Exploration of the effect of EphB2 inhibitors on cutaneous squamous cell carcinoma and their mechanisms of action based on Wnt/β-catenin signaling pathway / 中华皮肤科杂志

Objective:To screen small-molecule inhibitors of tyrosine kinase receptor B2 (EphB2) by using a molecular docking method, and to investigate their effect on cutaneous squamous cell carcinoma (CSCC) and possible mechanisms of action.Methods:The three-dimensional structure of EphB2 protein and its ligand binding sites were predicted by using the docking tool Schrodinger, and high-throughput virtual screening of EphB2 inhibitors was carried out by molecular docking. The anti-CSCC effect and mechanism of action of the screened EphB2 inhibitors kaempferitrin and aloe-emodin (AE) were verified in in vitro and in vivo experiments. In the in vitro experiments, human CSCC cell lines A431 and SCL-1, as well as the human immortalized keratinocyte HaCaT, were all divided into blank control group, dimethyl sulfoxide (DMSO) group, AE group and kaempferitrin group. Methyl thiazol tetrazolium (MTT) assay (AE at concentrations of 20, 40, 80, 160 μmol/L, kaempferitrin at concentrations of 12.5, 25, 50, 100 μmol/L), scratch and Transwell assays (AE at a fixed concentration of 80 μmol/L, kaempferitrin at a fixed concentration of 50 μmol/L) were performed to analyze the effect of EphB2 inhibitors on the proliferation, migration and invasion of CSCC cells. In the in vivo experiments, specific pathogen-free BALB/c female nude mice were subcutaneously injected with 0.2 ml of A431 cell suspension. After tumor growth, 24 tumor-bearing mice were randomly and equally divided into 4 groups: AE group and kaempferitrin group intraperitoneally injected with 20 mg·kg -1·d -1 AE and 25 mg·kg -1·d -1 kaempferitrin respectively, blank control group and DMSO group intraperitoneally injected with the same volume of sodium chloride physiological solution and DMSO respectively; the tumor size and body weight of nude mice were measured weekly; after consecutive treatment for 28 days, transplanted tumors were resected from the nude mice for hematoxylin and eosin (HE) staining, and real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to analyze the effect of AE and kaempferitrin on the mRNA and protein expression of E-cadherin, vimentin, glycogen synthase kinase 3β (GSK-3β), phosphorylated GSK-3β (p-GSK-3β) and β-catenin respectively. One-way analysis of variance and t test were used for comparisons between groups. Results:Two small-molecule compounds AA-504/20999031 (kaempferitrin) and AA-466/21162055 (AE) with high inhibitory activity against EphB2 were screened out. MTT assay showed that both AE and kaempferitrin exhibited strong cytotoxicity to SCL-1 and A431 cells compared with HaCaT cells, and their toxicity increased with the increase of their concentration ( F = 17.95, 11.34, respectively, both P < 0.001) ; after 48-hour treatment, the 50% inhibitory concentrations (IC50s) of AE against SCL-1 and A431 cells were 124.59 and 80.85 μmol/L respectively, and the IC50s of kaempferitrin against SCL-1 and A431 cells were 119.64 and 64.96 μmol/L respectively. Scratch assay showed that the migration distance of A431 cells was significantly shorter in the AE group and kaempferitrin group (36.7 ± 1.0 μm, 44.7 ± 3.5 μm, respectively) than in the DMSO group (88.1 ± 1.4 μm, F = 52.34, P < 0.001), while there was no significant difference in the migration distance of HaCaT cells among the above groups ( F = 1.73, P = 0.238). Transwell assay showed that the number of A431 cells crossing the Transwell membrane significantly decreased in the AE group and kaempferitrin group (145.0 ± 2.5, 94.7 ± 4.1, respectively) compared with the DMSO group (195.3 ± 5.7, F = 72.85, P < 0.001), while neither AE nor kaempferitrin showed significant inhibitory effects of on the number of HaCaT cells crossing the Transwell membrane ( F = 3.91, P = 0.055). The animal experiment revealed significantly decreased volumes of transplanted tumors in nude mice in the AE group and kaempferitrin group (407.42 ± 70.37 mm 3, 368.77 ± 62.7 mm 3, respectively) compared with the DMSO group (841.88 ± 84.63 mm 3, F = 73.78, P < 0.001). HE staining confirmed that AE and kaempferitrin could improve pathological changes of transplanted tumors. qRT-PCR and Western blot analysis showed that AE and kaempferitrin significantly up-regulated the mRNA and protein expression of E-cadherin and p-GSK-3β in tumor tissues (all P < 0.001), and down-regulated the mRNA and protein expression of vimentin, β-catenin and GSK-3β (all P < 0.001) . Conclusion:The small-molecule inhibitors screened by molecular docking can form a stable complex with EphB2, and inhibit the progression of CSCC by affecting the Wnt/β-catenin pathway-induced epithelial-mesenchymal transition.

