YAP activation attenuates toxicarioside G­induced lethal autophagy arrest in SW480 colorectal cancer cells.

Toxicarioside G (TCG), a natural product isolated from Calotropis gigantea, has been found to exhibit potent anticancer effects. The present study aimed to investigate the effect of TCG on the SW480 colorectal cancer cell line and the role of autophagy and Yes1 associated transcriptional regulator (YAP) in the TCG­mediated inhibition of cell proliferation and viability. Cell proliferation was detected using MTT, BrdU, colony formation and LDH release assays, while apoptosis was analyzed using flow cytometry and western blot analyses. Immunofluorescence and western blot analysis was used to determine TCG­induced autophagy and YAP activation. Pharmacological inhibition and siRNA was used to investigate the role of autophagy and YAP in TCG­mediated cell growth inhibition. The results revealed that TCG inhibited SW480 cell proliferation and viability, independent of apoptosis, and also induced autophagy. It was further demonstrated that TCG blocks autophagic flux, resulting in autophagy arrest in the SW480 cell line. The inhibition of autophagy restored the TCG­mediated inhibition of cell proliferation and viability, suggesting that TCG may induce lethal autophagy arrest in the SW480 cell line. Furthermore, TCG induced YAP activation in the SW480 cell line. Inhibition of YAP activity enhanced the TCG­mediated inhibition of cell proliferation and viability, suggesting that YAP may play a protective role in the TCG­induced effects. In conclusion, the findings of the present study indicated that TCG may induce lethal autophagy arrest and activate YAP, which serves a protective role in the SW480 cell line. These results suggested that the combined targeting of TCG and YAP may represent a promising strategy for TCG­mediated anticancer therapy.

Prevalence and risk factors of type 2 diabetes in the adults in haikou city, hainan island, china.

BACKGROUND: Type 2 diabetes mellitus (T2DM) occurs around the world with high prevalence and causes serious physical harm and economic burden to the afflicted. Haikou City is China's southernmost tropical island city, which has not been previously studied for its T2DM prevalence. The objective of the study in employing a cross-sectional survey is to discuss the epidemiologic status of T2DM in Haikou City and to analyze the possible determinants. METHODS: A total of 12,000 community residents over 18 years old from four districts in Haikou City were stratified-randomly sampled. A questionnaire survey and physical examination were conducted. Data entry and statistical analysis were performed using SPSS17.0 software. RESULTS: The prevalence of T2DM in Haikou City was 5.3% (5.15% for males and 5.46% for females). According to the multivariate analysis, the positive factors mainly associated with T2DM in the city included family history, Waist-to-Hip Ratio (WHR), triglycerides, low high-density lipoproteins (HDL), and blood pressure. For both men and women, family history was the highest independent risk factor associated with T2DM (OR= 47.128). The T2DM risk increased with increasing metabolic aggregate. CONCLUSION: The prevalence of T2DM for the community population of Haikou City was low. The possible risk factors included age, occupation, BMI, waist circumference, WHR, overweight, systemic obesity, central obesity, systolic blood pressure, diastolic blood pressure, total cholesterol, triglycerides, low-density lipoproteins, family history, and HDL.

Prokaryotic expression, identification and bioinformatics analysis of fbpB-esxA fusing gene from Mycobacterium tuberculosis.

OBJECTIVE: To obtain fbpB-esxA fusing gene of Mycobacterium tuberculosis (MTB), express the encoded fusing protein in Escherichia coli (E. coli), identify protein acquired, and predict the structure and function of the protein utilizing methods of bioinformatics. METHODS: fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR. The fbpB-esxA fusing gene ligated by (Gly(4)Ser)(3) linker was gained by means of Gene Splicing by Overlapping Extension PCR (SOE-PCR), and fusing gene was cloned into expression vector pET-30a. The recombinant plasmid was sequenced and expressed in E. coli BL21(DE3). The protein was identified by Western blot using anti-HIS antibody. Secondary structure and antigenic epitopes of the protein were predicting using tools of bioinformatics. RESULTS: The DNA sequences of fbpB-esxA were identical with that published by GenBank. The Ag85B-ESAT-6 fusion protein about 50 kDa comprised 485 amino acids was efficiently produced from expression system in E. coli BL21(DE3) under the induction of IPTG. Bioinformatics analysis showed the protein contained one transmembrane region and fourteen potential antigenic epitopes. CONCLUSIONS: The Ag85B-ESAT-6 fusion protein is successfully expressed with N-terminal HIS-tag. Gel filtration demonstrated that it exists as insoluble inclusion bodies mainly. The existence of linker doesn't affect immunogenicity of Ag85B and ESAT-6. It will allow for characterization in vitro and establish a foundation of further function research such as vaccine or diagnostic reagent.

Identification, structure and function of a novel tetraspaninhomologue from Spirometra erinaceieuropaei

OBJECTIVE@#To identify a full length c DNA sequence of a novel tetraspanin (TSP) homologue from Spirometra erinaceieuropaei and to predict the structure and function of its encoding protein using bioinformatics methods.@*METHODS@#Using the NCBI, EMBI, Expasy and other online sites, the open reading frame (ORF), conserved domain, physical and chemical parameters, signal peptide, transmembrane domain, epitope, topological structures of the protein sequences were predicted. And Vector NTI software was used for multiple sequence alignment and phylogenetic tree construction.@*RESULTS@#The target sequence was 1132 bp length with a 681 bpbiggest ORF encoding 226 amino acids protein with typical TSP conserved domain. It was confirmed as full length c DNA of TSP16 from Spirometra erinaceieuropaei and named as SeTSP16 (GenBank accession number: JF728872). The predicted molecular weight and isoelectric point of the deduced protein were 24 750.5 Da and 7.88 Da, respectively. Compared with TSP16s from Schistosoma japonicum and Schistosoma mansoni, it showed similarity of 59% and 59%, respectively. SeTSP16 contained four transmembrane domains (TM1-4), intracellular N and C-termini, one short small extracellular loop and one large extracellular loop. Four major epitopes that were significant different from the corresponding epitope regions of TSP16 from Schistosoma mansoni and Schistosoma japonicum were predicted.@*CONCLUSIONS@#The full length c DNA sequences of SeTSP16 are identified. It encodes a transmembrane protein which might be an ideal diagnosis antigen and target molecule for antiparasitic drugs.