Effects of dual-gene modification on biological characteristics of vascular endothelial cells and their significance as reserving cells for chronic wound repair.

bFGF is a commonly used and reliable factor for improving chronic wound healing, and hSulf-1 expression is abundant in surrounding cells of chronic wound tissue and vascular endothelial cells, which can reverse the effect of bFGF and inhibit the signalling activity of cell proliferation. In this study, an adenovirus, Ad5F35ET1-bFGF-shSulf1, was designed for establishing the dual-gene modified vascular endothelial cells, which were used as the repair cells for skin chronic wound. Ad5F35ET1-bFGF-shSulf1 infected ECV304 cells in vitro and mediated the overexpression of bFGF and the knockdown of hSulf-1, which effectively activated the AKT and ERK signal transduction pathways, facilitate cell proliferation and migration, with the cell viability to 128.29% at 72 h after infection, compared to 66.65%, 73.74%, 87.63%, 103.14% in the blank control, Ad5F35ET1-EGFP-shNC, Ad5F35ET1-shSulf1, Ad5F35ET1-bFGF groups, respectively. In the rat ear skin injury model, the wound healing was significantly accelerated in the Ad5F35ET1-rbFGF-shrSulf1 group compared to the blank control group (p = 0.0046), Ad5F35ET1-EGFP-shNC group (p = 0.0245), Ad5F35ET1-shrSulf group (p = 0.0426), and Ad5F35ET1-rbFGF group (p = 0.2853). The results demonstrated that this strategy may be a candidate therapy for chronic injury repair.

[Application of multislice computed tomography volume rendering and 3D printing technique of costal cartilage for auricular reconstruction].

Objective: To investigate the value of Multislice computed tomography volume rendering(VR) technique and 3D printing technique in auricular reconstruction. Methods: Six patients were enrolled for auricular reconstruction with costal cartilage,including 5 congenital microtia patients and 1 traumatic auricular defect patient. We harvest the three-dimensional reconstructive data of the contralateral sixth, seventh, eighth and ninth costal cartilage with VR technique.Three-dimensional solid models were 3D printed with nylon material according to the data exported in STL format.Preoperativesimulation was performed on the models, accordingly, we determined the strategies of costal cartilage harvest and framework fabrication, and operations were performed based on the pre-designed plan. Results: In all 6 patients, the actual costal cartilage harvest and framework fabrication process was consistent with the preoperative design and simulation results, and more scientific than before.The shapes of reconstructed ears were vivid and natural. No complications such as infection,absorption,distortion and chest deformity happened. Conclusions: Through costal cartilage VR and 3D printing technique, we could make more reasonable preoperative design and simulation. The results can be improved with reduced injury, while avoiding the risks of thoracic deformity.

IL-15 induces strong but short-lived tumor-infiltrating CD8 T cell responses through the regulation of Tim-3 in breast cancer.

IL-15 has pivotal roles in the control of CD8(+) memory T cells and has been investigated as a therapeutic option in cancer therapy. Although IL-15 and IL-2 share many functions together, including the stimulation of CD8 T cell proliferation and IFN-γ production, the different in vivo roles of IL-15 and IL-2 have been increasingly recognized. Here, we explored the different effects of IL-15 and IL-2 on tumor-infiltrating (TI) T cells from resected breast tumors. We found that neither IL-2 nor IL-15 induced intratumoral CD8 T cell proliferation by itself, but after CD3/CD28-stimulation, IL-15 induced significantly higher proliferation than IL-2 during early time points, at day 2, day 3 and day 6. However, the IL-15-induced proliferation leveled off at day 9 and day 12, whereas IL-2 induced lower but progressive proliferation at each time point. Furthermore, IL-15 caused an early and robust increase of IFN-γ in the supernatant of TI cell cultures, which diminished at later time points, while the IL-2-induced IFN-γ production remained constant over time. In addition, the IL-15-costimulated CD8 T cells presented higher frequencies of apoptotic cells. The diminishing IL-15-induced response was possibly due to regulatory and/or exhaustion mechanisms. We did not observe increased IL-10 or PD-1 upregulation, but we have found an increase of Tim-3 upregulation on IL-15-, but not IL-2-stimulated cells. Blocking Tim-3 function using anti-Tim-3 antibodies resulted in increased IL-15-induced proliferation and IFN-γ production for a prolonged period of time, whereas adding Tim-3 ligand galectin 9 led to reduced proliferation and IFN-γ production. Our results suggest that IL-15 in combination of Tim-3 blocking antibodies could potentially act as an IL-2 alternative in tumor CD8 T cell expansion in vitro, a crucial step in adoptive T cell therapy.

