《管道护理》CAI课件在外科护理临床教学中的应用评估

目的提高《管道护理》的临床教学效果。方法制作《管道护理》的CAI课件,并将课件应用于实习学生和临床护士的教学中。将实习学生及外科护士共210人分层随机分为传统教学组、课件教学组,对两组人员分别进行传统的授课教学和CAI教学比较。结果两组人员的课后考试成绩经统计学处理(t检验),P值<0.01,具有显著性差异;对课件教学组人员进行CAI教学效果调查,结果显示:课件教学组人员认为课件整体结构合理为91.3%,知识层次分明为96.5%,界面美观为95.7%,文字表达清楚为96.5%,操作简单、人机交互性好为92.2%,能帮助理解教学中的难点、重点为96.5%,能帮助增强记忆为91.3%,能使课堂注意力更集中为90.4%,能增加学习兴趣为96.5%。结论应用《管道护理》CAI课件教学解决了临床缺乏病例等难题;《管道护理》CAI课件教学增强了学生的学习兴趣和记忆,提高了《管道护理》的教学质量;《管道护理》CAI课件教学有助于临床护士的培训。     

Identification of nafamostat mesylate as a selective stimulator of NK cell IFN-γ production via metabolism-related compound library screening

Natural killer (NK) cells play important roles in controlling virus-infected and malignant cells. The identification of new molecules that can activate NK cells may effectively improve the antiviral and antitumour activities of these cells. In this study, by using a commercially available metabolism-related compound library, we initially screened the capacity of compounds to activate NK cells by determining the ratio of interferon-gamma (IFN-γ)⁺ NK cells by flow cytometry after the incubation of peripheral blood mononuclear cells (PBMCs) with IL-12 or IL-15 for 18 h. Our data showed that eight compounds (nafamostat mesylate (NM), loganin, fluvastatin sodium, atorvastatin calcium, lovastatin, simvastatin, rosuvastatin calcium, and pitavastatin calcium) and three compounds (NM, elesclomol, and simvastatin) increased the proportions of NK cells and CD3⁺ T cells that expressed IFN-γ among PBMCs cultured with IL-12 and IL-15, respectively. When incubated with enriched NK cells (purity?≥?80.0%), only NM enhanced NK cell IFN-γ production in the presence of IL-12 or IL-15. When incubated with purified NK cells (purity?≥?99.0%), NM promoted NK cell IFN-γ secretion in the presence or absence of IL-18. However, NM showed no effect on NK cell cytotoxicity. Collectively, our study identifies NM as a selective stimulator of IFN-γ production by NK cells, providing a new strategy for the prevention and treatment of infection or cancer in select populations.

     

创新特色护理服务模式的实施

目的 探讨开展特色护理服务的方法,提高护理服务质量和患者满意率.方法 对2003年12月来本院就诊的21 100例患者及家属开展创新特色护理服务,并每月随机抽取部分住院患者对护理服务质量及患者满意率进行问卷测评.结果 开展特色护理服务前后,护理服务质量提高6.3%,患者满意率提高5.8%.结论 特色护理模式的创新,有效改善了护惠沟通及传统宣教方式,保证了良好的护理服务质量,提高了患者满意率.     

抗癌消水膏治疗恶性胸腔积液的临床研究

探讨中药外敷法治疗恶性胸腔积液的临床疗效和特点.用黄芪、桂枝等研制成抗癌消水膏外敷恶性胸水患者的胸壁,与腔内免疫治疗法比较,对体力状况(PS)＞2的肿瘤病人并发恶性胸水者进行随机分组各25例.观察各组病人胸水、胸痛、生活质量的改善状况及胸水癌细胞核抗原(PCNA)弱阳性率的表达.结果提示抗癌消水膏治疗恶性胸水有效率为56%,优于腔内免疫治疗组(48%).特别是对胸痛、病人生活质量有明显改善作用,并可抑制胸水中癌细胞增殖.     

