Background: Wound healing is a dynamic and complex process that is regulated by a variety of factors and pathways. This study sought to identify the mechanisms of the four-herb Chinese medicine ANBP in enhancing wound repair. Material/Methods: By comparing the group treated with ANBP for 6 h (Z6h) with the corresponding control group (C6h), we used the new high-throughput differential acetylation proteomics method to explore the mechanism of ANBP treatment and analyse and identify new targets of ANBP for promoting wound healing. Results: ANBP promoted skin wound healing in mice; the wound healing process was accelerated and the wound healing time was shortened (P<0.05). The upregulated proteins were distributed mostly in the mitochondria to nuclear respiratory chain complexes and cytoplasmic vesicles. The dominant pathways for upregulated proteins were fatty acid metabolism, pyruvate metabolism, and tricarboxylic acid cycle. Pdha1 was upregulated with the most acetylation sites, while the downregulated Ncl, and Pfkm were most acetylated. Conclusions: The findings from our study showed that ANBP improved cell aerobic respiration through enhanced glycolysis, pyruvic acid oxidative decarboxylation, and the Krebs cycle to produce more ATP for energy consumption, thus accelerating wound repair of skin.

Context Having been used for thousands of years to treat gastrointestinal diseases, the natural isoquinoline alkaloid, berberine, has exhibited a wide spectrum of biochemical and pharmacological effects in studies of recent years. Objective The review intended to examine the many novel bioactivities of berberine, including antidiabetic, anticancer, neuroprotective, anti-inflammatory, and anti-atherosclerotic actions. Design The research team searched the MEDLINE database using PubMed, using different keyword combinations, including berberine AND diabetes, berberine AND cancer, berberine AND (neuron OR brain), berberine AND inflammation, and "berberine AND atherosclerosis to find studies evaluating the various effects exerted berberine. Conclusion Berberine is a promising multipotent agent to combat diabetes, cancer, Alzheimer's disease, and other diseases.

An injectable hydrogel is a powerful carrier for therapeutic bioactive molecules. Here, an injected concentrated conditioned medium (CCM)-silk nanofiber composite hydrogel was developed to achieve the sustained release of multiple proteins and better wound healing. All the encapsulated proteins showed slow delivery for more than 9 days in vitro. Bioactive molecules such as transforming growth factor-beta1 (TGF-β1), insulin-like growth factor binding protein-1 (IGFBP-1), and platelet-derived growth factor-AB (PDGF-AB) were successfully released from CCM-loaded hydrogels, and they induced the proliferation and migration of fibroblasts and endothelial cells in a dose- and time-dependent manner. The differentiation of fibroblasts into myofibroblasts was also inhibited, implying less scar formation in vivo. Skin wound regeneration results indicated that the CCM-loaded hydrogel enhanced neovascularization, accelerated wound closure, and promoted hair follicle regeneration. The injectable multiple protein delivery system shows a promising application in skin wound repair.

Background: Tissue contraction and the extracellular matrix deposition are part of the pathogenesis of hypertrophic scars. The transcriptional factor NFE2L2 inhibits fibroblast differentiation in idiopathic pulmonary fibrosis and promotes myofibroblast dedifferentiation. Our previous study showed that the transcription factor NFE2L2 was strongly induced on treatment with arsenic trioxide (ATO). Objective: The present study sought to investigate the effect of ATO on myofibroblast formation to determine its potential role in hypertrophic scar treatment. Methods: Small interfering RNA against NFE2L2 was used on treatment with ATO in human skin myofibroblasts. The expression levels of fibrosis markers were assessed by reverse transcription polymerase chain reaction, western blot, and immunofluorescence staining. The transforming growth factor-beta 1 (TGF-beta 1)/Smad2/3 signaling was detected by western blot. A rabbit ear model was used to evaluate the antifibrotic role of ATO. Results: At the cellular level, ATO abolished fibroblast differentiation in response to TGF-beta 1. ATO reduced TGF-beta 1-induced reactive oxygen species accumulation through increased expression of the antioxidant gene HO-1 in fibroblasts. In addition, ATO promoted the nuclear translocation of NFE2L2 and inhibited the phosphorylation of Smad2/3. In the rabbit ear model, ATO prevented the progression of hypertrophic scar formation. Conclusions: This study provides the first evidence implying that ATO inhibits the formation of myofibroblasts in vivo and in vitro and provides a possible treatment for hypertrophic scars.

