壳聚糖载体介导关节内基因转移：转染效率与基因产品的关系

目的:分析壳聚糖-DNA超微颗粒在关节内的转基因效应。方法:实验于2005-09/2006-06在上海交通大学医学院健康科学研究所骨科细胞与分子生物学实验室完成。实验材料:①模型制备:采用切断内侧副韧带,切除内侧半月板的方法制备骨关节炎兔模型。②基因产品:白细胞介素(interleukin,IL)1Ra基因、IL-10基因。实验分组:15只新西兰兔按随机数字表法分为3组:①空载体对照组(n=3),造模后5d两侧膝关节关节腔注射400μL壳聚糖-PcDNA3.1溶液,共3次,每48h1次。②IL-1Ra基因治疗组和IL-10基因治疗组,每组6只,造模后5d对照侧膝关节关节腔分别注射20μg裸DNA(PcDNA3.1-IL-1Ra或PcDNA3.1-IL-10),实验侧膝关节关节腔注射400μL壳聚糖-DNA超微颗粒(含20μgIL-1Ra或IL-10),注射次数及间隔时间同空载体对照组。实验评估:①采用酶联免疫吸附分析及免疫组织化学检测IL-1Ra和IL-10基因的表达和分布。②苏木精-伊红染色和甲苯胺蓝染色观察骨关节炎软骨组织学变化。结果:纳入新西兰兔15只,均进入结果分析。①IL-1Ra和IL-10基因在关节滑液中的表达:空载体对照组及IL-1Ra基因治疗组对照侧膝关节滑液中未检测到IL-1Ra表达,实验侧于第1次基因注射后7,14d检测到IL-1Ra表达。IL-10基因治疗组对照侧和实验侧均未检测到IL-10表达。②IL-1Ra基因在兔膝关节的分布:IL-1Ra基因治疗组兔软骨表层和中间层部分细胞内表达IL-1Ra,至少持续到第1次基因注射后14d。在滑膜组织中未观察到明显的IL-1Ra表达。③兔骨关节炎软骨组织学变化:空载体对照组呈早期骨性关节炎的典型性改变。苏木精-伊红染色显示软骨细胞坏死,蛋白多糖甲苯胺蓝染色不均一,软骨表层和中间层大部分区域失染,软骨细胞簇聚区域其周围深染。IL-1Ra基因治疗组在软骨损坏方面明显减轻,甲苯胺蓝部分失染。结论:①壳聚糖-DNA超微颗粒的转染效率与基因产品有关。②将IL-1Ra用关节腔直接注射壳聚糖-DNA超微颗粒的方法直接转移入关节腔能一定程度上减轻骨性关节炎的进程。     

应用质谱、红外光谱及核磁共振解析验证合成物9,10-二羟基硬脂酸的纯化性

目的：为获得大量的9,10-二羟基硬脂酸（Dihydroxy Stearic Acid,DHSA）,应用质谱、红外光谱及核磁共振解析验证合成物为DHSA。 方法：实验于2007-01/05在解放军总医院营养科实验室完成。在烧瓶中加入油酸后,再加入甲酸、过氧化氢,并不断搅拌。在整个反应过程中维持反应温度在40℃,反应四五小时后,反复水洗油层并收集。在油层中加入3mol/L NaOH溶液25mL,于100℃水浴加热1h,趁热加入3mol/L HCl50mL,剧烈搅拌,析出白色沉淀。用水反复清洗使清洗液pH值为中性,收集沉淀物。在沉淀物中加入体积分数为0.95的乙醇10mL,加热溶解,抽滤,将滤液于0℃析出结晶,抽滤,将抽滤物用体积分数为0.95的乙醇10mL加热溶解,于0℃结晶,抽滤。同样方法再结晶2次,最后得到白色结晶物,晾干,收率68.8%。将结晶物分别上FTS-65A红外光谱仪、ISONS EA1108型元素分析仪,Varian INOVA 600型核磁共振仪,安捷伦XCR-trap液相质谱（LC-MS）联用仪进行分析和结构鉴定。 结果：①LC-MS分析提示合成的DHSA经过多次重结晶后,纯度较高,可达95%。MS分析提示DHSA相对分子质量为315.1,与理论值316.48相近。②DHSA元素分析实验值C68.78%,H11.53%,与理论值C68.77%,H11.58%接近。③红外光谱分析提示DHSA为含有两个羟基的十八碳长链脂肪酸。④核磁共振的氢谱和碳谱均证实合成的化合物为9,10-二羟基十八碳脂肪酸。 结论：合成的DHSA经过3次重结晶后,纯度可95%以上,质谱、红外光谱及核磁共振解析结果证实合成的化合物是9,10-二羟基硬脂酸。     

