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.

Objective To investigate the effect of liver kinase B1(LKB1) on the radiosensitivity of subcutaneous xenograft tumor of lung cancer H460 cells in nude mice.Methods Human lung cancer H460 cells were implanted into female nude mice (BALB/c-nu) to establish a subcutaneous xenograft tumor model of lung cancer.A total of 24 female nude mice in which the model was successfully established were equally and randomly divided into four groups:pEGFP-Ctrl plasmid (empty vector plasmid) group, irradiation (IR)+pEGFP-Ctrl plasmid group, pEGFP-LKB1 plasmid (overexpressing LKB1) group, and IR+pEGFP-LKB1 plasmid group.The growth of xenograft tumors was observed and the tumor inhibition rate and enhancement factor (EF) were calculated.The expression of LKB1 in each group was measured by immunohistochemistry and Western blot to analyze the relationship between LKB1 and radiosensitivity.Results Compared with the pEGFP-Ctrl plasmid group, the IR+pEGFP-Ctrl plasmid group, pEGFP-LKB1 plasmid group, and IR+pEGFP-LKB1 plasmid group showed varying degrees of inhibition of tumor growth, particularly in the IR+pEGFP-LKB1 plasmid group, and the tumor inhibition rates were 31.30%, 14.78%, and 43.48%, respectively.The EF of LKB1 in the IR+pEGFP-LKB1 plasmid group was 1.18.The immunohistochemistry and Western blot showed that LKB1 could be effectively expressed in the pEGFP-LKB1 plasmid group and IR+pEGFP-LKB1 plasmid group, but not in the other two groups.Conclusions The subcutaneous xenograft tumor model of human lung cancer H460 cells has been successfully established in nude mice.LKB1 has a radiosensitizing effect on the subcutaneous xenograft tumor of lung cancer H460 cells in nude mice.

Panic disorder (PD) is an acute paroxysmal anxiety disorder with poorly understood pathophysiology. The dorsal periaqueductal gray (dPAG) is involved in the genesis of PD. However, the downstream neurofunctional changes of the dPAG during panic attacks have yet to be evaluated in vivo. In this study, optogenetic stimulation to the dPAG was performed to induce panic-like behaviors, and in vivo positron emission tomography (PET) imaging with F-flurodeoxyglucose (F-FDG) was conducted to evaluate neurofunctional changes before and after the optogenetic stimulation. Compared with the baseline, post-optogenetic stimulation PET imaging demonstrated that the glucose metabolism significantly increased (P < 0.001) in dPAG, the cuneiform nucleus, the cerebellar lobule, the cingulate cortex, the alveus of the hippocampus, the primary visual cortex, the septohypothalamic nucleus, and the retrosplenial granular cortex but significantly decreased (P < 0.001) in the basal ganglia, the frontal cortex, the forceps minor corpus callosum, the primary somatosensory cortex, the primary motor cortex, the secondary visual cortex, and the dorsal lateral geniculate nucleus. Taken together, these data indicated that in vivo PET imaging can successfully detect downstream neurofunctional changes involved in the panic attacks after optogenetic stimulation to the dPAG.