Ischemic heart disease within developed countries has been associated with high rates of morbidity and mortality. Cell‑based cardiac repair is an emerging therapy for the treatment of cardiac diseases; however, a limited source of the optimal type of donor cell, such as an autologous cardiomyocyte, restricts clinical application. The novel therapeutic use of induced pluripotent stem cells (iPSCs) may serve as a unique and unlimited source of cardiomyocytes; however, iPSC contamination has been associated with teratoma formation following transplantation. The present study investigated whether cardiomyocytes from mouse fibroblasts may be reprogrammed in vitro with four cardiac transcription factors, including GATA binding protein 4, myocyte‑specific enhancer factor 2C, T‑box transcription factor 5, and heart‑ and neural crest derivatives‑expressed protein 2 (GMTH). Cardiac‑specific markers, including α‑myosin heavy chain (α‑MHC), β‑MHC, atrial natriuretic factor, NK2 homeobox 5 and cardiac troponin T were observed within mouse fibroblasts reprogrammed with GMTH, which was reported to be more effective than GMT. In addition, Percoll density centrifugation enriched a population of ~72.4±5.5% α‑MHC+ induced cardiomyocytes, which retained the expression profile of cardiomyocyte markers and were similar to natural neonatal cardiomyocytes in well‑defined sarcomeric structures. The findings of the present study provided a potential solution to myocardial repair via a cell therapy applying tissue engineering with minimized risks of immune rejection and tumor formation.

Traditional Chinese medicine has great potential to improve wound healing. ANBP, the mixture of 4 Chinese herbs—Agrimoniapilosa, Nelumbonucifera, Boswelliacarteri, and Pollen typhae—is effective in trauma treatment while its mechanism is still elusive. In this study, quantitative proteomics and bioinformatics analyses were performed to decipher the possible roles of ANBP in accelerated wound healing of mouse skin. Among all 3171 identified proteins, 90, 71, 80, and 140 proteins were found to be differently expressed in 6 hours, 3 days, 7 days, and 14 days ANBP-treated tissues compared with corresponding control tissues, respectively. The result showed that different biological processes and pathways were activated at different healing stages. At the early healing stage, ANBP treatment mainly affected several biological processes, including immune and defense response, vascular system restoration, hemostasis and coagulation regulation, lipid metabolism and signal transduction, while muscle tissue, hair, epidermis, extracellular matrix and tissue remodeling related activities were the major events in ANBP promoted later wound healing. This is the first quantitative proteome study of ANBP-treated wound tissues, which provide a new perspective for the mechanism of ANBP accelerated wound healing and is of guiding significance for clinical application of ANBP in trauma disorders cure.

目的:探讨美国食品药品监督管理局(Food and Drug Administration,FDA)对医疗器械境外临床研究的监管要求,为我国相关从业人员提供参考和借鉴.方法:通过介绍美国法规中针对医疗器械境外临床研究的条款和FDA发布的涉及境外临床研究的指导原则,了解FDA对医疗器械境外临床研究的监管要求,从而在此方面为我国相关监管部门提出启示.结果:相关监管部门在接受医疗器械境外临床研究资料时,需考虑临床医疗条件、临床研究人群、临床研究监管要求等多方面的差异.结论:我国相关监管部门可借鉴FDA接受境外临床研究结果的经验,结合我国相关法规要求和实际情况,在确保产品安全有效的前提下,细化接受进口医疗器械提交境外临床试验资料的接受标准和要求.

This article introduces the situation of silver-contained wound dressings, considers the evaluation of their safety and effectiveness. It is hoped to be helpful for silver-contained wound dressings registration.

Resorbable adhesion barrier devices for use in abdominal and/or pelvic surgery are high-risk implantable medical devices. There are some difficulties in their clinical research. This article summarizes and analyzes the common problems of resorbable adhesion barrier devices for use in abdominal and/or pelvic surgery clinical studies from the perspective of technical review, in order to provide reference for Chinese manufacturers and investigators in the design of clinical studies and clinical research.

