部分脱乙酰甲壳质生物套管小间隙桥接修复周围神经损伤:免疫组织化学观察

目的:神经断端保留小间隙的静脉桥接模拟神经外膜形成神经再生室,为周围神经再生创造了良好的生理环境,从而保证神经束的良好对合。实验采用部分脱乙酰甲壳质作为套管材料,用小间隙桥接方法修复坐骨神经损伤,观察套管内的神经再生情况,并与传统外膜缝合法进行比较。方法:实验于2001-01/2002-10在北京大学人民医院创伤骨科实验室完成。①主要材料:实验所用中空圆柱形套管为北京大学人民医院与中国纺织科学院共同发明的一种部分脱乙酰甲壳质生物套管(专利号:01136314.2)。实验中所用套管尺寸为:管长4 mm,壁厚0.1 mm,内径1.5 mm。②实验动物:健康成年雄性SD大鼠20只,随机分成2大组,每组10只,每一大组全部10只动物的左腿坐骨神经为一组,右腿坐骨神经为另一组,每组10根坐骨神经。另取5只同样大鼠双侧坐骨神经未做处理作为正常对照组。③实验方法:外膜原位缝合组:切断坐骨神经,显微镜下神经外膜原位缝合;生物套管小间隙原位桥接组:切断坐骨神经,显微镜下小间隙套管桥接;断端旋转180°外膜缝合组:切断坐骨神经,显微镜下远端旋转180°后,神经外膜缝合;断端旋转180°生物套管小间隙桥接组:切断坐骨神经,显微镜下远端旋转180°后,小间隙套管桥接。④实验评估:术后第7,14,21,28,42天取坐骨神经,进行免疫组织化学染色及观察。结果:再生神经延套管中央走行,7 d时已有部分纤维通过2 mm间隙,14 d时有髓纤维数量明显多于近端。21 d后,套管组与原位外膜组新生有髓神经纤维数相近。再生纤维胞核数量较多,髓鞘纤细。套管结构完整。结论:此种部分脱乙酰甲壳质生物套管内的再生神经连续、整齐,髓鞘完整,其神经再生情况好于传统外膜缝合法。     

从兔股骨头中提取总RNA的方法特点

目的:建立一种高效、快速的骨组织总RNA提取方法。方法:实验于2005-01/2006-01在昆明医学院实验动物中心和中国科学院昆明动物研究所中科院细胞与分子进化重点实验室完成。取健康新西兰大白兔1只,截取其股骨头,迅速置于液氮罐中保存,于研钵中研磨,使骨组织始终保存于液氮中,继续研磨,如此重复3次,然后利用Trizol使骨细胞结构迅速破坏,将粉末转入离心管,室温静置5min。随后加入氯仿等有机溶剂处理、离心,使RNA与细胞DNA、蛋白质及其他成分分离从而得到总RNA。最后鉴定RNA的质量、纯度及产率,取2μL提取出的RNA在体积分数为0.008的甲醛变性琼脂糖凝胶上进行电泳,主要观察RNA的28S、18S及5S三个条带是否清晰,有无降解和DNA污染。以99μLDEPC水稀释1μLRNA样品,紫外分光光度计测量其吸光度(A)值,A260/A280之比值表示RNA的纯度,同时根据吸光度值计算其质量浓度。结果:①对提取的兔股骨头RNA进行琼脂糖凝胶电泳,可显示清晰的28S、18S两个条带,5S条带亦可见,表明了RNA是完整的。②用紫外分光光度法测定兔股骨头中提取出的RNAA260/A280,结果表明由本法提取的RNA纯度高,无DNA和蛋白质的污染。③经紫外分光光度计吸收定量,每毫克兔股骨头组织能提取1.0～1.2μg的总RNA。结论:本法提取骨组织总RNA方便、快捷,质量高,可用于骨组织的分子生物学研究。     

应用遥测肌电技术实测分析人体助行带的生物力学作用

目的:人体助行带为一种随身佩带的动力装置,应用遥测肌电技术测定和分析自行研制的人体助行带的生物力学作用机制。方法:实验于2005-12/2006-06在天津体育大学运动生物力学实验室进行。选取天津市天津医院30名受试对象,均对本实验知情同意。按照生物力学的测试要求,首先对受试者进行5次以上行走状态下的遥测肌电测定,然后在同等条件下对受试者佩带助行带实施对照研究。实验采用的助行带为自行研制的,由腰至足以双层弹力带作为动力装置的弹性结构。结果:行走摆动相大腿肌肌电:佩带助行带后肌内侧肌、肌二头肌、半腱肌肌电高于佩带前(P<0.01)。支撑相小腿肌肌电:佩带助行带后胫前肌、腓骨长肌、腓肠肌肌电高于佩带前(P<0.01)。结论:通过弹力带施加于下肢的外部载荷,能够起到增强下肢肌电以及产生助行的生物力学功效。     

