1.Application of Filamentous Phage Display Technology in Parasitology
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
This review focuses on the application of phage display technology in the prevention,diagnosis and treatment of parasitic diseases. It covers: ① an introduction of phage display technology,filamentous phage and the advantage of carrier. ② construction of phage antibody libraries of parasites,epitope mapping and mimotope,and their potential application in the development of novel diagnostic reagents and vaccines.
2.Effect of ADAM17-shRNA on proliferation of human breast cancer cell line MCF-7 in hypoxia
Guofu CHEN ; Lijun WU ; Xuepeng ZHANG ; Zhun CAI ; Xiangchao MENG
Chongqing Medicine 2017;46(1):29-32
Objective To investigate the effects of shRNA targeting a disintegrin and metalloproteinases 17(ADAM17)on the proliferation of human breast cancer MCF-7 cells in hypoxia environment.Methods The four specific ADAM17-shRNA se-quences aiming at ADAM17 were designed,transfected into MCF-7 cells by electroporation,and cultured in hypoxia environment. The experiment was divided into the control group (blank phosphate buffer solution,PBS),nonsense sequence group (transfected with ADAM17-shNC)and shRNA transfection group (transfected with ADAM17-shRNA,the highest silencing efficiency of shR-NA was selected for following experiments).Real-time PCR was used to detect the expression of ADAM17 mRNA.The prolifera-tion ability and cell cycle change of MCF-7 cells were detected by iCELLigence and flow cytometry (FCM),respectively.Results Compared with control group and nonsense sequence group,the four ADAM17-shRNA transfection groups all had the silence effect on ADAM17 gene expression (P <0.05 ),the difference was statistically significant(P <0.05 ),particularly shRNA1219 had the highest inhibitory rate (F =5.11,P <0.01 ).The cellular proliferation ability and cell growth speed in the shRNA transfection group were significantly decreased compared with the control group and nonsense sequence group (P <0.05).Most cells of shRNA transfection group remained in the G0/G1 phase (73.35 ± 2.45 ),which in the control group and nonsense sequence group was (62.56±2.35)and (62.68 ±1.20)respectively,the difference was statistically significant(P <0.05).The cell cycle progression was significantly delayed.Conclusion ADAM17-shRNA inhibits the proliferation of MCF-7 cells under hypoxic environment.
3.An ELISA Based on a Truncated Soluble ORF2 Protein for the Detection of PCV2 Antibodies in Domestic Pigs
Shuanghui YIN ; Shunli YANG ; Hong TIAN ; Jinyan WU ; Youjun SHANG ; Xuepeng CAI ; Xiangtao LIU
Virologica Sinica 2010;25(3):191-198
Postweaning multisystemie wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has at least three immunoreactive regions, and it can be a suitable candidate antigen for detecting the specific antibodies of a PCV2 infection. In the present study, an indirect enzyme-linked immunosorbent assay (TcELISA)based on a truncated soluble Cap protein produced in Escherichia coli (E.coli) was established and validated for the diagnostic PCV2 antibodies in swine. The TcELISA was validated by comparison with an indirect immunofluorescence assay (IIFA). The diagnostic sensitivity (DSN), specificity (DSP), and accuracy of the TcELISA were 88.6%, 90.7% and 89.4%, respectively. The agreement rate was 89.38% between results obtained with TcELISA and IIFA on 113 field sera. A cross-reactivity assay showed that the method was PCV2-specific by comparison with other sera of viral disease. Therefore ,the TcELISA will be helpful for the development of a reliable serology diagnostic test for large scale detection of PCV2 antibodies and for the evaluation of vaccine against PCV2 in swine.
4.Progress of model organism genome.
Journal of Biomedical Engineering 2004;21(3):506-511
This paper presents a description of the rules of construction and function of genome as well as evolutional relationship between organisms, and opens out the essence of life through introducing the progress in genome of model organism. The other purpose of this review is to highlight the status and function of model organism in the research of comparing genomics so as to provide the model of cycle for researches into high creature life, especially human beings life.