Glucagon-like peptide-1(GLP-1) is an incretin hormone released from intestinal L-cells in response to nutrients. GLP-1 stimulates insulin secretion from pancreatic β cells in a glucose-dependent manner. GLP-1 al?so has many other beneficial effects for glucose homeostasis such as slowing gastric emptying, suppressing appe?tite, reducing plasma glucagon, and stimulating glucose disposal. Therefore, there is increasing interest in incretin-based therapies for treatment of diabetes. In this review we discussed the current understanding of the effect of GLP-1 on pancreatic β cells.

Wound healing is a troublesome problem in diabetic patients. Besides, there is also an increased risk of postsurgical wound complications for diabetic patient. It has been revealed that traditional Chinese medicine may promote healing and inhibit scar formation, while the changes of morphology and physiology of wounds on such medicine treatment still remain elusive. In this study, we first used the ultralow temperature preparation method to produce mixed superfine powder from Agrimonia pilosa (A), Nelumbo nucifera (N), Boswellia carteri (B), and Pollen typhae (P), named as ANBP. Applying ANBP on 40 streptozotocin (STZ)-induced diabetic C57BL/6 mice (4-6 weeks, 20 ± 2 g), we observed that the wound healing process was accelerated and the wound healing time was shortened (14 days, P < .05). Pathological observation using hematoxylin–eosin staining indicated that inflammatory cells were reduced ( P < .05) while the thickness of granulation tissue and length of epithelial tongue were increased ( P < .05). The vascular density was increased on 7 and 14 days after ANBP treatment. Masson and Sirius red staining showed that, at the early stage of trauma, the expressions of Col I and Col III, especially Col III, were increased in the ANBP group ( P < .05). Studies in vitro demonstrated that tubular formation was significantly increased after ANBP treatment on human vascular endothelial cells in a dose-dependent way. Taken together, our studies revealed that ANBP treatment could accelerate wound healing, promote vascularization, and inhibit inflammation, suggesting the potential clinic application of ANBP for diabetes mellitus and refractory wounds.

Objective: To construct a recombinant lentiviral vector of short hairpin RNA(shRNA) targeting NLRP3 gene, and to identify its inhibitory effect in HepG2 cells. Methods: The NLRP3 shRNA sequence was designed and inserted into the lentiviral vector pWPT-U6-shRNA-CMV-GFP. The lentiviral vector was further packaged with accessory plasmids into lentivirus in 293T cells. The virus solution was concentrated and serially diluted into 9 levels, and transfected into 293T cells respectively for the titer determination. The resulted lentivirus was infect?ed HepG2 cell to stably suppress NLRP3 expression, and the inhibition effect was detected by quantitative real-time PCR(qRT-PCR) and Western blotting. Results: A lentiviral vector carrying NLRP3 shRNA was constructed. The virus titer was 2×108 TU/mL detected by dilution method. The NLRP3 expression in HepG2 cells was down-regulated significantly, confirming by the qRT-PCR and Western blotting. Conclusion: The constructed lentiviral vector targeting human NLRP3 gene can stably suppress NLRP3 expression in HepG2 cell lines.

The four-herb Chinese medicine ANBP is a pulverized mixture of four herbs including Agrimonia Eupatoria (A), Nelumbo Nucifera Gaertn (N), Boswellia Carteri (B) and Pollen Typhae Angustifoliae (P). The combination of the four herbs was first described in Chinese canonical medicine about 2000 years ago for treatment of various trauma disorders, such as hemostasis, antiinflammatory, analgesia, and wound healing, etc. However, the precise mechanisms of ANBP are still unclear. In our study, using rabbit ear hypertrophic scar models of full-thickness skin defect, we showed that local ANBP treatment not only significantly enhanced wound healing by relieving inflammation, increasing formation of granulation tissue and accelerating re-epithelialization, but also reduced scar formation by decreasing collagen production, protuberant height and volume of scars, and increasing collagen maturity. We demonstrated that these effects of ANBP are associated with transforming growth factor (TGF)-β1-mediated signalling pathways through Smad-dependent pathways. ANBP treatment significantly increased expression of TGF-β1 and Smad2/3 mRNA at the early stage of wound healing, and led to markedly decrease expression of TGF-β1 and Smad2/3 compared with the control group after 14 days post-wounding. Taken together, our results defined a bidirectional regulation role of ANBP for TGF-β1/Smad pathway in promoting wound healing and alleviating scar formation, which may be an effective therapy for human wounds at the earliest stage.

