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Article in Chinese | WPRIM | ID: wpr-933667


Objective:To explore the construction and mechanism of Mindin gene specific macrophage knockout mice in acute lung injury induced by lung ischemia-reperfusion injury(IRI).Methods:Mindin gene knockout mice were constructed by CRE-LOP system, Mice were divided into four groups of C57/B6 wild-type mice sham operation(n=10), C57/B6 mice operation(n=10), Mindin-/-macrophage-specific knockout mice operation(n=10)and C57/B6 mice operation + Mindin recombinant protein intervention(n=10). And lung ischemia-reperfusion injury model was established by clamping pulmonary portal.The effects of Mindin gene knockout and recombinant protein intervention on acute lung injury were observed in vivo and in vitro.t-test and ANOVA test were employed for data processing.Results:Mindin gene macrophage specific knockout mice was successfully constructed.Surgery(Mindin-/-)group significantly reduced pulmonary edema, release of inflammatory factors(IL1β: 2.73±0.19 vs. 5.81±0.61; IL-18: 6.52±0.63 vs. 11.03±0.34; TNF-α 2.18±0.14 vs. 4.76±0.20; HMGB1: 4.57±0.33 vs. 8.76±0.87), expression of NLRP3(2.07±0.27 vs. 4.91±0.22)and secretion of GSDMD(2.78±0.37 vs. 5.78±0.29)as compared with surgery group in vivo.In surgery(WT)+ Mindin group, the expression of lung IRI, inflammatory factors and cell pyroptosis were opposite, And the results were consistent in vitro and in vivo.As compared with surgery group, the above parameters were up-regulated in surgery(WT)+ Mindin protein group.And inter-group differences were statistically significant(all P<0.05). In vitro, the expressions of NLRP3(1.00±0.36, 0.41±0.06, 4.13±0.23), GSDMD(1.00±0.17, 0.34±0.16, 6.32±0.46)and integrin β4(1.00±0.11, 0.28±0.07, 3.53±0.17)were detected in different groups including hypoxia-recovery oxygen(HR), HR+ Mindin siRNA and HR+ Mindin protein groups in macrophage cell line(J774A); As compared with HR group, the above parameters were up-regulated in HR+ Mindin protein group and down-regulated in HR+ Mindin siRNA group.And the differences were statistically significant( P<0.05). The expressions of NLRP3(1.00±0.07, 1.13±0.11, 0.51±0.14)and GSDMD(1.00±0.09, 0.87±0.16, 0.37±0.12)were detected in Mindin, Mindin protein+ vehicle and Mindin protein+ integrin β4 knockout groups.The above parameters were down-regulated in Mindin protein+ integrin β4 knockout group as compared with Mindin protein and Mindin protein + vehicle groups.And the inter-group differences were statistically significant(all P<0.05). Conclusions:During pulmonary IRI, Mindin knockdown can alleviate pulmonary IRI.Mindin gene may promote the expression of inflammatory factors, NLRP3 and GSDMD protein by activating integrin β4 and aggravate cell pyroptosis to promote the development of pulmonary IRI.

Article in Chinese | WPRIM | ID: wpr-873543


@#Objective    To evaluate the safety, feasibility and short-term outcomes of single-direction gastric mobilization under 3D-laparoscopy in minimally invasive esophagectomy for the treatment of esophageal cancer. Methods    From February 2018 to December 2019, 118 consecutive patients who underwent minimally invasive McKeown esophagectomy for esophageal squamous cell carcinoma in our hospital were included. There were 94 males and 24 females with an average age of 53.7 (41–77) years. They were divided into two groups based on the methods of gastric mobilization: a traditional dissociation (TD) group (n=55) and a single-direction mobilization (MD) group (n=63). The clinical data of the two groups were compared. Results    Enbloc resection and a negative resection margin were obtained in all patients. There was no postoperative mortality or incision complication. The rate of postoperative complications was 22.9%. There was no significant difference in the spleen injury, gastric injury, conversion to open surgery, abdominal reoperation as well as cervical anastomotic leakage between the two groups (P>0.05). It took significantly less time in the MD group compared with the TD group (P<0.05). There was an obvious statistical difference in the incidence of gastric mobilization related complications between the MD group (1.6%, 1/63) and TD group (12.7%, 7/55, P<0.05). Conclusion    Application of single-direction gastric mobilization under 3D-laparoscopy in minimally invasive esophagectomy for the treatment of esophageal cancer is safe and easy to perform with a satisfactory short-term outcome.

Article in Chinese | WPRIM | ID: wpr-383105


Objective Pentamethylquercetin (PMQ) has a role in cardiovascular protection. We investigate the effects of 3,3' ,4' ,5,7-pentamethylquercetin, a derivative of PMQ, on intimal hyperplasia of the vein grafts in rats both in vivo and in vitro. Methods The proliferation of vascular smooth muscle cells ( VSMC ) was induced with Ang Ⅱ (0. 1μmol/L, 24 h)while PMQ was administrated at six different dosages (0. 1, 0.3, 1,3, 10 and 30 μmoL/L). Cell viability was identified with MTT; ROS was measured with DCFH-DA; and the expression of NADPH oxidase subunits Nox1, p47phox, and p22phox mRNA were measured with real-time PCR. For the experiment in vivo, 24 SD rats were randomly assigned to control group and PMQ groups, the latter was further divided into three different dosage groups. In the control group, solvent was administrated daily via gavage. In PMQ groups, PMQ ( 12.5 mg/kg, 25 mg/kg, 50 mg/kg) was administrated daily respectively in the same way.All SD rats received operation performed by one person. Reversed external jugular vein was implanted into the external carotid of the same side with interrupted suture. 4 weeks after operation, all vein grafts were harvested. Status of the vein grafts was observed and tissue sections were analyzed with HE staining. The intimal hyperplasia ( intima/media area index and intima/media thickness index) of the vein grafts was assessed. Results Cell viability and ROS of VSMC induced by Ang Ⅱ were suppressed by PMQ. Cell viability and ROS of VSMC were increased substantially when treated with Ang Ⅱ. The therapeutic effects of PMQ could be initially identified at dose of0. 3 μmol/L, with a peak at 3 μmol/L. The effects decreased from 30μmol/L to 10 μmol/L. PMQ at dose of 0.1 μmol/L had no effect on cell viability and ROS of VSMC induced by Ang Ⅱ. PMQ also downregulated the mRNA expression of NADPH oxidase subunits Nox1, p47phox and p22phox induced by Ang Ⅱ. A peak effect was observed at 3μmoL/L and decreased at 30 μmol/L. PMQ at o. 1 μmol/L had no effect on mRNA expression of NADPH oxidase subunits induced by Ang Ⅱ. As compared with control group, PMQ decreased intima/media area index ( 1. 64 ±0.20 in control, 0. 74 ±0.18 at 12.5 mg/kg, 1.09 ±0.17 at 25 mg/kg, 1.21 ± 0. 21 at 50 mg/kg) and intima/media thickness index ( 1.34 ± 0. 24 in control, 0.67 ± 0. 17 at 12.5 mg/kg, 0. 74 ± 0.14 at 25 mg/kg, 0.93 ± 0. 18 at 50mg/kg) at three dosages after implantation. Conclusion PMQ may suppress the proliferation of VSMC and inhibit neointima hyperplasia of vein grafts in rats. The effects may be attributed to the anti-oxidative activity and the downregulation of mRNA expression of NADPH oxidase subunits Noxl, p47phox and p22phox.