超声衰减成像技术评价肝脂肪变性的相关因素分析

目的探讨影响超声衰减系数（AC）变化的相关因素,并评估衰减成像（ATI）技术对非酒精性脂肪性肝病（NAFLD）定量诊断的临床实用性及测量的可重复性。 方法选取2019年9月至2020年12月在哈尔滨医科大学附属第一医院使用超声的ATI技术进行肝检测的114例受试者（NAFLD组56例,健康对照组58例）,分别获得二维超声和AC,同时收集患者相应的临床资料、血生化检测指标等,通过多因素线性回归分析确定AC的独立影响因素。使用常规二维超声检查对所有研究对象的肝脂肪变性程度进行四分制评分（S0~S3组）,采用方差分析检验各组间AC的差异性,使用Spearman相关分析方法评估AC与肝脂肪变性程度的相关性,并计算组内相关系数（ICC）,评估2名操作者间（AC-1组和AC-2组）ATI测量的可重复性。 结果多因素线性回归分析显示体质量指数（BMI）、肝脂肪变性程度是AC的独立影响因素（β=0.007,95%CI：0.001~0.013,P=0.017;β=0.083,95%CI：0.070~0.095,P＜0.001）。NAFLD组AC显著高于健康对照组［AC-1组：（0.716±0.098）dB/（cm·MHz）vs（0.539±0.058）dB/（cm·MHz）;AC-2组：（0.699±0.102）dB/（cm·MHz）vs（0.542±0.053）dB/（cm·MHz）］,且随着肝脂肪变性程度的加重,AC逐渐升高［S0组（58例）AC为（0.539±0.058）dB/（cm·MHz）,S1组（23例）AC为（0.640±0.063）dB/（cm·MHz）,S2组（20例）AC为（0.718±0.050）dB/（cm·MHz）,S3组（13例）AC为（0.845±0.059）dB/（cm·MHz）;F=122.27,P＜0.001］。AC与肝脂肪变性程度存在较强的相关性（r=0.840;P＜0.001）。不同肝脂肪变性程度AC的组内相关系数（ICC）分别为0.816（95%CI：0.708~0.881）、0.886（95%CI：0.301~0.967）、0.876（95%CI：0.479~0.960）、0.939（95%CI：0.819~0.981）,P均＜0.001。 结论AC与BMI、肝脂肪变性程度显著相关,且ATI技术的操作者间可重复性极好,结果较为稳定,有助于早期发现脂肪肝并对脂肪肝严重程度进行评估。     

Isolating and characterizing three alternatively spliced mu opioid receptor variants: mMOR‐1A,mMOR‐1O,and mMOR‐1P

Extensive alternative pre‐mRNA splicing of the mu opioid receptor gene, OPRM1, has demonstrated an array of splice variants in mice, rats and humans. Three classes of splice variants have been identified: full‐length seven transmembrane (TM) domain variants with C‐terminal splicing, truncated 6TM variants and single TM variants. The current studies isolates and characterizes an additional three full‐length C‐terminal splice variants generated from the mouse OPRM1 gene: mMOR‐1A, mMOR‐1O, and mMOR‐1P. Using RT‐qPCR, we demonstrated differential expression of these variants' mRNAs among selected brain regions, supporting region‐specific alternative splicing. When expressed in Chinese Hamster Ovary cells, all the variants displayed high mu binding affinity and selectivity with subtle differences in the affinities toward some agonists. [³⁵S]γGTP binding assays revealed marked differences in agonist‐induced G protein activation in both potency and efficacy among the variants. Together with the previous studies of mu agonist‐induced phosphorylation and internalization in several carboxyl terminal splice variants, the current studies further suggest the existence of biased signaling of various agonists within each individual variant and/or among different variants. Synapse 68:144–152, 2014 . © 2013 Wiley Periodicals, Inc.     

Neuroprotective effect of total glycosides from paeonies against neurotoxicity induced by strychnos alkaloids related to recovering the levels of neurotransmitters and neuroendocrine hormones in rat serum and brain