Cardiovascular disease is a leading cause of death in industrialized countries. Scientists are trying to generate cardiomyocytes in vitro and in vivo to repair damaged heart tissue. Pluripotent reprogramming brings an alternative source of embryonic-like stem cells, and the possibility of regenerating mammalian tissues by first reverting somatic cells to induced pluripotent stem cells, followed by redifferentiating these cells into cardiomyocytes. More recently, lineage reprogramming of fibroblasts directly into functional cardiomyocytes has been reported. The procedure does not involve reverting cells back to a pluripotent stage, and, thus, would presumably reduce tumorigenic potential. Interestingly, lineage reprogramming could be used for in situ conversion of cell fate. Moreover, zebrafish-like regenerative mechanism in mammalian heart tissue, which was observed in mice within the first week of postpartum, should be further addressed. Here, we review the landmark progresses of the two major reprogramming strategies, compare their pros and cons in cardiovascular regeneration, and forecast the future directions of cardiac repair.

The blastoderm cells isolated from the stage X embryos of chicken were cultured on feeder layer of mouse embryonic fibroblasts(MEF)with high glucose DMEM supplemented with 10 ng/mL bFGF,20 ng/mL hIGF-1,2 ng/mL mSCF,2 ng/mL hIL-11 and 1 000 U/mL LIF.Typical chicken embryonic stem cells(cESCs)colonies with high endogenous AKP activity were observed in the culture system.The cESCs derived from blastoderm cells expressed pluripotent markers SSEA-1 and Oct-4 as determined by immunocytological analysis.RT-PCR analysis further confirmed the expression of cENS-1,a gene specifically expressed in cESCs and early embryo.These results demonstrate the feasibility for using the culture system in isolating cESCs from the stage X chicken embryos and supporting the growth of cESCs with undifferentiating state in vitro.

BACKGROUND: The aim of myocardial tissue engineering is to repair or regenerate damaged myocardium with engineered cardiac tissue constructed by a combination of cells and scaffolds in vitro. However, this strategy has been hampered by the lack of cardiomyocytes and the significant cell death after transplantation in vivo.METHODS: In this study we explored the feasibility of in vitro construction of vascularized cardiac muscle using genetically modified mouse embryonic stem cells (ESCs) transfected by pMHC-neo/SV40-hygro. A stirred bioreactor was used to facilitate the formation of a large number of ESC-derived cardiomyocytes, which were then mixed with human umbilical vein endothelial cells (HUVECs) and mouse embryonic fibroblasts (MEFs) in a liquid collagen scaffold to construct highly vascularized cardiac tissue in vitro.RESULTS: The resulting tissue constructs were transplanted into dorsal subcutaneous sites of nude mice. Tumor formation was not detected in all samples and vascularized cardiac tissue could survive after transplantation. Vascularization of the implanted cardiac muscle was significantly enhanced by the addition of HUVECs and MEFs, which resulted in a thicker myocardium. The combination of genetically modified ESCs and stirred bioreactor cultivation not only benefited the large-scale production of pure ESC-derived cardiomyocytes, but also effectively controlled the potential risk of undifferentiated ESCs.CONCLUSIONS: Using liquid collagen as scaffold, the enriched cardiomyocytes derived from genetically modified ESCs mixed with HUVECs and MEFs in 3-dimensional culture resulted in highly vascularized cardiac tissues. J Heart Lung Transplant 2012;31:204-12 (C) 2012 International Society for Heart and Lung Transplantation. All rights reserved.

