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【Objective】 To explore the role and mechanism of TRPC in promoting extracellular matrix (ECM) deposition in rat glomerular mesangial cells (HBZY-1). Methods Immunofluorescence staining was performed to observe the distribution and expression of TRPC1 and TRPC6 in HBZY-1 cells. After AngⅡ stimulation, qRT-PCR and Western blotting were used to detect the mRNA and protein expressions of Gαq/PLCβ4/TRPC signaling pathway main proteins and ECM deposition indicators (α-SMA, collagenⅢ and fibronectin). By silencing the expressions of TRPC1 and TRPC6 by RNA interference, the expressions of ECM deposition indicators were detected. Changes in [Ca2+]i influx were determined through Fluo-4AM Ca2+ imaging. 【Results】 Both TRPC1 and TRPC6 were expressed in HBZY-1, and were mainly located in cell membrane and cytoplasm. After AngⅡ stimulation, Gαq/PLCβ4/TRPC signaling pathway was activated, and the mRNA and protein expressions of Gαq, PLCβ4, TRPC1 and TRPC6 were all increased (P<0.05). [Ca2+]i influx also increased (P<0.01), and the mRNA and protein expressions of ECM deposition indicators (α-SMA, ColⅢ and Fn) were upregulated (P<0.05). Silencing the expressions of TRPC1 and TRPC6 by RNA interference led to decreased [Ca2+]i influx (P<0.05), and downregulated mRNA and protein expressions of ECM deposition indicators in HBZY-1 cells (P<0.05). The results suggested that inhibition of TRPC expressions could inhibit AngⅡ induced ECM deposition in HBZY-1 cells, which might be associated with decreased [Ca2+]i influx. 【Conclusion】 TRPC may be a novel therapeutic target of renal fibrosis.
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Objective@#This study aims to analyze the outcome of free perforator flap for repairing soft tissue defects on the dorsum of foot.@*Methods@#Thirty-six patients with soft tissue defects on the dorsum of foot were treated at a single institution from March 2015 to September 2017. They were 20 males and 16 females, aged from 21 to 59 years old, with the mean age of 39.4 years. The causes of injury include crush injury (n=19), traffic injury (n=15), and electric injury (n=2). The injury site involves left foot (n=19) and right foot (n=17). The defect area of soft tissue was from 4.4 cm ×6.6 cm to 7.1 cm×16.2 cm in size. The vacuum sealing drainage dressing (VSD) was performed for all patients at the first stage of operation. The fracture and dislocated bone were fixed using Kirschner needle and plate screw. The ruptured tendon was repaired at the same time. The flap transplantation was performed at the second stage of operation. Twelve patients were treated with free anterolateral thigh perforator flap, 15 patients were treated with free sural artery perforator flap, and 9 were free peroneal perforator flap. The skin flap was from 4.9 cm× 7.2 cm to 7.9 cm × 17.8 cm in size.@*Results@#All 36 flaps survived. The wounds of both donor and recipient area primarily healed, without infection or skin necrosis. The venous crisis occurred in 1 patient of anterolateral thigh flap and 1 patient of free medial sural flap within 24 hours after surgery. Both flaps survived after stitches removed and blood cleaned. All patients were followed up for 6—22 months, with an mean follow-up period of 9.8 months. Except for the swollen anterolateral thigh perforator flap in 2 patients, the other patients were satisfied with the shape of flap and in wearing shoes. The sensation of flap was good, and the two-point discrimination was 11—16 mm. There was no obvious dysfunction occurred in the donor site.@*Conclusions@#The free anterolateral thigh perforator flap, free sural artery perforator flap and free peroneal perforator flap are suitable for repairing soft tissue defects on the dorsum of the foot. A reasonable surgical plan can help to obtain satisfactory clinical outcome.
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Objective: To establish the UPLC typical chromatogram and evaluate the quality of Jinji pills. Methods: The UPLC typi-cal chromatogram was performed on an Agilent proshell 120 C18column (150 mm×4. 6 mm,2. 7 μm) with gradient elution by the mobile phase of acetonitrile-0. 1% phosphoric acid aqueous solution system at a flow rate of 0. 8 ml·min-1. The detection wavelength was 240 nm. Results: The UPLC typical chromatogram included 15 common peaks when taking berberine hydrochloride as the reference peak, and 7 of them were identified by the comparison with the reference substances. Conclusion: The established methods have high specificity and good repeatability, and can be used for the quality control of the product.
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Objective:To identify toddalolactone and establish the analysis method for Jinji tablets.Methods: An Hypersil DBS C18column(250 mm×4.6 mm,5 μm) was used. The mobile phase consisted of acetonitrile-0.1% phosphoric acid solution. The flow rate was 0.8 ml·min-1and the detection wavelength was at 330 nm. Toddalolactone was identified by an LC-MS method and the re-sult was compared with that of the reference regent.TLC and HPLC analytical methods were used for analysis of toddalolactone.Re-sults:Totally 58 batches of Jinji tablets were tested,and the suspected samples were confirmed by HPLC-MS/MS.Toddalolactone was detected out from 20 batches of samples. Conclusion:The method is simple,rapid and accurate, which is able to detect toddalolac-tone quickly and effectively.