LncRNA TUG1 regulates FGF1 to enhance endothelial differentiation of adipose-derived stem cells by sponging miR-143.

Adipose-derived stem cells (ADSCs) have emerged as a cell source for regeneration medicine. ADSCs possess the capacity to differentiate into endothelial cells and serve an essential role in vascular development and function. LncRNA taurine upregulated gene 1 (TUG1) has recently been linked with angiogenesis in hepatoblastoma. However, the roles of TUG1 in endothelial differentiation of ADSCs remain unidentified. Human adipose-derived stem cells (hADSCs) were obtained and characterized by flow cytometry, Oil red O and Alizarin Red staining. HADSCs were maintained in the endothelial differentiation medium and the expressions of TUG1, miR-143, and FGF1 were examined by qRT-PCR. To assess endothelial differentiation, the expressions of CD31, von Willebrand factor (vWF), VE-cadherin were examined by Western blot analysis, qRT-PCR, and immunofluorescence. Tube formation in Matrigel was examined. The interactions between TUG1 and miR-143, miR-143 and FGF1 were validated by luciferase assays. During the endothelial differentiation process, TUG1 and FGF1 were upregulated, whereas miR-143 was downregulated. TUG1 overexpression downregulated miR-143, upregulated FGF1, CD31, vWF, and VE-cadherin, and enhanced capillary tube formation. Luciferase assays showed that TUG1 interacted with miR-143, and FGF1 was a direct target of miR-143. Furthermore, the enhancement of endothelial differentiation induced by TUG1 overexpression was abolished by miR-143 overexpression. Our findings implicated that lncRNA TUG1 promoted endothelial differentiation of ADSCs by regulating the miR-143/FGF1 axis.

Viability and Biomechanics of Bare Diced Cartilage Grafts in Experimental Study.

OBJECTIVE: To assess the viability and biomechanics of bare diced cartilage grafts. METHODS: Cartilage samples were collected from 1 ear in 15 rabbits as well as costal cartilage. Each rabbit was inserted bare diced- and single-strip costal-cartilage grafts, respectively, into paraspinal subcutaneous pockets: after euthanasia at 2 months, specimens were weighed, with diced cartilage grafts examined histomorphologically by hematoxylin-eosin staining, masson trichrome staining, and immunohistochemistry. Finally, biomechanical properties of grafts were assessed. RESULTS: Bare diced cartilage grafts were connected into an integrated mass after 2 months, and inward growth of fibrous tissues and angiogenesis were observed. Mean wet weights of diced cartilage grafts were 1.603 ±â€Š0.278 and 1.662 ±â€Š0.204 g pre- and postoperation, respectively; those of costal cartilage grafts were 0.053 ±â€Š0.008 and 0.058 ±â€Š0.008 g, respectively. In compression assays, mean modulus values of elasticity at yield in diced- and costal-cartilage grafts were 7.65 ±â€Š0.59 and 22.30 ±â€Š1.15 MPa, respectively (P < 0.05); mean stress values were 4.07 ±â€Š0.38 and 12.50 ±â€Š1.15 MPa, respectively (P < 0.05). In the tensile test, mean modulus values of elasticity at yield of diced- and costal-cartilage grafts were 4.70 ±â€Š0.78 and 10.59 ±â€Š1.39 MPa, respectively (P < 0.05), mean stress values were 0.82 ±â€Š0.05 and 1.76 ±â€Š0.21 MPa, respectively (P < 0.05). CONCLUSIONS: Diced cartilage grafts had favorable viability and growth. Despite reduced elasticity and stress values, they still can be served as substitute for supportive filling materials.

[Serum hydrogen sulfide levels in children with benign infantile convulsions associated with mild gastroenteritis].

OBJECTIVE: To study the changes and significance of serum hydrogen sulfide (H2S) levels in children with benign infantile convulsions associated with mild gastroenteritis (BICE). METHODS: Forty-two hospitalized children diagnosed with BICE were recruited to the observation group, and 46 children admitted due to acute gastroenteritis alone were recruited to the control group. Serum H2S levels were measured by a spectrophotometer. RESULTS: The serum H2S level in the observation group was significantly lower than in the control group (28±12 µmol/L vs 45±10 µmol/L; P<0.01). The patients with a number of convulsions greater than or equal to two had significantly lower serum H2S levels than those with a number less than two (P<0.05). The number of convulsions was negatively correlated with serum H2S level in BICE patients (r=-0.485, P=0.001). When a convulsion exceeded 5 minues in duration, the duration was negatively correlated with serum H2S level (r=-0.736, P=0.004). CONCLUSIONS: The reduction in endogenous H2S level might be one of the causes of convulsions in BICE patients. The degree of reduction in H2S level is associated with the number of convulsions and the duration of convulsion (when it exceeds 5 minues). Further investigation is needed to determine the clinical significance of these results.

