PCR—SSP基因分型技术在Rh血型鉴定中的应用研究

目的研究聚合酶链式反应-特异性序列引物（polymerase chain reaction-sequence specific primer,PCR-SSP）基因分型技术在Rh血型鉴定中的应用。方法收集广东省肇庆市无偿捐血者RhD阴性血型样本2例、RhD阳性样本1例,采用盐水法进行C、c、D、E、e抗原分型,对D抗原阴性的样本采用抗人球蛋白法进行确认。提取样本基因组DNA,并应用PCR-SSP基因分型技术特异性扩增RhD以及RhCE基因片段,并对D外显子进行检测。根据有无PCR产物以及产物长度,判断样本的基因型。结果1例RhD阳性样本以及ccdee样本的血清学分型与基因分型结果相符合,1例Ccdee样本的血清学分型与基因分型结果不相符。结论PCR—SSP基因分型技术在RhD血型鉴定中能识别血清意义上的假阴型,在临床输血实践中具有广阔的应用前景。     

PCR-SSP与血清学技术用于ABO血型分型的比较

目的 探讨ABO血型基因分型技术应用于临床检验的可行性.方法 随机抽样100例江西健康汉族个体,并收集20例正反定型不符的标本,采用血清学和序列特异性引物PCR法(PCR-SSP)鉴定血型.结果 100例健康个体的基因分型结果与血清学分型结果一致,19例正反定型不符标本的基因分型结果准确.分型结果表明江西地区以O型为多,100例健康个体中,O型36例(36%),A型32例(32%),B型24例(24%),AB型8例(8%),基因频率:p(A)基因为0.225 3,q(B)基因为0.175 3,r(O)基因为0.599 8,且符合Hardy-Weinberg平衡.结论 基因分型技术应用于临床检验具有可行性,在鉴定疑难血型方面可以作为血清学分型的补充.     

HLA-DQB1 PCR-SSP基因分型技术

HLA-Ⅱ(DR,DQ,DP)配型对提高异基因骨髓移植存活率减少GVHD有重要意义。既往HLA-DQ采用血清学技术检定表型,近年国外转向基因分型,不少实验室已将其列为常规。国内近年来有人报告聚合酶链反应-序列特异性寡核苷酸探针杂交(PCR-SSOP),限制性片断长度多态性     

PCR-SSP基因分型技术在ABO疑难血型定型中的应用

目的 研究聚合酶链式反应-特异性序列引物(PC-SSP)基因分型技术在ABO疑难血型鉴定中的应用.方法 收集8例来自广东省肇庆市无偿献血者的ABO疑难血型样本,采用正反定型实验、吸收放散试验等一系列血型血清学方法进行检测;提取基因组DNA,并用PCR-SSP基因分型技术,特异性扩增ABO基因片段,根据有无PCR产物以及产物长度,判断样本的基因型.结果 通过凝胶电泳检测PCR反应的特异性产物,判断样本1～6号ABO基因型为A/O型,样本7,8号为B/O型.8例疑难ABO基因分型结果与其血型的血清学分型结果吻合.结论 ABO疑难血型定型的解决是一个复杂的难题,而PCR-SSP基因分型是一种方便、快速、可靠的技术,对于解决ABO疑难血型鉴定非常有用.     

先证者为Rh阴性汉族家系RHD基因遗传方式研究

目的分析先证者为Rh阴性家系的RHD基因结构特征,了解Rh血型的基因遗传方式,探讨RHD基因组合和Rhesus盒子（Rh盒子）检测预测新生儿溶血病的意义。方法采用PCR-SSP方法对3例汉族Rh阴性家系的19份样本进行了RHD基因10个外显子的上游Rh盒子、下游Rh盒子和融合Rh盒子检测。结果基因型为RHD-／RHD一个体检测到融合Rh盒子,无上下游Rh盒子;基因型为RHD＋／RHD＋个体检测到上下游Rh盒子,无融合Rh盒子;基因型为RHD＋／RHD-的个体可同时检测到上下游Rh盒子和融合Rh盒子。结论用PCRSSP方法进行家系RHD基因结构分析,操作简便直观。本研究证实在Rh阴性家系中存在着非表达的RHD基因,其遗传方式呈单倍型遗传,符合孟德尔遗传规律。     

