1.Cloning of L-Methionine ?-lyase gene
Journal of Medical Postgraduates 2003;0(12):-
Objective:To identify,clone,sequence the L-Methionine ?-lyase gene from Trichomonas vaginalis(TVMGL1).Methods:Total RNA was prepared from primitive protozoon trichomonas vaginalis,cDNA fragment encoding Methionine ?-lyase gene was amplified by RT-PCR using specific primers,and then was cloned into pGEM-T vector.Inserted Methionine ?-lyase gene was sequenced by ABI 3730 DNA Sequencer.Results:Analysis indicates that the DNA fragment is 1 191 base pairs in length and has high nucleotide homology with that reported previously.Conclusion:L-Methionine ?-lyase gene was successfully cloned.
2.High expression of the L-Methionine ?-lyase gene in E. coli
Journal of Medical Postgraduates 2003;0(04):-
Objective: To identify and highly express the L-Methionine ?-lyase gene from Trichomonas vaginalis(TVMGL).Methods: The TVMGL gene was cloned into pGEX 4T-2,the recombinant protein expressed in E.coli DH5?and confirmed by SDS-PAGE.Results: Denaturing SDS-PAGE analysis confirmed the predicted size of the fusion protein(GST-TVMGL,68 000) and showed high purity after subsequent affinity chromatography on Glutathione sepharose 4B.The target proteins amounted to 34% of the total bacteria proteins.Conclusion: The L-Methionine ?-lyase gene was highly expressed in E.coli.
3.Molecular cloning and sequencing of apoA1 gene
Rong FANG ; Baikun MA ; Yufeng HUANG ;
Journal of Medical Postgraduates 2003;0(08):-
Objectives:Identification, clone and sequence of the chinese mature apoA1 peptide gene.Methods:Total RNA was prepared from Chinese fetal liver tissue, cDNA fragment encoding human apoA1 was amplified by RT PCR using specific primers, and cloned into pGEM T vector. Inserted apoA1 gene was sequenced by ABI 377 DNA Sequencer. Results:Analysis indicates that the DNA fragment is 739 base pairs in length and has 100% nucleotide homology with that reported previously. Conclusions:Human apoA1 gene was successfully cloned from fetal liver tissue.
4.The study of mannose-binding lectin gene polymorphism and serum concentration in patients with systemic lupus erythematosus
Aili WANG ; Qian CHENG ; Baikun MA ; Rong FANG
Chinese Journal of Rheumatology 2000;0(06):-
Objective To investigate the relationship between polymorphism of codon54of the man-nose-binding lectin(MBL)gene and serum concentration in patients with systemic lupus erythematosus(SLE).Methods The MBL gene54(GGC/GAC)polymorphism was analyzed by polymerase chain reaction and restrict fragment length polymorphism(PCR-RFLP)in44patients with SLE and45healthy controls.Serum levels of MBL were also determined by ELISA.Results The frequencies of the wild homozygons(GGC/GGC)and het-erozygotse(GGC/GAC)of MBL codon54did not differ significantly between patients(GGC/GGC71%,GGC/GAC27%,GAC/GAC2%)and healthy controls(GGC/GGC78%,GGC/GAC22%,GAC/GAC2%).The serum MBL level of the three genotypes was(1.8?0.6)mg/L(n=31),(1.5?0.6)mg/L(n=12),1.17mg/L(n=1)respective-ly in SLE group;(3.0?1.0)mg/L(n=35),(2.2?1.0)mg/L(n=10)in control group.The(GGC/GGC and GGC/GAC)genotypes in SLE group were significantly lower than the control group.Conclusion The condon54mutation of the MBL gene may be not associated with low serum level of MBL in patients with SLE.
5.A novel ROI extracting technique based on wavelet transform for the detection of micro-calcifications in mammograms.
Shunan LI ; Baikun WAN ; Zhenhe MA ; Ruiping WANG
Journal of Biomedical Engineering 2005;22(2):360-362
In order to preprocess mammograms for diagnosing the early cases of breast cancer and improving the computational efficiency in the computer-aided detection of micro-calcifications in mammograms, we have advanced a novel processing technique for the extraction of micro-calcification region of interest (MROI). The proposed method is based on a three-step procedure: (1) the mammogram is divided into sub-images of the same size; (2) the wavelet multi-resolution method is conducted on the sub-images, and the parameters related to wavelet transform and threshold T are discussed according to rho; (3) the classification of sub-images is determined by T. It is tested with 20 mammograms and the results show that the method can achieve a true positive rate as high as 89.7% with a false positive rate as low as 2.1%.
Breast Diseases
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diagnostic imaging
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pathology
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Breast Neoplasms
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diagnostic imaging
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pathology
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Calcinosis
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diagnostic imaging
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Diagnosis, Computer-Assisted
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Humans
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Mammography
6.Preliminary study on the treatment of ischemic heart disease with angiopoietin 1 gene transfection by adenovirus vector in rabbits
Shilin CHEN ; Hua JING ; Jieshou LI ; Long YI ; Baikun MA ; Rong FANG
Chinese Journal of Geriatrics 2000;0(06):-
Objective To study the potential effects of angiopoietin 1(Ang1) via adenovirus mediated gene transfer on ischemic heart disease in rabbits. Methods Forty-five male New Zealand white rabbits underwent high positioned double-ligation of the anterior descending left coronary artery, and were divided into three groups: Ang1 group (n=15) received direct myocardium injection of Ang1 recombinant adenovirus; DMEM group (n=15) and LacZ group (n=15) received only DMEM and LacZ recombinant adenovirus as controls respectively. The myocardial infarcted size was evaluated by N-BT macroscopic standing and the degree of angiogenesis was assessed by use of immunohistochemical analysis. The echocardiographic changes were measured on before operation and the 7 th, 14 th, and 28 th postoperative days. Results Animals in three groups had no significant difference in the percentage of infarction size of left ventricle at the 7 th, 14 th day and capillary density at the 7 th day. Animals in Ang1 group showed less infarcted size than DMEM group or LacZ group at the 28 th day and higher capillary density at the 14 th and the 28 th day. The results from echocardiographic measurements showed that animals in all three groups had no significant difference in the left ventricular systolic function before operation and at the 7 th , 14 th day, but the left ventricular systolic function in Ang1 group was better than that in DMEM group or LacZ group at the the 28 th day(P