1∶49枸橼酸钠抗凝剂在临床输血检验中的应用

目的:研究1∶49枸橼酸钠抗凝剂在血型鉴定、抗体筛查、临床交叉配血和输血传染性指标检测中的应用。方法:配制不同浓度的枸橼酸钠比例,确定最大抗凝比例,取68例标本分别用EDTA-K2、肝素钠和1∶49枸橼酸钠抗凝,分别用来鉴定血型和做抗体筛查,最后用聚凝胺法及微柱凝胶卡式法交叉配血检测不同抗凝剂对交叉配血的结果的影响。同时,取乙肝表面抗原(HBsAg)、丙肝抗体(HCV)、艾滋病抗体(HIV)、梅毒抗体(TP)各30例阳性标本,采用化学发光方法进行检测,观察1∶49枸橼酸钠抗凝管对检测HBsAg、HCV、HIV、TP的影响。结果:1∶49的枸橼酸钠抗凝管能达到抗凝效果,对血型鉴定、抗体筛查、交叉配血结果没有影响,与不抗凝采血管对HBsAg、HCV、HIV、TP阳性标本进行检测比较,2种采血管对结果的影响差异无统计学意义(P0.05)。结论:1∶49枸橼酸钠抗凝剂能提高临床交叉配血、输血前检查相关传染性指标检测的快速性和准确性。     

自制枸橼酸钠干粉抗凝管在EDTA依赖性假性血小板减少症中的应用研究

目的:制备最佳抗凝浓度的枸橼酸钠干粉抗凝管,使EDTA依赖性假性血小板减少症患者的血小板计数更加简便、准确。方法:1配置不同浓度枸橼酸钠干粉抗凝管,优选其最佳抗凝浓度、抗凝血量及检测时间范围。2使用不同抗凝管同时采集4种不同类型患者静脉血,进行血小板计数,对EDTA依赖性假性血小板减少症患者同时进行末稍血手工法血小板计数及抗凝血涂片染色观察血小板聚集情况。结果:1枸橼酸钠干粉抗凝管的最佳抗凝浓度为12.8mg/管,在抗凝血量200~800μl、上机检测时间10~120min的条件下计数血小板可得到较准确结果。2最佳抗凝浓度的枸橼酸钠干粉抗凝管在门诊体检者、血小板增多患者、真性血小板减少症患者中,血小板结果与EDTA-K2真空抗凝管血小板结果比较,差异无统计学意义;在EDTA依赖性假性血小板减少症患者中,血小板结果与末稍血手工法计数血小板结果比较,差异亦无统计学意义,枸橼酸钠干粉抗凝血涂片染色观察血小板呈单个散在分布。结论:自制的枸橼酸钠干粉抗凝管可简便、准确地纠正EDTA依赖性假性血小板减少症患者的血小板数量。     

恶性疟原虫乳酸脱氢酶的表达及免疫活性鉴定

目的　在大肠杆菌中表达恶性疟原虫乳酸脱氢酶 (LDHp)与谷胱甘肽S 转移酶 (GST)融合蛋白 ,测定重组蛋白的免疫活性。方法　采用PCR方法特异性扩增恶性疟原虫 (海南株 )乳酸脱氢酶基因 ,经双酶切后克隆入 pGEX 4T 1表达载体中 ,重组蛋白纯化后免疫小鼠制备特异性血清 ,并用琼脂双向扩散法检测效价 ,ELISA、Western bloting检测重组抗原的免疫活性。结果　得到了重组表达的蛋白抗原 ,表达产物能与兔抗恶性疟原虫血清发生反应 ,并能诱导小鼠产生特异性体液免疫应答 ,免疫琼脂扩散法抗体滴度为 1∶16。结论　LDHp在大肠杆菌中获得高效表达且表达产物具有良好的抗原性。     