Mesenchymal stem cells in the treatment of myocardial infarction: how long is the way from bench to bedside? / 中国组织工程研究

BACKGROUND: Mesenchymal stem cells, a kind of popular seed cells for cell transplantation after myocardial injury, have become a hot research topic in the treatment of ischemic heart diseases. OBJECTIVE: To review the new progress of mesenchymal stem cell transplantation in the treatment of ischemic heart diseases. METHODS: We searched Medline, Ovid Embase, PubMed Central, Cochrane Library, Chinese Periodical Full-Text Database, Chinese Biomedical Literature Database, VIP database, Wanfang Database for relevant articles in Chinese and English languages, and the retrieval time was from 1985 to 2015. Articles related to randomized controlled trials or clinical research on mesechymal stem cell transplantation for treatment of ischemic heart diseases were included, and relevant review were also included. Observational studies, casereports, study protocols with no standard or rational design, reviews with no normative and rigorous inclusion criteria, cross-test studies, cluster randomized trials, and studies that have nothing to do with the topic were excluded.RESULTS AND CONCLUSION: Through literature screening, we summarize the research progress in the mesenchymal stem cell transplantation, including cell source, safety, efficacy, action of mechanism and injection mode, and analyze the merits and demerits of mesenchymal stem cell transplantation. Bone marrowmesenchymal stem cell transplantation provides a new insight into the clinical treatment of myocardial infarction,but there are still many problems to be solved, such as cell quality, cell dose, transplantation mode, transplantation timing and patient selection, which so limit the clinical application of mesenchymal stem cell transplantation.

Roles of Nerve in Regeneration and Scar Healing of Skin Wound (review) / 中国康复理论与实践

Nerve is the foundation of the physiological function such as the skin feeling and thermoregulation, and also regulates the re-generation and scar healing of skin wound by multiple mechanisms. Though the denervated skin wound can heal spontaneously, the nerve might accelerate wound healing by activating the neurogenic inflammation and nerve trophism, increasing the blood supply around the wound, promoting the proliferation of fibroblasts and keratinocytes, stimulating the expression of collagen I and III, and interacting with im-mune system and releasing neuropeptides. If all kinds of repair cells, extracellular matrix, nerve, blood vessel and cutaneous appendages as-semble organically, the skin wound regenerates, and otherwise the scar heals. The nerve axon growth occurs mainly in the early stage of the wound healing and the scar rebuilding process, and the nerve growth rate is obviously slower than the granulation tissue formation and cica-trization. Furthermore, the axon growth of different nerve fibers are not entirely the same. The exogenous neuropeptides might promote the wound repair and the nerve regeneration. The antagonist of the neuropeptides or high selective nerve abscission might reduce the scar hyper-plasia. Therefore, it contribute to find methods to promote the regeneration of skin wound and peripheral nerve injury by understanding the effects and regulatory mechanism of both nerve and various repair elements in regeneration or scar healing of skin wound.

Advances in the research of modulation of dermal collagen fibrin assembly by decorin / 中华烧伤杂志

Formation of dermal collagen fiber is a complicated and sequential process with the progressive assembly of collagen. Collagen monomers form stepped and orderly protofibrils through longitudinal displacement. Subsequently, protofibrils or protofibrils and collagen are bonded by covalent bonds to form orderly lamellar structure of collagen fibers. Then collagen fibers are tightly wound into coarse collagen fiber bundles by covalent crosslinking. Decorin is a multifunctional small leucine-rich proteoglycan. It can prevent the aggregation of protofibrils by binding to the specific site of collagen with its core protein, and adjusting the spacing between the protofibrils with its glycosaminoglycan chain. Thus, by effecting the formation of collagen fibers with regulation of collagen assembly, decorin may help prevent scar formation and even promote regeneration.

Application of the repair in the facial skin and soft-tissue defects with local random flap / 国际外科学杂志

Objective To explore clinical efficacy of local random flap in reconstruction of facial skin and softtissue defects caused by lesions resection and injury.Methods Forty-six cases with facial skin and soft-tissue defects were cured by constructing and transplanting different local random flap that is designed according to parts,shapes and sizes of defect between July of 2008 and June of 2013.Results All the cases obtained satisfactory effects for full survivorship of flap,one-stage healing of wound,tiny scar and pigmentation,and good appearance by following up from 3 months to 6 months after operation.Conclusions Using local random flap in repair of facial skin defect could get satisfactoried functional and aesthetic efficacy,and it is worth being applicatied in the clinic.

