Chunbao LIU ; Xiao ZHANG ; Yiling SONG ; Fengzhen ZHANG ; Yingying ZHANG ; Yichun WANG ; Xiaoli LAN ; Yongxue ZHANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(6):346-350
Objective To investigate the imaging performance and feasibility of 99Tcm labeled scFv against VCAM-1(99Tcm-scFv-VCAM-1) on atherosclerosis model rabbits.Methods HYNIC was used as a chelator for 99Tcm labeling.The labeling efficiency and radiochemical purity of 99Tcm-scFv-VCAM-1 were measured by instant thin layer chromatography after PD-10 purification.New Zealand white rabbits were employed for establishing atherosclerotic animal models by endothelia immunity injury and high fat diet, and plaques at aorta lesions were examined by HE staining.Model rabbits were sacrificed after administration of 99Tcm-scFv-VCAM-1 at 1 or 2 h respectively, and tissue samples were measured with gamma counter and weighted to obtain in vivo biodistribution data.Planar imaging was performed 1 and 2 h after the injection of 99Tcm-scFv-VCAM-1 to investigate radioactivity of abdominal aorta.After imaging study, atherosclerosis plaque and VCAM-1 expression at aortas were confirmed by the immunohistochemistry (IHC) study.Two-sample t test was used to analyze data.Results 99Tcm-scFv-VCAM-1 was successfully synthesized.Its labeling efficiency was 75%-83%, radiochemistry purity was (98.54±1.03)% and specific activity was 216 MBq/nmol.Atherosclerosis plaque was confirmed at the aortas of experimental rabbits by HE staining, while no plaque was observed in controls.Biodistribution data indicated that the tracer was cleared mainly through the kidneys.Planar imaging showed that the tracer uptake in abdominal aorta of model rabbits was higher than that of control rabbits, the T/B ratios at 2 h of the model group and control group were statistically different (3.68±0.73 vs 2.42±0.39;t=2.950, P<0.05;n=5).Atherosclerosis plaque and high level of VCAM-1 expression were observed at aortas of model rabbits by IHC study.Conclusions It is feasible and effective to detect vulnerable plaques using 99Tcm-scFv-VCAM-1.It may provide a promising way for early diagnosis and accurate evaluation of atherosclerosis.
7.Rapid identification of Pseudomonas aeruginosa by proteinchip golden array
Chunbao XIE ; Daiwen XIAO ; Yongchang YANG ; Wei JIANG ; Hua LIU ; Wenfang HUANG
Chinese Journal of Microbiology and Immunology 2011;31(5):462-466
Objective To establish protein fingerprinting identification model of Pseudomonas aeruginosa (P. aeruginosa) and to lay a foundation for rapid identification of P. aeruginosa by proteinchip golden array. Methods Sixty-four P. aeruginosa and one hundred and ninety-nine control bacteria identified in our laboratory were collected and divided into training and testing group. Surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) and proteinchip golden array were used to detect the protein profiling of the bacteria. Data were automatically collected by Ciphergen Proteinchip Software and protein markers of P. aeruginosa were screened by BioMarker Wizard Software. Classification tree model was developed and validated by BioMarker Patterns Software. The model was blindly tested with twenty-nine P. aeruginosa and sixty-four control bacteria. Results Eighty protein peaks were detected between 3000 and 20 000, among which fifty-eight ones showed significantly difference between P. aeruginosa and the control bacteria (P<0.01). By BioMarker Patterns Software, one protein peak ( M/Z at 14 045.2) was chosen to develop a classification tree model. The results exhibited with sensitivity of 96. 55% and specificity of 100%. Conclusion Proteinchip golden array has the potential for rapid identification of P. aeruginosa.