Regulatory B Cell Function Is Suppressed by Smoking and Obesity in H. pylori-Infected Subjects and Is Correlated with Elevated Risk of Gastric Cancer.

Helicobacter pylori infection occurs in more than half of the world's population and is the main cause for gastric cancer. A series of lifestyle and nutritional factors, such as tobacco smoking and obesity, have been found to elevate the risk for cancer development. In this study, we sought to determine the immunological aspects during H. pylori infection and gastric cancer development. We found that B cells from H. pylori-infected patients presented altered composition and function compared to uninfected patients. IL-10-expressing CD24+CD38+ B cells were upregulated in H. pylori-infected patients, contained potent regulatory activity in inhibiting T cell pro-inflammatory cytokine secretion, and responded directly to H. pylori antigen stimulation. Interestingly, in H. pylori-infected smoking subjects and obese subjects, the number of IL-10+ B cells and CD24+CD38+ B cells were reduced compared to H. pylori-infected asymptomatic subjects. Regulatory functions mediated by CD24+CD38+ B cells were also impaired. In addition, gastric cancer positive patients had reduced IL-10-producing B cell frequencies after H. pylori-stimulation. Altogether, these data suggest that in H. pylori-infection, CD24+CD38+ B cell is upregulated and plays a role in suppressing pro-inflammatory responses, possibly through IL-10 production, a feature that was not observed in smoking and obese patients.

Quantitative analysis based on three-dimensional CT in the contouring surgery of craniofacial fibrous dysplasia / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the quantitative analysis based on three-dimensional computed tomography (3D-CT) in contouring surgery of complex craniofacial fibrous dysplasia (FD).</p><p><b>METHODS</b>14 patients with craniofacial FD underwent 3D-CT scan. Axial images of patients with craniofacial FD were reconstructed into 3D model by using Mimics 10.0. Anatomical landmarks were located and the coordinate of the landmarks obtained. The differences between the right landmarks and the left were calculated and analyzed. Quantitative contouring surgery was performed based on the quantitative analysis result.</p><p><b>RESULTS</b>With the detail data from the 3D-CT analysis, the surgery of contouring was more safe and accurate with less operation time, less bleeding and good results.</p><p><b>CONCLUSIONS</b>The method of 3D CT quantitative analysis can provide precise information in the diagnosis and treatment planning of craniofacial deformity. Based on the result of 3D-CT quantitative analysis, the operations can be performed more accurately and safely with good symmetric consequence.</p>

A highly infectious chimeric adenovirus expressing basic fibroblast growth factor exerts potent targeted therapy for rabbit ear chronic ischemic wounds.

BACKGROUND: Poor angiogenesis and impaired proliferation of cells responsible for the repair of chronic ischemic wounds result in impaired wound healing. The continuous and efficient expression of therapeutic factors by means of gene transfection is an ideal adjuvant treatment method to promote cell proliferation and angiogenesis. METHODS: A chimeric recombinant adenoviral vector, Ad5F35ET1-bFGF, was constructed that carried the basic fibroblast growth factor (bFGF) gene and used the endothelin-1 promoter to control the targeted expression of bFGF in endothelial cells and fibroblasts. Thus, the authors established a targeted gene therapy for chronic ischemic wounds. RESULTS: The chimeric adenovirus Ad5F35ET1-bFGF efficiently infected the endothelin-1-positive endothelial cells and fibroblasts, specifically expressed bFGF, and promoted cell proliferation. In the rabbit wound healing model, the chimeric recombinant adenovirus expressed a high level of bFGF in wound tissues, which continuously promoted angiogenesis and cell proliferation and thus accelerated wound healing. CONCLUSION: Targeted gene therapy that uses bFGF as a therapeutic gene provides an effective candidate strategy for the treatment of chronic ischemic wounds.