Effect of vacuum sealing drainage on nerve growth factor and microvessels in chronic wound / 中国组织工程研究

BACKGROUND:With the increase of aging degree, chronic wounds are increasing. There is few method to cure chronic wounds at present, and vacuum sealing drainage has been applied to the clinic. OBJECTIVE:To investigate the effect of vacuum sealing drainage on the expression of nerve growth factor and the change of microvascular number in the human chronic wounds, and to study the mechanism of accelerating chronic wound healing. METHODS:Ten patients with chronic wounds were included in this study, including one case of soft tissue defect of chest, one case of osteomyelitis with bone exposure, one case of large area skin defect after amputation stump, two cases of wound infection after skin avulsion injury, two cases of shinbone osteomyelitis, and three cases of postoperative wound infection. The skin around the wounds and granulation tissue within the wounds were col ected before vacuum sealing drainage treatment and at 7, 14 days after vacuum sealing drainage treatment. The nerve growth factor expressions and the microvascular number in the wounds were measured by the immunohistochemical technology. The wound healing was also observed. RESULTS AND CONCLUSION:The nerve growth factor expressions in fibroblasts and vascular endothelial cel s at 7 and 14 days after vacuum sealing drainage treatment were significantly higher than that before treatment (P<0.001). The microvascular number was also increased after vacuum sealing drainage treatment compared with pre-treatment (P<0.05-0.01). The vacuum sealing drainage treatment reduced necrotic tissue and purulent secretion within the wounds, as wel as swel ing around the wounds. Simultaneously, granulation tissue gradual y became fine granules, bright red, and bleeding easily, thus it is suitable for promoting wound healing.

Effects of amlodipine through percutaneous penetration on survival of ischemic random skin flap / 中华医学美学美容杂志

Objective To investigate the penetrative ability of amlodipine gels and to evaluate their effect on the survival of rat dorsal ischemic random skin flaps.Methods 0.5 ％,1.0 ％,1.5 ％,2.0 ％,and 2.5 ％ amlodipine gels were made.The accumulative penetrative quantities of amlodipine through rat skin were assayed with a modified Franz's diffusion cell in vitro.Pure gel without amlodipine,0.5 ％ and 1.5 ％ amlodipine gel were respectively applied on rat random ischemic skin flap once a day for 7 days.The viable area was measured on the seventh postoperative day and the quantities of amlodipine within skin flap were also detected at 2 and 6 hours after application of amlodipine gel.Results The accumulative penetrative quantities of amlodipine increased in time-and concentration-dependent manner (P＜0.05).Accumulative quantities of 0.5 ％ and 1.0 ％ amlodipine gel were lower than those of 1.5 ％,2.0 ％,and 2.5％ gel,respectively (P＜0.05).The quantities of amlodipine within flap tissue in 1.5 ％ amlodipine gel was higher than that of 0.5 ％ amlodipine gel (P＜0.05).The survival area of flap in 0.5 ％ amlodipine gel group (391.4±65.4) mm2 was higher than those of the pure percutaneous gel group (192.9±56.8) mm2 and the control group (191.0±50.2) mm2(P＜0.05),but no significant difference was seen between 1.5 ％ amlodipine gel group (265.7+88.3) mm2 and control group (P＞0.05).Conclusions Amlodipine could penetrate into skin tissues.0.5 ％ amlodipine gel could significantly increase survival area of ischemic random skin flap.

Effects of TGF-?_1 on activity of protein tyrosine kinase in scar tissue of rabbit ear / 中华医学美学美容杂志

Objective To observe the changes of protein tyrosine kinase (PTK) activity in scar tissue of rabbit ear induced by TGF-?_1 and to investigate the signal mechanism of TGF-?_1 in the scar proliferation. Methods TGF-?_1 (5 ng/ml) was applied on the wound and in the scar of rabbit ear. The PTK activity in the tissues from normal control, and 0, 14, 30, 45, and 60 d after wound epithelization was measured by phosphorus ( 32 P) incoporation. The scar changes were also observed. Results PTK activity in hypertrophic scar tissue [from (4.40?1.31) to (4.91?1.32)pmol?min -1 ?mg -1 ] was obviously elevated as compared with(2.87?0.96) pmol?min -1 mg -1 of normal skin ( P 0.05). Furthermore, PTK activity of hypertrophic scar tissue was obviously elevated as compared with that of normal scar tissue ( P 0.05) but could accelerate scar hypertrophy ( P