重组flap核酸内切酶1的表达及活性测定方法的建立

Flap核酸内切酶1(Flap endonuclease 1,FEN1)是一种能催化核酸侵入反应的核酸内切酶,可应用于信号放大检测方法,但该酶详细的表达纯化工艺尚无报道,并且活性难以准确测定,限制了其应用。通过合成嗜热古球菌Archaeoglobus fulgidus来源的FEN1基因序列,构建了p ET24a(+)-FEN1-His重组质粒,并通过优化表达条件,得到了FEN1最优表达条件为:37℃、200 r/min振荡培养8 h后,加入诱导剂IPTG至终浓度为0.05 mmol/L,再于37℃、200 r/min诱导表达11 h,最终经镍亲和层析成功纯化得到了分子量约为38 k Da的重组FEN1。同时建立了基于荧光标记探针的FEN1活性测定方法,准确测定了重组FEN1的活性,为建立基于该酶的核酸检测方法提供了可靠的酶活力依据。最终将重组FEN1用于实时荧光PCR偶联高特异核酸侵入信号扩增法检测了乙醛脱氢酶2基因(aldh2)的基因型,得到了准确的分型结果,表明重组FEN1能用于基因多态性的分型检测中,为发展基于核酸侵入反应的核酸检测方法提供了可靠的工具酶。     

Normalising surface EMG of ten upper-extremity muscles in handcycling: Manual resistance vs. sport-specific MVICs

Muscular activity in terms of surface electromyography (sEMG) is usually normalised to maximal voluntary isometric contractions (MVICs). This study aims to compare two different MVIC-modes in handcycling and examine the effect of moving average window-size. Twelve able-bodied male competitive triathletes performed ten MVICs against manual resistance and four sport-specific trials against fixed cranks. sEMG of ten muscles [M. trapezius (TD); M. pectoralis major (PM); M. deltoideus, Pars clavicularis (DA); M. deltoideus, Pars spinalis (DP); M. biceps brachii (BB); M. triceps brachii (TB); forearm flexors (FC); forearm extensors (EC); M. latissimus dorsi (LD) and M. rectus abdominis (RA)] was recorded and filtered using moving average window-sizes of 150, 200, 250 and 300 ms. Sport-specific MVICs were higher compared to manual resistance for TB, DA, DP and LD, whereas FC, TD, BB and RA demonstrated lower values. PM and EC demonstrated no significant difference between MVIC-modes. Moving average window-size had no effect on MVIC outcomes. MVIC-mode should be taken into account when normalised sEMG data are illustrated in handcycling. Sport-specific MVICs seem to be suitable for some muscles (TB, DA, DP and LD), but should be augmented by MVICs against manual/mechanical resistance for FC, TD, BB and RA.     

改良双侧V-Y皮瓣治疗Ⅳ期骶尾部压疮的疗效分析

摘要 目的：探讨改良双V-Y皮瓣治疗Ⅳ期骶尾部压疮的有效性和安全性。方法：回顾性分析2015年3月-2019年3月中国人民解放军总医院第七医学中心骨科收治骶尾部压疮Ⅳ期患者28例,其中男15例,女13例,年龄49-65岁,创面面积6 cm×7 cm~8 cm×10 cm。彻底清创后设计改良双侧V-Y推移皮瓣覆盖创面并行负压封闭引流,皮瓣供区减张缝合,皮瓣面积6 cm×7 cm~8 cm×10 cm。术后观察皮瓣整体血运、温度、水肿情况及VSD引流量,定期通过门诊、微信、电话随访。结果：手术后双侧V-Y皮瓣全部成活,1例术后切口周边皮肤表皮坏死,经换药后愈合,28例患者全部获得随访,随访时间12-24个月,平均18.2个月,全部压疮缺损修复成功,无创口裂开、皮瓣淤血、水肿、感染及压疮复发。结论：改良双侧V-Y推移皮瓣是治疗Ⅳ期骶尾部压疮的一种有效治疗方法。     

前臂皮瓣修复外伤性颌面部缺损的护理体会

目的:探讨颌面部外伤后遗留大面积软组织缺损采用血管化游离前臂皮瓣修复的护理方法。方法:采用前臂皮瓣血管吻合修复17例外伤导致的颌面部软组织缺损,围术期采取严格的护理措施,术前的心理护理、术后的严密观察,最大限度防止皮瓣血管危象发生,保证治疗效果。结果:2例出现血管危象,进行抢救,其中1例失败,另1例抢救成功,其余15例顺利成活。结论:术前进行心理护理、术后严密观察、及时抢救是确保手术成功的关键。     

马来酸桂哌齐特与腹部带蒂皮瓣对上肢烧伤组织缺损的修复效果

目的：探讨马来酸桂哌齐特联合腹部带蒂皮瓣治疗手及前臂皮肤软组织烧伤缺损的临床效果。方法：选取我院2011 年2 月 -2013 年2 月收治的68 例手及前臂皮肤软组织烧伤缺损患者作为观察组,行马来酸桂哌齐特结合腹部带蒂皮瓣治疗,另选择同 期接受股前外侧皮瓣修复治疗的50 例患者为对照组。观察并比较两组患者皮瓣修复效果以及神经功能烧伤缺损评分。结果：观 察组患者皮瓣存活率高于对照组,差异具有统计学意义(P<0.05);观察组皮瓣感染率低于对照组,差异具有统计学意义(P<0.05); 观察组皮瓣断蒂时间及神经功能烧伤缺损评分均低于对照组,差异具有统计学意义(P<0.05)。结论：马来酸桂哌齐特结合腹部带 蒂皮瓣治疗具有皮瓣存活率高、神经功能恢复良好,术后恢复快等特点,值得临床推广应用。     

