Objective To establish a stable HCV full-length genome replication cell model which is labeled with reporter gene and easyly to quantify intracellular HCV proteins and RNA level. Methodsneo gene was inserted into Luc-JC1 to make Luc-JC construct. Luc-JC RNA was obtained by in vitro transcription and then delivered into Huh7 cells by transfection. G418-resistant clones of Huh7 cells were obtained by selection. Clones of HCV full-length genome replication cell were confirmed by luciferase activity assay, Western blot and cleaveage of eYFP-MAVS by HCV NS3/4A protease. Then, HCV replication cell colonies were treated by different dose IFN-α in order to observe the change of luciferase activity, HCV protein and RNA level. Results At 3-4 weeks post-transfection, visible colonies were selected and stained by crystal violet. Luciferase activity and HCV NS3, NS5A protein were detected by luciferase activity assay and Western blot, respectively. Subcellular localization of eYFP-MAVS transferred from mitochondria to cytoplasms by cleavage of NS3/4A protease in cell colonies. Luciferase activity, HCV protein and RNA diminished obviously after IFN-α treatment. Conclusion A stable HCV full-length genome replication cell model labeled by reporter gene was successfully established and reporter activity can be used to indicate level of HCV proteins and RNA in cells. This cell model is a useful tool for the study on HCV pathogenesis and the screening of antiviral drugs.

OBJECTIVETo evaluate the alternations of degree centrality (DC) in the brain of a patient with alternating Horner's syndrome (AHS).

METHODSConventional magnetic resonance imaging (MRI), 3D structure reconstruction and resting-state functional MRI were performed in a patient with AHS and 8 healthy adults. The DC of brain functional connectivity was calculated and statistically analyzed to evaluate the changes in the nodes in the brain default network of the patient.

RESULTSIn the AHS patient, the DC at onset of left eye involvement was located mainly in the bilateral anterior middle frontal gyri, frontal operculum and opercula insulae; the brain regions with a DC greater than the mean DC of the whole brain were found mainly in the bilateral occipital lobes, temporal lobes and cingulate gyri, and the brain regions with a significantly decreased DC included mainly the left inferior temporal gyrus, postcentral gyrus, right superior temporal gyrus, lateral occipital gyri and bilateral paracentral lobules (P<0.05). At the onset of right eye involvement, the DC was mainly located in the anterior part of the bilateral superior and middle frontal gyri, parietal lobes, middle cingulate gyri and medial occipital gyri; the brain regions with a DC greater than the mean DC of the whole brain included the bilateral occipital lobers, temporal lobes, cingulate, orbital gyri and gyrus rectus, and the brain regions with a significantly decreased DC included the left supramarginal gyrus, right lateral occipital gyrus, paracentral lobule and bilateral superior temporal gyrus.

CONCLUSIONThe AHS patient exhibited a decreased DC of functional connectivity in multiple brain regions.

Adult ; Brain ; physiopathology ; Case-Control Studies ; Female ; Horner Syndrome ; physiopathology ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged