detection-of-rare-mutations-in-ctdna-using-next-generation-sequencing

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing

The analysis of circulating tumor DNA (ctDNA) using next-generation sequencing (NGS) has become a valuable tool for the development of clinical oncology. ...

Geneplus-Beijing Institute

Pericytes play a crucial role in the homeostasis and maturation of newly formed vessels, but their precursor and the process of their differentiation remain unclear. In this study, we show that in vivo, pericytes in human granulation tissue taken from burn patients expressed CD13 and collagen I, which are cell markers of peripheral blood fibrocytes (PBFCs). Mouse PBFCs, ex vivo labeled by PKH-26 and administered intravenously back to mice, formed lumens in the granulation tissue and expressed pericyte marker NG2. Furthermore, in cell culture, human PBFCs in co-culture with human umbilical vascular endothelial cells or human dermal microvascular endothelial cells (HDMECs) expressed pericyte markers desmin and myosin heavy chain of smooth muscle cell, and some migrated to the membrane of human umbilical vascular endothelial cells or HDMECs to form PBFC/vascular endothelial cell clusters, or formed round PBFC only clusters with their nuclei aligned along a circle. The formation of PBFC/vascular endothelial cell clusters and round PBFC clusters were inhibited by an antibody against monocyte chemotactic protein-1. PBFCs cultured with homogenate of granulation tissue also expressed desmin and myosin heavy chain of smooth muscle cell. Our findings support that the PBFC is a precursor of pericytes, the differentiation of PBFCs is induced by vascular endothelial cells in a paracrine fashion.

Differentiation of the pericyte in wound healing: The precursor, the process, and the role of the vascular endothelial cell.

Dysregulation of microRNA-21 plays critical roles in tumor initiation and progression. The purpose of this study was to investigate the status of microRNA-21 expression in human cutaneous malignant melanoma and determine its clinical significance. TaqMan(®) real-time RT-PCR assay was performed to examine the expression of microRNA-21 in 10 cases of dysplastic nevi, 86 cases of primary cutaneous melanomas, 10 cases of melanoma metastases. The correlation of microRNA-21 expression with clinicopathological factors or prognosis of patients with cutaneous melanoma was statistically analyzed. Additionally, the effects of microRNA-21 expression on growth, apoptosis and chemo- or radiosensitivity of melanoma cells were also investigated by transfection of microRNA-21 inhibitor. We firstly showed that increased levels of microRNA-21 expression were shown from dysplastic nevi to primary cutaneous melanomas to melanoma metastases. Moreover, high miR-21 expression was found to be correlated with Breslow thickness and advanced clinical stage. Patients with high microRNA-21 expression showed shorter 5-year disease-free or overall survival than those with low microRNA-21 expression. Furthermore, multivariate regression analysis showed that the status of microRNA-21 expression was an independent prognostic factor for overall survival of patients. Antisense-mediated microRNA-21 inhibition could significantly suppress growth, increase apoptosis and enhance chemo- or radiosensitivity of human cutaneous melanoma cells by inducing the increased Bax/Bcl-2 ratio. Thus, the status of microRNA-21 might be an independent prognostic factor for patients with cutaneous melanoma, and microRNA-21 has the potential of being a novel molecular target for the treatment of human cutaneous melanoma.

The status of microRNA-21 expression and its clinical significance in human cutaneous malignant melanoma.

A refined minimally invasive procedure for radical treatment of axillary osmidrosis: combined tumescent liposuction with subcutaneous pruning through a small incision.

The potential of acellular dermal matrix (ADM) to improve cosmetic and functional outcomes has been demonstrated; however, there have been few clinical comparative studies assessing the long-term morphological, histological and functional changes after ADM placement. This study was designed to retrospectively evaluate the long-term outcomes of the cograft acellular dermal matrix with autologous thin split-thickness skin for the coverage of wounds in extensively burned patients. Thirty burn patients treated with a composite graft of ADM with autologous split-thickness skin from January 2007 to December 2009 were enrolled in this study. Another group of thirty patients who received only an autogenous split-thickness skin implant served as the control. Our study revealed that the collagen in the dermis treated with ADM were ordered, and the proportion of collagen III/I was much higher in the control group than in the ADM group. The basement membrane was prominent and continuous. Meanwhile, the VBSS (Vancouver Burn Skin Score) was used to evaluate skin quality, which shows a significant differences between the two group (P<0.001). Then the functional level was evaluated by the BI (Barthel Index), and the ADM group was much better than the control group (P=0.005). Based on these results, we concluded that the composite graft of ADM with autologous thin split-thickness skin was suitable for repairing the defects in functional areas after a burn. This technique might facilitate wound management with acceptable esthetic outcomes, good functional recovery and less scar hyperplasia at the donor site.

Human acellular dermal matrix allograft: A randomized, controlled human trial for the long-term evaluation of patients with extensive burns.

This paper reports the fabrication of biomimetic nanofibrous matrices via co-electrospinning of polycaprolactone (PCL)/cellulose acetate (CA) and layer-by-layer self-assembly (LBL) of positively charged chitosan (CS) and negatively charged Type Ⅰ collagen on the nanofibrous matrix. FE-SEM images indicate that the average fiber diameter increased from 392 to 541 nm when the coating bilayers varied from 5 to 20.5. Besides, the excellent biocompatibility and enhanced attachment and spreading of normal human dermal fibroblasts (NHDFs) of prepared nanofibrous mats are confirmed by MTT and SEM results. Furthermore, the LBL structured (CS/collagen)n nanofibrous mats greatly improve the cell migration in vitro, promote re-epithelialization and vascularization in vivo, and up-regulate the expression of collagen Ⅳ and α-tubulin, as well as the Integrin β1 and phosphorylation of focal adhesion kinase (FAK) at Tyr-397. The levels of expressed protein are significantly enhanced with increasing coating bilayers via immunohistochemistry and western blotting analyses. Collectively, these results suggest that the LBL structured biomimetic nanofibrous matrices may enhance cell migration and further promote the skin regeneration by up-regulating the secretion of ECM protein and triggering Integrin/FAK signaling pathway, which demonstrate the potential use of the nanofibrous mats to rapidly restore the structural and functional properties of wounded skin.

Biomimetic LBL structured nanofibrous matrices assembled by chitosan/collagen for promoting wound healing.

Published studies have generated inconsistent results related to the contribution of CRR9 rs401681 C allele to the risk of developing non-melanoma skin cancer (NMSC), and it is the inconsistency that promoted us to undertake a meta-analysis to identify the degree of impact the C allele has on NMSC.

Association of CRR9 locus with elevated risk of squamous cell carcinoma and basal cell carcinoma.

Split-thickness skin graft (STSG) donor site dressing has been controversial until now. Our study aimed to assess the patient comfort and wound-healing efficacy with the application of thin split-thickness skin grafts regrafting on STSG donor sites.

Wound-healing improvement by resurfacing split-thickness skin donor sites with thin split-thickness grafting.

Hypertrophic scarring (HS) is an overcompensation of wound healing that increases the risk of cosmetic disfigurement and functional impairment. No gold standard has been established for the treatment or prevention of HS. Our study aims to elucidate the expression and function of lumican in the pathogenesis of HS as well as the underlying mechanism involved in this procedure. An animal model of HS (rabbit ear) was established, and the Ad-lumican vectors were locally injected. Primary fibroblasts isolated from patients with hypertrophic burn scars were used in vitro. Histological and molecular changes in HS pathogenesis were evaluated. The results showed that lumican is significantly reduced in HS tissues and fibroblasts from HS patients as compared to normal skin or cells. Lumican levels were further suppressed in response to TGF-β stimulation. However, lumican upregulation effectively thinned the scar area and inhibited fibroblast proliferation and the cell cycle. Meanwhile, Ad-lumican administration suppressed the deposition of extracellular matrix, such as collagen and CTGF. Ad-lumican injected animals or fibroblasts presented comparable integrin α2β1 expression while greatly reduced phosphorylation of FAK compared to the negative control. Moreover, Ad-lumican administration largely enhanced the binding of lumican to integrin α2β1 and may thus inhibit the signaling propagation of collagen-integrin α2β1. Overall, the restoration of lumican levels contributed to suppressing the HS progression by inhibiting collagen-integrin α2β1-FAK signaling.

Lumican alleviates hypertrophic scarring by suppressing integrin-FAK signaling.

Although microRNA-301a has been reported to function as an oncogene in many human cancers, the roles of miR-301a in malignant melanoma (MM) is unclear. The present study aims to investigate the functional roles of miR-301a in MM and its possible molecular mechanisms.

Xiaoxing Lv

Geneplus-Beijing Institute

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing