GFAP and BrdU immunoreactivity were seen in the migratory stream.

<p>(<b>A</b>) Schematic illustration showing the migration routes of the cells from the ependymal layers to the ipsilateral nucleus ambiguus (NA) at 14 days after the recurrent laryngeal nerve (RLN) avulsion injury. (<b>B</b>) The CM-DiI+ cells (red) were observed in the region between the ependymal layer and the ipsilateral NA, whereas no migration was observed in the control animals (<b>C</b>). A CM-DiI+/GFAP+/BrdU+ cell (white frame) was observed in the migratory stream, indicating that this neural progenitor cell (NPC) was differentiating into an astrocyte and proliferating while migrating to the NA. CM-DiI+/GFAP+ cell (star) can also be observed, indicating that it is not proliferating. Moreover, CM-DiI+ cells with neuronal morphology were observed in the migratory stream, indicating that the NPCs can differentiate into a neuronal fate. Some of the CM-DiI+ cells (arrow) showed BrdU immunoreactivity, indicating that they are proliferating, but others (arrowhead) are not. (<b>B</b>) shows the merged image of (<b>D</b>) CM-DiI (red), (<b>E</b>) BrdU (green), (<b>F</b>) GFAP (violet) and (<b>G</b>) DAPI (blue). Bars in A-F  = 25 µm. Unop =  unoperated animals; dpi =  days post injury, 4V =  the fourth ventricle, NA =  nucleus ambiguus.</p

Histogram illustrating the numbers of migrating cells in the ipsilateral brainstem after injury.

<p>(<b>A</b>) CM-DiI+ cells in the migratory stream increased beginning at 6 h post-injury. The maximal immunoreactivity of CM-DiI was observed at 14 days post-injury, followed by a decline at 28 days post-injury, compared with control animals. (<b>B</b>) In contrast to the control animals, the CM-DiI+/GFAP+ cells in the migratory stream were observed at 14 days post-injury. These cells decreased in number in injured animals at later survival times. (<b>C</b>) An increase in immunoreactivity of CM-DiI+/DCX+ cells was observed in the migratory stream from 7 to 14 days post-injury and declined from 14 to 28 days after the injury. (<b>D</b>) The CM-DiI+/NeuN+ cells in the migratory stream were observed at 21 days post-injury and increased at 28 days post-injury, compared with unoperated animals. (<b>E</b>) Quantitation of the CM-DiI+/GFAP+ cells in the NA ipsilateral to the injury showed an increase from 14 to 28 days post-injury. Values are the mean±SD, * <i>P</i><.05, ** <i>P</i><.01. Unop =  unoperated animals.</p

Synergetic Effect of Blended Alkylamines for Copper Complex Ink To Form Conductive Copper Films

Cu­(II) complex ink consisting of copper formate (Cuf) and a primary alkylamine could yield highly conductive copper films at low heating temperatures without a reducing atmosphere. A synergetic effect of the blended alkylamines on the formation of conductive films was observed. It was found that blending two types of amines with different alkyl chain lengths as ligands could improve the conductivity of copper films, compared with using one of these amines alone. The decomposition mechanism of the Cuf–amine complex and the role of amines with different alkyl chain lengths were investigated. It was found that the decrease in the decomposition temperature and the formation of copper films were attributed to the activating effect and capping effect of the amine, and these two effects were dependent on the alkyl chain length. The relative intensity of the dual effects determined the decomposition rate of the complex and the nucleation and growth of particles. The use of blended amines with different alkyl chain lengths as ligands could balance the two effects and lead to appropriate nucleation and growth rates, so that densely packed copper films with low resistivity could be obtained at low heating temperature in a short time. The Cuf–butylamine–octylamine (Cuf–butyl–octyl) ink with 1:1 molar ratio of the amines showed the best performance. The understanding of the synergetic effect could provide guidance to the design of copper complex inks to control the morphology of the films

CM-DiI+/NeuN+ cells were seen outside the ipsilateral nucleus ambiguus (NA) at 28 days after injury.

