ObjectiveTo explore the clinical role of island flap nourished by sural nerve nutrition blood vessel for repairing of the deep burn wounds of the lower extremities.MethodsIsland flap nourished by sural nerve nutrition blood vessel were performed to repair the deep burn wounds of the lower extremities below the knee in 22 cases,aged 10 ～58 years old,average aged 34 years old,including flame burns in 6 cases,electrical burns in 10 cases,chemical burns in 4 cases,scalding hot water in 2 cases.All were deep skin damage,The island flaps,including anterograde and retrograde flaps,was from 5cm ×4cm to 13cm × 10cm in size,The defect at the donor site was sutured primarily or repaired by a skin graft.ResultsAll the flaps survived well,18 patients were follow-up postoperatively for 4 ～24 months.The color,texture,shape of the flaps were satisfactory,but the sensory recovery was unsatisfactory.ConclusionSural neurovascular flap had a constant anatomy and reliable blood supply without the sacrificeof the major blood vessels.The procedure was simple and easy,and an effective method of repairing the skin and soft tissue defects below the knee.

Objective To evaluate the normalized correction effect of the ROX reference fluorescence on the real-time quantitative RT-PCR assessment for the Nrf2 mRNA.Methods The Taq-man probe was utilized in the real-time quantitative RT-PCR method to assess the Nrf2 mRNA; The reaction specificity of the Nrf2 PCR products obtained from the rat cDNA was assessed using gel electrophoresis and sequencing identification.The PCR calibrators with different concentrations varied at 10 gradients(from 2.0 × 109 to 2.0 × 100) were prepared for real-time quantitative RT-PCR assessment,and then the influence of the ROX reference fluorescence correction on the linear range of the PCR standard curve was investigated experimentally.The recombinant plasmids with different concentration levels(e.g,high,medium or low concentration) were assessed by the real-time RT-PCR and 20 replicate measurements were performed for all the samples in both intra- and inter-assays.The repeatability differences of both the inter- and intra-assays were analyzed systemically with/without the ROX reference fluorescence correction.Results The primers and probes required for Nrf2 mRNA real-time quantitative RT-PCR assessments were constructed,and the reaction system and conditions were also established.The length of the amplified fragment agreed well with the expected length(122 bp),and the sequencing analysis showed the amplified fragment was verified by the sequencing identification.After applying the ROX reference fluorescence correction,the linear range ofthe PCR standard curve was measured as 2.0 × 109 -2.0 × 100 copies/μl(R2 ＞ 0.99),which was 100times wider than that measured without ROX reference fluorescence correction(2.0 × 109 - 2.0 × 102copies/μl,R2 ＞ 0.99).Without the ROX reference fluorescence correction,the intra-assay CVs of the Ct value were measured as 4.1%,2.7% and 2.1% for the recombinant plasmids with high-,medium-,and low-level concentration,respectively.The inter-assay CVs of the Ct value were 4.3% 、3.0% 、2.4%.By contrast,with the ROX reference fluorescence correction,the intra-assay CVs of the Ct value became 0.7%,0.5%,0.4% and the inter-assay CVs of the Ct value turned to 1.0%,0.8% and 0.7%.The discrete degree of the Ct value with the correction was lower than that without the correction.The intra- and interassay Ct values of the high- and medium-level samples exhibited statistic significance for the measurements with/without the correction.In the high-level group,the t values of the intra- and inter-assays were 4.843 and 2.566,with P＜0.05.In the medium-level group,the t values of the intra- and inter-assays were 4.293 and 4.423,with P＜0.05.However,no statistic significance was observed the low-level group(the t values of the intra- and inter-assays were 0.753 and 1.279,with P ＞.0.05).Conclusions The utilization of the ROX reference fluorescence in the real-time quantitative PCR assessment for Nrf2 mRNA could widen the linear range of the standard PCR calibration curve and improve the repeatability of the assessment,which should be quite helpful for accurately assessing the expression level of the Nrf2 mRNA.

Objective To explore the effect of elastic outside distraction on microvessel density and to illustrate the mechanism correcting inverted nipples. Methods Three female 3-month-old minipigs,with 12 nipples each,were employed.4 nipples of each minipig were remained without distraction were control group,and the other 8 nipples were continuously distracted with inverted nipple correction instruments.Corresponding nipples were excised at 2,4,8 and 12 weeks after traction.The excised nipple specimens were employed with immunohistochemical staining to detect the expression of vascular endothelial cells growth factor (VEGF) and CD34 in those tissues,and the percentageof positive cells and microvessel density (MVD) changes were counted.Results The expression of CD34 and VEGF in the experimental groups increased at 2 weeks after tracted,and reached at the peak at 4 weeks,then gradually decreased during 8 to 12 weeks after tracted.While those were weakly positive in control group.Compared with the control group,the integral optical density of VEGF-positive cells and MVD were significantly different (P＜0.01),and there was a significantly positive correlation between MVD and VEGF.Conclusions Continuous elastic outside distraction can promote the expression of VEGF and CD34 in female minipig nipple and enhance angiogenesis,which increases oxygen supply locally,and therefore those local tissue proliferation may be one of key mechanisms of correcting the inverted nipple by continuous elastic outside distraction.