HLA-A、B、DR血清学分型与基因分型的比较研究

目的比较研究HLA-Ⅰ、Ⅱ血清学分型与基因分型结果,分析HLA-A、B、DR血清学分型误定规律,提高移植配型的准确性。方法应用聚合酶链反应-序列特异性引物(PCR-SSP)技术,对240名骨髓资料库中已用血清学分型的自愿者进行HLA-A、B、DR基因分型,并对血清学分型与基因分型结果进行低分辨水平的比较研究。结果HLA-A特异性血清学分型错误率在纯合子与杂合子中分别占30.65%与11.52%,总错误率14.35%;HLA-B特异性血清学分型错误率在纯合子与杂合子中分别占42.22%与16.15%,总错误率18.85%;HLA-DR特异性血清学分型错误率在纯合子与杂合子中分别占37.50%与14.58%,总错误率18.63%。HLA-A特异性血清学分型假阴性16.55%,假阳性1.44%,错误指认特异性7.42%,HLA-B分别为20.32%,1.84%和13.56%,HLA-DR分别为13.33%,2.21%和10.05%。结论HLA-A、B、DR纯合子血清学分型错误率明显高于对应的杂合子分型,HLA-B,DR特异性血清学分型错误率显著高于HLA-A特异性;为了提高移植配型的准确性,骨髓资料库中自愿者HLA-A、B、DR位点须重新用基因分型方法分型。     

100例江西地区健康汉族人群ABO血型基因分型的研究

目的研究PCR-SSP法进行ABO血型基因分型的技术,探讨其应用于临床检验的可行性。方法 2006年10月至2011年10月,随机抽取江西地区健康汉族个体100例,采用血清学和PCR-SSP两种方法鉴定血型。结果 100例基因分型结果与血清学结果一致,江西地区人群以O型为多：O〉A〉B〉AB,基因频率：A基因为0.2253,B基因为0.1753,O基因为0.5998,共检出4种等位基因,9种基因型。结论基因分型技术应用于江西地区临床检验具有可行性,可以作为弥补血清学技术不足的一种快速实验方法。     

HLA的PCR-SSP分型技术

PCR-SSP是采用PCR技术,针对被检基因设计一系列具有等位基因多态性的序列特异性引物扩增被检标本,分析PCR产物判定HLA型别。HLA-DR1～DR1l8可以采用19对引物检定;采用两步法以79对引物可以检定DR全部基因型。HLA-DQ采用20条引物不同组合,除检定与血清学型别一致的特异性外还可分析部份基因型;以22对引物则可检定DQB1全部基因型。PCR-SSP技术比PCR-SSOP及PCR-RFLP简易快速,适于临床常规。     

HLA的PCR—SSP分型技术

ＰＣＲ－ＳＳＰ是采用ＰＣＲ技术，针对被检基因设计一系列具有等位基因多态性的序列特异上物扩增被检标本，分析ＰＣＲ产物判定ＨＬＡ型别，ＨＬＡ－ＤＲＩ￣ＤＲ１８可以采用１９对引物检定，采用两步法以７９对引物可以检定ＤＲ全部基因型。ＨＬＡ－ＤＱ采用２０条引物不同组合，陈检定与血清学型别一致的特异性还可分析部份基因型；以２２对引物则可检定ＤＱＢ１全部基因型。ＰＣＲ－ＳＳＰ技术比ＰＣＲ－ＳＳＯＰ及ＰＣＲ－ＲＥ     

徐州地区汉族人群血小板HPA基因多态性的研究

目的 采用序列特异性引物聚合酶链反应(PCR-SSP)检测徐州地区汉族人群血小板特异性抗原(HPA)基因多态性.方法 2010年期间对徐州地区100名无血缘关系的汉族成年人进行HPA1～17基因及新等位基因Caba+分型研究.根据HPA基因分型试剂盒的要求,调整DNA模板浓度;根据人类生长激素基因(HGH)的保守片段设计内参引物进行PCR-SSP,在包含引物混合液的96孔反应板中,按照相同的循环条件扩增PCR产物;用x2检验比较基因分布期望值与观察值,以验证是否符合Hardy-Weinberg遗传平衡.结果 在徐州地区汉族人群的HPA1～17系统中,呈多态性分布的等位基因是HPA1a,HPA2a,HPA3a,HPA4a,HPA5a,HPA6a和HPA15a,其频率分别为0.995 0,0.935 0,0.590 0,0.9950,0.980 0,0.975 0和0.575 0,HPA7～14,16～17及新等位基因Caba+呈单线性分布.结论 徐州地区汉族人群HPA的基因有地区特点,人类血小板抗原HPA的基因具有民族多态性.     

Evaluation of two assessment techniques for adaptation to stress

Two conceptual nursing models, Roy's adaptation model and Erickson and Swain's adaptive potential assessment model are explained, and knowledge is identified within these two assessment techniques for adaptation to stress. The purpose is to identify common, noteworthy areas of nursing science as well as areas where further development in nursing knowledge is needed. A case study is used to compare and contrast these models. When assessing adaptation to stress, one similarity between the approaches appears to be a propositional linkage that supports the influence of developmental level on basic need satisfaction. A divergent area between the two models identifies a need for further development in nursing knowledge regarding the adaptive potential status. This includes information pertaining to what an individual can reasonably do or be expected to do when contending with stressors.     

