A diagnostic ceiling for exome sequencing in cerebellar ataxia and related neurological disorders

Genetic ataxias are associated with mutations in hundreds of genes with high phenotypic overlap complicating the clinical diagnosis. Whole‐exome sequencing (WES) has increased the overall diagnostic rate considerably. However, the upper limit of this method remains ill‐defined, hindering efforts to address the remaining diagnostic gap. To further assess the role of rare coding variation in ataxic disorders, we reanalyzed our previously published exome cohort of 76 predominantly adult and sporadic‐onset patients, expanded the total number of cases to 260, and introduced analyses for copy number variation and repeat expansion in a representative subset. For new cases (n = 184), our resulting clinically relevant detection rate remained stable at 47% with 24% classified as pathogenic. Reanalysis of the previously sequenced 76 patients modestly improved the pathogenic rate by 7%. For the combined cohort (n = 260), the total observed clinical detection rate was 52% with 25% classified as pathogenic. Published studies of similar neurological phenotypes report comparable rates. This consistency across multiple cohorts suggests that, despite continued technical and analytical advancements, an approximately 50% diagnostic rate marks a relative ceiling for current WES‐based methods and a more comprehensive genome‐wide assessment is needed to identify the missing causative genetic etiologies for cerebellar ataxia and related neurodegenerative diseases.     

Identification of potential metabolic biomarkers in Yin deficiency syndrome using LC–MS

Yin and Yang are the two counter-balancing aspects in ancient Chinese philosophy. In traditional Chinese medicine, Yin deficiency syndrome (YDS) is a common sub-health state with complex causes. While the syndrome may be treated to various degrees of effectiveness with traditional Chinese medicine, efficient modern methods are yet to be developed for diagnosing and treating the YDS. Here we performed a metabolomics study on YDS in rats. Serum metabolites in rats were analyzed using ultra-performance liquid chromatography-mass spectrometry (UPLC–MS) method to identify potential biomarkers for YDS. The rats were divided randomly into the healthy control group, the untreated YDS group, and the anemarrhena treated YDS group. Compared with the control group, significant increase in the metabolites such as dihydrotestosterone (DHT) and 5β-DHT, 4-imidazolone-5-propanoate, 4-(L-alanin-3-yl)-2-hydroxy-cis,cis-muconate 6-semialdehyde, and 5-(L-alanin-3-yl)-2-hydroxy-cis,cis-muconate 6-semialdehyde were observed in the serum of untreated YDS group, which returned to normal in the anemarrhena treated group. Therefore, these metabolites may serve as potential biomarkers for YDS, and may facilitate the diagnosis and treatment of YDS.     

三维可视化技术在胸椎黄韧带骨化症手术治疗中的应用

目的 探讨三维可视化技术辅助设计手术治疗胸椎黄韧带骨化症的临床应用价值。方法 回顾性分析2013年11月—2016年12月郑州市骨科医院脊柱一科采用椎板切除治疗的44例胸椎黄韧带骨化症的临床资料。其中,男25例、女19例,年龄40～76岁。术前均采用Mimics软件对胸椎CT扫描的DICOM数据进行三维重建,在胸椎椎板和骨化黄韧带的三维可视化模型上观察骨化灶的立体结构以及与椎弓根、关节突、椎板的位置关系,计算数字模拟骨化灶体积;按照“分区椎板切除”的方式在三维可视化数字模型上进行骨化黄韧带的模拟切除;按照术前模拟切除步骤,实施手术操作。观察手术时间、术中出血量及并发症发生情况。术后12个月采用日本骨科协会(JOA)评分评估神经功能,根据术前和术后JOA评分计算JOA改良率,评估手术疗效。结果 三维重建模型44个,重建骨化灶103个,骨化灶体积为(1 831±443)mm³。44例患者均顺利完成手术,手术时间65～180(96.7±19.6)min;术中出血量230～1 350(432±83.5)mL。患者术后无脊髓神经损害症状加重者,术后神经症状均逐步好转。44例患者均获随访,随访时间13～46(25±10.3)个月。随访期间无迟发性感染、神经症状加重、内固定失败等并发症发生。术后12个月JOA评分(8.8±1.8)分,明显高于术前的(5.3±2.0)分,差异有统计学意义(t=11.566, P＜0.01);JOA评分改善率为64.2%±21.7%,疗效评价优13例、良21例、可10例,优良率77.3%(34/44)。结论 应用三维可视化技术进行术前评估,能够立体、全面了解骨化黄韧带的形态,通过模拟手术,设计手术切除范围,可以提高手术精准度、安全性和有效性,为胸椎黄韧带骨化症的诊疗提供了一种新的术前影像学辅助方法。     