超重及肥胖与脂肪肝关系的病例对照分析

目的:分析超重及肥胖与脂肪肝的关系,探索肥胖与其他危险因素在脂肪肝发病中的交互作用。方法:选择2002-09/2004-12期间某社区人群,按分层多级整群抽样原则进行抽样调查,共调查2458例。根据1∶1配对病例对照研究的设计原理,选择其中确诊的425例脂肪肝患者作为研究病例,纳入标准:要求所有病例均有B超诊断报告,且年龄≥18岁,并从该人群的非脂肪肝患者中选择425例条件相似的作为对照,病例与对照在年龄、性别、居住地区方面均衡可比。对全体调查对象进行当面访谈调查和体格检查。结果:纳入调查对象850例,均进入结果分析。超重及肥胖者比正常体重者有显著升高的脂肪肝患病危险(OR=4.07,OR95%CI为2.99～5.54),OR值随体质量指数的增大而增大,呈明显的剂量反应关系(趋势χ2=104.25,P<0.0001)。腰臀比偏高的腹型肥胖者比非腹型肥胖者脂肪肝的发病危险更大。交互作用分析提示,肥胖与饮酒同时存在能显著增加疾病的危险性,其超额相对危险度为8.16,归因比为0.5767。结论:超重及肥胖与脂肪肝的发病关系密切,肥胖与饮酒在脂肪肝的发病中存在协同交互作用,控制体质量是降低人群脂肪肝患病率的重要措施。RA     

藏族糖尿病患者代谢功能改善与面对面教育的效应

目的:观察面对面随访方式的糖尿病教育对藏族患者代谢功能改善的作用。方法:①选择2005-08/2006-08在西藏军区总医院心内科住院和门诊就诊的糖尿病患者113例。男69例,女44例;年龄48～71岁。纳入糖尿病患者均符合1999年世界卫生组织糖尿病诊断标准,且对检测项目知情同意;均为离解放军西藏军区总医院最近的娘热乡藏族糖尿病患者。②测量身高、体质量、收缩压、舒张压,计算体质量指数=体质量/身高2。采用日立7600型全自动生化分析仪测定血清总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇水平。采用TosohHLC-723GHbV型高压液相色谱仪测定糖化血红蛋白。③所有参试者得到一份本院编写的藏汉对照糖尿病教育手册,并在分组前统一讲解,以上大课的形式进行普及教育。根据每个患者不同情况,由内分泌专业医师给予不同处方,包括饮食、用药、生活方式调整。随后将患者随机分为教育组56例(男35例,女21例)和对照组57例(男34例,女23例)。对照组主要采用患者主动咨询(电话咨询或医院复查)和每一两个月一次普及教育的方式,但医生不采用面对面主动干预;教育组的干预方式除上述普及教育以外,每2周进行一次面对面、一对一随访,并及时调整处方。半年后比较两组的代谢指标差异。④计量和计数资料的差异比较分别采用方差分析和χ2检验。结果:藏族糖尿病患者113例均完成随访并进入结果分析。①组间比较:干预前两组体质量指数、收缩压、舒张压、总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、糖基化血红蛋白比较,差异不明显(P>0.05)。教育半年后教育组的体质量指数、收缩压、舒张压、总胆固醇、三酰甘油、低密度脂蛋白胆固醇和糖化血红蛋白均明显低于对照组(t=2.0156～4.7592,P<0.05～0.01),高密度脂蛋白胆固醇明显高于对照组(t=3.8031,P<0.01),其中以糖基化血红蛋白差异最明显。②组内比较:教育组糖尿病教育半年后血清总胆固醇、三酰甘油、低密度脂蛋白胆固醇和糖化血红蛋白均较干预前明显降低(t=1.9832～3.6446,P<0.05～0.01),高密度脂蛋白胆固醇较干预前升高(t=2.8219,P<0.01),教育干预前后体质量指数无明显变化(P>0.05)。对照组糖尿病教育半年后,体质量指数、总胆固醇、三酰甘油、低密度脂蛋白胆固醇和糖化血红蛋白、高密度脂蛋白胆固醇与教育前相近(P>0.05)。结论:藏族糖尿病患者采取面对面的随访方式进行糖尿病教育较一般形式的糖尿病教育更好的改善其代谢功能。     

Physician adoption of hyperbaric oxygen therapy in the treatment of chronic wounds