Semen Strychni, a classical traditional Chinese medicine, has been widely used for its anti-tumor, analgesic and anti-inflammatory angiogenesis effects. However, taking an overdose of Semen Strychni might result in extreme neurotoxicity. Strychnos alkaloids are the main toxic constituents of Semen Strychni. Total glycosides from paeonies are considered to have neuroprotective effects. In this study, twelve potential endogenous biomarkers in rat serum and brain were monitored to investigate the protective effect of total glycosides from the paeony against strychnos alkaloids-induced neurotoxicity. A sensitive liquid chromatography-tandem mass spectrometry method was developed and validated to monitor eight neurotransmitters including glutamate, γ-aminobutyric acid, acetylcholine serotonin, dopamine, norepinephrine, tryptophan and tyrosine. An enzyme-linked immunosorbent assay method was selected for determination of four neuroendocrine hormones including thyrotrophin-releasing hormone, corticotrophin-releasing hormone, antidiuretic hormone and prolactin. Results showed that continuous administration of strychnos alkaloids for 15 days caused significant changed levels of the biomarkers (especially the four neuroendocrine hormones). Meanwhile, total glycosides from paeony pretreated rats (administrated with total glycosides from the paeony for 15 days before exposure to strychnos alkaloids) showed recovered levels of these biomarkers. The results suggested that the neurotransmitters and neuroendocrine hormones in serum and brain might play potential roles as biomarkers. This study provides the possibility of alleviating the Semen Strychni-induced neurotoxicity in clinic by pre-protection with total glycosides from paeonies.

Eight neurotransmitters and four neuroendocrine hormones in rat serum and brain were quantified to investigate the neuroprotective effect of total glycosides from paeony against neurotoxicity induced by strychnos alkaloids.     

Circulating cell‐free mitochondrial deoxyribonucleic acid is increased in coronary heart disease patients with diabetes mellitus

Aims/Introduction

Circulating cell‐free mitochondrial deoxyribonucleic acid (ccf‐mtDNA) is presumably derived from injured tissues or cells in the body and has been suggested to be potential biomarker in several diseases. The present study explored whether mtDNA could be used as a biomarker to evaluate disease in coronary heart disease (CHD) patients with or without diabetes mellitus (DM).

Materials and Methods

A total of 50 CHD patients with type 2 diabetes, 50 CHD patients without type 2 diabetes, and 50 age‐ and sex‐matched patients without CHD and DM (non‐CHD‐DM) were recruited. Ccf‐mtDNA levels were assessed by measuring the nicotinamide adenine dinucleotide dehydrogenase 1 gene using quantitative real‐time polymerase chain reaction. Receiver operating characteristic curve analysis of plasma mtDNA in CHD with or without DM was also determined. Multivariate logistic regression analyses were carried out to determine the correlation between the mtDNA levels and traditional CHD risk factors.

Results

The plasma ccf‐mtDNA levels were significantly elevated in CHD patients with DM compared with those without and non‐CHD‐DM. The area under the receiver operating characteristic curves of mtDNA in CHD patients with DM vs non‐CHD‐DM was 0.907%. Correlation analyses of the mtDNA levels and traditional CHD risk factors showed that the mtDNA levels were significantly correlated with fasting blood glucose in CHD patients with DM.

Conclusions

Ccf‐mtDNA levels can be used as a biomarker in CHD patients with DM.     

颅痛定对心肺复苏术后患者的影响及其机制探讨

目的探讨分析颅痛定对心肺复苏术后患者的影响及其机制。方法选择32例心肺复苏术后患者并随机分为观察组与对照组各16例,所有患者均予以常规治疗,观察组在此基础增加颅痛定治疗,比较两组的临床疗效。结果观察组治疗总有效率显著高于对照组;观察组治疗后SOD显著高于对照组,MDA、TXB2显著低于对照组;治疗后第1天观察组NO显著高于对照组,第2天和第3天观察纽显著低于对照组;治疗后第1天观察纽CO显著低于对照组;以上差异均有统计学意义（尸〈0．05）。结论颅痛定可以有效改善心肺复苏术后患者意识,降低记忆力及智力减退损害,改善缺血一再灌注损伤。     

儿童咯血介入栓塞治疗后复发8例

【摘要】 目的 分析儿童咯血经导管支气管动脉栓塞术（BAE）后复发的原因,为预防复发提供支持。方法 回顾性分析2014年6月至2018年6月46例咯血患儿经BAE治疗后复发的8例临床资料,通过支气管动脉再次造影综合分析复发原因。结果 BAE术后8例咯血复发患儿再次造影发现,复发咯血由原责任血管再通所致2例,异常供血支气管动脉遗漏栓塞所致2例,病灶新生侧支循环供血动脉建立所致3例,血管变异超选失败所致1例。二次BAE治疗后随访6个月至3年。7例患儿未再出现咯血症状,1例再次出现咯血,给予外科右肺下叶切除后治愈。结论 BAE可有效治疗咯血,完全栓塞是关键。但术中造影应仔细,避免遗漏,再通和新发动脉也值得注意,是咯血复发原因之一。     

