[Construction and package of a recombinant adenoviral vector to inhibit miR-21 expression and its effects on HepG2 cells in vitro].

AIM: To construct the recombinant adenoviral vector which can efficiently inhibit mature miR-21 expression and explore its effects and the underlying mechanisms on hepatoma cell line HepG2. METHODS: Based on miRNA-sponge technology, we synthesized 8 duplicated fragments, fully complementary with the miR-21 sequence and cloned them into a shuttle vector pAdTrack-CMV. The constructed plasmid was sequenced and linearized for homologous recombination with pAdEasy-1 vector in BJ5183 bacteria, then transfected into 293A cells. The recombinant adenovirus was used to challenge HepG2 cells. Mature miR-21 level was detected by real-time PCR. Apoptosis and proliferation of the HepG2 cells were detected by Hochest 33258 staining, Western blotting and MTT assay. RESULTS: The restriction enzyme digestion, DNA sequencing and detection of GFP expression demonstrated that recombinant adenoviral vector was constructed successfully. The recombinant adenovirus inhibited the expression of miR-21 in HepG2 cells, and also depressed the proliferation of HepG2 cells and promoted the apoptosis. CONCLUSION: The recombinant adenoviral vector we successfully constructed could efficiently reduce the expression of miR-21 in HepG2 cells and depress the proliferation of HepG2 cells.

[Construction of a recombined adenovirus vector carrying pri-miR-21 gene and research on it's target gene TLR4].

AIM: To construct the recombined adenovirus vector carrying pri-miR-21 gene, which can express mature miR-21 efficiently, and to study the interaction of miR- 21 with its target gene TLR4. METHODS: Using healthy mouse's gDNA as template, the primary miR-21 coding sequence was amplified by PCR and cloned into a shuttle vector pAdTrack-CMV. Constructed plasmid was sequenced and linearized for homologous recombination with pAdEasy-1 vector in BJ5183 bacteria. The recombined adenovirus vector carrying pri-miR-21 gene was used to challenge HeLa cell. The candidate target gene of miR-21 was determined by miRNA analysis databases. The expression level of TLR4 protein was detected by western blotting. RESULTS: Through the PCR, restriction enzyme digestion, DNA sequencing and expression of GFP, recombinant adenoviral vector pri-miR-21 gene was constructed successfully. Bioinformatic analysis suggested a few possible binding sites between miR-21 and TLR4. Results showed that miR-21 down-regulated TLR4 at protein levels. CONCLUSION: The recombinant adenoviral vector containing pri- miR-21 was successfully constructed. miR-21 gene interfered with the expression of TLR4 target gene.

[Interaction between polyamide and zinc salt].

X-ray diffraction and FTIR spectroscopic method have been used to investigate the interaction between the amide group of nylon 6 and zinc chloride. The nylon sample obtained by adding formic acid solution of nylon into aqueous solution saturated by zinc chloride is significantly different from the nylon precipitated after mixing formic acid solution of nylon and water. X-ray diffraction results demonstrate that nylon precipitated from water has crystallization while the sample acquired from saturated solution of ZnCl2 remains amorphous. FTIR spectroscopic study shows that the interaction between zinc ion and the amide group of nylon 6 brings about significant variation of amide I and II bands and prevents the crystallization of nylon. The interaction between nylon and zinc ions may be helpful in the development of a new method of manufacturing new nylon material with superior performance.

Cyano-Bridged 4f-3d Coordination Polymers with a Unique Two-Dimensional Topological Architecture and Unusual Magnetic Behavior.

Rings of 24 atoms form part of the two-dimensional structure of the novel compounds [NdM(bpym)(H2 O)4 (CN)6 ]⋅3 H2 O (M = Fe (1), Co (2); bpym = 2,2'-bipyrimidine). These compounds are the first two-dimensional cyano-bridged species containing lanthanide and transition metal centers. The structures and magnetic properties are investigated.