The complete mitochondrial genome sequence of Phlaeoba albonema Zheng Was determined by using L-PCR and conserved primers walking sequencing.The obtained genome sequence is 15657 bp in size.containing 13 protein-coding genes,2 ribosomal RNA and 22 transfer RNA genes.All the 37 genes are conserved in the same orientations as observed in Locusta migratoria.11202 bp of the mtDNA are coding for proteins,1486 bp for tRNAs,1312 bp for rRNA large subunit(1rRNA),and 844 bp for rRNA small subunit(srRNA).The A+T-rich region is 728 bp in size.The genes overlapping sequences are 41 bp in total and are spreading over 9 locations(1-8 bp at each site).A total of 126 bp intergenic spacer sequences are scattered in 21 regions at the size of 1 to 20 bp,where the largest 20 bp region iS located between the tRNALys and ATP8 genes.The predicted secondary structures of both srRNA and lrRNA were compared with that of Ruspolia dubia,and the patterns of base pairs in tRNA anticodon stem and A/T,C/G bias of protein-coding genes in different strands were discussed.

Objective To over-express human trefoil factor 2 (hTFF2) by Escherichia coli system and an-alyze its activities in promoting migration and anchorage-independent growth in SW480 colonic cancer cells. Meth-ods hTFF2 gene encoding mature peptide was obtained by RT-PCR, and the recombinant expression vector pET32a-hTFF2 was constructed. Then pET32a-hTFF2 was transformed into E. coli BL21-32a and TrxA-hTFF2 fu-sion protein was induced to over-express. The expressed product was isolated by Ni-NTA affinity chromatography, purified by dialysis and identified by Western blotting. The activities of the recombinant hTFF2 in promoting SW480 cells migration and anchorage-independent growth were analyzed by MicroChemotaxis Chamber migration assay and Soft-agar assay,respectively. Results The TrxA-hTFF2 fusion protein was expressed to 220 mg/L at high purity. In vitro model demonstrated that recombinant hTFF2 obviously enhanced SW480 cell migration activity and anchor-age-independent growth. Conclusion The recombinant hTFF2 can be expressed in E. coli with high production, purity and biological activities. And its roles in cell migration and anchorage-independent growth suggest that up-regulation of TFF2 in colonic cancer might be involved in cancer invasion and metastases.

OBJECTIVETo investigate the expression profile of serum micro (mi)RNAs in cholangiocarcinoma (CCA) and investigate the regulatory contribution of miRNAs to the invasive and metastasis.

METHODSMicroarray analysis was carried out using serum samples collected from 30 patients with CCA, bile duct cancer tissues and the corresponding normal tissues collected from 10 patients, and serum samples from 50 healthy volunteers. The miRNAs identified as dysregulated in CCA were verified by RT-PCR. Focused analysis on miR-224 was carried out using the human CCA cell lines HCCC-9810 and RBE to investigate the role of this miRNA in IL-6 expression (using IL-6 induction), cell growth, invasiveness and metastasis (using miR-224 mimic transfection). The one-way ANOVA test was used for statistical analysis.

RESULTSForty-three miRNAs were dysregulated in CCA (vs. non-CCA, P<0.01), of which 22 were upregulated and 21 were downregulated. RT-PCR data showed that the miR-224 was significantly upregulated in serum as well as in cancer tissue from CCA patients. Induction of HCCC-9810 and RBE cells with IL-6 showed a time-dependent upregulation of miR-224. Furthermore, the HCCC-9810 and RBE cells transfected with miR-224 mimic showed enhanced cell growth, invasiveness and migratory ability.

CONCLUSIONIL-6 may promote the invasive and metastatic properties of CCA through upregulated miR-224. Studies of the differentially expressed serum miRNAs in CCA may help to further elucidate the pathogenic processes of this disease and aid in the development of a novel and effective therapeutic strategy.

Bile Ducts, Intrahepatic ; Cell Line, Tumor ; Cell Proliferation ; Cholangiocarcinoma ; Down-Regulation ; Humans ; Interleukin-6 ; MicroRNAs ; Microarray Analysis ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Transfection ; Up-Regulation

Objective:To summarize the construction experience of the cardiorespiratory rehabilitation specialist nurse practice base under the integration of Shenzhen and Hong Kong, so as to provide excellent practice bases for cardiorespiratory rehabilitation specialist nurses.Methods:This study constructed a training course system for practice bases, set up clinical practice courses, and improved the professional literacy and practical skills of cardiorespiratory rehabilitation specialty nurses through theoretical assessment, mind mapping, case reporting, and simulation assessment.Results:Cardiopulmonary rehabilitation specialist nurses all passed the operational assessment of the Nursing Committee of the Chinese Association of Rehabilitation Medicine and obtained completion certificates. The care report performance of cardiorespiratory rehabilitation specialist nurses improved compared to the training period, with a statistically significant difference ( P<0.001) . Conclusions:The standardized practice base training lays a solid foundation for cultivating excellent and high-quality cardiorespiratory rehabilitation specialist nurses, promoting the development of cardiorespiratory rehabilitation in China.