重叠延伸PCR技术定点突变TFEB原核表达载体的构建及体外诱导表达和纯化

目的：基于重叠延伸PCR (SOE PCR)技术构建转录因子EB (TFEB)丝氨酸114位点突变体原核表达质粒,并进行体外诱导表达和纯化。方法：根据SOE PCR技术原理设计突变引物,以pGEX-6p-1-TFEB质粒为模板,分别采用外侧引物F和R及突变引物Fn和Rn进行第1步PCR扩增,获得含突变位点的产物1和产物2。经第2步PCR退火延伸对产物1和产物2进行重叠拼接,以拼接后DNA片段为模板,采用外侧引物F和R进行第3步PCR扩增获得含突变位点的目的 DNA;将其克隆至pGEX-6p-1载体,采用Bam HⅠ和SalⅠ进行酶切鉴定,DNA测序验证突变结果。于大肠杆菌(E. coli)中分别采用不同浓度异丙基-β-D-硫代半乳糖苷(IPTG)在不同条件下诱导TFEB及其突变体重组蛋白表达。Glutathione-Sepharose 4B琼脂糖凝珠分离纯化蛋白,SDS-PAGE凝胶电泳检测纯化产物蛋白浓度和相对分子质量。结果：第1步PCR扩增获得均含有突变位点的2条DNA片段,经第2步PCR退火延伸和第3步PCR扩增后获得大量含有突变位点的完整DNA片段。双酶切鉴定,连接的DNA片段...     

自体鼻中隔软骨联合耳廓软骨的鼻中隔延伸物移植术在鼻尖整形中的应用

目的:探讨运用鼻中隔软骨联合耳廓软骨构建鼻中隔延伸物复合体重置鼻尖位置和重塑鼻尖形态的可行性。方法:2018年9月至2019年10月,以中国医学科学院整形外科医院17例低鼻患者为研究对象,17例患者中男2例,女15例;年龄19~39岁,平均27岁。17例患者均为初次鼻整形病例。术中在全身麻醉下行鼻整形术。切取鼻中隔软骨...     

Soft-Shelled Turtle Peptides Extend Lifespan and Healthspan in Drosophila

In traditional Chinese medicine, soft-shelled turtle protein and peptides serve as a nutraceutical for prolonging the lifespan. However, their effects on anti-aging have not been clarified scientifically in vivo. This study aimed to determine whether soft-shelled turtle peptides (STP) could promote the lifespan and healthspan in Drosophila melanogaster and the underlying molecular mechanisms. Herein, STP supplementation prolonged the mean lifespan by 20.23% and 9.04% in males and females, respectively, delaying the aging accompanied by climbing ability decline, enhanced gut barrier integrity, and improved anti-oxidation, starvation, and heat stress abilities, while it did not change the daily food intake. Mechanistically, STP enhanced autophagy and decreased oxidative stress by downregulating the target of rapamycin (TOR) signaling pathway. In addition, 95.18% of peptides from the identified sequences in STP could exert potential inhibitory effects on TOR through hydrogen bonds, van der Walls, hydrophobic interactions, and electrostatic interactions. The current study could provide a theoretical basis for the full exploitation of soft-shelled turtle aging prevention.     

终腱斜行游离延长治疗陈旧性锤状指畸形

正2013年10月~2015年9月,我们采用改良术式终腱斜行游离延长方法治疗14例陈旧性锤状指畸形患者,疗效满意,报道如下。1材料与方法1.1病例资料本组14例,男10例,女4例,年龄20~65岁。畸形手指:食指3例,中指5例,环指5例,小指1例。伤后至手术时间:7~12 d 11例,21 d以上3例。     

Expressman导引延伸导管辅助主动迎接技术成功治疗冠状动脉慢性完全闭塞病变一例报道

逆向介入途径是治疗冠状动脉慢性完全闭塞病变(CTO)的主要技术之一,而逆向导丝体外化是CTO患者经皮冠状动脉介入治疗(PCI)成功的关键。随着延伸导管等新器械的出现及主动迎接技术(AGT)等新技术的不断发展,复杂CTO治疗成功率逐渐升高。本文报道了1例Expressman导引延伸导管辅助AGT成功治疗的冠状动脉CTO患者,以为该技术的临床应用提供参考。     

Calvarial bone cavernous hemangioma with intradural invasion: An unusual aggressive course—Case report and literature review

IntroductionCavernous hemangioma of the skull is a rare pathological diagnosis, accounting for 0.2% of bone tumors and 7% of skull tumors. Usually calvarial bone cavernous hemangioma are associated with a benign clinical course and, despite their enlargement and subsequent erosion of the surrounding bone, the inner table of the skull remains intact and the lesion is completely extracranial.Presentation of a caseThe authors present the unique case of a huge left frontal bone cavernous malformation with intradural extension and brain compression determining a right hemiparesis.DiscussionCalvarial cavernous hemangiomas are benign tumors. They arise from vessels in the diploic space and tend to involve the outer table of the skull with relative sparing of the inner table. More extensive involvement of the inner table and extradural space is very unusual and few cases are reported in literature. To the best of our knowledge, intradural invasion of calvarial hemangioma has not been previously reported.ConclusionOur case highlights the possibility of an aggressive course of this rare benign pathology.     

屈伸检查在颈椎病X线诊断中的应用

目的探讨前屈-后伸位功能摄片在颈椎病X线诊断中的应用价值.方法对本院100例颈椎病患者进行前屈及后伸颈椎侧位片检查,测量其前屈角、后伸角及相对角(前屈角-后伸角=相对角),观察椎体向前及/或向后滑移情况和棘突分离或相互靠近情况.结果前屈角为85°～154°,平均130°.后伸角为42°～87°,平均62°.相对角32°～98°,平均67°.前屈位见颈椎椎体前移者71例,后移者5例,部分椎体前移部分椎体后移者2例;后伸位见颈椎椎体后移者82例;前屈位见局部棘突不能分离者13例;后伸位见局部棘突不能靠近者51例.结论屈伸检查能发现常规颈椎正侧斜位片所未能显示的颈椎失稳和功能改变;有时还能发现常规片中未能显示或显示不清楚的解剖结构性改变;对X线观察颈椎运动,记录局部韧带损害,颈椎失稳及用以估计治疗后功能恢复的程度有重要作用.