急性心肌梗死后外周血单个核细胞表面CD147与基质金属蛋白酶-9 mRNA表达的相关性

目的：基质金属蛋白酶在急性心肌梗死后的心室重构中起着重要作用，但其调节机制目前尚未明确。实验拟通过动物模型的建立及体外细胞培养，观察急性心肌梗死后单个核细胞表面CD147与心肌成纤维细胞基质金属蛋白酶-9 mRNA表达的关系。 方法：实验于2006—08／2007-06在河北省人民医院临床实验中心完成。实验材料：SD大鼠及SD仔鼠（出生1～3d）购自河北医科大学试验动物中心。实验过程中对动物处置符合动物伦理学标准。实验方法：①将30只大鼠随机分为急性心肌梗死组（n=15）和假手术组（n=15），假手术组只过线不结扎。流式细胞分析法检测大鼠术后24h外周血单个核细胞表面CD147表达。②选择SD仔鼠制备心肌成纤维细胞。将单个核细胞与心肌成纤维细胞以细胞数0．5：1，1：1，2：1混合培养24h后，半定量反转录一聚合酶联反应法检测基质金属蛋白酶-9 mRNA表达。当单核细胞与心肌成纤维细胞2：1混合时，加入CD147单克隆抗体1，2，4μL/L，培养24h后检测基质金属蛋白酶-9 mRNA表达。 结果：①急性心肌梗死后外周血单个核细胞表面CD147表达明显增加。②单个核细胞与心肌成纤维细胞混合培养，随着单个核细胞比例的增加，心肌成纤维细胞基质金属蛋白酶-9 mRNA表达增加。③在单个核细胞与心肌成纤维细胞2：1混合培养体系中，随着加入CD147单克隆抗体浓度的增加，基质金属蛋白酶-9 mRNA生成减少。 结论：急性心肌梗死后单个核细胞表面CD147表达明显增加，对心肌成纤维细胞基质金属蛋白酶-9生成起上游调节作用。     

包膜活性炭吸附血液灌流清除人血浆中毒鼠强的实验

目的:观察包膜活性炭对血浆中毒鼠强的清除率及吸附规律。方法:实验于2004-05/2006-04在军事医学科学院毒物药物研究所国家重点实验室完成。采用包膜活性炭灌流器对毒鼠强血浆样进行血液灌流吸附,在灌流的1,2,3h分别取样,经乙酸乙酯萃取后,用气相色谱氮磷检测器法(GC/NPD)测定其含量并计算清除率。结果:活性炭对毒鼠强的吸附作用在血液灌流1h最高,灌流2h后毒鼠强质量浓度无明显变化。400,200μg/L毒鼠强血液灌流1h清除率分别为(57.83±1.85)%,(48.18±1.81)%。结论:用包膜活性炭吸附剂进行血浆的灌流吸附,能清除大部分毒物,迅速降低血浆中毒鼠强的质量浓度。     

肾移植前后严重焦虑对肾功能衰竭患者病情的影响：1例报告

目的:观察焦虑情绪对肾功能衰竭患者肾移植前后病情转归的影响,并探讨其治疗对策。方法:患者为36岁男性,2005-11入华中科技大学同济医学院附属同济医院肾内科,入院诊断为慢性肾小球肾炎,慢性肾功能不全,失代偿期,患者知情同意。因严重焦虑引起肾功能迅速恶化,在1个多月内从代偿期迅速进展到尿毒症期,并经肾活检证实病情不可逆转,接受血液透析治疗,并行肾移植。观察其接受血液透析前后和肾移植前后的焦虑状态评分及肾功能。汉密顿焦虑量表包括14个项目,总评分超过29分,可能为严重焦虑;小于6分,患者没有焦虑症状。结果:①焦虑状态的评估结果:血液透析前患者的焦虑状态评分为31分,属于严重焦虑。血液透析后和肾移植前后多次评分,患者焦虑状态均完全解除。②随访结果:肾移植术后1个月,患者重新工作,随访至今,复查各项指标基本正常。结论:严重焦虑可使慢性肾小球肾炎患者肾功能在短期内迅速恶化,对病情的发展和转归有明显影响。在治疗过程中除了积极治疗原发疾病外,还应重视心理干预治疗。     

Discomfort and factual recollection in intensive care unit patients

Introduction

A stay in the intensive care unit (ICU), although potentially life-saving, may cause considerable discomfort to patients. However, retrospective assessment of discomfort is difficult because recollection of stressful events may be impaired by sedation and severe illness during the ICU stay. This study addresses the following questions. What is the incidence of discomfort reported by patients recently discharged from an ICU? What were the sources of discomfort reported? What was the degree of factual recollection during patients' stay in the ICU? Finally, was discomfort reported more often in patients with good factual recollection?

