肾病患儿45例血清甲状腺激素水平及肝素治疗效果观察

目的：探讨肾病患儿血清甲状腺激素水平及肝素治疗的疗效。方法：45例肾病患儿随机分为肝疗组（27例）和常规组（18例），用ACCESS化学发光免疫分析系统检测甲状腺激素水平、反向间接血凝法检测治疗前后的尿FDP水平，并结合临床对肝素治疗效果做出判断。结果：血清TT3、TT4、FT3和FT4水平肝疗组与常疗组之间无差异（P＞0．05），两治疗组与对照组比较差异显著（P＜0．01）；治疗前尿FDP含量肝疗组与常疗组之间无显著差异，（P＞0．05），治疗后两组差异非常显著（P＜0．01）；疗效评定为肝疗组完全缓解23例，占85％；常疗组完全缓解10例，占56％，两组比较差异显著（P＜0．05）。结论：肾病患儿血清甲状腺激素水平低下与尿蛋白大量丢失和血中白蛋白低下有关，尿FDP检测可反映肾小球损伤状况，肝素治疗可减轻肾脏损害程度，缓解肾小球硬化进程。     

乙酰肝素酶的表达与胰腺癌临床病理参数及肿瘤血管生成的关系

目的：检测胰腺癌标本中乙酰肝素酶及CD34表达，探讨乙酰肝素酶的表达与胰腺癌临床病理参数及与肿瘤血管生成之间的关系。方法：免疫组化SP法检测78例胰腺癌中乙酰肝素酶及CD34的表达，分析乙酰肝素酶表达与胰腺癌临床病理特征及与肿瘤微血管密度（MVD）的相关性。结果：乙酰肝素酶表达与胰腺癌转移相关，有淋巴结或远处转移者表达明显升高（P〈O．05）；同TNM分期相关，Ⅲ～Ⅳ期阳性率较Ⅱ期升高（P〈O．01）。胰腺癌中乙酰肝素酶表达阳性者MVD值明显高于乙酰肝素酶表达阴性者（P〈0．01）。结论：乙酰肝素酶在胰腺癌中高表达，同胰腺癌转移、分期及血管生成密切相关，乙酰肝素酶的表达促进了胰腺癌的转移及血管生成。     

CVVH治疗两种抗凝方式的比较

目的：比较两种抗凝方式的优劣。方法运用全身肝素化、低分子肝素抗凝及体外枸橼酸抗凝等方式进行抗凝治疗,并对 CVVH 治疗的患者进行回顾性研究。结果肝素组死亡11例（47.83％）,出血发生率51.35％,堵管发生率10.81％;低分子肝素组死亡6例（35.29％）,出血发生率3.33％,堵管发生率13.33％。结论进行普通肝素抗凝时需按照个体化的情况而定,不可追求统一的抗凝剂量;低分子肝素抗凝方法可大大降低治疗的并发症、易于掌握,治疗效果优于肝素钠组。     

胶体金免疫层析法快速检测非法添加磺胺甲噁唑的研究

目的探索快速检测止咳平喘类中成药和保健食品中非法添加磺胺甲嗯唑的方法。方法采用胶体金免疫层析技术,吸取适量待测溶液,滴加于检测卡中,以T线和C线的颜色对比显示检测结果。结果收集样品40批,检出14批阳性样品,漏检率0,误检率0,正确率100％。结论本快速检测方法操作简便,灵敏度高,可作为现场监督检查非法添加磺胺甲嗯唑的有效手段。     

低分子肝素对急性缺血性脑卒中凝血机制的影响

目的:研究低分子肝素对急性缺血性脑卒中患者高凝状态的影响。方法:观察50例缺血性脑卒中患者应用低分子肝素后凝血和抗凝血机制变化。结果:抗凝血酶活性在用药前为(0.870±0.064)IU药后10h为(1.115±1.104)IU(t=3.23,P<0.05,有显著性差异;组织型纤溶酶原激活酶活性在用药前、后分别为(1.51±0.21)、(2.30±1.03)IU,有极显著性差异t=8.43P<0.01。结论:低分子肝素可改善急性的缺血性脑卒中患者高凝状态,且没有出血倾向。     

