To explore the effect of high glucose on the NF-κB/IκB signal pathway in cultured rat glomerular mesangial cells. The results showed that high glucose increased the degradation of IκB-α and the translocation to nucleus of NF-κB. These changes could be reverted mostly by MG132, a proteasome inhibitor. It suggests that the activation of the NF-κB signal pathway by high glucose concentration may probably be via the ubiquitin-proteasome pathway.

PURPOSE: Circulating free DNA (cfDNA) in plasma is promising to be a surrogate for tumor tissue DNA. However, not all epidermal growth factor receptor (EGFR) mutations in tumor tissue DNA has been detected in matched cfDNA, at least partly due to inefficient cfDNA extraction method. The purpose of this study was to establish an efficient plasma cfDNA extraction protocol. MATERIALS AND METHODS: The yield of plasma cfDNA extracted by our modified phenol-chloroform (MPC) method from non-small-cell lung cancer (NSCLC) patients was compared with that by QIAamp MinElute Virus Spin kit (Qiagen kit) as control, using the Wilcoxon rank-sum test. TaqMan quantitative polymerase chain reaction (qPCR) assays were used to quantify the plasma cfDNA extracted. Both Mutant-enriched PCR (ME-PCR) coupled sequencing and DxS EGFR mutation test kit were used to evaluate the impact of extraction method on EGFR mutation analysis. RESULTS: MPC method extracted more plasma cfDNA than Qiagen kit method (p=0.011). The proportion of longer fragment (> or =202 bp) in cfDNA extracted by MPC method was significantly higher than by Qiagen kit method (p=0.002). In the sequencing maps of ME-PCR products, a higher mutant peak was observed on plasma cfDNA extracted by MPC method than by Qiagen kit method. In DxS EGFR mutation test kit results, plasma cfDNA extracted by MPC method contained more tumor-origin DNA than by Qiagen kit method. CONCLUSION: An improved plasma cfDNA extraction method of MPC is provided, which will be beneficial for EGFR mutation analysis for patients with NSCLC.
Base Sequence ; Carcinoma, Non-Small-Cell Lung/*genetics ; Chloroform ; DNA Mutational Analysis/*methods ; DNA, Neoplasm/*blood/*isolation & purification ; Genetic Testing/methods ; Humans ; Lung Neoplasms/*genetics ; Molecular Sequence Data ; Phenol ; Polymerase Chain Reaction/methods ; Receptor, Epidermal Growth Factor/*genetics

Objective To provide anatomical information and clinical application of periosteal perforator bone-skin flap of proximal lateral tibia. Methods From March, 2015 to March, 2017, 15 fresh cadavers who underwent injected with imaging technology and dissected with layer by layer. The origins, branches, distribution and anastomosis of periosteal perforator vessels in the proximal lateral tibial were observed. Sixteen patients of composite tissue defect in hands and feet were repaired with the method of free transplantation of this flap from March, 2015 to March, 2017. Injured area was from 3.0 cm × 0.8 cm to 6.0 cm × 5.5 cm. Bony defect size was from 1.7 cm × 1.5 cm × 1.0 cm to 5.0 cm × 1.0 cm × 1.0 cm. The bone-skin flap size ranged respectively from 3.0 cm × 0.8 cm to 6.0 cm × 5.5 cm and 1.6 cm ×1.0 cm × 0.8 cm to 5.0 cm×1.0 cm × 1.0 cm. Postoperative followed-up was done termly. Results The diameter and superficial length of the main perforators respectively were 0.5 to 1.2 mm and from 2.5 to 4.3 cm. The followed-up time was from 6 to 24 months in 14 cases, with the results of the bone-skin flaps presented favourable contours and good functions. The healing time of bone flap was 2 to 4 months. The function of shank was normal. Conclusion The periosteal perforator of proximal lateral tibia has favourable appearance, constant vascular pedi-cle, reliable blood supply and large diameter. The free transplantation of this flap offers a satisfactory alternative for repairing the small and medium-sized area of composite tissue defects of hands and feet.

Objective: To report the clinical effect of skin grafting in small wounds with exposed vascular anastomosis. Methods: From January, 2011 to May, 2018, 16 small wounds with anastomotic vascular exposure were treated by full-thickness skin grafting. Of which, 4 performed after replantation, 9 after reconstruction and 3 after flap transplantation. Thirteen wounds were on hand and 3 in foot. After anastomosing the vessels, 3 arterial anastomoses, 9 venous anastomoses and 4 arterial-and-venous anastomoses were left exposure in wounds. Sizes of artery exposed in wound were 0.8 to 2.3 mm with an average of 1.0 mm. Sizes of vein exposed in wound were 0.8 to 2.5 mm with an average of 1.2 mm. The areas of soft tissue defect were 1.0 cm×1.5 cm to 2.6 cm×6.0 cm, and the areas of grafted skins were 1.0 cm×1.5 cm to 2.6 cm×6.0 cm. Grafted skin were covered without package nor pressurization. Donor areas were directly sutured. Postoperative follow-up was conducted to observe the postoperative effect. Results: Fourteen grafted skin completely survived, one partially survived and healed after immobilization of the limb and change of dressing, and one developed necrosis. All patients were followed-up for 6-24 months (mean 14.4 months). CDU, HHD or CTA were used at the final follow-up. Vascular anastomoses were patency in 15 patients, and 1 patient had embolism developed. No pigmentation was found on the grafted skin. All grafted skin was soft and wearable with two point discrimination at 7-10 mm. The pulse of anastomotic artery could be felt on the grafted skins. Only linear scars were left in the donor sites. Conclusion The operation of full-thickness skin grafting in small wounds with exposed vascular anastomosis was easy to perform and with high survival rate. The effect of operation is satisfactory. The exposure of anastomosed vessels does not affect the patency of anastomotic vessels, and has considerable clinical values.