In vitro selection of a sodium-specific DNAzyme and its application in intracellular sensing

Over the past two decades, enormous progress has been made in designing fluorescent sensors or probes for divalent metal ions. In contrast, the development of fluorescent sensors for monovalent metal ions, such as sodium (Na⁺), has remained underdeveloped, even though Na⁺ is one the most abundant metal ions in biological systems and plays a critical role in many biological processes. Here, we report the in vitro selection of the first (to our knowledge) Na⁺-specific, RNA-cleaving deoxyribozyme (DNAzyme) with a fast catalytic rate [observed rate constant (k_obs) ∼0.1 min⁻¹], and the transformation of this DNAzyme into a fluorescent sensor for Na⁺ by labeling the enzyme strand with a quencher at the 3′ end, and the DNA substrate strand with a fluorophore and a quencher at the 5′ and 3′ ends, respectively. The presence of Na⁺ catalyzed cleavage of the substrate strand at an internal ribonucleotide adenosine (rA) site, resulting in release of the fluorophore from its quenchers and thus a significant increase in fluorescence signal. The sensor displays a remarkable selectivity (>10,000-fold) for Na⁺ over competing metal ions and has a detection limit of 135 µM (3.1 ppm). Furthermore, we demonstrate that this DNAzyme-based sensor can readily enter cells with the aid of α-helical cationic polypeptides. Finally, by protecting the cleavage site of the Na⁺-specific DNAzyme with a photolabile o-nitrobenzyl group, we achieved controlled activation of the sensor after DNAzyme delivery into cells. Together, these results demonstrate that such a DNAzyme-based sensor provides a promising platform for detection and quantification of Na⁺ in living cells.Metal ions play crucial roles in a variety of biochemical processes. As a result, the concentrations of cellular metal ions have to be highly regulated in different parts of cells, as both deficiency and surplus of metal ions can disrupt normal functions (1–4). To better understand the functions of metal ions in biology, it is important to detect metal ions selectively in living cells; such an endeavor will not only result in better understanding of cellular processes but also novel ways to reprogram these processes to achieve novel functions for biotechnological applications.Among the metal ions in cells, sodium (Na⁺) serves particularly important functions, as changes in its concentrations influence the cellular processes of numerous living organisms and cells (5–8), such as epithelial and other excitable cells (9). As one of the most abundant metal ions in intracellular fluid (10), Na⁺ affects cellular processes by triggering the activation of many signal transduction pathways, as well as influencing the actions of hormones (11). Therefore, it is important to carefully monitor the concentrations of Na⁺ in cells. Toward this goal, instrumental analyses by atomic absorption spectroscopy (12), X-ray fluorescence microscopy (13), and ²³Na NMR (14) have been used to detect the concentration of intracellular Na⁺. However, it is difficult to use these methods to obtain real-time dynamics of Na⁺ distribution in living cells. Fluorescent sensors provide an excellent choice to overcome this difficulty, as they can provide sensitive detection with high spatial and temporal resolution. However, despite significant efforts in developing fluorescent metal ion sensors, such as those based on either genetically encoded probes or small molecular sensors, most fluorescent sensors reported so far can detect divalent metal ions such as Ca²⁺, Zn²⁺, Cu²⁺, and Fe²⁺ (15–21). Among the limited number of Na⁺ sensors, such as sodium-binding benzofuran isophthalate (22), Sodium Green (23), CoroNa Green/Red (24, 25), and Asante NaTRIUM Green-1/2 (26), most of them are not selective for Na⁺ over K⁺ (22–25, 27, 28) or have a low binding affinity for Na⁺ (with a K_d higher than 100 mM) (25, 27–31). Furthermore, the presence of organic solvents is frequently required to achieve the desired sensitivity and selectivity for many of the Na⁺ probes (32–34), making it difficult to study Na⁺ under physiological conditions. Therefore, it is still a major challenge to design fluorescent sensors with strong affinity for Na⁺ and high selectivity over other monovalent and multivalent metal ions that work under physiological conditions.To meet this challenge, our group and others have taken advantage of an emerging class of metalloenzymes called DNAzymes (deoxyribozymes or catalytic DNA) and turned them into metal ion probes. DNAzymes were first discovered in 1994 through a combinatorial process called in vitro selection (35). Since then, many DNAzymes have been isolated via this selection process. Among them, RNA-cleaving DNAzymes are of particular interest for metal ion sensing, due to their fast reaction rate and because the cleavage, which is catalyzed by a metal ion cofactor, can easily be converted into a detectable signal (36–38). Unlike the rational design of either small-molecule or genetically encoded protein sensors, DNAzymes with desired sensitivity and specificity for a metal ion of interest can be selected from a large library of DNA molecules, containing up to 10¹⁵ different sequences (35, 39). A major advantage of DNAzymes as metal ion sensors is that metal-selective DNAzymes can be obtained without prior knowledge of necessary metal ion binding sites or specific metal–DNA interaction (40, 41). In addition, through the in vitro selection process, metal ion binding affinity and selectivity can be improved by tuning the stringency of selection pressure and introducing negative selection against competing metal ions (39, 40). Finally, DNA is easily synthesized with a variety of useful modifications and its biocompatibility makes DNAzyme-based sensors excellent tools for live-cell imaging of metal ions. As a result, several metal-specific DNAzymes have been isolated and converted into sensors for their respective metal ion cofactors, including Pb²⁺ (35, 42, 43), Cu²⁺ (44, 45), Zn²⁺ (46), UO₂²⁺ (47), and Hg²⁺ (48). They have recently been delivered into cells for monitoring UO₂²⁺ (41, 49), Pb²⁺ (50), Zn²⁺ (51), and histidine (52) in living cells.However, in contrast to the previously reported DNAzymes with divalent metal ion selectivity, no DNAzymes have been reported to have high selectivity toward a specific monovalent metal ion. Although DNAzymes that are independent of divalent metal ions have been obtained (53–55), including those using modified nucleosides with protein-like functionalities (i.e., guanidinium and imidazole) (56–58), no DNAzyme has been found to be selective for a specific monovalent metal ion over other monovalent metal ions. For example, the DNAzyme with the highest reported selectivity for Na⁺ still binds Na⁺ over K⁺ with only 1.3-fold selectivity (54). More importantly, those DNAzymes require very high concentrations of monovalent ions (molar ranges) to function and display very slow catalytic rates (e.g., 10⁻³ min⁻¹) (53–55). The poor selectivity, sensitivity, and slow catalytic rate render these DNAzymes unsuitable for cellular detection of Na⁺, due to interference from other monovalent ions such as K⁺ (which is present in concentrations about 10-fold higher than Na⁺), and the need to image the Na⁺ rapidly.In this study, we report the in vitro selection and characterization of an RNA-cleaving DNAzyme with exceptionally high selectivity (>10,000-fold) for Na⁺ over other competing metal ions, with a dynamic range covering the physiological Na⁺ concentration range (0.135–50 mM) and a fast catalytic rate (k_obs, ∼0.1 min⁻¹). This Na⁺-specific DNAzyme was transformed into a DNAzyme-based fluorescent sensor for imaging intracellular Na⁺ in living cells, by adopting an efficient DNAzyme delivery method using a cationic polypeptide, together with a photocaging strategy to allow controllable activation of the probe inside cells.     

