Pristimerin enhances recombinant adeno-associated virus vector-mediated transgene expression in human cell lines in vitro and murine hepatocytes in vivo

OBJECTIVE:In the present study,we systemically evaluated the ability of two bioactive compounds from traditional Chinese medicine,celastrol and pristimerin,to enhance recombinant adeno-associated virus(rAAV) serotype vector-mediated transgene expression both in human cell lines in vitro,and in murine hepatocytes in vivo.METHODS:Human cell lines were infected with rAAV vectors with either mock treatment or treatment with celastrol or pristimerin.The transgene expression,percentage of nuclear translocated viral genomes and the ubiquitination of intracellular proteins were investigated post-treatment.In addition,nonobese diabetic/severe combined immunodeficient gamma(NSG) mice were tail vain-injected with rAAV vectors and co-administered with either dimethyl sulfoxide,celastrol,pristimerin or a positive control,bortezomib.The transgene expression in liver was detected and compared over time.RESULTS:We observed that treatment with pristimerin,at as low as 1 umol/L concentration,significantly enhanced rAAV2 vector-mediated transgene expression in vitro,and intraperitoneal coadministration with pristimerin at 4 mg/(kgd) for 3 d dramatically facilitated viral transduction in murine hepatocytes in vivo.The transduction efficiency of the tyrosine-mutant rAAV2 vectors as well as that of rAAV8 vectors carrying oversized transgene cassette was also augmented significantly by pristimerin.The underlying molecular mechanisms by which pristimerin mediated the observed increase in the transduction efficiency of rAAV vectors include both inhibition of proteasomal degradation of the intracellular proteins and enhanced nuclear translocation of the vector genomes.CONCLUSION:These studies suggest the potential beneficial use of pristimerin and pristimerincontaining herb extract in future liver-targeted gene therapy with rAAV vectors.     

肘关节后外侧旋转不稳定韧带损伤的载线影像学特点研究

目的 ：探讨肘关节外侧韧带复合体对肘关节后外侧旋转不稳定的作用及韧带损伤时肘关节侧位X线影像学中桡肱率变化特点。方法：取冷冻保存的新鲜成人肘关节标本20侧,将标本制成肘关节＂骨-韧带＂标本,随机分为两组。利用自行设计的维持肘关节后外侧旋转活动度的设备固定标本,A组按次序切断外侧结构：A1组,外侧韧带完整的标本组;A2组,切断桡侧尺副韧带组;A3组,增加切断环状韧带组;A4组,最后切断桡侧副韧带组。B组分为：B1组,外侧韧带完整的标本组;B2组,切断桡侧副韧带组;B3组,再切断环状韧带组;B4组,最后切断桡侧尺副韧带组。分别摄肘关节侧位X线片,在PACS系统中测量桡肱率,比较各组的差异,并统计分析。结果：A组,统计分析各组数据,各组间桡肱率增加差异有统计学意义。B组,统计分析前2次切断韧带,各组间桡肱率增加差异无统计学意义。切断桡侧尺副韧带（B4组）,与前3组比较差异有统计学意义。结论：桡侧尺副韧带是限制肘关节后外侧旋转脱位的主要结构,环状韧带、桡侧副韧带为次要结构;并根据肘关节后外侧旋转不稳定X线影像学特点将其分为4个等级。     

The effect of collagen-binding NGF-β on the promotion of sciatic nerve regeneration in a rat sciatic nerve crush injury model

Nerve growth factor plays a critical role in peripheral nerve regeneration. However, the lack of efficient NGF delivery approach limits its clinical application. It has demonstrated in our previous work that the native human NGF-β (NAT-NGF) fused with a collagen-binding domain (CBD) could bind to collagen specifically. Since collagen is the major component of nerve extracellular matrix, we speculated that the collagen-binding NGF would target to nerve cells and improve their regeneration. In this report, we found that the fusion protein could specifically bind to endogenous collagen of the rat sciatic nerves and maintain NGF activity both in vitro and in vivo. In the rat sciatic nerve crush injury model, we found that collagen-binding NGF could be retained and concentrated at the nerve injured site to promote nerve repair and enhance function recovery following nerve damage. Thus, the collagen-binding NGF could improve the repair of peripheral nerve injury.     

运用UV打印图片和仿真模型对肠造口患者造口周围皮炎自我护理能力的影响

目的 探讨术前运用UV打印图片和仿真模型对肠造口患者术后造口周围皮炎自我护理能力的效果,以寻找一种更有效的健康宣教模式。 方法 选择2019年1—8月在温州医科大学附属第二医院择期行肠造口术患者共80例,采用随机数字法分为对照组和观察组,各40例,对照组给予一般护理,观察组在对照组基础上术前1周采用UV打印图片和仿真模型进行健康教育,观察2组患者护理知识评分和达标率、自我护理能力评分以及造口周围皮肤受损程度。 结果 出院前1 h观察组护理知识得分高于对照组[(85.6±9.5)分vs.(77.6±10.2)分,t=3.630,P<0.001],观察组的达标率高于对照组(85.0%vs. 65.0%,χ2=4.267,P=0.039);观察组1个月和3个月自我意愿、自护知识和自护技能得分均明显高于对照组(P<0.05);观察组造口皮肤评估工具(DET)评分明显低于对照组[出院前:(4.1±0.6)分vs.(7.8±1.5)分,出院1个月:(2.6±0.5)分vs.(5.3±1.2)分,出院3个月:(1.4±0.3)分vs.(3.1±0.8)分],均P<0.05。 结论 术前运用UV打印图片和仿真模型较常规健康教育能够显著提高肠造口患者造口周围皮炎的自我护理能力,同时能增强护理知识水平,减少造口周围的皮肤伤害,值得推广。