Animals
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Chromosome Mapping
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Genes
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genetics
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Genome
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Genomics
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methods
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trends
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Human Genome Project
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Humans
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Models, Animal
5.A Simple and Rapid Colloidal Gold-based Immunochromatogarpic Strip Test for Detection of FMDV Serotype A
Tao JIANG ; Zhong LIANG ; Weiwei REN ; Juan CHEN ; Xiaoying ZHI ; Guangyu QI ; Xiangtao LIU ; Xuepeng CAI
Virologica Sinica 2011;26(1):30-39
A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test, affinity purified polyclonal antibodies from Guinea pigs which were immunized with sucking-mouse adapted FMD virus (A/AV88(L) strain) were conjugated to colloidal gold beads and used as the capture antibody, and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted FMD virus (A/CHA/09 strain) were used as detector antibody. On the nitrocellulose membrane of the immunochromatographic strip, the capture antibody was laid on a sample pad, the detector antibody was printed at the test line(T) and goat anti-guinea pigs IgG antibodies were immobilized to the control line(C). The lower detection limit of the test for a FMDV 146S antigen is 11.7ng/ml as determined in serial tests after the strip device was assembled and the assay condition optimization. No cross reactions were found with FMDV serotype C, Swine vesicular disease (SVD), Vesicular stomatiti svirus (VSV) and vesicular exanthema of swine virus (VES) viral antigens with this rapid test. Clinically, the diagnostic sensitivity of this test for FMDV serotypes A was 88.7% which is as same as an indirect-sandwich ELISA. The specificity of this strip test was 98.2% and is comparable to the 98.7% obtained with indirect-sandwich ELISA. This rapid strip test is simple, easy and fast for clinical testing on field sites;no special instruments and skills are required, and the result can be obtained within 15 min. To our knowledge, this is the first rapid immunochromatogarpic assay for serotype A of FMDV.
6.Screening of cDNA Clone for Putative RNA Polymerase Subunit of Cysticercus cellulosae
Xuenong LUO ; Yadong ZHENG ; Yongxi DOU ; Junlin HOU ; Zhizhong JING ; Xuepeng CAI
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(03):-
Objective To obtain related genes of Cysticercus cellulosae from spliced leader (SL) cDNA library. Methods Spliced leader library of Cysticercus cellulosae was constructed using SL specific primer and oligo (dT)15 with M13M4 primer, and positive clones were then screened randomly, identified with enzyme restriction, followed by sequencing and homologous analysis. Results The amino acid sequence, encoded by the positive clone with a poly (A) 22 tail and a complete open reading frame (ORF), was with homology of RNA polymerase subunit genes of human, B. napus, fission yeast, A. thaliana, C. elegans and fruit fly up to 71.6%. Conclusion The protein, RNA polymerase subunit encoded putatively by the clone, is high conservative in different species.
7.CT Diagnosis of Renal Leiomyosarcoma
Fan XU ; Zhaoxia XU ; Xuepeng GONG ; Chaoyang JIA ; Dongshun CAI ; Huimin LI
Chinese Medical Equipment Journal 2003;0(10):-
Objective To discuss the CT manifestations of renal leiomyosarcoma. Methods 9 cases of renal leiomyosarcoma proved by pathology were analyzed retrospectively. Results CT imaging showed homogeneous hyperattenuation in two cases. Heterogeneous in density with necrosis inside was observed on the CT scan in six cases. After contrast medium administration heterogeneous enhancement with cystic and necrotic areas could be revealed in all cases on both CT scan. Conclusion Large renal mass with intra-tumor necrosis,cystic and hemorrhagic change is the characteristic manifestations of renal leiomyosarcoma. But differential diagnosis from renal carcinoma by CT clinical setting is difficult before surgery, and final diagnosis should depend on pathology examination.
8.Suckling mouse model establishment of myocarditis induced by foot-and-mouth disease vi rus
Keshan ZHANG ; Jijun HE ; Jiajun WU ; Yongjie LIU ; Weiwei CHENG ; Youjun SHANG ; Xiangtao LIU ; Xuepeng CAI
Chinese Journal of Zoonoses 2014;(10):987-989,996
Interactions between FMDV and cardiac cells are multifaceted and complex ,these interactions leads to pro-teins alterations in cardiac cells inevitably .To understand the pathogenesis of myocarditis after FMDV infection in mice ,the suckling mouse model for myocarditis induced by foot-and-mouth disease virus (FMDV) was established in this study .Suckling mice within 3 days old was selected to infect by FMDV .Myocarditis caused by FMDV in suckling mice was confirmed with clinical symptom monitor .The observation of Hematoxylin and eosin stain (H&E stain) and transmission electron microscopy (TEM) were performed after samples processing .According to conventional polymerase chain reaction (PCR) methods ,prim-ers of VP1 gene was designed ,synthesised and specific FMDV VP1 gene was amplified from the heart muscle of suckling mice . The results indicated that suckling mice appeared low spirit condition ,dyspnea ,and dull reaction within 36 hours after chal-lenge with FMDV .Infiltration of inflammatory cells and dissolution of myocardial fibers were observed with H&E stain and TEM .Special target gene of FMDV was amplified from the heart of infected group .Obvious inflammation in the heart of suck-ling mice caused by FMDV was observed .It's suggested that suckling mouse model for myocarditis induced by FMDV was es-tablished successfully ,which would lay the foundation for researches of myocarditis mechanism in young cloven-hoofed ani-mals .