西咪替丁抗疟作用的研究Ⅰ．西咪替丁对体外恶性疟原虫生长的影响

在体外培养条件下观察了西咪替丁对恶性疟原虫生长的影响。当西咪替丁的浓度为５×１０－３～４×１０－２ｍｏｌ／Ｌ时，具有明显的直接杀灭疟原虫及抑制其生长的作用。进一步观察西咪替丁对恶性疟原虫红内期发育的影响，结果表明，该药对环状体期的虫体最为敏感，可致原虫数目减少，并抑制其发育成滋养体。     

长期口服抗凝剂对老年人消化性溃疡复发的影响

老年人因防治心脑血管疾病发生常长期口服抗凝剂。该药长期口服对消化道粘膜有一定损伤。本文对50例长期口服抗凝剂的老年人消化性溃疡复发做观察分析。 1 临床资料 1.1 观察对象与分组 Ⅰ组(观察组)50例,原有消化性溃疡的高粘度血症50例,男44例,女6例。平均年龄67.5岁(60～75岁)。经钡透或胃镜检查确诊。原胃溃疡9例,十二指肠球部溃疡13例,慢性胃炎28例(浅表性10例,萎缩性16例,肥厚性2例)。病史18～31年。近10年病情稳定。未服药治疗。Ⅱ组(对照     

瑞香素杀疟原虫裂殖体的作用

[目的 ]研究中药瑞香素在体外和体内的杀裂殖体作用。 [方法 ]在恶性疟原虫FCCl株常规体外培养中测试瑞香素杀裂殖体活性 ,并按“四天抑制性试验”在感染伯氏疟原虫ANKA株的小鼠中测定不同剂量瑞香素的体内抗疟活性。 [结果 ]体外试验中瑞香素在 1～ 10 μmol L剂量范围内有明显杀灭裂殖体作用 ,而体内试验中按D4减虫率与感染鼠在 30d内的平均生存天数评价 ,5 0或 10 0mg kg .d- 1 × 4d瑞香素灌胃以及 10 ,5 0或 10 0mg kg.d- 1 × 4d瑞香素腹腔注射给药在伯氏鼠疟原虫ANKA株感染鼠中的抗疟作用与 10mg kg .d- 1 × 4d氯喹 (CQ)灌胃的疗效相似。 [结论 ]瑞香素在体外和体内均有一定的杀裂殖体作用。     

蚊虫基因对疟原虫发育的影响

疟原虫在传播至脊椎动物宿主之前 ,必须在其传播媒介按蚊体内经历一个复杂的发育过程。蚊吸食了带有疟原虫配子体的血液后 ,配子体即开始在蚊体内发育 :动合子穿过蚊胃上皮细胞后转变为卵囊。疟原虫入侵蚊胃的同时 ,蚊的先天免疫系统被激活 ,但是 ,迄今为止没有公开发表的证据表明蚊虫的基因能够影响疟原虫的发育。本文通过基因抑制的方法发现 :在冈比亚按蚊体内 ,存在着对疟原虫发育具有保护作用的C型凝集素 (CTLs)及对其有拮抗作用的富含亮氨酸的蛋白质 (LRR)。     

大蒜素体外抗疟作用的研究(摘要)

目的研究大蒜素对恶性疟原虫体外生长活性的影响.方法用新鲜配制或储存的疟疾完全培养基大蒜素稀释液在不同浓度(100μg/mL～5μg/mL)处理恶性疟原虫,实验过程中在12h,24h换液或者不换液.分别作用12h,24h和48h后,检测虫体感染率的变化分析药物的作用效果.同时以大蒜素处理正常红细胞观察对虫体宿主细胞的影响.结果以新鲜配制的大蒜素处理疟原虫时,作用12h后50μg/mL的浓度即可清除虫体,当作用48h后在低至10μg/mL的浓度也可杀灭原虫.而使用储存大蒜素换液或者新鲜配制的大蒜素不换液则需要较高的浓度.大蒜素处理正常红细胞时,在高于50μg/mL的浓度对宿主红细胞有破坏作用,而该浓度下虫体生长可被有效抑制.结论大蒜素在体外具有强烈的抗疟作用.30μg/mL是可以有效抑制疟原虫生长但对宿主红细胞没有损伤的适宜浓度.     

