AIM: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer. METHODS: Extract total RNA from tissue of local cancer metastasis...AIM: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer. METHODS: Extract total RNA from tissue of local cancer metastasis lymph nodes of patients with colorectal cancer. RT-PCR was used to amplify the heavy chain Fd and light chain kappa and the amplification products were inserted successively into the vector pComb3 to construct the human libraries of Fab antibodies. They were then panned by phage display technology. By means of Dot immunoblotting and ELISA, the libraries were identified and the Fab phage antibodies binding with antigens of colorectal cancer were selected. RESULTS: The amplified fragments of Fd and kappa gained by RT-PCR were about 650 bp. Fd and kappa PCR products were subsequently inserted into the vector pComb3, resulting in a recombination rate of 40% and the volume of Fab phage display library reached 1.48 x 10(6).The libraries were enriched about 120-fold by 3 cycles of adsorption-elution-multiplication (panning). Dot immunoblotting showed Fab expressions on the phage libraries and ELISA showed 5 clones of Fab phage antibodies which had binding activities with antigens of colorectal cancer. CONCLUSION: The natural immune Fab antibody phage display libraries of colorectal cancer were constructed. They could be used to select the relative antibodies of colorectal cancer.展开更多
Although the gut microflora is thought to be an essential factor in the development of ulcerative colitis (UC), the entire gut microflora occurring in UC remains unknown. Most studies use feces to represent the microf...Although the gut microflora is thought to be an essential factor in the development of ulcerative colitis (UC), the entire gut microflora occurring in UC remains unknown. Most studies use feces to represent the microflora distribution;however, here we analyzed the bacterial diversity in colonic mucus from UC patients receiving colectomy surgery and control patients. The diversity of microflora was investigated using a combination of terminal restriction fragment length polymorphism (T-RFLP) and clone library analyses of the 16S rRNA gene sequences. In the T-RFLP analysis, the number of terminal restriction fragments (T-RFs) decreased significantly in UC patients when compared to control samples. Also in the clone library analysis, the number of operational taxonomic units (OTU) and the Shannon diversity index were reduced significantly in UC patients. These molecular analyses reveal an overall dysbiosis in UC patients. No specific pathogen was found, and a strong negative correlation in relative abundance of bacterial populations was observed between the phyla Bacteroidetes and Firmicutes in the UC patients. This is the first report showing a significant correlation between these two phyla, which may be important characteristics in the pathogenesis of UC.展开更多
Digital library is a process of exploiting more scientific methods to construct a social knowledge network. It represents an “extension” from a traditional library. The core and essence is to build a favorable surro...Digital library is a process of exploiting more scientific methods to construct a social knowledge network. It represents an “extension” from a traditional library. The core and essence is to build a favorable surroundings for knowledge accumulation, knowledge transfer and knowledge communication. Management communication covers information transfer and organization cooperation. Compared with traditional library, the management communication in digital library shoulders the commitment to support knowledge innovation. On the basis of analysis of the connotation of digital library, this paper analysed the aspects of management communication, including communicators, communication styles, contents, channels and outer communication, all of which tries to construct an integrated model of management communication in a digital library and turns a library into an actual “knowledge fountain”.展开更多
Screening humanized antibodies from a human Fab phage display library is an effective and quick method to obtain beta-amyloid oligomers. Thus, the present study prepared amyloid-beta 42 oli- gomers and constructed a h...Screening humanized antibodies from a human Fab phage display library is an effective and quick method to obtain beta-amyloid oligomers. Thus, the present study prepared amyloid-beta 42 oli- gomers and constructed a have human Fab phage display library based on blood samples from six healthy people. After three rounds of biopanning in vitro, a human single-domain antibody that spe- cifically recognized amyloid-beta 42 oligomers was identified. Western blot and enzyme-linked im- munosorbent assay demonstrated this antibody bound specifically to human amyloid-beta 42 tetramer and nonamer, but not the monomer or high molecular weight oligomers. This study suc- cessfully constructed a human phage display library and screened a single-domain antibody that specifically recognized amyloid-beta 42 oligomers.展开更多
A test items knowledge library system of for adaptive learning is proposed in this paper. The first step is to carry out the quantity and quality analysis of the test items by using the Bloom's revised taxonomy and s...A test items knowledge library system of for adaptive learning is proposed in this paper. The first step is to carry out the quantity and quality analysis of the test items by using the Bloom's revised taxonomy and scale anchoring respectively to produce the characteristics for test items. A smoothing method of arbitrary anchoring revised from scale anchoring is first proposed to make tests more accurate in distinguishing test levels. In addition, raised three dimensional indicators based on the Bloom's revised taxonomy are adopted to validate test contents and therefore it concretely describes the examining function of items. The items obtained have the precise and concrete properties; an item knowledge library is therefore constructed combining teaching materials and items using the technologies of ontology and knowledge management. Finally, a knowledge library system of test items is established to achieve the purpose of adaptive learning for learners.展开更多
