液相芯片技术联合等位基因特异性引物延伸反应检测结核分枝杆菌耐多药突变

目的建立一种耐多药结核分枝杆菌检测方法。方法设计并构建含有kat G315、rpo B526、rpo B531耐药基因突变位点的质粒;通过PCR扩增及纯化、等位基因特异性引物延伸反应(ASPE)、荧光微球杂交反应及液相芯片系统Luminex 200检测;并对该方法反应条件进行探索及方法学评价。结果 ASPE反应中Tsp DNA聚合酶最适浓度为0.025 U/μL,杂交荧光微球浓度为每μL 100个,链霉亲和素藻红蛋白浓度及杂交时间分别为2μg/m L和20 min;所建立方法的检测灵敏度最低可达1ng/m L;当质粒浓度为1.5×105ng/m L时,批间和批内变异系数(CV)分别为6.48%~12.15%和0.35%~6.92%;当质粒浓度为2 ng/m L时,批间和批内变异系数(CV)分别为5.73%~10.77%和0.97%~8.91%;特异性引物探针与其他突变位点均无交叉反应。结论液相芯片技术联合等位基因特异性引物延伸反应检测耐多药结核分枝杆菌灵敏且具有成本低、高通量等特点,极具临床应用潜力。     

悬浮阵列技术在研究与临床中的应用

悬浮阵列技术是一种以荧光编码微球为核心,集流式细胞、激光分析、高速数字信号处理等多种技术于一体的多指标并行分析技术平台,可一次同时准确定量检测100种不同的生物分子;具有高通量、高灵敏度,并行检测等特点;常用于免疫分析、核酸研究、酶学分析、受体、配体识别分析等研究.     

Automation of biochip array technology for quality results.

Background: Proteomics' requirement for simultaneous measurement of multiple markers is now possible with biochip array technology. Many laboratories utilise in-house, manual procedures for biochip fabrication and sample testing. Reproducibility and standardisation of biochip processes is vital to ensure quality of results and offer the best tool for elucidation of complex relationships between multiple proteins in diseased conditions. Methods: Various novel control checks have been implemented in biochip fabrication, reagent manufacture, automation and imaging processes for the Evidence analyser. Reference spots enable location of discrete test regions on the surface of the biochip and simultaneous quantification of multiple markers. Performance and standardisation methods are presented. Results: Formulation of dispense solution for discrete test regions had a direct effect on their shape, stability and integrity on the biochip surface. Assays for fertility hormones and drugs of abuse demonstrated excellent precision, stability and comparison with other commercial methods. Conclusion: Control processes employed in the manufacture and analysis of Evidence components ensure reproducibility of assays for a range of routine and novel markers.     

DNA array technology and diagnostic microbiology

Near instantaneous detection of pathogens from clinical material, combined with simultaneous prediction of their antimicrobial resistance profiles, would revolutionize the impact of microbiology on the management of infection. Array-based assays allow a range of characteristics to be rapidly and simultaneously determined. At present these systems have found their primary role as research tools for the monitoring of mRNA expression in the form of DNA microarrays or 'chips'. As fabrication costs reduce and validated targeted arrays are developed, it is inevitable they will be used for more routine applications. Microfluidics offers the exciting possibility of combining purification, amplification and detection in a single disposable device; microarrays are particularly suitable for use within these systems. Arrays will become an important tool for clinical diagnostics.     

移动式悬吊牵引轮椅的制作及应用

目的 探讨自行设计制作的移动式悬吊牵引轮椅的临床应用效果。方法 选取我科脊柱中心2017年9月—2018年8月收治的125例青少年特发性脊柱侧凸住院患者,采用随机数字表法分为试验组67例,对照组58例。试验组采用自行设计制作的移动式悬吊牵引轮椅进行术前牵引治疗,对照组采用常规床尾固定牵引法进行牵引治疗。比较两组脊柱柔韧性、治疗耐受时间和坚持治疗意愿、舒适度、局部皮肤情况。结果 试验组治疗后脊柱柔韧性优于对照组（t=5.317,P<0.001）;试验组治疗耐受时间为（557.45±23.28） min,对照组（312.46±29.85） min,两组比较,差异有统计学意义（t=51.493,P<0.001）;试验组坚持治疗意愿（98.5%）明显高于对照组（36.2%）;试验组舒适度和局部皮肤情况均优于对照组（P<0.05）。结论 使用移动式悬吊牵引轮椅可改善青少年脊柱侧凸患者脊柱柔韧性,提高其术前牵引治疗的耐受时间、治疗依从性和舒适度,避免皮肤不适,使用方便且并发症少。     

Dual-spectra encoded suspension array using reversed-phase microemulsion UV curing and electrostatic self-assembling

