异丙酚激活蛋白激酶C对心肌缺血再灌注损伤的保护作用

目的:观察异丙酚对大鼠离体心肌缺血再灌注的影响及蛋白激酶C激活的作用。方法:实验于2004-04/2005-03在河北省医学科学院药研室完成。48只大鼠离体心脏随机分为6组,每组8只。分别为:正常对照组,持续灌注Lock液65min;缺血再灌注模型组,用含脂肪乳对照的灌流液灌注15min后,以钳夹主动脉灌注管造成全心常温缺血25min后,恢复再灌注30min,灌流液与预灌时相同;异丙酚15,30,60μmol/L组,缺血前和再灌期的灌流液中分别含相应浓度的异丙酚,余同缺血再灌注模型组;异丙酚60μmol/L 蛋白激酶抑制剂5μmol/L组,灌流液中含5μmol/L的chelerythrine的灌流液,其余同异丙酚60μmol/L组。实验评估:①Powerlab/8s仪记录各组平衡末、缺血前及再灌30min时心率、左室发展压、左室舒张末压、左室压力变化速率、冠状动脉流量等心功能指标。②测定冠状动脉流出液中乳酸脱氢酶、磷酸肌酸激酶活性。③透射电镜观察心肌细胞超微结构变化。④差速离心提取心肌线粒体,测定线粒体超氧化物岐化酶、谷胱甘肽过氧化物酶、ATP酶活性和丙二醛含量。结果:48只大鼠均进入结果分析。①平衡灌注末、缺血前各组间心功能指标差异无显著性(P>0.05),再灌注30min末,异丙酚30,60μmol/L组左室发展压、左室压力变化速率、冠状动脉流量明显高于缺血再灌注模型组(P<0.05),左室舒张末压明显低于缺血再灌注模型组(P<0.05)。异丙酚60μmol/L 蛋白激酶C抑制剂5μmol/L组左室发展压、左室压力变化速率、冠状动脉流量明显低于单纯异丙酚60μmol/L组(P<0.05),但仍高于与缺血再灌注模型组(P<0.05)。②心肌缺血再灌后,冠状动脉流出液中乳酸脱氢酶和肌酸激酶活性明显高于正常对照组(P<0.05)。异丙酚30,60μmol/L组、异丙酚60μmol/L 蛋白激酶C抑制剂5μmol/L组乳酸脱氢酶、肌酸激酶活性明显低于缺血再灌注模型组(P<0.05),异丙酚60μmol/L 蛋白激酶C抑制剂5μmol/L组乳酸脱氢酶和肌酸激酶活性与异丙酚60μmol/L组无明显差异。③与缺血再灌注模型组相比,异丙酚组心肌损伤明显减轻,尤其是60μmol/L组,心肌纤维排列均匀,线粒体膜结构完整,仅轻度水肿,嵴清晰,糖原可见。异丙酚60μmol/L 蛋白激酶C抑制剂5μmol/L组心肌超微结构显示损伤程度重于异丙酚60μmol/L组。④异丙酚30,60μmol/L、异丙酚60μmol/L 蛋白激酶C抑制剂5μmol/L组丙二醛含量低于缺血再灌注模型组(P<0.05),ATP酶、超氧化物歧化酶、谷胱甘肽过氧化物酶活性明显高于缺血再灌注模型组(P<0.05)。异丙酚60μmol/L组与异丙酚60μmol/L 蛋白激酶C抑制剂5μmol/L组相比,上述指标无明显差异。结论:异丙酚对离体大鼠心肌缺血再灌注损伤有保护作用,可能与其抗脂质过氧化和激活蛋白激酶C有关。     

