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Objective: To study the expression of Toll-like receptors (TLRs) mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase (IDO) mRNA expression in response to TLR ligand stimulation., Methods: The expressions of TLRs and IDO mRNA in human HTR-8/SVneo cells were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR., Results: IDO and TLR1-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γ increased its expression., Conclusions: The expression of IDO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.

To investigate molecular mechanism of traditional Chinese medicine Rheum offcinale against Yersinia pestis, whole genome DNA microarray that contains 4005 annotated genes of Y. pestis was used. The minimal inhibition concentration (MIC) of R. offcinale extract against Y. pestis was determined by liquid dilution method. The gene expression profile of Y. pestis was performed after exposured to R. offcinale extract at a concentration of 10 X MIC for 30 and 60 minutes. The total RNA extracted and purified from Y. pestis were reverse-transcribed to cDNA and labeled by Cy3-Cy5 dye. The labeled probes were hybridized to the microarray and the results were obtained by a laser scanner and analyzed by the SAM software. The microarray data was confirmed by RT-PCR. The platform of the DNA microarray-based bacteria transcriptional profiling was eshtablished. The results revealed general gene expression changes of Y. pestis were a global phenomenon. Down-regulation of genes encoding proteins involved in ribosome protein synthesis was a remarkable change. Genes encoding cell envelope and transport/binding proteins were the major changed genes of the Y. pestis in response to R. offcinale.

To understand the function of Mayven and investigate the pathogenesis of multiple sclerosis, the gene sequences of different truncated Mayven were amplified from the gene library of human brain. These truncated fragments, including fragment P1 (1-902 bp), fragment P2 (1-523 bp), fragment P3 (507-182 bp) and fragment P4 (887-1782 bp), were cloned into pGEX-4T-2 vector to construct recombinant plasmids. The recombinant plasmids were transformed into E. coli BL21(DE3) and induced to express by IPTG. The expressed proteins were detected by SDS-PAGE and Western blot, and were purified by GST purifying system. The results showed that recombinant express vectors of different truncated GST-Mayven were successfully constructed and were expressed in soluble form protein induced by IPTG. The fusion proteins have good reactivity to GST antibody. The construction of recombinant express vectors of different truncated GST-Mayven lays a basis for further function study on Mayven.