ABSTRACT
AIM:To investigate the expression of transmembrane protein 16A(TMEM16A) in Fischer rat thy-roid follicular epithelial ( FRT) cells and its electrophysiologic properties .METHODS: The eukaryotic expression vector of pUB6/V5-TMEM16A was constructed and transfected into FRT cells by liposome-mediated transfection .In order to ob-tain the high efficiency of gene transfection and expression , the quantity and ratio of lipid/DNA complexes were optimized . The FRT cells stably expressing TMEM16A were gained by the selection with blasticidin and confirmed by the techniques of RT-PCR and immunofluorescence .The expression and location of TMEM 16A in the FRT cells were observed under an in-verted fluorescence microscope .TMEM16A protein was associated with calcium-dependent chloride current , as measured with halide-sensitive fluorescent protein and patch-clamp technique .RESULTS: The results of double digestion and se-quencing indicated that TMEM16A was cloned into pUB6/V5.The results of RT-PCR and immunofluorescence confirmed that TMEM16A was expressed in the FRT cells after transfection with TMEM16A.The classical calcium-activated chloride channel currents were recorded in the FRT cells stably expressing TMEM 16A by the technique of patch-clamp and halide-sensitive fluorescent protein YFP-H148Q/I152L.CONCLUSION:The protein expression of TMEM16A in the FRT cells was observed.TMEM16A is the molecular identity of calcium-activated chloride channels .
ABSTRACT
Objective To build the prognostic model of STBI and provide guidance for making clinical decision .Methods A total of 486 patients with STBI was collected .Six months after injury ,according to GOS ,the patients were divided into five levels :good , mild-to-moderate disability ,serious disability ,persistent vegetative state and death .Ordinal regression analysis were used to find out the main factors of different populations .Results There were significant differences between different levels in nine factors .The re-sult of ordinal regression analysis showed that GCS ,age ,rescue time and reaction of pupil to light stood as independent variables in prognosis of patients with STBI(P<0 .05) .The outcomes from the prognostic model were pretty much exactly the same as the ac-tual results(Kappa= 0 .640 ,P= 0 .026) .Conclusion Though using the prognostic model of STBI ,the present study found out prognosis was closely related to GCS ,age ,rescue time and reaction of pupil to light in patients with STBI ;thus it could provide guidance for prognosis evaluation .