聚合酶链反应在恶性疟疾普查中的应用

作者应用滤纸片干血液分离恶性疟原虫DNA进行聚合酶链反应的方法对所罗门群岛的489份血样品进行检查。结果表明此方法具有1）用血量少，经自然干燥后，易于保存和运输。2）DNA分离的方法简便、快捷。3）PCR的检测极限为1．5个原虫／μl血。其次，其结果与姬姆萨染色方法的结果进行了比较。结果提示姬姆萨染色方法的结果与其检查者的技术水平密切相关。在大面积普查时容易出现漏诊及误诊、此外，也证明单片段基因扩增同样容易出现漏诊及误诊。     

微滴度板杂交试验在疟疾种属特异性诊断的应用

本实验应用113份指尖取血制作的滤纸片干血滴样品分离疟原虫DNA进行微滴度板杂交试验。结果表明：1．滤纸片干血滴一微滴度板杂交实验用血量少、经过自然干燥后，样品易于保存和运输，而且提取DNA的方法简便。2．此方法特异性高、敏感性强、其检测极限可达到1．5个原虫／μl血。3．此方法可在进行疟疾诊断的同时进行种属鉴定，并可对多种疟原虫同时感染进行诊断。     

丁酸钠对TNBS结肠炎模型大鼠肠黏膜修复的影响

背景：肠道黏膜的炎症损伤和修复是炎症性肠病（IBD）的重要特征之-。丁酸盐为肠上皮细胞的主要能量来源,参与了肠道黏膜内稳态的维持并具有抗炎效应。目的：观察丁酸钠对结肠炎模型大鼠肠黏膜炎症损伤和修复的影响．探讨其治疗IBD的可能机制。方法：以三硝基苯磺酸（TNBS）诱导大鼠结肠炎模型并予丁酸钠或美沙拉嗪治疗,同时设置正常对照组和结肠炎模型组。于急、慢性炎症期分批处死大鼠,行结肠组织学检查和评分,免疫组化方法检测结肠组织中与黏膜修复相关的转化生长因子-β1（TGF-β1）、血管内皮生长因子（VEGF）表达,ELISA方法检测血浆促炎细胞因子白细胞介素．8（IL-8）、抗炎细胞因子IL-10水平。结果：与结肠炎模型组相比,丁酸钠治疗组一般情况和结肠组织学表现有所改善．美沙拉嗪治疗组则有明显改善。丁酸钠治疗组和美沙拉嗪治疗组结肠组织TGF—β1阳性表达率显著高于结肠炎模型组（P〈0．05）,VEGF阳性表达率无明显变化．慢性炎症期血浆IL_8水平显著降低（P〈0．05）．IL-10水平显著增高（P〈0．05）。结论：丁酸钠对TNBS结肠炎模型大鼠的肠黏膜修复有-定促进作用,该作用可能与其增加TGF-β1表达和调节促炎细胞因子与抗炎细胞因子平衡有关。     

瑞香素抗红外期疟原虫作用的研究

目的　研究瑞香素 (DPNT)抗红外期疟原虫的作用。方法　于ICR小鼠腹腔注射约氏疟原虫子孢子后 0 5h灌胃给药 ,连续 4d。不同剂量的DPNT及DPNT伍用伯氨喹 (PQ)的抗疟作用 ,分别以d7ICR小鼠阴性率及d1 1 或d1 2 ICR小鼠每千个红细胞被原虫感染数作评价 ,并观察DPNT对ICR小鼠血红蛋白浓度的影响。结果　DPNT的剂量范围为每天 10～ 10 0mg/kg ,连服 4d ,d7原虫阴性小鼠数及d1 1 红细胞被感染程度与对照组相比其差异均无显著性 ;DPNT每天 5 0mg/kg和每天PQ5mg/kg配伍组的d7小鼠阴性率与PQ每天 10mg/kg组相当。ICR小鼠DPNT每天 5 0mg/kg组与对照组血红蛋白浓度在d8天有差异。结论　DPNT单独用药 ,无明显抗红外期疟原虫作用 ,但DPNT每天 5 0mg/kg与PQ每天 5mg/kg伍用的抗疟效果与PQ每天 10mg/kg相当。DPNT在短期内可致小鼠贫血。     

