Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are still occurring, particularly in reconstructive flap surgery. Multiple experimental flap models have been developed to analyze underlying causes and mechanisms and to investigate treatment strategies to prevent ischemic complications. The limiting factor of most models is the lacking possibility to directly and repetitively visualize microvascular architecture and hemodynamics. The goal of the protocol was to present a well-established mouse model affiliating these before mentioned lacking elements. Harder et al. have developed a model of a musculocutaneous flap with a random perfusion pattern that undergoes acute persistent ischemia and results in ~50% necrosis after 10 days if kept untreated. With the aid of intravital epi-fluorescence microscopy, this chamber model allows repetitive visualization of morphology and hemodynamics in different regions of interest over time. Associated processes such as apoptosis, inflammation, microvascular leakage and angiogenesis can be investigated and correlated to immunohistochemical and molecular protein assays. To date, the model has proven feasibility and reproducibility in several published experimental studies investigating the effect of pre-, peri- and postconditioning of ischemically challenged tissue.     

Investigation of flap flexibility of β-secretase using molecular dynamic simulations

Flap motif and its dynamics were extensively reported in aspartate proteases, e.g. HIV proteases and plasmepsins. Herein, we report the first account of flap dynamics amongst different conformations of β-secretase using molecular dynamics simulation. Various parameters were proposed and a selected few were picked which could appropriately describe the flap motion. Three systems were studied, namely Free (BACE_Free) and two ligand-bound conformations, which belonged to space groups P6₁22 (BACE_Bound1) and C222₁ (BACE_Bound2), respectively and four parameters (distance between the flaps tip residue, Thr72 and Ser325, d₁; dihedral angle, ? (Thr72-Asp32-Asp228-Ser325); TriCα angles, θ₁ (Thr72-Asp32-Ser325), and θ₂ (Thr72-Asp228-Ser325)) were proposed to understand the change in dynamics of flap domain and the extent of flap opening and closing. Analysis of, θ₂, d₁, θ₁ and ? confirmed that the BACE_Free adopted semi-open, open and closed conformations with slight twisting during flap opening. However, BACE_Bound1 (P6122) showed an adaptation to open conformation due to lack of hydrogen bond interaction between the ligand and flap tip residue. A slight flap twisting, ? (lateral twisting) was observed for BACE_Bound1 during flap opening which correlates with the opening of BACE_Free. Contradictory to the BACE_Bound1, the BACE_Bound2 locked the flap in a closed conformation throughout the simulation due to formation of a stable hydrogen bond interaction between the flap tip residue and ligand. Analyses of all three systems highlight that d₁, θ₂ and ? can be precisely used to describe the extent of flap opening and closing concurrently with snapshots along the molecular dynamics trajectory across several conformations of β-secretase.     

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。     

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。     

Fibrinogen Induces Microglia-Mediated Spine Elimination and Cognitive Impairment in an Alzheimer’s Disease Model

1. Download high-res image (279KB)
2. Download full-size image

     

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。     

Neurovascular coupling and oximetry during epileptic events

Epilepsy is an abnormal brain state in which a large population of neurons is synchronously active, causing an enormous increase in metabolic demand. Recent investigations using high-resolution imaging techniques, such as optical recording of intrinsic signals and voltagesensitive dyes, as well as measurements with oxygen-sensitive electrodes have elucidated the spatiotemporal relationship between neuronal activity, cerebral blood volume, and oximetry in vivo. A focal decrease in tissue oxygenation and a focal increase in deoxygenated hemoglobin occurs following both interictal and ictal events. This “epileptic dip” in oxygenation can persist for the duration of an ictal event, suggesting that cerebral blood flow is inadequate to meet metabolic demand. A rapid focal increase in cerebral blood flow and cerebral blood volume also accompanies epileptic events; however, this increase in perfusion soon (>2 s) spreads to a larger area of the cortex than the excitatory change in membrane potential. Investigations in humans during neurosurgical operations have confirmed the laboratory data derived from animal studies. These data not only have clinical implications for the interpretation of noninvasive imaging studies such as positron emission tomography, single-photon emission tomography, and functional magnetic resonance imaging but also provide a mechanism for the cognitive decline in patients with chronic epilepsy.