Nuclear factor I-C inhibits platelet-derived growth factor-induced enhancement of dermal fibroblast sensitivity to TGF-β / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of nuclear factor I-C (NFI-C) on platelet-derived growth factor (PDGF)-induced up-regulation of TGF-β receptor II (TβRII) in dermal fibroblasts.</p><p><b>METHODS</b>A lentiviral vector containing NFI-C sequence (Lenti-GFP-NFI-C) was transfected into a human foreskin fibroblast cell line (HFF-1). Cultured HFF-1 cells, cells transfected with Lenti-GFP-NFI-C, and cells transfected with a negative virus were stimulated with PDGF-BB, and Western blotting and RT-qPCR were used to detect the expression levels of TβRII in the treated cells.</p><p><b>RESULTS</b>PDGF treatment significantly increased the expression level of TβRII in HFF-1 cells (P<0.05). The cells transfected with Lenti-GFP-NFI-C expressed a significantly lower level of TβRII than non-transfected cells in response to PDGF stimulation (P<0.05), but the negative virus showed no such inhibitory effect (P>0.05). No significant difference was found in the expression level of TβRII protein between cells transfected with Lenti-GFP-NFI-C-transfection before PDGF stimulation and the blank control cells.</p><p><b>CONCLUSION</b>NFI-C can inhibit PDGF-induced up-regulation of TβRII and thus reduce the sensitivity of the dermal fibroblasts to TGF-β.</p>

Effects of blocking two sites of transforming growth factor-β/Smads signaling on the formation of scar-related proteins in human skin fibroblasts / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the effects of blocking two sites of TGF-β/Smads signaling on the formation of scar-related proteins in human skin fibroblasts.</p><p><b>METHODS</b>Two lentivirus vectors encoding soluble TGF-β receptor II (sTβRII) and mutant Smad 4-Smad 4ΔM4 were respectively transfected into human skin fibroblast cell line human foreskin fibroblast 1 (HFF-1) cells with the optimum multiplicity of infection (MOI) of 50. The protein expressions of sTβRII and Smad 4ΔM4 of the two types of transfected cells were determined by Western blotting so as to compare with those of the untransfected cells. The HFF-1 cells were divided into 6 groups as named below according to the random number table, with 6 dishes in each group, 1×10(4) cells per dish. Co-transfection group, transfected with the two previous lentivirus vectors, mixed with the ratio of 1:1 and MOI of 50, and then stimulated with 5 ng/mL TGF-β1 for 72 h; sTβRII group, transfected with lenti-sTβRII with MOI of 50, with the other treatment as above; Smad 4ΔM4 group, transfected with lenti-Smad 4ΔM4 with MOI of 50, with the other treatment as above; negative virus group, transfected with empty lentivirus vector, with the other treatment as above; positive control group, stimulated with 5 ng/mL TGF-β1 for 72 h; and blank control group, conventionally cultured without any other treatment. After stimulation, Western blotting and real-time fluorescent quantitative RT-PCR were respectively used to determine the protein and mRNA expressions of fibronectin in cells of each group. ELISA and Sircol collagen assay were respectively used to determine the protein expressions of connective tissue growth factor (CTGF) and total collagen in the cell culture supernate of each group. Data were processed with one-way analysis of variance and SNK-(q test).</p><p><b>RESULTS</b>(1) HFF-1 cells transfected with lenti-sTβRII and HFF-1 cells transfected with lenti-Smad 4ΔM4 respectively expressed higher levels of sTβRII protein and Smad 4ΔM4 protein compared with those of untransfected cells, confirming that HFF-1 cells transfected with the two lentivirus vectors can efficiently express the target proteins. (2) There were statistically significant differences in the protein and mRNA expressions of fibronectin in cells of the 6 groups (with F values respectively 53.536 and 24.365, P values below 0.001). The protein and mRNA expressions of fibronectin in cells of positive control group (respectively 1.60 ± 0.18 and 1.99 ± 0.40) were similar with those of negative virus group (respectively 1.60 ± 0.15 and 1.94 ± 0.28, with q values respectively 0.091 and 0.419, P values above 0.05), and they were significantly higher than those of the rest 4 groups (with q values from 5.245 to 18.228, P values below 0.05). The protein and mRNA expressions of fibronectin in cells of co-transfection group (respectively 0.60 ± 0.05 and 0.70 ± 0.11) were significantly lower than those of sTβRII group (respectively 0.89 ± 0.13 and 1.24 ± 0.17) and Smad 4ΔM4 group (respectively 0.91 ± 0.14 and 1.28 ± 0.19, with q values from 3.964 to 4.294, P values below 0.05). (3) There were statistically significant differences in the protein expressions of CTGF and total collagen in the cell culture supernate of the 6 groups (with F values respectively 107.680 and 38.347, P values below 0.001). The protein expressions of CTGF and total collagen in the cell culture supernate of positive control group were similar with those of negative virus group (with q values respectively 1.106 and 0.491, P values above 0.05), and they were significantly higher than those of the rest 4 groups (with q values from 6.414 to 26.420, P values below 0.05). The protein expressions of CTGF and total collagen in the cell culture supernate of co-transfection group were significantly lower than those of sTβRII group and Smad 4ΔM4 group (with q values from 3.424 to 7.143, P values below 0.05).</p><p><b>CONCLUSIONS</b>In human skin fibroblasts, blockage of two sites of TGF-β/Smad signaling can reduce the expression of scar related proteins which are up-regulated by TGF-β1 to a greater extent than that of blocking one single site.</p>

Ultrasound-targeted microbubble destruction combined with polyethylenimine mediated Bax gene silence in H9c2 cells / 中华超声影像学杂志

Objective To silence Bax gene in H9c2 cells by ultrasound-targeted microbubble destruction(UTMD) combined with polyethylenimine.Methods The shRNA expression plasmid targeting Bax gene was co-treated with microbubbles and polyethylenimine.Then the mixture added to H9c2 cells were irradiated by ultrasound.Grouping:① plasmid group;② Lipofectamine2000 + plasmid group; ③ UTMD + plasmid group;④ PEI + UTMD + plasmid group;⑤ PEI + SonoVue + plasmid group (without ultrasound irradiation).The transfection efficiency was assessed by fluorescence microscope and FCM.The expression of Bax mRNA and protein were detected by RT-PCR and Western Blot.Results Bax shRNA wasn't destroyed by ultrasound exposure.The transfection efficiency of PEI + UTMD + plasmid group was higher than any other groups obviously (P ＜0.05).The result of RT-PCR and Western Blot showed inhibition efficiency of Bax mRNA and protein expression in PEI + UTMD + plasmid group were (69.41 ± 4.91) ％ and (64.09 ± 2.38)％ separately,which were higher than any other groups significantly (P ＜0.05).Conclusions UTMD combined with polyethylenimine can inhibit the expression of Bax gene in H9c2 cells effectively.

Prospect of artificial muscle in medical applications / 国际生物医学工程杂志

Artificial muscle can be sorted into two classes. Electron transported materials were powered by electron field at high voltage including electric field responsive elastomer, piezoelectric and ferroelectric polymer. Other new materials are phase electrolytic phase transformation actuators, liquid crystal, molecular actuator, ets. Ion transported materials works at low voltage, pH response gels, ionic polymer metal composites, conductive polymers, as well as carbon nanotubes are belong to this type. Some of the artifrcial muscle designs showed potential application in assay sampling, eye muscle, limb prosthesis muscles, and artificial heart materiales.