应用间断流动式血细胞分离机采集外周血造血干细胞的效果分析

目的:为寻找采集外周血造血干细胞更合适的手段,探讨间断流动式血细胞分离机采集外周血造血干细胞的效果。方法:选择中山大学第五附属医院血液风湿科2004-2006年12例行外周血造血干细胞移植住院患者。①实验对象:供者5例,男1例,女4例,一般状况良好,与患者关系为同胞妹妹3例,同胞弟弟1例,同胞姐姐1例;患者7例,男3例,女4例,23～62岁,7例为自体移植,5例为异基因移植。血液系统疾病患者9例(非霍奇淋巴瘤3例,急性淋巴细胞白血病2例,急性髓细胞白血病2例,慢性粒细胞白血病1例,骨髓增生异常综合征1例);自身免疫性疾病3例(重型系统性红斑狼疮2例,难治复发性类风湿关节炎合并干燥综合征1例)。②实验过程:每位供/患者均知情同意并签署知情同意书。对于自体患者,根据患者的疾病类型采取不同的干细胞动员化疗方案,联合化疗后7～10d,待白细胞下降至最低点,一般为≤1.0×109 L-1时,给予粒细胞集落刺激因子5μg/kg皮下注射,白细胞升至3.0×109 L-1时开始采集;对于allo-PBSCT供者直接给予粒细胞集落刺激因子5μg/kg,皮下注射,1次/d,共5d,同时应用流式细胞仪检测CD34 细胞数,至白细胞升至≥20.0×109 L-1或当CD34 细胞升高>20个/μL时采用增强型多功能血细胞分离机PBSC程序卡采集健康供者和患者外周血造血干细胞。③实验评估:分析采集效率、血液学参数、不良事件发生率、以及供、受者ABO血型不合的患者回输干细胞溶血反应等情况。结果:12例供者、患者均进入结果分析。①共进行了24次采集,其中1次采集1例,2次采集7例,3次采集3例,平均循环次数(25±5)次,采集时间(228±32)min,处理血量(7234±1205)mL,复方枸橼酸钠溶液用量为(623±96)mL,干细胞收集量为(102±21)mL,CD34 细胞采集效率为(54.3±30.7)%,细胞计数示白细胞为(156±34)×109 L-1,单个核细胞为(79.7±13.2)×109 L-1,流式细胞仪检测CD34 细胞为(10.30±4.38)×106/kg受者体质量。②不良反应轻微,24次采集过程中出现不良反应6次均为枸橼酸盐所致低钙血症反应。血红蛋白和血小板与采集前相比分别下降9.36%和11.10%。供、受者ABO血型不合的3例患者在输注造血干细胞悬液后均未出现溶血反应。③12例均获造血功能重建,无移植相关死亡。结论:用间断流动式血细胞分离机采集外周血造血干细胞效果良好,不良反应轻微,能有效的减少被采集者红细胞、血小板的丢失,对供、受者ABO血型不合者不需去除造血干细胞悬液中的红细胞,值得临床应用。     

Prevention of iron deficiency with carbonyl iron in female blood donors

The effectiveness of elemental, nontoxic carbonyl iron in replacing iron lost at blood donation was examined. In a randomized double-blind design, 99 women, aged 18 to 40, were given placebo or low-dose carbonyl iron (100 mg orally) at bedtime for 56 days after phlebotomy. Compliance was equivalent for the two regimens. Mild side effects were slightly greater with carbonyl iron. At Day 56, estimated net iron absorption from therapy or diet, or both, was sufficient to replace iron in 85 percent of those receiving carbonyl iron but in only 29 percent of those taking placebo (p less than 0.001). The rates of deferral from repeat donation were 8 percent in the carbonyl iron group and 36 percent in the placebo group (p less than 0.01), and the positive predictive value of routine screening in identifying participants without iron deficiency was 83 versus 13 percent (p less than 0.01). It can be concluded that short-term carbonyl iron supplementation in female blood donors can replace the iron lost at phlebotomy, protect the women against iron deficiency, and enhance their ability to give blood.     

Postural Fall in Blood Pressure in the Elderly in Relation to Drug Treatment and Other Lifestyle Factors

In a study of 843 independent-living men and women aged between60 and 87 in Perth, Western Australia, stepwise multiple regression,after correction for initial levels of systolic blood pressure,showed that postural fall in systolic blood pressure was positivelyrelated to alcohol intake of more than 20 ml/day, the use ofsleeping tablets and higher levels of anxiety on the Spielbergerstate-trait scale, and negatively related to body mass index.Postural fall in blood pressure was not significantly relatedto treatment for hypertension, age, sex, patterns of usual physicalactivity, tea or coffee drinking, or the diagnosis of diabetesmellitus. This analysis is the first to examine the relationshipbetween lifestyle factors and the magnitude of the fall in systolicblood pressure on standing after adjustment for the associationbetween the change in a variable and its initial level. Ouranalysis suggests the need for further study of the possiblerole of lifestyle factors such as the use of sleeping tabletsand alcohol in postural hypotension in the elderly.     