带蒂皮瓣移植在小腿开放性骨折中的临床研究

目的:探讨不同带蒂皮瓣移植术式在小腿开放性骨折中的临床应用效果。方法:35例小腿开放性骨折患者,完善术前准备后,根据小腿皮肤软组织的缺损部位、面积、深度分别选择腓肠神经营养皮(肌)瓣、隐神经营养逆行皮瓣、局部旋转皮(肌)瓣修复创面。结果:所有移植皮瓣中,1处移植失败(坏死面积>1/2),2处皮瓣远端1-2.5cm坏死,经清创换药后愈合,余患者皮瓣均成活。从皮瓣成活的优良率和出血量上对比,三组间无明显差别,从手术时间上对比,局部旋转皮(肌)瓣组优于另两组(P<0.05)。结论:在小腿开放性骨折伴皮肤软组织缺损的治疗中,完善的术前准备及根据皮损形态选择适当的皮瓣是获得满意疗效的基础。     

Electromyography study of six parts of the latissimus dorsi during reaching tasks while seated: A comparison between healthy subjects and stroke patients

PurposeTo compare the excitation of the six different segments of the latissimus dorsi (LD) while reaching different distances and in different directions in stroke patients and healthy controls.MethodSurface electromyography was used to measure the excitation of the LD segments (LD1-LD6) in 12 chronic stroke patients and 11 healthy controls during reaching tasks. A target was placed in the sagittal and scapular planes at arm’s length, 125% of arm's length, and maximum reaching distance. The clinical trial registration number is NCT04181151 (date of registration November 25, 2019).ResultsThe excitation of the LD segments during the arm’s length reaching task was similar between the groups (p greater than 0.05). The excitation of LD1, LD2, and LD5 in the sagittal plane and of LD1, LD2, LD3, and LD5 in the scapular plane was higher during the reaching 125% of arm’s length task compared to the controls (p < 0.05). During the maximum reaching task, the excitation of LD1 was higher in the stroke patients in both the sagittal and scapular planes (p < 0.05).ConclusionThe excitation of the LD segments was influenced by the direction and distance of the reaching in the stroke patients. The results of this study may help us to better understand how the LD behaves after stroke and to design rehabilitation approaches with a greater focus on the LD.     

Host-induced effects on the morphology of Ostertagia circumcincta in grazing sheep

Waller P. J. and Thomas R. J. 1978. Host-induced effects on the morphology of Ostertagia circumcincta in grazing sheep. International Journal for Parasitology8: 365–370. Morphological changes are described in adult Ostertagia circumcincta recovered from lambs slaughtered monthly from June to November. Comparisons were made between continuously grazed principal lambs exposed to infection for increasingly long periods, and tracer Iambs exposed for only four weeks. Lambs were either slaughtered immediately off pasture or held 2 weeks under worm free conditions before slaughter. Although no resistance to establishment of infection occurred, shown by comparable worm burdens of principals and tracers, a rapid parasite population turnover was observed in the principal lambs which ceased once the lambs were removed from the infected pasture. Significantly fewer adult female Ostertagia with fully developed vulval flaps were recorded in principals compared to tracers. Such differences are considered to be primarily host induced responses which rapidly come into play, evidenced by the entire populations of developing female worms in tracers slaughtered immediately off pasture showing arrested vulval flap development after they had completed development in those tracers held for a further two weeks. A progressive reduction in the proportion of adult male worms also occurred in the principal lambs. Population density was shown to have an inverse correlation with worm size. By analogy, it is suggested that seasonal change in the presence of cuticular inflations in Haemonchus contortus females with linguiform vulval flaps may be primarily host induced, rather than a change in the relative fitness of discrete genotypes.     

Flap flexibility amongst plasmepsins I,II, III,IV, and V: Sequence,structural, and molecular dynamics analyses

Herein, for the first time, we comparatively report the opening and closing of apo plasmepsin I – V. Plasmepsins belong the aspartic protease family of enzymes, and are expressed during the various stages of the P. falciparum lifecycle, the species responsible for the most lethal and virulent malaria to infect humans. Plasmepsin I, II, IV and HAP degrade hemoglobin from infected red blood cells, whereas plasmepsin V transport proteins crucial to the survival of the malaria parasite across the endoplasmic reticulum. Flap‐structures covering the active site of aspartic proteases (such as HIV protease) are crucial to the conformational flexibility and dynamics of the protein, and ultimately control the binding landscape. The flap‐structure in plasmepsins is made up of a flip tip in the N‐terminal lying perpendicular to the active site, adjacent to the flexible loop region in the C‐terminal. Using molecular dynamics, we propose three parameters to better describe the opening and closing of the flap‐structure in apo plasmepsins. Namely, the distance, d₁, between the flap tip and the flexible region; the dihedral angle, ?, to account for the twisting motion; and the TriCα angle, θ₁. Simulations have shown that as the flap‐structure twists, the flap and flexible region move apart opening the active site, or move toward each other closing the active site. The data from our study indicate that of all the plasmepsins investigated in the present study, Plm IV and V display the highest conformational flexibility and are more dynamic structures versus Plm I, II, and HAP. Proteins 2015; 83:1693–1705. © 2015 Wiley Periodicals, Inc.