肝素对微血栓大鼠作用的实验研究

经静脉注射高分子右旋糖酐,复制循环微血栓及心电图表现异常的大鼠模型,观察肝素对微血栓的治疗作用。结果发现,肝素治疗组心电图改善,血粘度下降,循环微血栓数目减少,心肌细胞膜上Na~ —K~ —ATP酶活性较盐水对照组略有上升,但统计学上无显著差异。提示肝素可减少实验动物循环血中微血栓的形成,改善心电图异常变化,但心肌细胞膜的Na~ —K~ —ATP酶活性恢复较慢。     

低分子量肝素治疗不稳定心绞痛

目的：观察低分子量肝素治疗不稳定心绞痛的治疗效果；方法：选择加用低分子量肝素钙治疗不稳定心绞痛32例，与常规治疗组30例，进行疗效比较；结果：前者急性事件发生率显著降低。结论：低分子量肝素钙明显减少急性事件发生，更少发生出血事件，应用较为方便，且安全。     

分离大鼠原代肝细胞的改进方法

目的：建立一种简便、经济、高效分离原代大鼠肝细胞的方法，适用于体外药物代谢研究。方法：改进方法采用低浓度胶原酶“软消化”方式，并增加预孵育过程。比较传统分离方法和改进方法在细胞产量、活率以及胶原酶用量上的差异，并将改进方法所得细胞分别与CYP2C、CYP2E、CYP3A典型底物及新药17-AAG共孵育，检测CYP450酶活性及其实际应用价值。结果：改进方法分离的肝细胞在细胞产量尤其是活率上明显高于传统方法，胶原酶用量大大降低，典型底物甲苯磺丁脲、氯唑沙宗和睾酮经细胞代谢检测到典型羟基化代谢产物，17-AAG经孵育后与体内样品检测结果一致。结论：改进方法可明显提高细胞产量和活率，所得肝细胞具备良好的CYP450酶活性，适用于体外代谢研究，预测药物体内过程。     

噻唑蓝比色法中应采用分析纯二甲基亚砜作为甲月替溶解剂

噻唑蓝（ＭＴＴ）可被哺乳动物活细胞线粒体中的琥珀酸脱氢酶还原成蓝紫色的甲月替颗粒（Ｆｏｒｍａｚａｎ）,且甲月替颗粒生成量与活细胞数量及细胞活化状态呈线性关系。基于此建立的ＭＴＴ比色法,快速简便、能同时处理大量样品,并可定量检测,已广泛应用于细胞毒测定、淋巴因子活性、肿瘤细胞药敏试验等多个方面。ＭＴＴ比色法中,甲月替溶剂是一个重要影响因素。二甲基亚砜（ＤＭＳＯ）较传统酸化异丙醇等溶剂溶解力强、溶解甲月替颗粒时间短,仅需１０ｍｉｎ左右,有很强的应用前景。然而有报道［１］,ＤＭＳＯ与ＭＴＴ有颜色反…     

动脉内联合灌注尿激酶,丹参及肝素的溶栓效果观察

目的：评估经导管动脉内联用尿激酶、丹参及肝素溶栓的临床效果。材料和方法：经股动脉穿刺，将５Ｆ导管插入至血栓近心端，经导管注入尿激酶、丹参、术中术后配合肝素抗凝，然后行血管造影，评估治疗效果，结果：血管通畅者１０例，通而不畅者１例，无效得１例，总有效率为９２％。结论：经导管动脉内联有尿激酶、丹参、配合肝素抗凝治疗外周动脉栓塞是一种有效、安全的治疗方法。     

肿瘤转移关键酶:人乙酰肝素酶氨基端亚基的表达纯化及鉴定

目的:乙酰肝素酶在肿瘤转移的过程中起重要作用,其氨基端小亚基蛋白(Mr为8×103)对其功能活性是不可缺少的。得到此蛋白将有助于深入研究乙酰肝素酶的功能活性。方法:我们从人胎盘cDNA文库中分离乙酰肝素酶蛋白全长编码基因,将乙酰肝素酶氨基端小亚基的基因克隆入pGE-X2TK,转化大肠杆菌进行融合表达。经发酵,纯化,复性得到乙酰肝素酶的氨基端亚基蛋白。结果:得到6.9g纯度>90%的氨基端小亚基融合蛋白。GSTpull-down实验表明,此融合蛋白可与乙酰肝素酶羧基端大亚基蛋白结合。复性后的融合蛋白可被激酶切割为GST蛋白和乙酰肝素酶氨基端亚基蛋白。结论:所得的乙酰肝素酶的氨基端小亚基蛋白为进一步研究该亚基蛋白在细胞内的免疫亚定位及功能奠定了基础。     