<p>(<b>A</b>) Schematic illustration showing the routes of the cells migrating from the ependymal layers to the ipsilateral NA at 28 days after the recurrent laryngeal nerve (RLN) avulsion injury. (<b>B</b>) A decrease in the number of motor neurons stained with NeuN was observed in animals with RLN avulsion injuries at 28 days post-injury (dotted circle), in contrast to the control animals(<b>C</b>). CM-DiI+/NeuN+ cells (arrow) were observed outside the ipsilateral NA at 28 days post-injury (<b>B</b>), compared with the control animals (<b>C</b>). This indicated that the neural progenitor cells (NPCs) were differentiating into neurons, but they did not reach the ipsilateral NA. Merged images of (<b>D</b>) CM-DiI (red), (<b>E</b>) NeuN(violet) and (<b>F</b>) DAPI (blue) are shown as (<b>B</b>). Bars in A-E = 75 µm. Unop =  unoperated animals; dpi =  days post injury, 4V =  the fourth ventricle, NA =  nucleus ambiguus.</p

Migration of the CM-DiI+ cells from the ependymal layer after injury.

<p>(<b>A</b>) Schematic illustration showing the CM-DiI+ cells migrating from the ependymal layer to the ipsilateral nucleus ambiguus (NA) at 14 days after the recurrent laryngeal nerve (RLN) avulsion injury. Animals subjected to RLN avulsion injury (<b>B</b>) show BrdU immunoreactivity (green, star) was expressed in the ependymal cells labeled with CM-DiI (red), indicating that the ependymal cells were proliferating, in contrast to the control animals (<b>C</b>). CM-DiI+/GFAP+ cells were observed migrating from the ependymal layer, indicating that the neural progenitor cells (NPCs) were migrating from the ependymal layer and differentiating into astrocytes. Some of these cells (arrow) were positive for BrdU (green), indicating that they were proliferating. However, others (arrowhead) showed no immunoreactivity for BrdU. CM-DiI+ cells (asterisk) with neuronal morphology were also observed outside the ipsilateral ependymal layer, indicating that NPCs may be differentiating into neuronal cells while migrating from the ependymal layer. Merged images of (<b>D</b>) CM-DiI (red), (<b>E</b>) BrdU (green), (<b>F</b>) GFAP (violet) and (<b>G</b>) DAPI (blue) are shown as (<b>B</b>). Bars in A-F =  25 µm. Unop =  unoperated animals; dpi =  days post injury, 4V =  the fourth ventricle, NA =  nucleus ambiguus.</p

CM-DiI+/GFAP+ cells were observed in the ipsilateral nucleus ambiguus (NA) at 28 days after injury.

<p>(<b>A</b>) Schematic illustration showing the routes of the cells migrating from the ependymal layers to the ipsilateral NA at 28 days after the recurrent laryngeal nerve (RLN) avulsion injury. (<b>B</b>) The CM-DiI+/GFAP+ cells (white frame) were observed in the ipsilateral NA at 28 days post-injury, which was not observed in the control animals (<b>C</b>). The (<b>D</b>) CM-DiI-labeled (red) cells were colabeled with (<b>E</b>) GFAP (violet) and (<b>F</b>) DAPI(blue), indicating that the neural progenitor cells migrated into the ipsilateral NA and differentiated into astrocytes. Bars in A-E = 25 µm. Unop =  unoperated animals; dpi =  days post injury, 4V =  the fourth ventricle, NA =  nucleus ambiguus.</p

Cell immunofluorescence for the observation of surface protein expression.

<p>Both vocal fold fibroblasts (VFFs) (A) and differentiated adipose-derived mesenchymal stem cells (dADSCs) (B) can express vimentin and fibronectin. Scale bar  =  50μm.</p

Diagram of the layered structure of the vocal fold (Hoechst staining).

<p>The canine vocal fold includes the epithelium (E), lamina propria (LP) and muscle (M). Scale bar  =  100μm.</p

Validation of 5′-leader region expression by RT-PCR.

<p>Electrophoresis of PCR amplicons of 7 5′-leader regions using 3% agarose gels. Lane M: Marker, Lane 1-7: SYPCC_Ir_01, SYPCC_Ir_02, SYPCC_Ir_03, SYPCC_Ir_04, SYPCC_Ir_05, SYPCC_Ir_06, and SYPCC_Ir_07.</p

The 7 selected candidate 5′-leader regions for RT-PCR validation.

<p>The 7 selected candidate 5′-leader regions for RT-PCR validation.</p