8.Therapeutic effect of chemokine SLC combined with immune adjuvant CpG-ODN in treatment of implanted mouse melanoma
Xiangfan XU ; Zhenzhu XU ; Lihua TANG ; Anna LI ; Xianhui XU ; Chunbao LIU
Chinese Journal of Cancer Biotherapy 2010;17(1):25-29
Objective: To study the therapeutic effect of secondary lymphoid tissue chemokine (SLC) combined with CpG oligodeoxynucleotide (CpG-ODN) in treatment of implanted mouse melanoma and the possible mechanism. Methods: SLC-Fc fusion protein was prepared and its chemotaxis of lymphocytes was detected by chemotaxis assay. Implanted melanoma mouse models were established and randomly divided into 4 groups: control group, SLC-Fc group, CpG-ODN group, and SLC-Fc+CpG-ODN group. The growth of implanted tumors in each group was observed after treatment. Subtype and infiltration of lymphocytes in implanted tumor tissues were examined by flow cytometry. Results: SLC-Fc protein was successfully prepared, and it dose-dependently attracted lymphocytes (0.03, 0.3, and 3 μg/L). Intra-tumor injection SLC-Fc and CpG-ODN alone or in combination significantly inhibited growth of B16-implanted tumors. Tumor size in SLC-Fc+CpG-ODN group was significantly smaller than that in control group (P<0.01), and animals in SLC-Fc+CpG-ODN group survived longer. Tumor-infiltrated CD4~+ T, CD~8+ T, and dendritic cells (DCs) in SLC-Fc+CpG-ODN group were markedly increased as compared with those in control group (P<0.05), and tumor draining lymph nodes were dramatically enlarged. Conclusion: SLC combined with CpG-ODN can inhibit the growth of implanted melanoma by attracting CD4~+ T and CD8~+ T and promoting proliferation of DCs.
9.Rapid identification of four common bacteria by SELDI-TOF MS protein fingerprints
Daiwen XIAO ; Yongchang YANG ; Hua LIU ; Hua YU ; Chunbao XIE ; Hangfeng ZHANG ; Wenfang HUANG
Chinese Journal of Microbiology and Immunology 2012;32(6):566-570
Objective To establish protein fingerprints of common bacteria in clinics and to lay a foundation for rapid identification of bacteria.Methods Strains of Escherichia coli,Klebsiella pneumoniae,Pseudomonas aeruginosa and Staphylococcus aureus were detected by surface enhanced laser desorption and ionization time-of-flight mass spectrometry (SELDI-TOF MS).Stable expression protein peaks were screened and the data was input into the self-constructed Fingerwave software for identification of target bacteria by protein fingerprint comparison.Two hundred and fifty-six clinical isolates,including E.coli,K.pneumoniae,P.aeruginosa and S.aureus were detected and the data was compared with constructed database to evaluate its diagnostic value.Results The protein fingerprints including four common bacteia was used to identify the target bacteria with identification rate of 93.1% (54/58) for E.coli,87.2% (75/86) for K.pneumoniae,96.2% (60/63) for P.aeruginosa and 96.2% (51/53) for S.aureus,respectively.Conclusion Common bacteria can be rapidly identified by using the protein fingerprint comparison,which provides a powerful tool for bacterial identification.
10.Preliminary study on carbapenem resistance mechanism of clinical isolate of Raoultella planticola
Chunbao XIE ; Hua YU ; Daiwen XIAO ; Yongchang YANG ; Wei JIANG ; Hua LIU ; Wenfang HUANG
Chinese Journal of Laboratory Medicine 2014;(6):459-462
Objective To investigate the mechanism of one carbapenems resistant Raoultella planticola( R.planticola) isolate.Methods This is an experimental study.R.planticola was isolated from a patient′s drainage fluid from orthopedic department in November 2010 in Sichuan Provincial People′s Hospital.Minimum inhibitory concentration of R.planticola to 13 antibiotics was determined by using the agar dilution method.Modified Hodge test was used to detect carbapenemase .EDTA synergistic test was performed to research metallo-beta-lactamase.The genes coded the β-lactamase were amplified by polymerase chain reaction ( PCR ) , including class A carbapenemase ( KPC ) , class B carbapenemases (NDM, IMP, VIM, SIM), extended spectrum beta-lactamases[ESBL(CTX, TEM, SHV)], and AmpCβ-lactamases ( FOX, EBC, ACC, DHA, CIT, MOX).Results The susceptibility test showed that R.planticola was resistant to 9 antibiotics.MIC value of meropenem for R.planticola was up to 32 mg/L.R.planticola kept intermediary to imipenem , whereas it was susceptible to cefepime , amikacin and polymyxin B.Modified Hodge test and EDTA synergistic test were positive in R.planticola.Class B carbapenemase (IMP) gene and two extended spectrum β-lactamases(CTX, SHV) genes were positive by PCR.The genes were conformed as IMP-4, CTX-M3 and SHV-12 by sequencing and compared with GenBank.Other resistant genes were negative.Conclusion IMP-4 was identified in R.planticola, the combined produce IMP-4 and ESBLs might be the main mechanism of R.planticola resistant to carbapenems.