9.Codon Usage Bias and Determining Forces in Taenia solium Genome.
Xing YANG ; Xusheng MA ; Xuenong LUO ; Houjun LING ; Xichen ZHANG ; Xuepeng CAI
The Korean Journal of Parasitology 2015;53(6):689-697
The tapeworm Taenia solium is an important human zoonotic parasite that causes great economic loss and also endangers public health. At present, an effective vaccine that will prevent infection and chemotherapy without any side effect remains to be developed. In this study, codon usage patterns in the T. solium genome were examined through 8,484 protein-coding genes. Neutrality analysis showed that T. solium had a narrow GC distribution, and a significant correlation was observed between GC12 and GC3. Examination of an NC (ENC vs GC3s)-plot showed a few genes on or close to the expected curve, but the majority of points with low-ENC (the effective number of codons) values were detected below the expected curve, suggesting that mutational bias plays a major role in shaping codon usage. The Parity Rule 2 plot (PR2) analysis showed that GC and AT were not used proportionally. We also identified 26 optimal codons in the T. solium genome, all of which ended with either a G or C residue. These optimal codons in the T. solium genome are likely consistent with tRNAs that are highly expressed in the cell, suggesting that mutational and translational selection forces are probably driving factors of codon usage bias in the T. solium genome.
Animals
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Base Sequence
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Codon/*genetics
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Evolution, Molecular
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*Genome, Helminth
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Helminth Proteins/*genetics
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Molecular Sequence Data
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Taenia solium/*genetics
10.Characteristic profiles of biofilm, enterotoxins and virulence of Staphylococcus aureus isolates from dairy cows in Xinjiang Province, China
Yehui WU ; Jing LI ; Mengfan QIAO ; Dan MENG ; Qingling MENG ; Jun QIAO ; Xingxing ZHANG ; Lixia WANG ; Kuojun CAI ; Jinsheng ZHANG ; Zaichao ZHANG ; Weiwei YU ; Xuepeng CAI
Journal of Veterinary Science 2019;20(6):e74-
As an important zoonotic pathogen, Staphylococcus aureus has led to serious mastitis and endometritis in infected dairy cows. In this study, a total of 164 strains of S. aureus were isolated from dairy cows in Xinjiang Province, China, and subjected to assays to determine drug susceptibility and biofilm (BF) formation ability. Enterotoxin-related genes were detected, and the transcription levels of genes related to BF formation were determined by using reverse transcription-quantitative polymerase chain reaction. Moreover, the pathogenicity of isolates with different BF formation abilities was determined by measuring their hemolysis activity, half lethal dose (LD₅₀) and organ bacterial load. The results showed that 86.0% of S. aureus isolates could form BF. Among them, 42.1% of the strains had weak BF formation ability, and most strains with a strong BF formation ability were ica gene carriers. The S. aureus isolates displayed multidrug resistance and their drug resistance was positively correlated with their BF formation ability. Moreover, 96.3% of the S. aureus isolates carried enterotoxin genes. Among them, the detection rates of the novel enterotoxin genes were higher than those of conventional enterotoxin genes. Furthermore, isolates with a strong BF formation ability had higher LD50 but lower hemolysis ability and organ bacterial load than those of the isolates with weak or no BF ability. However, isolates without BF ability produced more severe pathological changes than those of isolates with strong BF formation ability. These findings suggest that higher BF ability and presence of novel enterotoxin genes are important characteristics of S. aureus isolates from dairy cows in Xinjiang Province, China, and such isolates may pose potential threats to food safety.
Bacterial Load
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Biofilms
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China
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Drug Resistance
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Drug Resistance, Microbial
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Drug Resistance, Multiple
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Endometritis
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Enterotoxins
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Female
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Food Safety
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Hemolysis
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Lethal Dose 50
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Mastitis
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Polymerase Chain Reaction
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Staphylococcus aureus
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Staphylococcus
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Virulence