Probing nanoparticle translocation across the permeable endothelium in experimental atherosclerosis

Therapeutic and diagnostic nanomaterials are being intensely studied for several diseases, including cancer and atherosclerosis. However, the exact mechanism by which nanomedicines accumulate at targeted sites remains a topic of investigation, especially in the context of atherosclerotic disease. Models to accurately predict transvascular permeation of nanomedicines are needed to aid in design optimization. Here we show that an endothelialized microchip with controllable permeability can be used to probe nanoparticle translocation across an endothelial cell layer. To validate our in vitro model, we studied nanoparticle translocation in an in vivo rabbit model of atherosclerosis using a variety of preclinical and clinical imaging methods. Our results reveal that the translocation of lipid–polymer hybrid nanoparticles across the atherosclerotic endothelium is dependent on microvascular permeability. These results were mimicked with our microfluidic chip, demonstrating the potential utility of the model system.Improving the design of nanomedicines is key for their success and ultimate clinical application (1). The accumulation of such therapeutic or diagnostic nanomaterials primarily relies on enhanced endothelial permeability of the microvasculature in diseased tissue (2). This holds true for a wide range of pathological conditions, including inflammation, atherosclerosis, and most notably, oncological disease (3–6). Although attributed to the enhanced permeability and retention (EPR) effect, the exact mechanism by which nanoparticles accumulate in tumors continues to be a topic of research (7, 8). The “leaky” vasculature of tumors, which facilitates the extravasation of nanoparticles from microvessels (9), is a heterogeneous phenomenon that varies between different tumor models and even more so in patients. Moreover, the exploitation of nanomedicines in other conditions with enhanced microvessel permeability has only recently begun to be studied in detail. For example, in the last 5 y, a small but increasing number of preclinical studies that apply nanoparticle therapy in atherosclerosis models has surfaced (10). Although several targeting mechanisms have been proposed (4), the exact mechanism by which nanoparticles accumulate in atherosclerotic plaques remains to be investigated, but is likely facilitated by highly permeable neovessels that penetrate into the plaque from the vasa vasorum (Fig. 1A), a network of microvessels that supplies the wall of larger vessels (11).Open in a separate windowFig. 1.Development of an endothelialized microfluidic device to probe nanoparticle translocation over a permeable microvessel. (A) Schematics of continuous normal capillaries surrounding the vessel wall as well as permeable capillaries that penetrate into the atherosclerotic plaque from the vasa vasorum. (B) Schematic of an endothelialized microfluidic device that consists of two-layer microfluidic channels that are separated by a porous membrane (3 μm pore) on which ECs are grown. (C) TEER was dynamically measured across the endothelial layer on the membrane between the upper and lower channels. (D) A well-established monolayer of the microvascular endothelium is formed at TEER ∼400 (Ω·cm²). (E) The monolayer becomes highly permeable when stimulated with the inflammatory mediator, TNF-α, as well as with shear stress, with disruption of intercellular junctional structures (i.e., adherens junctions) between ECs, as evidenced by patchy expression of VE–cadherin (green) in the image on the right versus the left. Blue depicts nuclei stained with DAPI. (Scale bar, 20 μm.) (F) FITC–albumin translocation through the endothelial monolayer increases when the chip is treated with TNF-α. (G) The chip with endothelium cultured in different culture media [base, +FBS, +growth factors (GFs)] for 6 h shows a decrease in TEER with increased FITC–albumin translocation. No cell indicates the membrane only. TEER was normalized to the level with no cells (membrane only). (H) Schematic and TEM image of PEGylated lipid-coated nanoparticles encapsulating PLGA-conjugated AuNCs and Cy5.5. The average size was 69.7 ± 14 nm, which was measured from TEM images. (Scale bar, 100 nm.) Details on labeling, synthesis, characterization, and large-scale production procedures can be found in Materials and Methods and Fig. S2.Advances in biomedical imaging allow the study of plaque-targeting nanoparticles in a dynamic fashion with exceptional detail (12, 13). Microchip technology has the potential to monitor nanoparticle behavior at the (sub)cellular level. Microfluidic chips in which endothelial cells (ECs) are grown in the channels can serve as unique in vitro test systems to study microvascular function and associated disorders (14–18). They allow the isolation of specific biological hallmarks relevant to nanoparticle accumulation, such as the leaky endothelium. In the current study, we validate the potential utility of our microchip technology to study nanoparticle translocation over the endothelium and combine this with in vivo and ex vivo multimodality imaging studies on a rabbit model to better understand nanoparticle targeting of atherosclerotic plaques.