Analysis of Gene Expression through RNA Sequencing in Prefrontal Cortex of Nicotine-Treated Rats / 天津医药

Objective To investigate the effects of nicotine on the gene expression profile in prefrontal cortex (PFC) region of rats. Methods Twenty-four adult male F344 rats were randomly divided into treatment group and control group, with 12 animals in each group. Animals in treatment group were injected with nicotine solution while those in control group were given physiological saline for 17 days. At the end of drug administration, the tissue of PFC region was sliced from the brain of each animal. RNA sequencing (RNA-Seq) was utilized to analyze the gene expression profile in PFC of each animal. Bioinformatics tools including Bowtie, Tophat and Cufflinks were used to map the sequencing reads to the reference genome of rat, and to measure the relative expression level of each transcript. Genes whose expression levels were significantly differ-ent between the two groups were identified. The functional categories and the biochemical pathways associated with these genes were further analyzed. Results By comparing the expression level of each transcript between two groups, 885 signifi-cantly differentially expressed genes were identified. These genes were enriched in multiple Gene Ontology Biological Pro-cess categories (e.g., G protein coupled receptor protein signaling pathway, protein ubiquitination, and neurotransmitter up-take) and biological pathways (e.g., insulin signaling pathway, Alzheimer's disease, and long term potentiation). Conclu-sion Nicotine treatment may regulate signaling transduction, energy metabolism and other biological processes in PFC of rat brain.

Study on the interaction between hepatitis virus C nonstructural protein 4A and calcium modulating cyclophipin ligand by in vivo coimmunoprecipitation / 中华传染病杂志

Objective To prove the interaction between hepatitis virus C(HCV)nonstruetural protein 4A(HCV NS4A)and calcium modulating cyclophilin tigand(CAML)with yeast-two hybrid- ization and coimmunoprecipitation.Methods The gene encoding CAML was cloned,and subcloned into the yeast expression vector pGADT7 and eucell expression vector pcDNA3.1/His-A.The back- cross test between HCV NS4A and CAML was performed in yeast cells.After that,the pCMV-Myc/ NS4A plasmid and pcDNA3.1/His-A-CAML plasmid were co transfected into 293 cells and,then, coimmunoprecipitation and Western blot were performed.Results The gene encoding CAML was cloned sucessfully,and then the gene was subcloned into yeast expression vectors,pGADT7.After the interaction between NS4A and CAML was ensured in yeast cells,the eukaryotic expression vec- tors of NS4A and CAML were constructed and their interaction was ensured again by Co-immunopre- cipitation.Conclusions The interaction between HCV NS4A and CAML is proved.CAML is one of the proteins involved in Ca~(2+)signaling,which suggests that the interaction of HCV NS4A and CAML may be a new clue of the chronic mechanism of HCV infection.Future studies will be required to de- fine the physiologic significance of this interaction.

Free fatty acids and insulin resistance / 中国药理学通报

Insulin resistance ,a key component of type 2 diabetes,is recognized as the pathological basis of the metabolic syndrome. H owever,its underlying mechanisms are still not fully understood.Recently,eleva ted levels of free fatty acids (FFA)have been observed in many insulin-resist ance states,which has gradually drawn attention.Not only do FFA interfere in se veral steps involved in glucose metabolism,but also play an important role in i nsulin signal transduction,resulting in decreased insulin-stimulated glucose t ransport. The thiazolidinediones,as insulin-sensitizing drugs,have entered th e market. A prominent effect of these agents is the lowering of circulating FFA levels and it is believed that targeting FFA metabolism may therefore offer wide r expectations in therapy for insulin resistance and type 2 diabetes.