Increased expression of heat shock protein 70 and heat shock factor 1 in chronic dermal ulcer tissues treated with laser-aided therapy.

BACKGROUND: Chronic dermal ulcers are also referred to as refractory ulcers. This study was conducted to elucidate the therapeutic effect of laser on chronic dermal ulcers and the induced expression of heat shock factor 1 (HSF1) and heat shock protein 70 (HSP70) in wound tissues. METHODS: Sixty patients with 84 chronic dermal ulcers were randomly divided into traditional therapy and laser therapy groups. Laser treatment was performed in addition to traditional therapy in the laser therapy group. The treatment efficacy was evaluated after three weeks. Five tissue sections of healing wounds were randomly collected along with five normal skin sections as controls. HSP70-positive cells from HSP70 immunohistochemical staining were counted and the gray scale of positive cells was measured for statistical analysis. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were performed to determine the mRNA and protein expressions of HSF1 and HSP70. RESULTS: The cure rate of the wounds and the total efficacy in the laser therapy group were significantly higher than those in the traditional therapy group (P < 0.05, P < 0.01, respectively). Immunohistochemical staining revealed that the HSP70-positive cell count was significantly higher in laser therapy group than those in the traditional therapy group and controls (P < 0.01), and the gray scale of the cell signal was obviously lower than traditional therapy group and controls (P < 0.05). By contrast, the traditional therapy group and the control group were not significantly different. The RNA levels of HSF1 and HSP70 were higher in the laser therapy group by RT-PCR, but very low in normal skin and the traditional therapy group. The analysis on the gray scale of the Western blot bands indicated that the expression of HSF1 and HSP70 in the laser therapy group was significantly higher than in the traditional therapy group and the control group (P < 0.01), and the expression in the traditional therapy group was also higher than in the control group (P < 0.05). CONCLUSION: Laser-aided therapy of chronic dermal ulcers plays a facilitating role in healing due to the mechanism of laser-activated endogenous heat shock protection in cells in wound surfaces.

Research Advances in the Treatment of Melanoma by Treat Melanoma.

Melanoma is a highly malignant tumor. Prognoses of melanoma patients are often unsatisfactory due to poor operational and chemoradiational efficacy. Recently, researches for melanoma treatment have found multipeptide vaccines a favorite and possible breakthrough as they are stable in chemical property and easy to be synthesized, have no carcinogenecity and dispense with virus vector. Studies have shown that the immunogenicity of multipeptide vaccines could be enhanced by use of immunoadjuvants, joining dendritic cells (DCs), full-length or epitope-superposited antigen peptides, costimulatory molecules and cellpenetrating peptides fusion, thereby improving anti-tumor effect. Certain achievements have been obtained in clinical treatment of melanoma by multipeptide vaccines, but problems including poor immunogenicity and human leukocyte antigen (HLA) phenotype restriction may require further study.

[Killing activity of cytotoxic T lymphocytes stimulated by dendritic cell vaccine loaded with autologous cervical cancer antigen].

BACKGROUND & OBJECTIVE: Dendritic cells (DCs), the strongest antigen-presenting cells (APCs), can present antigens to T lymphocytes in vivo and in vitro, and induce cytotoxic T lymphocyte (CTL) reaction. This study was designed to investigate the killing activity of CTLs stimulated by Dcs loaded with autologous cervical cancer antigen in vitro. METHODS: Tumor antigens were made from frozen-thawed cervical cancer cells from patients after operation. DCs were isolated from peripheral blood mononuclear cells of patients with cervical cancer, cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4), loaded with tumor antigen to prepare DC vaccine, and used to stimulate autologous T lymphocytes to prepare antigen-specific CTLs. The killing activities of CTLs on autologous cervical cancer cells and HeLa, HepG2, MCF7, A549, and MGC803 cells were observed. RESULTS: CTLs stimulated by the DC vaccine had high killing activity on autologous cervical cancer cells, with killing rates of 79.32%-89.27% which were obviously higher than that of lymphokine-activated killing cells (t> or =2.89, P<0.05). The killing activity of CTLs was significantly weaker on HeLa cells (40.35%-58.09%) than on autologous cervical cancer cells (t> or =2.97, P<0.05). The specific CTLs had no obvious killing activity on HepG2, MCF7, A549, and MGC803 cells. CONCLUSIONS: CTLs stimulated by autologous cervical cancer antigen-loaded DCs have highly efficient and specific immune activity on autologous cervical cancer cells. It may be used in biotherapy for cervical cancer.