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Objective:To evaluate regulatory effects of fibroblast growth factor receptor 3 (FGFR3) and human papillomavirus type 2 (HPV2) E2 protein on the differentiation of an immortalized human keratinocyte line HaCaT and a normal human epidermal keratinocyte line NHEK.Methods:In both HaCaT and NHEK cells, HPV2 E2-stably transfected cell lines (HPV2 E2-transfected groups) were established by using the lentivirus transfection method, wide-type FGFR3-overexpressing cells (FGFR3-WT transfected groups) and FGFR3-K650E mutant-overexpressing cells (FGFR3-K650E transfected groups) were constructed by using the plasmid transfection method, and cells transfected with blank vectors served as control groups (blank vector control groups). Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of HPV2 E2, and Western blot analysis to determine the protein expression of HPV2 E2, FGFR3, and keratinocyte differentiation markers including loricrin, filaggrin, as well as involucrin. Laser scanning confocal microscopy was conducted to observe the spatial localization of HPV2 E2 and FGFR3 in HaCaT cells. Statistical analysis was carried out by using two-independent-sample t test for the comparison between two groups, one-way analysis of variance for the comparison among multiple groups, and Dunnett t-test for multiple comparisons. Results:The HPV2 E2-stably transfected cell lines were successfully constructed, and the expression of HPV2 E2 FLAG protein was significantly higher in the HPV2 E2-transfected groups than in the blank vector control groups in both HaCaT and NHEK cells ( t = 13.71, 25.91, respectively, both P < 0.001) ; both FGFR3-WT and FGFR3-K650E were successfully overexpressed in both HaCaT and NHEK cells, and the FGFR3 protein expression was significantly higher in the FGFR3-WT transfected groups and the FGFR3-K650E transfected groups than in the blank vector control groups ( F = 473.90, 579.90, respectively, both P < 0.001). In both HaCaT and NHEK cells, the expression of keratinocyte differentiation markers including loricrin, filaggrin, and involucrin was significantly upregulated in the HPV2 E2-transfected groups, the FGFR3-WT transfected groups, and the FGFR3-K650E transfected groups than in the blank vector control groups (all P < 0.05). In the HPV2 E2-stably transfected HaCaT and NHEK cells, the expression of loricrin, filaggrin, and involucrin was significantly down-regulated in the HPV2 E2 + FGFR3-WT transfected groups and the HPV2 E2 + FGFR3-K650E transfected groups than in the HPV2 E2 + blank vector groups (all P < 0.05). Laser scanning confocal microscopy showed the spatial co-localization of HPV2 E2 and FGFR3 in the nuclear membrane and cytoplasm of HaCaT cells. Conclusion:HPV2 E2 and FGFR3 could both induce the differentiation of HaCaT and NHEK cells, while FGFR3 could inhibit HPV2 E2-induced differentiation trend of HaCaT and NHEK cells, which may be related to the cellular spatial co-localization of HPV2 E2 and FGFR3.
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Objective:To investigate clinical and dermoscopic manifestations of nail damage secondary to hand, foot and mouth disease.Methods:Clinical data were collected from 12 patients with nail damage secondary to hand, foot and mouth disease in Department of Dermatology, Shandong Provincial Hospital from June to November 2015, and characteristics of nail damage were analyzed.Results:The clinical manifestations of nail damage following hand, foot and mouth disease included dryness of periungual skin (12 cases) , nail plate cavities (11 cases) , nail fractures (11cases) and Beau′s lines (4 cases) . Dermoscopy showed nail discoloration (12 cases) , nail delamination (12 cases) , horizontal streaks (10 cases) , longitudinal streaks (8 cases) , brown background of the nail matrix (11 cases) , telangiectasia (7 cases) , periungual desquamation (12 cases) and red background (10 cases) .Conclusion:The clinical manifestations of nail damage following hand, foot and mouth disease are mainly dryness of periungual skin, nail plate cavities and fractures, and its dermoscopic manifestations include nail discoloration, delamination, horizontal and longitudinal streaks, brown background of the nail matrix, telangiectasia, periungual desquamation and red background.
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BACKGROUND:In three-dimensional CT reconstruction model, fracture virtual reduction and internal fixation cannot be achieved, and reasonable operation scheme cannot be formulated. It cannot fuly meet the needs of clinical orthopedics physicians. OBJECTIVE:To explore the methods to perform three-dimensional reconstruction, reduction on proximal tibial fracture and to design digital plate using software. METHODS:Nine specimens of knee joint were built to be A1-3, B1-3 and C1-3 types of proximal tibial fracture according to the AO standard. Al specimens were treated with CT scan. Three-dimensional reconstruction and reduction on nine types of bone fracture models were performed in Mimics. The digital plates and screws were designed in Solidworks. Virtual internal fixation on the reduced three-dimensional models was performed. RESULTS AND CONCLUSION:Al bone fracture models received three-dimensional reconstruction and reduction, and the design for internal fixational plates and screws as wel as virtual internal fixation were accomplished. A3, B3 and C3 types were presented. These results suggest that the digital plates for proximal tibial fracture could be designed in Mimics and Solidworks with a personal computer. Our experience could be used in the real operation.
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Neurons in the laterodorsal tegmentum (LDTg) and pedunculopontine tegmental nucleus (PPTg) play important roles in central autonomic circuits of the kidney. In this study, we used a combination of retrograde tracers pseudorabies virus (PRV)-614 and fluorescence immunohistochemistry to characterize the neuroanatomic substrate of PPTg and LDTg innervating the kidney in the mouse. PRV-614-infected neurons were retrogradely labeled in the rostral and middle parts of LDTg, and the middle and caudal parts of PPTg after tracer injection in the kidney. PRV-614/TPH double-labeled neurons were mainly localized in the rostral of LDTg, whereas PRV-614/TH neurons were scattered within the three parts of LDTg. PRV-614/TPH and PRV-614/TH neurons were located predominantly in the caudal of PPTg (cPPTg). These data provided direct neuroanatomical foundation for the identification of serotonergic and catecholaminergic projections from the mid-brain tegmentum to the kidney.
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Objective To analyze the features of dynamic contrast-enhanced CT of blunt hepatic injury in rabbits. Methods The model of blunt hepatic injury was established in 40 New Zealand white rabbits with a steel ball falling down to the xiphoid process of the animals. Plain CT scan and dynamic contrast-enhanced CT scan (Hispeed spiral CT/2i, GE, America) of the liver were performed. Arterial,portal and balanced phases were respectively at 8-10 s, 35-40 s and 120-150 s after initiation of the contrast medium injection. The non-enhanced and enhanced images were compared in aspects of location and range of injury, tear of the liver capsule, active bleeding, involvement of the main hepatic veins and CT features of abdominal hemorrhage, which was further compared with the results of gross anatomy. Results The rate of plain CT scan was obviously lower than that of dynamic contrast-enhanced CT scan,which defined single tear in 13 patients, multiple lacerations in 18, liver subcapsular hematoma in seven,liver hematoma in nine, liver coated gap in 17, active bleeding in nine and main hepatic vein injury in five, with coincidence rates with the results of gross observation for 13/13,18/18,7/9,9/9,25/30,9/5and 5/4 respectively. According to Moore' s classification, CT/Laparotomy performed from grade Ⅰ to grade Ⅵ were 5/4 patients at grade Ⅰ , 15/13 at grade Ⅱ, 9/11 at grade, 5/6 at grade Ⅳ, 1/2 at grade Ⅴ, O at grade respectively. Conclusion Dynamic contrast-enhanced CT scan, especially at portal and balanced phases, is of great value for diagnoses of liver injuries and determination of injury severity.