Median nerve determinations: analysis of two techniques

The objective of this study was to compare two techniques of obtaining median motor and sensory nerve determinants. One method utilized premeasured electrode placement, while the other used anatomic landmarks. It was postulated that increased accuracy could be achieved by more precise measuring techniques. The right median nerve in 50 able-bodied subjects was investigated. Each subject was tested by both anatomic and premeasured methods. Skin temperatures were maintained between 31C and 33C. Latencies were measured to onset and peak, and amplitudes were gauged from baseline to peak. Latency studies were evaluated. Results concluded that mean distal motor latencies, stimulating 8cm from active recording electrode, was 3.1msec +/- 0.4 and amplitude was 11.1mV +/- 3.0. Stimulating at distal wrist crease, mean latency was 2.6msec +/- 0.38 and amplitude was 11.3mV +/- 3.6. Sensory latencies to onset of response when stimulating 14cm from active ring electrode with reference 4cm distally were 2.1msec +/- 0.25. Sensory latency when stimulating at wrist crease and ring electrodes over proximal interphalangeal and distal interphalangeal joints was 2.1msec +/- 0.25 to onset and 2.8msec +/- 0.32 to peak and amplitude was 36.1 +/- 14.6. The authors concluded that there was no statistically significant difference in accuracy between the premeasured and anatomic methods.     

两种麻醉方法在声带息肉摘除术中的效果比较

目的比较瑞芬太尼静脉复合麻醉与芬太尼静吸复合麻醉在声带息肉摘除术中的临床效果。方法ASAI-II级择期行声带息肉摘除术患者60例,随机分为瑞芬太尼-丙泊酚组（I组）和芬太尼．异氟烷组（II组）,每组30例。I组：瑞芬太尼诱导插管,瑞芬太尼一丙泊酚维持麻醉;II组：以芬太尼诱导插管后以异氟烷持续吸入维持麻醉。观察两组麻醉诱导及维持期间血流动力学变化和苏醒拔管时间及苏醒质量评分。结果I组麻醉术中循环波动较小,且显著低于II组。I组呼吸恢复、拔管、呼之睁眼及定向能力恢复时间均早于II组,且I组在拔管后5min的OAAS评分高于II组,组间差异有显著性。结论瑞芬太尼麻醉应用于声带息肉摘除术,不仅能有效地抑制气管插管和置人支撑喉镜时的应激反应,维持血流动力学稳定,而且具有苏醒迅速的优点。     

An electromyographic analysis of two techniques for squat lifting and lowering.

The purpose of this study was to examine the effects of two different alignments of the pelvis and three different loads on electromyographic (EMG) activity of the erector spinae and oblique abdominal muscles during squat lifting and lowering. Each of 15 healthy subjects lifted and lowered loads with the pelvis aligned both in an anterior tilt and in a posterior tilt. Based on total duration, both the lift and the lower were divided into two equal phases. The EMG activity of each muscle was quantified for each half of both the lift and the lower and was normalized to the total EMG produced by the muscle during a maximal voluntary isometric contraction. The results indicate (1) that the EMG activity of the erector spinae muscles was greater when subjects maintained an anterior tilt than when they maintained a posterior tilt; (2) that the EMG activity of the oblique abdominal muscles was greater in the first half of the lift than in the second half for both lifting styles, although the opposite was true for both lowering styles; and (3) that the EMG activity increased with increasing loads. The results suggest that the greater trunk muscle activity occurring with the anterior tilt position may ensure optimal muscular support for the spine while handling loads, thereby reducing the risk for low back injury.     

Upper-limb biomechanical analysis of wheelchair transfer techniques in two toilet configurations

BackgroundUsing proper technique is important for minimizing upper limb kinetics during wheelchair transfers. The objective of the study was to 1) evaluate the transfer techniques used during toilet transfers and 2) determine the impact of technique on upper limb joint loading for two different toilet configurations.MethodsTwenty-six manual wheelchair users (23 men and 3 women) performed transfers in a side and front wheelchair-toilet orientation while their habitual transfer techniques were evaluated using the Transfer Assessment Instrument. A motion analysis system and force sensors were used to record biomechanical data during the transfers.FindingsMore than 20% of the participants failed to complete five transfer skills in the side setup compared to three skills in the front setup. Higher quality skills overall were associated with lower peak forces and moments in both toilet configurations (−0.68 < r < −0.40, p < 0.05). In the side setup, participants who properly placed their hands in a stable position and used proper leading handgrips had lower shoulder resultant joint forces and moments than participants who did not perform these skills correctly (p ≤ 0.04). In the front setup, positioning the wheelchair within three inches of the transfer target was associated with reduced peak trailing forces and moments across all three upper limb joints (p = 0.02).InterpretationTransfer skills training, making toilet seats level with the wheelchair seat, positioning the wheelchair closer to the toilet and mounting grab bars in a more ideal location for persons who do sitting pivot transfers may facilitate better quality toilet transfers.     