阿魏酸钠对血小板源生长因子和内皮素1诱导的血管平滑肌细胞迁移的影响

为探讨阿魏酸钠对血小板源生长因子二聚体和内皮素 1诱导的血管平滑肌细胞迁移的影响 ,采用组织块外生法体外培养血管平滑肌细胞 ,采用改良的Boyden微孔膜双槽法进行细胞迁移实验 ,荧光染料Fura 2 /AM法测定细胞内游离钙离子浓度。结果发现 ,血小板源生长因子二聚体和内皮素 1均可诱导血管平滑肌细胞迁移 ,作用峰值浓度分别为 10 μg/L和 10 - 7mol/L。阿魏酸钠 (10 - 7～ 10 - 3mol/L)呈浓度依赖性抑制上述物质诱导的细胞迁移 ,10 - 3mol/L阿魏酸钠对血小板源生长因子二聚体和内皮素 1诱导的细胞迁移的抑制率分别为 85 .0 4 %和 81.92 %。血小板源生长因子二聚体和内皮素 1促进细胞内游离钙离子浓度升高 (P <0 .0 5 ) ,作用峰值浓度分别为 10μg/L和 10 - 8mol/L。阿魏酸钠明显抑制该作用 ,峰抑制率分别为 80 .14 %和 76 .6 9%。以上提示 ,阿魏酸钠可能通过抑制细胞内游离钙离子浓度的升高来抑制上述物质诱导的血管平滑肌细胞迁移。     

Effect of Plasma Storage on Erythrocyte Sodium Pump Measurements

The possibility that sodium handling in red cells may be an indicator of abnormalities associated with hypertension has encouraged many investigators to study erythrocyte sodium transport. The results have often been in conflict, perhaps because of the variety of laboratory techniques and procedures employed. Investigation into the effect of storage of red cells in plasma showed larger changes when cells were stored at 0°C than at room temperature (19°). At 0°C, sodium efflux via the pump increased 22.1% (p < 0.001) after 2 hours storage and 54.2% (p < 0.005) after 4 hrs of storage. At 19°C, sodium efflux values were more scattered after storage, but the mean value was not significantly different even after 4 hours. Intracellular sodium increased 17.3% in cells stored 4 hrs at 0°C; significant changes were not observed when cells were stored 2 hrs at 0°C or 2-4 hours at 19°C, The number of Na, K-ATPase sites per red blood cell did not change with either storage condition. For samples processed within one hour of drawing, or stored at room temperature, there were close correlations between the sodium efflux per site and the intracellular sodium concentration as well as between the intracellular sodium and the sites per red cell. These relationships were not evident for cells stored at 0°C. These data indicate that red cells to be used in pump measurements should be stored at room temperature and should be processed within 2 hours.     

Inhibitory Effects of Sildenafil Citrate on the Tonus of Isolated Dog Internal Anal Sphincter