Embryonic stem cells (ESCs) can propagate unlimitedly in vitro and differentiate into cardiomyocytes, which have been proposed as unlimited cell sources for cardiac cell therapy. This was limited by difficulties in large-scale generation of pure cardiomyocytes. In this study, we used stirred bioreactors to optimize the differentiation condition for mass production of embryoid bodies (EBs) derived from genetically modified mouse ESCs. Stirred suspension culture could more efficiently produce EBs and have a more uniform EB population without large necrotic centers, compared with the conventional static culture. Importantly, the cardiac-specific gene expressions (GATA binding protein 4, α-cardiac myosin heavy chain and myosin light chain-2v) were increased within EBs cultured in stirred bioreactor. Stirred suspension culture significantly increased the proportion of spontaneously contracting EBs, yielded a greater percentage of α-sarcomeric actinin-positive cells detected via flow cytometry, and harvested relatively more cardiomyocytes after G418 selection. Stirred suspension culture provided a more ideal culture condition facilitating the growth of EBs and enhancing the cardiogenic differentiation of genetically modified ESCs, which may be valuable in large-scale generation of pure cardiomyocytes.

Pluripotent embryonic stem cells (ESC) have the ability to differentiate into a variety of cell lineages in vitro , including cardiomyocytes. Successful applications of ESC-derived cardiomyocytes in cell therapy and tissue engineering were limited by difficulties in selecting the desired cells from the heterogeneous cell population. We describe a simple method to generate relatively pure cardiomyocytes from mouse ESCs. A construct comprising mouse cardiac α-myosin heavy chain (MHC) promoter driving the neomycin resistance gene and SV40 promoter driving the hygromycin resistant gene designated pMHCneo/ SV40-hygro, was stably transfected into mouse ESCs. The transgenic ESC line, designated MN6 retained the undifferentiated state and the potential of cardiogenic differentiation. After G418 selection, more than 99% of cells expressed α-sarcomeric actin. Immunocytological and ultrastructural analysis demonstrated that, the selected cardiomyocytes were highly differentiated. Our results represent a simple genetic manipulation used to product essentially pure cardiomyocytes from differentiating ESCs. It may facilitate the development of cell therapy in heart diseases. Key words: Embryonic stem cells, α-myosin heavy chain promoter, cardiomyocytes, differentiation, genetic enrichment.

Objective: To study the role of LRP16 in the signal transduction pathways of ionizing radiation(IR)-induced nuclear factor-κB(NF-κB) activation.Methods: The effects of LRP16 on NF-κB response reporter(3×κB-Luc) and the down streaming target gene XIAP of NF-κB were measured by dual-luciferase report assay system and Western blotting respectively in HeLa cells.Results: The dual-luciferase report assay system showed that LRP16 overexpression increased the IR-induced relative luciferase activities of NF-κB response reporter and the inhibition of LRP16 expression decreased the IR-induced relative luciferase activities of NF-κB response reporter.Western blot also demonstrated that LRP16 overexpression strengthened the expression of the anti-apoptotic gene XIAP induced by NF-κB and the inhibition of LRP16 expression weakened the expression of the anti-apoptotic gene XIAP induced by NF-κB.Conclusion: LRP16 can enhance NF-κB transcriptional activity induced by IR and affect the expression of NF-κB-targeting anti-apoptotic gene XIAP induced by IR,which may provide a foundation for studying the molecular mechanism of IR-induced NF-κB transcriptional activation.

Objective: To establish a mammalian inducible expression system with the non-toxic biotin as an inducer.Methods: This system comprised three key components: E.coli biotin ligase(BirA),streptavidin-tetracyclin repressor(SA-TetR) and biotinylation signal-VP16 transactivation domain(AT-VP16).The three genes were introduced into a tricistronic expression vector,and co-transfected 293f cells together with the response vector.The EGFP taken for a reporter gene,EGFP fluorescent intensities were detected under different biotin concentrations.Results: With the increasing of biotin concentrations,EGFP displayed sequential OFF-ON-OFF expression profiles,the fluorescence intensity of EGFP in the induced state was about 3 times that in the inhibited state.Conclusion: The expression of the target gene could be reversibly regulated by adjusting the concentration of biotin.This is a promising inducible expression system.