汉语纯失读症患者的评价与初步分析

目的：从语言、认知、影像学角度对1例纯失读症患者进行分析。方法：利用汉语版的西方失语症成套测验和波士顿诊断性失语症检查法、综合功能评定法(CFE)、其他认知能力评定法以及CT、MRI、SPECT等对纯失读症患者进行分析。结果：该纯失读症患者朗读、阅读理解、计算功能部分受损，抄写功能轻度受损。影像学提示左枕及左侧脑室后角旁损害。结论：左枕叶损害不伴胼胝体压部同时受损同样可导致纯失读症，症状和以往报道相似。     

两种麻醉方法用于取卵术的临床效果分析

目的研究不间麻醉方式下取卵手术效果及对获卵率的影响。方法将体外受精-胚胎移植、卵胞浆内单精子注射手术的患者随机分为全麻组（异丙酚静脉推注，80例）和局麻组（度冷丁肌注＋利多卡因宫颈旁阻滞，80例），观察两组患者的麻醉效果、循环系统的变化及获卵率。结果术中两组麻醉效果比较无显著性差异（P〉0．05），局麻组术后镇痛效果明显优于全麻组（P〈0．05）；术中两组患者血压比较有非常显著性差异（P〈0．01）、心率有显著性差异（P〈0．05）；两组获卵率比较无显著性差异（P〉0．05）。结论两种麻醉方法对取卵手术过程中的镇痛效果及获卵率没有影响，但是术后局麻镇痛效果明显优于全麻；全麻组多数患者在手术中出现一过性血压和心率下降。     

2种糖化血清蛋白自动分析法评价

目的　对酶法和硝基四氮唑盐 (NBT)法测定糖化血清蛋白的自动化分析法进行评价。方法　分别使用Genzyme公司酶法试剂盒和Roche公司NBT试剂盒 ,在SYNCHRONCX9全自动生化分析仪上建立自动化分析程序 ,评价 2种方法的重复性、线性范围、回收率及相关性。结果　重复性 :酶法批内和批间变异系数 (CV)分别为 0 .6 5 %～ 0 .98%和 1 .98%～ 2 .84 % ,NBT法分别为 1 .1 1 %～ 1 .2 6 %和 2 .73%～ 3.1 5 %。线性范围 :酶法 30～ 1 5 0 0 μmol/L ,NBT法 30～ 1 0 0 0 μmol/L。回收率 :酶法 99.7%～ 1 0 2 .4 % ,NBT法 99.1 %～ 1 0 3.2 %。灵敏度 (A/ 1 0 0 μmol/L) :酶法 0 .0 1 82 ,NBT法 0 .0 2 5 9。相关性 :YNBT法 =1 .0 0 36X酶法 +46 .3,r =0 .9776。正常参考范围 ( x± 2s) :酶法 1 2 3.4～ 2 38.2 μmol/L、NBT法 1 98.2～ 2 84 .6 μmol/L。 结论　 2种方法均具有较高的精密度和准确度 ,测定结果呈直线正相关 ,应用时必须使用各自的参考范围     

2种糖化血清蛋白自动分析法评价

目的对酶法和硝基四氮唑盐(NBT)法测定糖化血清蛋白的自动化分析法进行评价.方法分别使用Genzyme公司酶法试剂盒和Roche公司NBT试剂盒,在SYNCHRON CX9全自动生化分析仪上建立自动化分析程序,评价2种方法的重复性、线性范围、回收率及相关性.结果重复性酶法批内和批间变异系数(CV)分别为0.65%～0.98%和1.98%～2.84%,NBT法分别为1.11%～1.26%和2.73%～3.15%.线性范围酶法30～1 500 μmol/L,NBT法30～1 000 μmol/L.回收率酶法99.7%～102.4%,NBT法99.1%～103.2%.灵敏度(A/100 μmol/L)酶法0.018 2,NBT法0.025 9.相关性YNBT法=1.003 6X酶法+46.3,r=0.977 6.正常参考范围(±2s)酶法123.4～238.2 μmol/L、NBT法198.2～284.6 μmol/L.结论 2种方法均具有较高的精密度和准确度,测定结果呈直线正相关,应用时必须使用各自的参考范围.