二乙酰吗啡干预离体乳鼠心肌细胞动作电位和钙离子电流的变化

文题释义：二乙酰吗啡：半合成的阿片受体纯激动剂,由吗啡和醋酸酐为原料经化学反应制成,其镇痛作用强于吗啡,镇痛效果为吗啡的4-8倍,其成瘾速度快,戒断难度大。长期使用后不仅会对人体的免疫功能造成严重破坏,还可导致心、肝、肾等重要脏器的功能损害。钙离子：作为细胞内第二信使,主要负责维持细胞膜正常电生理功能,以及确保机体多种生物信号转导途径正常进行。在心肌缺血缺氧过程中,当缺血引起心肌异常的L型钙电流逐渐恢复正常,钾通道活性程度降低,导致动作电位时程相对延长,L型钙电流在激活和失活的重叠区形成“窗流”,导致心肌细胞膜电位不稳而形成电震荡;当细胞内Ca2+大量积聚,增加 Ca2+-CaM复合物生成,心肌细胞易造成早期和晚期后除极,导致室性心律失常概率增加。 背景：阿片类药物可调控心肌细胞膜电位和Ca²⁺电流的变化,但是目前二乙酰吗啡是否诱导心肌节律、细胞动作电位和钙离子电流改变尚未见报道。 目的：探讨二乙酰吗啡对离体SD乳鼠心肌细胞动作电位和钙离子电流的影响。方法：①体外培养SD乳鼠心肌细胞,采用5个浓度二乙酰吗啡(0,10^-2,10^-3,10^-4,10^-5 mol/L)和20 mol/L维拉帕米作用于心肌细胞;②将细胞分为对照组、二乙酰吗啡组、二乙酰吗啡+维拉帕米组。后2组分别以二乙酰吗啡、二乙酰吗啡+维拉帕米(浓度20 μmol/L)联合干预心肌细胞;对照组加入等量PBS。实验于2018-05-21经新疆医科大学第一附属医院动物实验医学伦理委员会批准,批准号IACUC201805-K1。 结果与结论：①当不同浓度二乙酰吗啡干预心肌细胞24 h后,心肌细胞形态异常数量亦随其浓度的改变而明显增加,呈剂量依赖性改变。当二乙酰吗啡浓度逐渐升高,细胞存活数量亦逐渐减少,细胞胞质体积缩小,胞膜收缩,伪足减少,细胞核结构模糊。②与对照组相比,当加入二乙酰吗啡干预心肌细胞后,心肌细胞自发性搏动频率和节律差异有显著性意义。静息膜电位负值降低,动作电位中动作电位时程显著增加,动作电位幅度显著减小。③与对照组相比,二乙酰吗啡组中心肌膜电位改变数量明显增加,当加入维拉帕米后,心肌膜电位改变细胞数量有所降低。与对照组相比,心肌膜电位改变细胞数量有所升高。④提示二乙酰吗啡可致离体SD乳鼠心肌细胞形态异常,心肌自发性搏动频率和节律显著增加,心肌细胞膜电位和动作电位发生改变,钙通道阻滞剂维拉帕米对二乙酰吗啡引起的心肌细胞节律异常有一定改善作用。  ORCID: 0000-0002-5113-6050(苏丽萍) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