Hyperbaric oxygen therapy, a treatment alternative for chronic wounds, has been used for several decades yet little is known about factors that influence physicians to incorporate this therapy into practice. To assess wound care physician knowledge of, attitudes toward, and adoption of hyperbaric oxygen therapy and to identify factors associated with physician adoption of this therapy, a 23-item questionnaire, based on Rogers' diffusion of innovation model, was developed by the authors and distributed to 653 American Academy of Wound Management Board-certified physicians. Of the 246 (43%) physicians in the study population who responded to the study survey, 167 (68%) reported they had used or referred patients for hyperbaric oxygen therapy during the past 12 months. More than half of the respondents reported a relatively high level of familiarity with and a positive attitude toward the effects of hyperbaric oxygen therapy on wound healing. Physician adoption of hyperbaric oxygen therapy was significantly associated with a community of >100,000 residents (adjusted odds ratio = 2.29, 95% confidence interval = 1.05 to 5.04); patient request for hyperbaric oxygen therapy (adjusted odds ratio = 5.38, 95% confidence interval = 2.50 to 11.56); positive attitude toward (adjusted odds ratio = 3.38, 95% confidence interval = 1.49 to 7.66) and high level of familiarity with hyperbaric oxygen therapy (adjusted odds ratio = 5.33, 95% confidence interval = 1.72 to 6.49); and practice location in either Florida or Texas (adjusted odds ratio = 3.44, 95% confidence interval = 1.24 to 9.54). Although the majority of the respondents reported adoption of hyperbaric oxygen therapy, most adopters are concentrated only in a few geographic areas. Despite the limitations of this study, especially the potential effects of sampling and response bias, the results help explain factors that have facilitated and hindered the adoption of this technology into practice.     

Clinical characteristics,treatment intent,and outcome in a consecutive 10-year cohort of oral cancer patients aged 75 years and older

The prevalence of oral squamous cell carcinoma (OSCC) in the elderly is expected to increase by nearly a third in the next decade. Its management in older patients is potentially more challenging due to their pre-existing medical comorbidities, frailty, reduced life expectancy, and social issues. The aim of this retrospective review was to report on treatment given to patients aged 75 years and over, case mix, and survival. All patients aged 75 years and over who were diagnosed with OSCC in Merseyside between 1 January 2007 and 31 December 2016, and treated with either curative or palliative intent, were included. Their hospital notes were reviewed. Fisher’s exact test and Kaplan-Meier analysis were used for data analysis. There were 236 patients (median (IQR) age 81 (78-86) years); 67% were treated curatively and 33% palliatively. Factors associated with palliation included older age, advanced tumour stage, cognitive impairment, and residence in a nursing or residential home. Of the 165 patients who were offered curative treatment, six (4%) declined due to personal or family reasons. Overall survival for palliative patients was 12% at one year and 7% at two years, whereas for patients treated curatively it was 74% at one year, 56% at two years, and 34% at five years. Patients over 85 years of age were less likely to have composite free flaps and postoperative radiotherapy. Perioperative mortality was 2.6%. Improvements in surgical techniques and perioperative management have enabled clinicians to offer treatment with curative intent to older frail patients, and with careful case selection outcomes can be very good.     

样品固相净化及反相高效液相色谱法测定血浆中茶碱

本文用氧化铝固相净化血样以消除血中杂质及某些合并用药的干扰,用Zorbax-C18柱及甲醇-(0.05 mol/L)醋酸钠缓冲液(50:50)为流动相,254 nm波长检测,咖啡因为内标测定了茶碱人体血浆浓度的变化。该法样品处理简便、快速,净化回收率好(80～86%)茶碱的检测限为0.2ng(信噪比3),在血浆中的最低检测浓度为20 ng/ml。标准曲线r为0.9995,日内变异系数1.89%,日间变异系数为2.41%,方法平均回收率为98.44±0.89%。     

对青少年特发性脊柱侧弯患者的体感诱发电位检查

本体感觉神经传导异常被认为与青少年特发性脊柱侧弯有关。用体感诱发电位检查在青少年特发性脊柱侧弯患者中是否合并存在本体感觉传导通道的功能异常。研究包括147例青少年特发性脊柱侧弯患者及31位同年龄分布正常对照。对每一位受试者检查胜后神经体感皮质诱发电位，电信号缺失、传导时间延长或双侧传导时间不对称为本体感觉传导通道结构性异常诊断指标。在脊柱侧弯患者中有7人电信号单或双侧缺失，其余140例脊柱侧弯患者中10例传导时间延长，其中4例双侧延长，6例单侧延长。结果证实，部分青少年特发性脊柱侧弯患者同时有本体感觉传导异常，提示青少年特发性脊柱侧弯可进一步分为有本体感觉传导异常及无异常两组。     

N-(4-乙氧羰基-4-取代丁酰基)-N-取代甘氨酸的合成

本文报道9个估计具有血管紧张素转化酶抑制活性的N-(4-乙氧羰基-4-取代丁酰基)-N-取代甘氨酸及其相应叔丁酯的合成和鉴定。药理初试结果,化合物Ⅶ₇和Ⅶ₉显示明显降压活性。