The rapid growth of demand for high-throughput multiplexed biochips from modern biotechnology has led to growing interest in suspension array based on multi-channel encoded microbeads. We prepare dual-spectra encoded PEGDA microbeads (DSEPM) by reversed-phase microemulsion UV curing method and layer-by-layer electrostatic self-assembly method. Excitation of the synthesized DSEPM results in two spectra, including fluorescence spectra from quantum dots and laser induced breakdown spectra from nanoparticles with specific elements. With further surface modification and bio-probes grafting, we use DSEPM to carry a series of detection experiments of biomolecules. The adsorption experiment to two types of anti-IgG in mixture sample has demonstrated the availability of DSEPM in multiplexing. Then, the contrast experiment has verified the specificity of DSEPM in detection. Finally, we carry out the concentration gradient experiment and obtain the response curve to show the performance of DSEPM in quantitative analysis. The results indicate our method provide an effective way to improve multiplexed biochips with more coding capacity, accuracy and stability.

The rapid growth of demand for high-throughput multiplexed biochips from modern biotechnology has led to growing interest in suspension array based on multi-channel encoded microbeads.     

液相芯片法检测终末期肾病患者胞外Th1/Th2细胞因子与促炎细胞因子

目的探讨终末期肾脏病（ESRD）患者胞外Th1/Th2细胞因子的状况和临床意义。方法选取12例ESRD患者为实验组,12例健康人为对照组,运用液相芯片技术（SA）测定各实验组血清中Th1/Th2和促炎因子的水平,同时用酶联免疫吸附测定法（ELISA）定量检测细胞因子白细胞介素-8（IL-8）的水平。通过用ELISA和液相芯片法同时检测IL-8的相关性来评估液相芯片法的稳定性和准确性。结果ESRD患者血液透析前IL-10水平与对照组相比有所上升;血液透析后IL-1β水平与对照组相比降低;而血液透析前后肿瘤坏死因子-β（TNF-β）和IL-8水平均降低。ESRD患者血液透析后IL-1β、IL-4、IL-8和IL-10水平与血液透析前相比均有下降。在检测IL-8时,液相芯片法和ELISA表现出显著的相关性（P〈0.001,r=0.946）。结论血液透析后ESRD患者体内存在着细胞因子的变化,即Th2细胞因子IL-10上升而Th1细胞因子TNF-β和促炎因子IL-8降低。血液透析影响ESRD患者的细胞因子谱。     

急性冠脉综合征患者超敏C-反应蛋白、脂蛋白a、脂联素的悬浮芯片技术集成检测及其意义

目的:检测急性冠脉综合征(ACS)患者血中超敏C-反应蛋白(hs-CRP)、脂蛋白a[Lp(a)]及脂联素(APN)3种蛋白因子的浓度并探讨其临床意义.方法:用悬浮芯片技术集成检测116例ACS患者(ACS组)hs-CRP、Lp(a)及APN 3种蛋白因子的浓度,并与30例单纯稳定型心绞痛患者(SAP组)及35例正常人群(NC组)进行对照.结果: ACS组hs-CRP、Lp(a)较SAP组及NC组浓度均增高,APN值则明显降低(均P ＜ 0.05),其中ACS组中AMI 组hs-CRP、Lp(a)浓度高于UAP组,APN值低于UAP组(均P ＜ 0.05).ACS各亚组中预后不良患者hs-CRP、Lp(a)水平高于预后良好者(均P ＜ 0.001),而APN浓度则显著低于预后良好患者(P ＜ 0.01).hs-CRP、Lp(a)的含量与斑块不稳定程度呈显著正相关关系(r=0.592,P ＜ 0.001;r=0.385,P ＜ 0.001),APN的含量与斑块不稳定程度则呈显著负相关关系(r=-0.627,P ＜ 0.001).结论:hs-CRP、Lp(a)、APN可能参与ACS病变全过程,与ACS临床类型及其预后高度相关,因此可用来预测ACS的病变稳定程度和心血管事件发生的危险度,从而判断预后.     

Nursing care planning. Overview and application to the patient in shock

The systematic nursing process is an essential prerequisite for providing effective care for patients in shock. Nurses use assessment skills to identify actual and potential collaborative problems and nursing diagnoses. Once collaborative problems and nursing diagnoses are identified, the nurse develops appropriate individualized patient outcomes and plans care to accomplish the outcomes and resolve the problems. Nursing actions from the three domains may be included in the care plan. The evaluation phase assesses the adequacy of nursing actions in accomplishing the stated goals and returns the process to the stage of assessment. Application of the nursing care planning process to the patient in shock can be somewhat difficult because of the critical nature of the illness, complex assessment data, and rapidly changing interventions. It is indeed a challenge for the critical care nurse to integrate sound principles of nursing care planning into the busy intensive care environment, but the outcome of improved patient care is worth the effort.     