Photophobia and phonophobia in tension-type and cervicogenic headache

Light and sound-induced discomfort and pain thresholds were measured in 26 patients with cervicogenic headache, in 40 patients with tension-type headache, and in 100 headache-free controls. Neither headache group was significantly different as to photophobia and phonophobia, but both were significantly more sensitive to light and sound than controls ( p <0.0001), even when patients were tested in the headache-tree period ( p <0.05). Episodic and chronic tension-type headache had similar photo- and phonophobia thresholds ( p ≥0.7). Tension-type headache patients were more photo- and phonophobic during headache than outside attack ( p <0.05), but this was not true for cervicogenic headache ( p ≥0.56). In cervicogenic headache patients, photophobia ( p <0.05) but not phonophohia ( p =0.28) was greater on the symptomatic side than on the non-symptomatic side.     

Moving beyond the conventional stratified analysis to estimate an overall treatment efficacy with the data from a comparative randomized clinical study

For a two-group comparative study, a stratified inference procedure is routinely used to estimate an overall group contrast to increase the precision of the simple two-sample estimator. Unfortunately, most commonly used methods including the Cochran-Mantel-Haenszel statistic for a binary outcome and the stratified Cox procedure for the event time endpoint do not serve this purpose well. In fact, these procedures may be worse than their two-sample counterparts even when the observed treatment allocations are imbalanced across strata. Various procedures beyond the conventional stratified methods have been proposed to increase the precision of estimation when the naive estimator is consistent. In this paper, we are interested in the case when the treatment allocation proportions vary markedly across strata. We study the stochastic properties of the two-sample naive estimator conditional on the ancillary statistics, the observed treatment allocation proportions and/or the stratum sizes, and present a biased-adjusted estimator. This adjusted estimator is asymptotically equivalent to the augmentation estimators proposed under the unconditional setting. Moreover, this consistent estimation procedure is also equivalent to a rather simple procedure, which estimates the mean response of each treatment group first via a stratum-size weighted average and then constructs the group contrast estimate. This simple procedure is flexible and readily applicable to any target patient population by choosing appropriate stratum weights. All the proposals are illustrated with the data from a cardiovascular clinical trial, whose treatment allocations are imbalanced.     

大豆甙元—PVP共沉淀物的制备及其体外溶出度的研究

以乙醇为溶媒用溶剂蒸发法制备了大豆甙元—PVP共沉淀物。X-衍射分析和偏光显微镜观察的结果表明,形成共沉淀物所需的PVP比例应大于5:1。体外溶出度试验结果表明,共沉淀物可以显著提高大豆甙元的体外溶出度。大豆甙元:PVP(1:10)的共沉淀物制成片剂后,在1 h内,90%以上的大豆甙元释放于蒸馏水或人工肠液中。     

不同产地和品种淫羊藿中淫羊藿苷的HPLC分析

目的 :对中药淫羊藿中淫羊藿苷的含量分析进行方法学研究 ,并测定淫羊藿的不同产地、不同品种、不同药用部位的淫羊藿苷的含量。方法 :采用RP HPLC技术对 7个品种 2 3个样品进行测定。以PERKIN EIMER SH/5C18柱为分析柱 ,流动相为乙腈 水 36%乙酸 ( 2 5∶73.5∶1 .5)流速 1 .0ml/min。UV检测波长 2 70nm。结果 :本方法测定淫羊藿苷含量在 0 .0 2 1 2～ 0 .1 2 7μg ,0 .53～ 1 .696μg范围内呈良好的线性关系。不同产地、不同品种、不同药用部位的淫羊藿其淫羊藿苷的含量为 0 .0 0 30 7%～ 1 .55%。同一植株不同部位中淫羊藿苷的含量分布叶 >叶茎 >茎 >根。结论 :淫羊藿由于产地不同、品种不同其淫羊藿苷的含量相差悬殊。本测定方法为筛选优良品种和扩大药源提供了简便易行的方法