乙酰肝素酶信号肽对其活性功能的影响研究

目的:乙酰肝素酶在肿瘤转移过程中起关键作用,其信号肽在该酶翻译后处理过程中起重要作用.本研究旨在探索乙酰肝素酶在哺乳动物细胞中的表达特性及其信号肽对此酶功能活性的影响.方法:从人胎盘cDNA文库中扩增乙酰肝素酶全长编码基因,克隆入pGEM-T载体中,测序鉴定序列完全正确后,将此基因的全长和不含信号肽基因序列分别克隆入真核表达载体pcDNA4.1/Myc-His中构建pcDNA4.1/Myc-His full HPA和pcDNA4.1/Myc-His part HPA,转化COS-7细胞进行瞬时表达,分析这两种乙酰肝素酶蛋白的表达特性及功能活性.结果:在转染了乙酰肝素酶全基因的COS-7细胞裂解液中检测到该酶的功能活性及大小约53×103(Mr)的目的蛋白免疫印迹表达带,为切割激活后乙酰肝素酶羧基端大亚基与标签蛋白的融合体.在转染了敲除信号肽的乙酰肝素酶基因的COS-7细胞裂解液中测到未被切割激活的大小约65×103(Mr)的目的蛋白免疫印迹表达带,未能检测到该酶的功能活性.结论:在COS-7细胞中表达的完整的乙酰肝素酶蛋白和不含信号肽的乙酰肝素酶蛋白均位于细胞内,没有或很少被分泌到细胞外,提示该酶在正常状态下主要分布在细胞内.含信号肽的酶蛋白在翻译后处理过程中被细胞内的蛋白酶切割成2个亚基而被激活,具有功能活性.而不含信号肽的酶蛋白没有被切割而成为有活性的酶,提示乙酰肝素酶的信号肽对其翻译后加工、激活过程有重要的影响.     

乙酰肝素酶、组织蛋白酶D在宫颈癌的表达

目的探讨乙酰肝素酶(Hps)和组织蛋白酶D(CathD)与宫颈癌的发生、发展及预后的关系。方法取我院宫颈癌住院手术患者组织标本89例;宫颈上皮内瘤变(CIN)41例,正常宫颈组织35例。采用免疫组化SP法对乙酰肝素酶和组织蛋白酶D的表达进行检测。结果宫颈癌组织中乙酰肝素酶和组织蛋白酶D的免疫积分明显高于宫颈CIN组织及正常宫颈组织。而在CIN和正常宫颈组织间比较无统计学差异。乙酰肝素酶和组织蛋白酶D的表达与宫颈癌的临床分期、血管浸润和淋巴结转移有关。结论乙酰肝素酶和组织蛋白酶D可能在宫颈癌的发生、发展及预后中起着重要作用。     

重组人乙酰肝素酶在大肠杆菌中的表达、纯化

目的：表达纯化重组的人乙酰肝素酶（heparanase,HPA）。方法：以表达乙酰肝素酶全长质粒pcDNA3.1-HPA为模板,扩增不包括信号肽的乙酰肝素酶片段,将其插入pET-28a（＋）载体,转化到大肠杆菌BL21（plysS,DE3）中,在起始诱导菌密度、诱导温度、诱导时间、诱导剂浓度4个方面对诱导条件进行了优化,实现了重组人乙酰肝素酶的可溶性表达。采用HisTrapTMcrude亲和层析对目的蛋白进行了初步纯化。结果：得到了初步纯化的HPA蛋白,Western印迹证实表达产物可被乙酰肝素酶抗体和His标签抗体识别,证明表达产物具有乙酰肝素酶的免疫学特性。结论：表达纯化的人HPA蛋白为特异性单抗的制备和鉴定提供了实验材料。     

人体胃癌细胞的分离

目的：探讨人体胃癌细胞的分离方法。方法：应用免疫磁珠分选法（MACS）分离人体胃癌细胞，并用免疫组化方法鉴定。结果：免疫磁珠分选的胃癌细胞活力达到90％，且经鉴定证实为上皮源性细胞胃癌细胞。结论：免疫磁珠分选法能分离出高活性的人体胃癌细胞。     

Removal of liver activity contamination in teboroxime dynamic cardiac SPECT imaging with the use of factor analysis