基金Supported by the Natural Science Foundation of Guangdong Province,China, No. 980120the Foundation of Excellent Youth Teacher,China,2001
文摘AIM: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer. METHODS: Extract total RNA from tissue of local cancer metastasis lymph nodes of patients with colorectal cancer. RT-PCR was used to amplify the heavy chain Fd and light chain kappa and the amplification products were inserted successively into the vector pComb3 to construct the human libraries of Fab antibodies. They were then panned by phage display technology. By means of Dot immunoblotting and ELISA, the libraries were identified and the Fab phage antibodies binding with antigens of colorectal cancer were selected. RESULTS: The amplified fragments of Fd and kappa gained by RT-PCR were about 650 bp. Fd and kappa PCR products were subsequently inserted into the vector pComb3, resulting in a recombination rate of 40% and the volume of Fab phage display library reached 1.48 x 10(6).The libraries were enriched about 120-fold by 3 cycles of adsorption-elution-multiplication (panning). Dot immunoblotting showed Fab expressions on the phage libraries and ELISA showed 5 clones of Fab phage antibodies which had binding activities with antigens of colorectal cancer. CONCLUSION: The natural immune Fab antibody phage display libraries of colorectal cancer were constructed. They could be used to select the relative antibodies of colorectal cancer.
文摘Although the gut microflora is thought to be an essential factor in the development of ulcerative colitis (UC), the entire gut microflora occurring in UC remains unknown. Most studies use feces to represent the microflora distribution;however, here we analyzed the bacterial diversity in colonic mucus from UC patients receiving colectomy surgery and control patients. The diversity of microflora was investigated using a combination of terminal restriction fragment length polymorphism (T-RFLP) and clone library analyses of the 16S rRNA gene sequences. In the T-RFLP analysis, the number of terminal restriction fragments (T-RFs) decreased significantly in UC patients when compared to control samples. Also in the clone library analysis, the number of operational taxonomic units (OTU) and the Shannon diversity index were reduced significantly in UC patients. These molecular analyses reveal an overall dysbiosis in UC patients. No specific pathogen was found, and a strong negative correlation in relative abundance of bacterial populations was observed between the phyla Bacteroidetes and Firmicutes in the UC patients. This is the first report showing a significant correlation between these two phyla, which may be important characteristics in the pathogenesis of UC.
文摘Digital library is a process of exploiting more scientific methods to construct a social knowledge network. It represents an “extension” from a traditional library. The core and essence is to build a favorable surroundings for knowledge accumulation, knowledge transfer and knowledge communication. Management communication covers information transfer and organization cooperation. Compared with traditional library, the management communication in digital library shoulders the commitment to support knowledge innovation. On the basis of analysis of the connotation of digital library, this paper analysed the aspects of management communication, including communicators, communication styles, contents, channels and outer communication, all of which tries to construct an integrated model of management communication in a digital library and turns a library into an actual “knowledge fountain”.
基金supported by the National Natural Science Foundation of China,No.30600099(FD)
文摘Screening humanized antibodies from a human Fab phage display library is an effective and quick method to obtain beta-amyloid oligomers. Thus, the present study prepared amyloid-beta 42 oli- gomers and constructed a have human Fab phage display library based on blood samples from six healthy people. After three rounds of biopanning in vitro, a human single-domain antibody that spe- cifically recognized amyloid-beta 42 oligomers was identified. Western blot and enzyme-linked im- munosorbent assay demonstrated this antibody bound specifically to human amyloid-beta 42 tetramer and nonamer, but not the monomer or high molecular weight oligomers. This study suc- cessfully constructed a human phage display library and screened a single-domain antibody that specifically recognized amyloid-beta 42 oligomers.
文摘A test items knowledge library system of for adaptive learning is proposed in this paper. The first step is to carry out the quantity and quality analysis of the test items by using the Bloom's revised taxonomy and scale anchoring respectively to produce the characteristics for test items. A smoothing method of arbitrary anchoring revised from scale anchoring is first proposed to make tests more accurate in distinguishing test levels. In addition, raised three dimensional indicators based on the Bloom's revised taxonomy are adopted to validate test contents and therefore it concretely describes the examining function of items. The items obtained have the precise and concrete properties; an item knowledge library is therefore constructed combining teaching materials and items using the technologies of ontology and knowledge management. Finally, a knowledge library system of test items is established to achieve the purpose of adaptive learning for learners.