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@#[摘 要] 目的:探讨恶性肿瘤中Dickkopf-1(DKK1)表达对CD4+ T细胞极化的影响及其作为肿瘤免疫治疗靶点的潜在价值。方法:利用生物信息学方法分析DKK1在多种类型肿瘤组织和癌旁组织中的表达水平,分析DKK1表达与肿瘤患者预后及肿瘤微环境免疫浸润间的相关性。利用流式细胞术检测DKK1蛋白对CD4+ T细胞表型变化的影响。构建黑色素瘤B16F10细胞小鼠皮下移植瘤模型,观察阻断DKK1对小鼠移植瘤生长和移植瘤组织中免疫细胞浸润与表型的影响。结果:DKK1 mRNA表达水平在多种肿瘤组织中显著高于癌旁组织,DKK1高表达与多数肿瘤患者的不良预后相关且在多数肿瘤中DKK1对CD4+ T细胞抗肿瘤免疫应答功能有重要负性调节作用(P<0.05或P<0.01)。流式细胞术检测结果显示,DKK1蛋白刺激可显著降低CD4+ T细胞中T-bet、IFN-γ及CD107a表达水平(均P<0.01)。在小鼠皮下黑色素瘤模型中发现,阻断DKK1可以显著抑制小鼠移植瘤的生长(P<0.01),有效改善抗肿瘤免疫应答,表现为Th1细胞(T-bet+ CD4+)占比显著升高(P<0.001),效应性CD8+ T细胞(CD44+ CD62L-)占比显著升高(P<0.01),Th2细胞(GATA3+ CD4+)与Treg细胞占比显著下降(均P<0.01)。结论:阻断DKK1可有效促进CD4+ T细胞向Th1型极化,DKK1具有作为肿瘤免疫治疗靶点的潜在价值。
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@#Objective To compare the patency of the postoperative vessel bridge grafts between the notouch technique and the conventional technique for saphenous vein harvest in the coronary artery bypass grafting. Methods A total of 70 patients underwent coronary artery bypass grafting from January 2014 to January 2015 in the First Affiliated Hospital of Zhengzhou University. They were randomly divided into a conventional technique group (saphenous vein obtained by traditional technique) and a no-touch technique group (saphenous vein obtained by notouch technique). There were 34 patients in the conventional technique group, including 16 males and 18 females with an average age of 61.6±6.4 years. There were 36 patients in the no-touch technique group, including 16 males and 20 females with an average age of 62.2±6.6 years. The 1-year postoperative coronary CT angiography (CTA) results were compared. Results Seventy patients underwent coronary CTA examination 1 year postoperatively. The patency rate of the conventional technique group and no-touch technique group were 83/116 (71.6%) and 113/122 (92.6%). Logistic regression analysis showed a statistical difference between the non-touch technique and the conventional technique (P<0.05). Conclusion The short-term patency rate of vessel bridge grafts is higher when the no-touch technique is used.
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@#Objective To analyze the efficacy of no-touch technique and traditional technique in drawing great saphenous vein during coronary artery bypass grafting. Methods We searched the literatures on no-touch technique versus traditional technique in drawing great saphenous vein during coronary artery bypass grafting in PubMed, Cochrane Library, Chinese Journal Full-text Database, Wanfang database between January 1997 and November 2017. Jadad scale was used for quality verification. RevMan 5.0 was used for analysis. Results Six studies were included. Jadad scale for both 6 randomized controlled trials was 5 points. Meta-analysis showed that there was no statistical difference in postoperative mortality between the two groups with RR=0.68 and 95%CI 0.11 to 4.11(P=0.67). There was no statistical difference in leg wounds with RR=1.46 and 95%CI 0.23 to 9.16 (P=0.68). There was no statistical difference in short-term cardiogenic death with RR=0.33, 95%CI 0.10 to 1.03 ( P=0.06). The no-touch group’s long-term cardiogenic death was significantly lower than the traditional group withRR=0.36, 95%CI 0.16 to 0.79(P=0.01). Postoperative no-touch group’s short-term patency rate was significantly better than that of the traditional group with MD=3.08 and 95%CI 1.58 to 5.99 (P<0.01). The long-term patency rate in the no-touch group was also higher than that of the traditional group withMD=3.08 and 95%CI 1.58 to 5.99 (P<0.01). Conclusion Compared with the traditional group, the no-touch group increases postoperative long-term survival rate, short-term and long-term patency rate.
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Background Lysozyme plays a crucial role in innate immunity with its well-recognized bacteriolytic activity. In this study, the influence of expression parameters (inoculation volume, culture volume, growth time, induction temperature and time, initial pH and methanol concentration) on human lysozyme (HLZ) production in recombinant P. pastoris SMD1168 was investigated through Plackett-Burman (PB) design and response surface methodology (RSM). Results It was revealed that induction temperature, induction time and culture volume had significant influence (P < 0.01) on HLZ expression level, which were elected for further optimization with three-dimensional response surface designs for enhanced HLZ production. The highest lysozyme activity reached 3301 U/mL under optimized conditions (at 23.5°C for 90 h with culture volume of 48 mL) in shake flask, which increased 2.2 fold compared with that achieved with the standard protocol (Invitrogen). When high-cell-density fermentation of the recombinant Pichia pastoris was performed in a 15 L fermenter under optimized conditions, the extracellular lysozyme activity reached 47,680 U/mL. SDS-PAGE analysis of the product demonstrated that HLZ was produced as a single major protein with a molecular weight of approximately 14.7 kDa, consistent with its expected size. Conclusions The results indicated that the optimized culture conditions using PB design and RSM significantly enhanced the expression level of HLZ, and the Pichia expression system for HLZ production was successful and industrially promising.