Methods

All ICU patients older than 18 years who had needed prolonged (>24 hour) admission with tracheal intubation and mechanical ventilation were consecutively included. Within three days after discharge from the ICU, a structured, in-person interview was conducted with each individual patient. All patients were asked to complete a questionnaire consisting of 14 questions specifically concerning the environment of the ICU they had stayed in. Furthermore, they were asked whether they remembered any discomfort during their stay; if they did then they were asked to specify which sources of discomfort they could recall. A reference group of surgical ward patients, matched by sex and age to the ICU group, was studied to validate the questionnaire.

Results

A total of 125 patients discharged from the ICU were included in this study. Data for 123 ICU patients and 48 surgical ward patients were analyzed. The prevalence of recollection of any type of discomfort in the ICU patients was 54% (n = 66). These 66 patients were asked to identify the sources of discomfort, and presence of an endotracheal tube, hallucinations and medical activities were identified as such sources. The median (min–max) score for factual recollection in the ICU patients was 15 (0–28). The median (min–max) score for factual recollection in the reference group was 25 (19–28). Analysis revealed that discomfort was positively related to factual recollection (odds ratio 1.1; P < 0.001), especially discomfort caused by the presence of an endotracheal tube, medical activities and noise. Hallucinations were reported more often with increasing age. Pain as a source of discomfort was predominantly reported by younger patients.

Conclusion

Among postdischarge ICU patients, 54% recalled discomfort. However, memory was often impaired: the median factual recollection score of ICU patients was significantly lower than that of matched control patients. The presence of an endotracheal tube, hallucinations and medical activities were most frequently reported as sources of discomfort. Patients with a higher factual recollection score were at greater risk for remembering the stressful presence of an endotracheal tube, medical activities and noise. Younger patients were more likely to report pain as a source of discomfort.

     

Oxidative metabolism of the human eosinophil

We have compared the oxidative metabolism of human eosinophils (80%-90% purity) to that of neutrophils. Hexose monophosphate (HMP) shunt activity of eosinophils was higher than that of neutrophils under either resting or phagocytizing conditions. Eosinophil HMP shunt activity also was stimulated by phorbol myristate acetate, a membrane- active agent. Eosinophils showed a marked incorporation of 125I into trichloroacetic acid-insoluble material under resting conditions, which increased markedly during phagocytosis. Eosinophils likewise showed a greater reduction of nitroblue tetrazolium dye during phagocytosis than did neutrophils. Measurement of other parameters of oxidative metabolism indicated that eosinophils generated superoxide anion following phagocytosis and also elicited a burst of chemiluminescence similar to that observed during phagocytosis by neutrophils. Measurement of NADPH oxidase activity demonstrated that this enzyme was 3-6 times more active in fractions isolated from eosinophils than in corresponding fractions isolated from neutrophils; this was observed over a range of substrate concentrations. The eosinophil enzyme sedimented differently than the neutrophil enzyme with differential centrifugation; neither showed sedimentation characteristics of peroxidase. These data indicate that eosinophils possess a similar, although in some ways more potent, oxidative burst than neutrophils and are consistent with a role for NADPH oxidase in the initiation of that burst.     

微创经皮钢板置入内固定治疗胫腓骨骨折的技术操作特点

目的:总结微创经皮钢板置入内固定治疗胫腓骨骨折的技术操作特点,并观察材料及宿主反应。方法:2004-06/2006-10在济宁医学院附属金乡医院对18例胫腓骨骨折患者在微创手术下进行了手术置入内固定材料。男13例,女5例,年龄17～73岁。术后按Johner-Wruhs方法评价测试各大关节功能,分为优、良、中、差。全部病例进行临床随访。术前、术后1周、6周、3个月、半年及1年分别摄X线片与健侧对比测量患肢外观、成角、旋转和短缩情况,并观察材料及宿主反应。结果:①本组病例切口均顺利愈合,术后3～14d出院。②全部获得随访,平均随访时间14个月;骨愈合时间3～10个月。③按Johner-Wruhs方法评价功能,优13例,良4例,中1例,差0例,以优良为满意标准,本组病例总体满意率94.4%。④术后1例出现跛行步态,中度疼痛,骨成角畸形15°,可能因为患者对不锈钢材料有排斥反应造成固定不牢所致。结论:微创经皮钢板置入内固定材料治疗胫腓骨骨折具有手术创伤小、骨折愈合快、功能恢复好的特点;内固定材料未出现特殊材料反应与宿主反应。