结肠镜检查等待时间对患者肠道准备质量的影响研究

目的 了解结肠镜检查患者肠道准备现状并分析其影响因素,评价肠道准备完成至结肠镜检查开始的间隔时间对肠道准备质量的影响。 方法 收集2019年4月—9月在福州市3所三级甲等医院消化内镜中心行结肠镜检查门诊患者的人口统计学与临床资料。采用卡方检验筛选对肠道准备质量影响显著的因素（P<0.10）,应用倾向值匹配分析法消除混杂因素后,对比间隔时间（2,3] h、（3,4] h、（4,5] h、（5,6] h、（6,7] h、（7,8] h、（8,∞） h组的肠道准备质量差异。 结果 共有401例患者纳入研究,经匹配新生成316组样本数据,并检验证实已排除混杂因素影响。方差分析得出：不同间隔时间组之间的肠道准备质量存在差异,间隔（2,3] h肠道准备质量显著优于（4,5] h、（6,7] h,间隔（3,4] h肠道准备质量显著优于（6,7] h,（2,3] h与（3,4] h两组之间无显著差异,（4,5] h、（6,7] h、（7,8] h、（8,∞） h任意两组之间无显著差异。 结论 肠道准备完成至开始结肠镜检查间隔时间在2~4 h内,肠道准备质量可以保持在最佳清洁状态。     

不同无损伤针摆放方法对胸壁完全植入式输液港患者维护效果的研究

目的 观察无损伤针2种摆放方法对肿瘤患者胸壁完全植入式输液港维护效果的影响。 方法 通过体外实验对无损伤针正向摆放组和背向摆放组冲洗量的差异进行比较,然后选取193例首次植入输液港的肿瘤患者作为研究对象,随机分为正向摆放组和背向摆放组,比较两组输液速度和导管堵塞程度,以及胸壁输液港座不同位置时,2种摆放方法对无损伤针敷贴固定效果和患者舒适度的影响。 结果 体外实验背向摆放组冲洗量为（4.28±0.67） ml,显著低于正向组（6.51±0.92） ml,差异有统计学意义（P<0.05）。临床试验中正向摆放组输液速度（183.96±9.23）滴/min,显著高于背向组（151.02±10.11）滴/min,两组差异有统计学意义（P<0.001）。输液港底座靠近腋窝处时,正向摆放组无损伤针敷贴的固定效果优于背向组（P<0.001）,正向摆放组患者的舒适度也优于背向组（P<0.001）。输液港底座靠近锁骨中线处,两组无损伤针敷贴固定效果相似（P=0.416）,患者舒适度比较,差异无统计学意义（P=0.457）。 结论 针对肿瘤患者胸壁完全植入式输液港的维护,无损伤针背向摆放组在体外所需要的冲洗量较正向摆放组更少。当输液港底座靠近腋窝处时,正向摆放组敷贴的固定效果更好,患者的舒适度更高,输液速度更快。     

乳腺癌相关淋巴水肿患者运动指导方案的证据总结

目的 评价并总结乳腺癌相关淋巴水肿运动指导的相关证据,为医护人员对淋巴水肿患者采取运动干预提供依据。方法 系统检索BMJ Best Practice、UpToDate、Cochrane Library、Joanna Briggs Institute（JBI）循证卫生保健国际合作中心图书馆、JBI在线临床治疗及护理证据网络、英国国家卫生与临床优化研究所网站、美国国立指南库、国际指南协作网、苏格兰校际指南网络、美国国立综合癌症网络、加拿大安大略注册护士协会网站、国际淋巴水肿网站、国际淋巴水肿协会网站、PubMed、CINAHL、Embase、Web of Science、Physiotherapy Evidence Database、中国指南网、中国知网、万方、中国生物医学文献服务系统、维普数据库关于乳腺癌相关淋巴水肿运动方面的指南、专家共识、证据总结、系统评价等证据,检索时间为建库至2019年7月,由2名具有循证护理知识的研究人员独立对文献进行质量评价和证据级别评定。结果 共纳入19篇文献,包括1篇临床决策,1篇推荐实践,1篇证据总结,5篇指南,3篇专家共识,8篇系统评价,从运动禁忌证、运动前评估、运动前测试、运动强度、运动安全性、运动方案、注意事项、健康教育8个方面汇总23条最佳证据。 结论 运动干预可改善淋巴水肿症状,医护人员需结合临床情境,考虑患者的意见,为淋巴水肿患者提供个性化的运动指导,发挥运动疗效,以进一步提高患者的生活质量。     