PURPOSE Although the exact pathogenesis of anal fissure is not known, hypertonicity of the internal anal sphincter might be involved in its pathogenesis as main event. To gain information about possible usefulness of the novel, smooth-muscle–relaxing drug, sildenafil, in chronic anal fissure, we investigated the effect of sildenafil citrate on acetylcholine-induced contractility of internal anal sphincter isolated from dogs.METHODS Internal anal sphincter strips were taken from German shepherd dogs and suspended in a tissue bath filled with Krebs solution at 37°C (pH 7.4) continuously bubbled with 95 percent oxygen and 5 percent carbon dioxide, and isometric contractions were recorded. Contractions were evoked by 10 μM acetylcholine, and the effects of different concentrations of sildenafil citrate (0.1, 0.3, and 1 mM) on the isometric tension of each internal anal sphincter strip were examined. The statistical significance was analyzed by one-way analysis of variance.RESULTS Pretreatment with sildenafil citrate (0.1 mM) attenuated contractile response to acetylcholine (n = 3), which were significantly weak compared with the maximum contractile response to the acetylcholine alone (610 ± 110 mg vs. 2,825.17 ± 416 mg; n = 12; P < 0.05). Sildenafil citrate also significantly inhibited the acetylcholine-induced contractions in a dose-dependent manner when applied after.CONCLUSIONS This experimental in vitro study showed that sildenafil citrate relaxes acetylcholine stimulated contractions of isolated dog internal anal sphincter. This may be of importance for raising the possibility that sildenafil cit-rate may have future potential in the treatment of chronic anal fissure. Further studies are needed for a conclusive decision on possible usefulness of sildenafil citrate in patients with chronic anal fissure.Poster presentation at the Cukurova Colo-Proctology Course and Symposium, Adana, Turkey, June 3 to 6, 2003.     

磷制剂对砷染毒细胞增殖抑制作用影响的实验研究

目的 探讨磷制剂对NaAsO2染毒的Vero细胞体外增殖抑制的影响.方法 用不同浓度的NaAsO2处理细胞后,在培养液中加入不同剂量的磷制剂,通过噻唑蓝还原法(MTT)观察对细胞增殖抑制的影响,并用维生素E作为标准对照.结果 一定剂量NaAsO2对Vero细胞增殖有抑制作用;207.00 mg/L,103.50 mg/L,51.75 mg/L三种剂量的磷酸钠及53.3 mg/L,26.65 mg/L的卵磷脂对砷染毒的细胞具有保护作用,可拈抗砷对细胞的增殖抑制作用,与维生素E的拮抗作用相比,高剂量的磷酸钠与维生素E相当,中、低剂量的磷酸钠及高、中剂量的卵磷脂的保护作用次于维生素E,而次磷酸钠的作用相反,与砷的毒性具有协同作用.结论 磷制剂对对砷染毒细胞增殖抑制作用是双向的,与磷的不同价态、不同化合物在体内代谢不同有关.     

Increased Red Cell Sodium Lithium Countertransport Activity,Total Exchangeable Sodium,and Hormonal Control of Sodium Balance in Normoalbuminuric Type 1 Diabetes

The relationship between erythrocyte sodium lithium countertransport activity (SLC), total exchangeable sodium (NaE), and hormonal control of renal function was examined in 40 normotensive, normoalbuminuric, non-neuropathic Type 1 diabetic subjects, of whom 8 had elevated SLC (> 0.40 mmol Li h^?1l^?1 rbc). Eleven health controls with normal SLC, who were of comparable age, body mass, and blood pressure were also studied. By contrast with healthy controls, SLC in Type 1 diabetes was not associated with plasma renin activity (PRA), aldosterone, systolic blood pressure or lean body mass. SLC was also unrelated to atrial natriuretic peptide (ANP) (Type 1 diabetes only) and NaE. NaE was not correlated with any other variables. The relationships between PRA and aldosterone in healthy controls were retained in Type 1 diabetes (R² 0.37 supine, p = 0.00001, and 0.27 ambulant, p = 0.0005), as were respective direct and inverse relations between vasopressin and ANP and both PRA (r_s 0.54 to 0.57, r_s ?0.43 to ?0.53), and aldosterone (r_s 0.78 to 0.80, r_s ?0.71 to ?0.80). Fasting free serum insulin and vasopressin were both inversely related to ANP (r_s ?0.91 and ?0.71, respectively). In the absence of autonomic dysfunction, hypertension or early nephropathy in Type 1 diabetes, increased SLC or exchangeable sodium were unrelated to each other or with hormonal control of sodium balance, but the homeostatic factors controlling hormonal interaction appear to be maintained. The interaction between insulin and hormonal control of sodium and water balance may be modified by circulating free insulin concentrations.