BACKGROUND: One of the major problems associated with technetium 99m teboroxime cardiac imaging is the high concentration of activity in the liver. In some cases it is impossible to diagnose defects on the inferior wall because of the finite resolution and scatter that cause images of the inferior wall and the liver to overlap. METHODS AND RESULTS: The least-squares factor analysis of dynamic structures method, with correction for non-unique solutions, was used to remove the liver activity from the image. The method was applied to dynamically acquired Tc-99m teboroxime data. The liver activity removal method was tested through use of computer simulations and tomographically acquired canine and patient cardiac studies. In all studies the least-squares factor analysis of dynamic structures method was able to extract the liver activity from the series of dynamic images, thereby making it possible to remove it quantitatively from the entire series. The method was used successfully to remove the liver activity that partially overlapped the inferior wall in normal hearts. The method tends to increase the contrast between defects and normal myocardial tissue in abnormal hearts. CONCLUSIONS: The method presented can be used to assist in diagnosis of cardiac disease when dynamically acquired teboroxime data are used. Because the contrast between the defect and normal myocardial tissue can be changed, the processed image cannot be used by itself to make an accurate diagnosis. However, with the liver activity removed, the image provides additional information that is very useful in the imaging of patients whose liver activity overlaps the inferior heart wall.     

Development and implementation of a systems-based practice project requirement for radiology residents

RATIONALE AND OBJECTIVES: Radiology residents must now participate in a learning activity that involves identification and analysis of system errors and implementation of potential solutions. Evidence of participation must be documented in the resident's learning portfolio. MATERIALS AND METHODS: An activity based on adult learning principles was designed for residents to increase their knowledge of health care systems and problems; gain experience in writing a proposal, conducting an investigation, and reporting results; organize and present information in a supportive environment; and document participation in their portfolio. Postgraduate year 2 residents identify a systems problem. A written proposal is required and must include a "plan" that includes a hypothesis, a methods section that explains what they will "do" as an intervention, a "study" that involves collection and analysis of relevant data, and a systems change or "act." Proposals approved by the program director are presented to the radiology department. Residents complete projects, evaluate their effectiveness, and present their results by spring of postgraduate year 3. RESULTS: Goals, objectives, and guidelines were written. Assessment tools were identified. From 2006 to 2008, nine projects were proposed and conducted by 12 residents. Local and national systems issues were selected, and interventions involved residents, faculty, and medical students. A timeline was imposed to promote sustained effort. CONCLUSION: Opportunities of daily work can be used to teach and learn systems-based practice concepts. Residents can use the scientific method to test effects of change on health care systems. Relevant and practical projects enable residents to learn while improving their clinical and learning environments.     

Development of a Scottish physical activity questionnaire: a tool for use in physical activity interventions

OBJECTIVES: Three studies were undertaken to establish the reliability and validity of the Scottish physical activity questionnaire (SPAQ), developed to aid seven day recall of leisure and occupational physical activity. METHODS: To establish reliability, SPAQs (n = 34) were completed on a Monday and the following Wednesday. Thus each questionnaire measured four identical days. To establish concurrent validity, 94 participants completed a SPAQ and an adapted stage of exercise behaviour change questionnaire. Responses to SPAQ were then analysed by stage of exercise behaviour change. In a further study of criterion validity, 30 volunteers wore a Caltrac motion sensor for four consecutive days, after which they completed a SPAQ. RESULTS: In the first study, total physical activity had a coefficient of repeatability (R) of 53 minutes. Occupational physical activity showed a similar variance (R = 54.6 minutes) but leisure physical activity was more reliable (R = 29.3 minutes). The main variation in occupational physical activity was found to be walking (R = 39.8 minutes). In study 2, a one way analysis of variance showed the expected relation between physical activity and stage of exercise behaviour change, confirming the concurrent validity of SPAQ with the stage of exercise behaviour change model. In study 3, several erroneous recordings affected both SPAQ and the Caltrac results (kcal). After relevant corrections had been made, the correlation between the two measurement devices was 0.52 (p < 0.05). CONCLUSIONS: SPAQ has been shown to be reliable and to hold strong concurrent validity and limited criterion validity. The main limitation in SPAQ appears to be the measurement of occupational walking. It is therefore recommended that further work be conducted to refine the measurement of this physical activity component. It is evident nonetheless that SPAQ can be used with confidence to measure outcomes in physical activity interventions when account is taken of its limitations.