0.05).The level of serum EGF in the tumor group(0.426?0.084 ng/ml～0.456?0.095 ng/ml)was significantly higher than that in the control group(0.243?0.086 ng/ml,P

During recent years, some new methods for phage display panning have been developed by using complex biological systems instead of purified antigens, antibodies or receptors as selector targets. These complex biological systems include living cells, viruses, mouse tissues and tumors etc. The peptides identified by using tumors as selector molecules can selectively home to blood vessels of experimental tumors and may have potential applications in targeted anti-tumor treatment.

One of the aims of biotechnological pharmaceutics is to educate innovative highquality medical talents.The second classroom activities can be used as a complementation to ensure the teachiug effectiveness of the course.This study focuses on the experience of carrying out the second class of biotechnological pharmaceutics and tends to provide references for teachers who are going to carry out the second class.

Objective To develop the purification method of rhIL 18 expressed in E coli Methods The harvested cells were disrupted using lysozyme and sonication The rhIL 18 inclusion bodies were extracted by centrifugation Washed inclusion bodies were solubilized in 8 mol/L urea The solubilized inclusion bodies were first purified by ion exchange chromatography under denaturing conditions After renaturation, gel filtration chromatography was used for further purification Samples were analyzed by SDS PAGE, HPLC, Western blotting and N terminal amino acid sequencing The biological activity of purified rhIL 18 was also measured Results It was found that rhIL 18 was expressed intracellularly in E coli as insoluble inclusion bodies The purity of rhIL 18 in extracted inclusion bodies was above 80% The final purified rhIL 18 was of high purity and exhibited the mass of molecule 19?10 3 by SDS PAGE The sequence of 15 amino acid residues form NH2 terminus of the purified rhIL 18 was consistent with the predicted sequence The purified rhIL 18 also induced IFN ? production of Con A stimulated human PBMC Conclusion The established method for purifying rhIL 18 is simple and effective and might be useful in the development of the large scaled purification process of rhIL 18

Objective To evaluate the role of mitochondrial pathway in the apoptosis of gastric epithelial cells induced by H. pylori in Mongolian gerbils. Methods A total of 48 Mongolian gerbils were randomly divided into H. pylori infection and without H. pylori infection groups. Eight animals from each group were killed at 1, 3 and 6 months, and histopathological changes in their stomachs were examined. A flow cytometry was used to measure apoptosis, mitochondrial membrane potential and intracellular free Ca2+ . Results The incidences of intestinal metaplasia and dysplasia in the gastric mucosa with H. pylori infection were significantly higher than those without H. pylori infection (P

MicroRNAs (miRNA) are a newly discovered class of endogenous,evolutionarily conserved small noncoding RNAs involved in posttranscriptional gene regulation. Functionally speaking,miRNAs act as key regulators in a wide variety of biological processes,including cell proliferation,cell differentiation,apoptosis,metabolism,developmental timing,signal transduction and tumor. Recent publications have provided compelling evidence that a range of miRNAs are involved in the regulation of immunity,including the innate immunity,proliferation of monocytes and neutrophils,the development and differentiation of B and T cells,infection and immunity,and the release of inflammatory mediators. In this review,we examine what is presently known of the function and mechanism of these miRNAs in the regulation of the innate and acquired immune response.

Objective To clone hpaA gene of H.pylori and construct its eukaryotic expression plasmid. Methods Gene hpaA amplificated from genome of H.pylori 11637 strain by PCR was subcloned into pMD18-T vector. Then hpaA cut down from the vector with restriction enzyme was inserted to pTCAE, and the product confirmed by restriction enzyme digestion and sequence was transformed to E.coli DH5?. pT-hpaA was transfected into CHO cell by electroporation, and HpaA protein was detected by Western blot. Results hpaA gene was inserted into pTCAE and the immunological reaction band with anti-HpaA serum at MW 30 000 was detected by Western blot. Conclusion Eukaryotic expression plasmid of hpaA of H.pylori is successfully constructed. Western blot analysis demonstrates that the expression of HpaA protein can be detected in culture supernatants of transfeced CHO cells, which lays the foundation for the further study.

Objective To construct the recombinant adenovirus encoding human IL-24 gene for future gene therapy. Methods The human IL-24 gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria. Then the recombinant adenovirus was transfected into 293T cells using Lipofectine DOTAP. The target gene was detected by polymerase chain reaction (PCR). The titer and its infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. The expression of target protein was measured by the method of immunohistochemistry. Results Restriction endonuclease and PCR analysis confirmed that the human IL-24 gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.2?10 10 pfu/ml. The adenovirus has a strong effect on A549 cells and human IL-24 can express in it. Conclusion The recombinant adenovirus containing human IL-24 gene was successfully constructed by the method of homogenous recombination in bacteria.

Objective To construct the recombinant adenovirus encoding human p16 gene for future gene therapy.Methods The human p16 gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria.Then the recombinant adenovirus was transfected into 293T cells using Lipofectine DOTAP.The target gene was detected by polymerase chain reaction(PCR).The titer and its infection rate were determined using the green fluorescent protein(GFP) expression in the shuttle plasmid.Results Restriction endonuclease and PCR analysis confirmed that the human p16 gene was successfully inserted into the adenovirus vector.The titer of the recombinant adenovirus was 6.1?10~(10)pfu/ml.The adenovirus has a strong effect on human fibroblast cells.Conclusion The recombinant adenovirus containing human p16 gene was successfully constructed by the method of homogenous recombination in bacteria.

Objective To investigate and compare the humoral immune response induced by eukaryotic expression plasmid pTCAEureB (pTureB)and pTCAEhpaA (pThpaA) injected intramuscularly in mice. Methods The ureB gene fragment was inserted into a eukaryotic expression vector pTCAE, and pTCAEureB was constructed. A total of 60 BALB/c mice were randomly assigned to be immunized by intramuscular injection of blank plasmid pTCAE, plasmid pTureB and pThpaA (n=20 in each group). Titers of specific IgG antibody in serum of each group were detected through ELISA respectively on week 2, 4, 6 after immunization. Results Specific IgG were observed in pTureB and pThpaA immunized groups and titers of antibody increased as time prolonged. IgG level was significantly higher in pT-ureB immunized group than in pT-hpaA group on week 6 after immunization. Conclusion Plasmid pT-ureB possesses good immunogenicity and induces a higher immune response than pT-hpaA.