护理结局分类系统的概况及应用

当今,许多国家的健康服务系统重视从健康服务费用和服务后病人的结局两个方面评价健康服务系统的有效性,护理学作为一个单独学科,必须建立护理学科领域独立语言系统,确立受本学科影响的病人结局.     

应用液相芯片技术检测拉米夫定耐药相关性HBV变异

法较     

便捷悬挂医用引流袋的制作与应用

现临床应用的一次性医用引流袋,其悬挂设置为一长不足3 cm的固定塑料管系带,使用时,需另外在病床的床框边缘拴系其他绳索来固定引流袋.此法存有以下缺点:病人变更体位、卧位时,引流袋需因病人体位的变更而变更其悬挂位置,这样,就需解开拴系引流袋的绳索,重新拴系到其他位置,操作麻烦,增加护理人员工作量;若病人下床活动时,引流袋则无处悬挂,如引流袋内液体稍多,解系引流袋移动时也不易操作.     

静脉输液针头悬挂环的设计与应用

目的观察排气时使用针头悬挂环的效果。方法将186例输同种药物的患者随机分为实验组和对照组,各93例,实验组使用针头悬挂环进行排气,对照组采用临床常规排气方法。结果实验组排气时间为（7．1±0．56）s,对照组为（15．8±0．56）s,两组比较差异有统计学意义（t=-26．817,P〈0．01）;实验组损失液量为（0．03±0．01）ml,对照组损失液量为（0．21±0．06）ml,两组比较差异有统计学意义（t=-8．084,P〈0．01）。结论针头悬挂环制作简单,可以缩短排气时间、提高一次排气成功率、避免浪费药液,使护理人员工作更省时省力,具有临床推广意义。     

医学模拟教育的应用概况

分析了医学模拟教育在临床技能训练和医学教育中的优势,认为其应用前景可观,但医学模拟教育也有其局限性,不能完全取代临床教学.当前我国医学模拟教育应用尚处于起步阶段,在未来一定会有蓬勃的发展,更好地为医学教育服务.     

Application of a multiplex suspension array for rapid and simultaneous identification of clinically important mold pathogens

We have developed a microsphere-based suspension array (MSA) for the identification of 23 medically important mold pathogens including Aspergillus spp., Fusarium spp., Mucor spp., Rhizopus spp., Rhizomucor pusillus, Penicillium marneffei, Saksenaea vasiformis, Apophysomyces elegans, Lichtheimia corymbifer, and Syncephalastrum racemosum. Twenty-one oligonucleotide probes were designed based on the internal transcribed spacer (ITS2) region for species level identification of molds. Among the 21 probes, 2 probes are shared by more than one species due to low or absence of sequence variability, i.e. Rpam for Rhizopus azygosporus/Rhizopus microsporus and Fumop for Fusarium moniliforme/Fusarium oxysporum/Fusarium pallidoroseum. No cross reactivity was identified except for probes of Mucor racemosus (Murac) which cross react with Mucor hiemalis and Mucor ramosissimus. The sensitivity of MSA is 100 fg–1 ng. The whole procedure including DNA extraction and PCR amplification can be finished within 5 h. The MSA is simple, rapid, specific, high-throughput and capable of multiple-species detection in one reaction tube.     

组织芯片制备与原位分析技术

目的 建立组织芯片制备技术并观察在各种原位分析中的应用。方法 1．在传统病理学技术的基础上制备各种组织蜡块作为组织供体，再用组织阵列仪按照预先设计制作各种组织阵列蜡块，经切片和烤片后即成为各种组织芯片。2．用各种组织芯片按照常规方法进行HE染色、免疫组化(IHC)、原位杂交(ISH)和原位PCR(IS-PCR)实验分析。结果 (1)用本方法可分别制备低、中和高密度组织芯片，而且组织芯片排列整齐，外形美观，组织点大小一致且不易脱落。(2)HE染色结果均一，核浆对比明显，便于进行诊断和比较诊断；IHC和免疫荧光染色清晰，便于观察和诊断；ISH和IS-PCR实验效果较好，可用于各种基因扩增分析。结论 (1)本方法是一种比较成熟的组织芯片制备技术。(2)用本方法制备的组织芯片可以进行HE染色和各种原位分析。     

多媒体技术在手术室护生带教中应用

目的：讨论多媒体技术运用于手术室护生带教的优越性。方法：选取2008年6月-2009年6月在我院手术室实习的148名护理专业实习生，随机分为多媒体组和传统组各74名，多媒体组实施多媒体技术带教，传统组按传统方法带教，比较两组的教学效果。结果：多媒体组护生的出科前综合考试成绩、带教老师和手术医师的满意度明显提高，两组的教学效果比较差异有统计学意义（P〈0．05），同时多媒体组教学需要时间明显减少，减少了带教老师重复工作。结论：多媒体技术是一种科学、实用、适合手术室的新型教学方法。