Axin1调节骨骼肌GLUT4蛋白表达的作用研究

目的：探讨体轴发育抑制因子（Axin1）调节骨骼肌葡萄糖转运蛋白4(GLUT4)蛋白表达的作用及其机制。方法：利用RNAi干扰的Axin1腺病毒（Ad-siAxin1）感染C2C12小鼠骨骼肌细胞敲低Axin1,荧光显微镜和MTS实验明确Ad-siAxin1最佳感染浓度和时间。分别用携带绿色荧光蛋白的腺病毒（Ad-GFP）、Ad-siAxin1、载体质粒（vector）和Axin1质粒转染C2C12小鼠骨骼肌细胞敲低或过表达Axin1蛋白,Western印迹检测Axin1、端锚聚合酶蛋白（TNKS）和GLUT4蛋白水平。结果：荧光显微镜和MTS实验结果显示,在C2C12小鼠骨骼肌细胞中Ad-siAxin1作用的最佳浓度和时间分别为160μL和48 h。Western印迹结果显示,与Ad-GFP组相比,Ad-siAxin1组中Axin1蛋白降低（t=6.746,P<0.01）。Ad-siAxin1组TNKS蛋白水平降低（t=4.019,P<0.05）,GLUT4蛋白水平下调（t=3.248,P<0.05）。与vector组相比,转染Axin1质粒后,Axin1蛋白水平上...     

胎盘生长因子串联scFv质粒构建表达及亲和力测定

目的:构建并表达胎盘生长因子串联scFv,提升对胎盘生长因子的亲和力。方法:设计不同长度的柔性连接子连接的串联scFv,构建串联scFv质粒,并分别将串联scFv与亲本scFv于原核表达系统进行表达。通过间接ELISA分别测定串联scFv及对应亲本scFv亲和力,分别对数据进行单因素ANOVA方差分析,并使用Dunnett-t检验进行组间比较。结果:柔性连接子长度的不同,会对串联scFv亲和力造成差别。单因素ANOVA方差分析结果表明,各组亲和常数间存在明显的统计学差异,Dunnett-t检验组间比较结果可知,当串联scFv间柔性连接子（GGGGS）n长度n=4和n=5时,亲和力较亲本提升不明显（P＞0.05）;但亲和力在n=6（t335=33.90,t7A10=32.00,均P<0.000 1）及n=7（t335=91.68,t7A10=90.71,均P<0.000 1）时较两亲本均有明显提升。结论:串联scFv可以明显提高单体scFv亲和力。     

外泌体微RNA治疗骨关节炎研究进展

骨关节炎是骨科常见的顽固性疾病之一,因其高发病率、高致残率,给患者和社会带来沉重的负担。随着组织工程领域的飞速发展,外泌体治疗骨关节炎的潜力已得到证实。其中,外泌体特异性微RNA（miRNA）可抑制软骨降解和滑膜炎症、促进软骨下骨重塑,从而缓解甚至逆转骨关节炎的病理改变。本文对外泌体miRNA治疗对骨关节炎患者软骨、滑膜、软骨细胞外基质及软骨下骨的影响进行综述,以期为临床上应用外泌体miRNA治疗骨关节炎提供参考。     

基因诊断技术在脊柱结核诊断中的应用现状

脊柱结核是一种常见的肺外结核病。近年来,随着脊柱结核患者逐年增多,不典型脊柱结核患者亦逐渐增多,而不典型脊柱结核诊断较为困难,需要与一般细菌感染、肿瘤以及非结核分枝杆菌感染相鉴别。基因诊断技术是一项诊断脊柱结核的重要工具,对临床不典型脊柱结核的诊断有较高价值。本文将对脊柱结核的流行现状、诊断及基因诊断技术进行综述,以帮助临床工作者对脊柱结核做出更加准确的诊断。     