Inhibition of hypoxia-induced proliferation of pulmonary arterial smooth muscle cells by a mTOR siRNA-loaded cyclodextrin nanovector

The proliferation of pulmonary arterial smooth muscle cells (PASMCs) is a key pathophysiological component of vascular remodeling in pulmonary arterial hypertension (PAH), an intractable disease, for which pharmacotherapy is limited and only slight improvement in survival outcomes have achieved over the past few decades. RNA interference provides a highly promising strategy to the treatment of this chronic lung disease, while efficient delivery of small interfering RNA (siRNA) remains a key challenge for the development of clinically acceptable siRNA therapeutics. With the aim to construct useful nanomedicines, the mammalian target of rapamycin (mTOR) siRNA was loaded into hybrid nanoparticles based on low molecular weight (Mw) polyethylenimine (PEI) and a pH-responsive cyclodextrin material (Ac-aCD) or poly(lactic-co-glycolic acid) (PLGA). This hybrid nanoplatform gave rise to desirable siRNA loading, and the payload release could be modulated by the hydrolysis characteristics of carrier materials. Fluorescence observation and flow cytometry quantification suggested that both Ac-aCD and PLGA nanovectors (NVs) may enter PASMCs under either normoxia or hypoxia conditions as well as in the presence of serum, with uptake and transfection efficiency significantly higher than those of cationic vectors such as PEI with Mw of 25 kDa (PEI25k) and Lipofectamine 2000 (Lipo 2k). Hybrid Ac-aCD or PLGA NV containing siRNA remarkably inhibited proliferation and activated apoptosis of hypoxic PASMCs, largely resulting from effective suppression of mTOR signaling as evidenced by significantly lowered expression of mTOR mRNA and phosphorylated protein. Moreover, these hybrid nanomedicines were more effective than commonly used cationic vectors like PEI25k and Lipo 2k, with respect to cell growth inhibition, apoptosis activation, and expression attenuation of mTOR mRNA and protein. Therefore, mTOR siRNA nanomedicines based on hybrid Ac-aCD or PLGA NV may be promising therapeutics for diseases related to hypoxic abnormal growth of PASMCs.     

Optical imaging of fibrin deposition to elucidate participation of mast cells in foreign body responses

Mast cell activation has been shown to be an initiator and a key determinant of foreign body reactions. However, there is no non-invasive method that can quantify the degree of implant-associated mast cell activation. Taking advantage of the fact that fibrin deposition is a hallmark of mast cell activation around biomaterial implants, a near infrared probe was fabricated to have high affinity to fibrin. Subsequent in vitro testing confirmed that this probe has high affinity to fibrin. Using a subcutaneous particle implantation model, we found significant accumulation of fibrin-affinity probes at the implant sites as early as 15 min following particle implantation. The accumulation of fibrin-affinity probes at the implantation sites could also be substantially reduced if anti-coagulant – heparin was administered at the implant sites. Further studies have shown that subcutaneous administration of mast cell activator – compound 48/80 – prompted the accumulation of fibrin-affinity probes. However, implant-associated fibrin-affinity probe accumulation was substantially reduced in mice with mast cell deficiency. The results show that our fibrin-affinity probes may serve as a powerful tool to monitor and measure the extent of biomaterial-mediated fibrin deposition and mast cell activation in vivo.