早期空肠内营养在重症急性胰腺炎综合治疗中的疗效观察

背景：营养支持治疗是重症急性胰腺炎（SAP）综合治疗的重要组成部分，其方式和方法的选择对改善SAP患者全身状况、防治肠道功能衰竭、减轻全身性炎症反应、减少感染机会以及改善预后尤为重要。目的：探讨早期空肠内营养支持疗法对SAP患者病程、一般情况和预后的影响。方法：38例SAP患者随机分为肠内营养（EN）组和全胃肠外营养（TPN）组；EN组（18例）于发病早期（3～5d）胃镜下放置空肠营养管，并给予空肠内要素饮食；TPN组（20例）常规给予静脉营养支持治疗。所有患者冶疔前后定期检测血常规、血糖、肝功能、肾功能、血清白蛋向和前白蛋白水平等，并予腹部超声、CT或磁共振成像（MRI）检查以评估胰腺和腹腔病变情况等变化。结果：两组治疗前肝功能、肾功能、外周血向细胞汁数等均无显著差别。治疗2，3周后EN组白细胞恢复正常时间显著短于TPN组（P〈0.01）。EN组治疗后血清白蛋门水平和前白蛋白水平均显著高于其治疗前水平（P〈0.01和P〈0．001）和TPN组治疗后水平（P〈0．01和P〈0．001）：EN组的并发症发生率、感染率和死亡率均低于TPN组。但无显著性差异（P〉0．05）。结论：早期空肠内营养支持疗法可促进SAP患者肝脏蛋白合成，提高血清白蛋白和前白蛋白水平，并有可能降低感染率和并发症的发生率，改善SAP患者的预后。     

医学监护在联苯胺作业膀胱癌高发人群中应用的意义

本文报道了对联苯胺作业膀胱癌高发人群实行医学监护的方法和结果。指出定期进行尿液脱落细胞检查,并对其中巴氏分级Ⅲ级以上者做膀胱镜检查,不但可以早期发现膀胱肿瘤,同时也可以对尿路粘膜上皮组织炎症及增生等病变作出一定的诊断。有利于早期治疗。尿液脱落细胞检查方法简便易行,对病人安全无痛苦,并可重复检验,诊断肿瘤的阳性率较高,可以作为普查和职业人群医学监护的主要手段。