银离子、铜离子和锌离子对常见念珠菌的抗菌性能比较

目的 探讨银离子(Ag⁺)、铜离子(Cu²⁺)、锌离子(Zn²⁺)对5种常见念珠菌(白色念珠菌、热带念珠菌、近平滑念珠菌、克柔念珠菌和光滑念珠菌)的抗菌作用。方法 使用去离子水分别将硝酸银、氯化铜和氯化锌配制为0.1 mol·L^-1的水溶液。取保存的白色念珠菌标准菌株ATCC-60193、近平滑念珠菌、光滑念珠菌、热带念珠菌、克柔念珠菌,加入溶菌肉汤培养液,使用比浊仪调配菌液浊度为0.5麦氏单位,然后使用无菌去离子水将菌液稀释至10⁶ CFU·L^-1。取配制好的0.1 mol·L^-1的硝酸银、氯化铜和氯化锌溶液,分别倍比稀释为6个浓度(10^-2、10^-3、10^-4、10^-5、10^-6、10^-7 mol·L^-1),分别取各浓度的金属离子溶液100μL与预制好的5种念珠菌悬液100μL进...     

Triptolide inhibits IL-12 production and T cell-stimulatory capacity of dendritic cells

Extracts of Tripterygium wilfordii Hook F.(TWHF ) are effective in traditional Chinesemedicine for treatment of autoimmune diseasessuch as rheumatoid arthritis, systemic lupus ery thematosus, nephritis and asthma [1, 2]. Trip tolide, a diterpenoid triepoxide, is derived fromTWHF and is responsible for most of the immuno suppressive and anti inflammatory effects ofTWHF. Triptolide has been shown to be effectivein the treatment of autoimmune diseases, …     

The Distribution of Synaptotagmin II in RBL-2H3 and Its Regulation on Exocytosis of Lysosomes in RBL-2H3

Synaptotagmin(Syt)constitutes a family of membrane-trafficking proteins,so far nearly 20 Syts have beendiscovered.Extensive work showed that synatotagmins were a potential Ca~(2+) sensor for regulated exocytosis.Thisstudy was to investigate the expression and location of synaptotagmin Ⅱ(Syt2)in RBL-2H3(RBL)and its role inregulating exocytosis of RBL.The expression of Syt2 in RBL was confirmed by Western blot.The recombinantexpression vector pEGFP-N1-Syt2 was constructed and transfected into RBL by electroporation,the stabletransfectant RBL-Syt2-S expressing fusion protein Syt2-EGFP were obtained and Syt2 was highly concentrated atplasma membrane with little detected in cytoplasm.To analyze the role of Syt2 during exocytosis of RBL,therelease of cathepsin D was assayed by immunoblotting.Compared with control,the release of cathepsin D byRBL-Syt2-S was markedly decreased.The results indicated that Syt2 played a negative regulation in exocytosis oflysosomes in RBL.Cellular & Molecular Immunology.2005;2(3):205-209.     

The Effects of Extracellular Matrix on Tissue Engineering Construction of Cartilage in Vitro

In the study of bone and cartilage diseases in our laboratory, chondrocyte stereo-differentiation mod-el was cultured into cartilage tissue. The cultured cartilage tissue was closely similar to natural tissue inboth histological condition and biochemical metabolism. The model mimicked the chondrocyte differenti-ation and metabolism in vivo completely〔1〕. We studied some essential extracellular matrices, includingcollagen, proteoglycan(PG) and hyaluronic acid(HA) on the construction of grow…     

亚定点医院在重大突发公共卫生事件中的实践探索

探索重大突发公共卫生事件中以亚定点医院为代表的医疗管理模式。从上海新国际博览中心W1亚定点医院的实际运行情况出发,对其医疗供需、收治标准、运行效果、存在问题等逐一分析,结合本医疗队的经验,提出新的工作模式和管理思路。亚定点医院为普通型和有基础疾病的新冠病毒阳性感染者提供及时有效的救治,缓解了定点医院的运行压力,但也暴露出人员配置、信息化支撑、院感防控等方面的问题。采取“两级缓冲、双向转诊”的工作模式和“三个快、三个准、三个稳、三个全”的管理方案,可为患者提供及时、有效、适宜的医疗服务,缓解定点医院运行压力。