实时细胞电子分析技术用于水蛭、三七质量评价的初步研究

采用实时细胞电子分析技术测定水蛭、三七不同样本的时间剂量依赖性细胞反应曲线(TCRPs),对不同产地样本的TCRPs进行分析,并对生物活性强弱进行排序.同时,采用中国药典2010年版中的化学检测法和凝血酶滴定法分别对水蛭、三七各样本进行同步分析,比较不同检测方法得出结果的相关性.可见以实时细胞电子分析技术为基础的细胞生物活性测定结果与另两法所得结果基本一致.说明本技术能反映中药生物活性,有望为中药质量标准研究提供一种新方法.     

一测多评法同时测定水蛭中4种有效成分的含量

目的：建立同时测定水蛭药材中4 种主要成分含量的方法。方法：采用高效液相色谱法,以次黄嘌呤为内标,计算其对尿嘧啶、黄嘌呤和尿苷的相对校正因子(RCF),通过RCF计算上述3种成分的含量;以外标法测定的含量作实测值,比较计算值与实测值的差异。结果：尿嘧啶、次黄嘌呤、黄嘌呤和尿苷检测质量浓度线性范围分别为1.60～32.00 mg/ml、4.05～81.00 mg/ml、1.50～30.00 mg/ml、1.25～25.00 mg/ml(r>0.999);精密度、稳定性、重复性试验的RSD＜2.0％;加样回收率试验的RSD 分别为95.05％～99.45％(RSD＝1.73％,n＝6)、95.89％～99.38％(RSD＝1.52％,n＝6)、94.00％～97.88％(RSD＝1.74％,n＝6)、96.07％～99.35％(RSD＝1.18％,n＝6)。计算值与实测值差异无统计学意义(P＞0.05)。结论：该方法操作简便,精密度、稳定性、重复性良好,可用于水蛭药材中4种有效成分含量的同时测定。     

水蛭及其养殖基地农药与重金属残留分析

目的：通过对水蛭药材、养殖基地土壤、养殖用水的重金属与农药残留的比较研究,为水蛭的安全规范化生产提供依据和实践。方法:采用等离子发射光谱仪、原子荧光分光光度计和气相色谱仪对水蛭、基地土壤和养殖用水的重金属与农药残留进行了比较分析。结果与结论:除水蛭药材中铅超出国家现行有关标准外,水蛭药材,养殖基地土壤和养殖用水的六六六、DDT和重金属残留各项指标均符合国家标准规定。建议今后在制订国家有关标准时将类似水蛭类等动物的安全限量单独列出。     

The Hospital for the Ruptured and Crippled: William Bradley Coley, Third Surgeon-in-Chief 1925–1933

In January 1925, the Board of Managers of the New York Society for the Relief of the Ruptured and Crippled appointed William Bradley Coley, M.D., age 63, Surgeon-in-Chief of the Hospital for the Ruptured and Crippled (R & C) to succeed Virgil P. Gibney who submitted his resignation the month before. It would be the first time a general surgeon held that position at the oldest orthopedic hospital in the nation, now known as Hospital for Special Surgery (HSS). Coley had been on staff for 36 years and was world famous for introducing use of toxins to treat malignant tumors, particularly sarcomas. A graduate of Yale College and Harvard Medical College, Coley interned at New York Hospital and was appointed, soon after, to the staff of the New York Cancer Hospital (now Memorial Sloan Kettering Cancer Center) located at that time at 106th Street on the West Side of New York. With his mentor Dr. William Bull, Coley perfected the surgical treatment of hernias at R & C. He was instrumental in raising funds for his alma maters, Yale, Harvard and Memorial Hospital. His crusade in immunology as a method of treatment for malignant tumors later fell out of acceptance in the medical establishment. After his death in 1936, an attempt to revive interest in use of immunotherapy for inoperable malignancies was carried out by his daughter, Helen Coley Nauts, who pursued this objective until her death at age 93 in 2000. Coley's health deteriorated in his later years, and in 1933, he resigned as chief of Bone Tumors at Memorial Hospital and Surgeon-in-Chief at R & C, being succeeded at Ruptured and Crippled as Surgeon-in-Chief by Dr. Eugene H. Pool. William Bradley Coley died of intestinal infarction in 1936 and was buried in Sharon, Connecticut.     

蚂蟥药材头部与尾部抗凝活性比较研究

目的比较中药蚂蟥药材头部与尾部抗凝血活性的强度差别。方法通过考察蚂蟥药材头部及尾部对家兔全血凝固时间的影响,并以生理盐水为空白对照,比较其头部、尾部抗凝活性的差别。结果与空白对照组相比,蚂蟥的头部能明显的延长家兔全血的凝固时间（P〈0.01）,而尾部反而使凝固时间缩短。结论蚂蟥头部的抗凝活性强于尾部,其抗凝血物质可能在其头部分布较多,在蚂蟥的尾部可能存在某种能够促进血液凝固的物质。     

水蛭醇提物与水提物对肺缺血再灌注模型大鼠的保护作用

目的：研究水蛭醇提物与水提物对肺缺血再灌注模型大鼠的保护作用,并比较两者的作用效果。方法：夹闭阻断大鼠左肺门30min,开放后再灌注120min以复制肺缺血再灌注损伤模型。32只SD大鼠随机分为假手术（等容生理盐水）组、模型（等容生理盐水）组、水蛭醇提物（400mg／kg）组、水蛭水提物（400mg／kg）组。于术前3d腹腔注射给药,每天1次,连续3d。放射免疫法测定大鼠血浆血栓素B2（TXB2）和6酮前列腺素F1a（6-keto-PGF1a）含量[可反映血栓素A2（TXA2）、前列环素（PGI2）水平];酶联免疫吸附法测定大鼠血清一氧化氮（NO）含量与一氧化氮合酶（NOS）活性。结果：与假手术组比较,模型组大鼠血浆中TXB2含量显著增加,TXB2／6．keto-PGF1a比值显著降低,血清中NO含量显著增加,NOs活性显著增强（P〈0．01或P〈0．05）;与模型组比较,水蛭醇提物组、水蛭水提物组大鼠血浆TXB2、6-keto-PGF1a含量显著减少,TXB2／6．keto—PGF1a比值显著升高,血清N0含量显著减少,NOS活性显著减弱（P〈0．01或P〈0．05）,且以上指标水蛭醇提物组均好于水蛭水提物组,但差异无统计学意义（P〉0．05）。结论：水蛭醇提物与水蛭水提物均可降低肺缺血再灌注模型大鼠TXA2、PGI2、NO含量和NOS活性,且前者效果好于后者。     

蚂蟥与螺蛳耗氧率、窒息点相关性及最适投喂方式的研究

采用静水呼吸室法测定不同质量蚂蟥和螺蛳的耗氧量、耗氧率和窒息点,生物量测定法研究不同投喂方式对蚂蟥生长的影响。结果表明:25℃条件下,蚂蟥和螺蛳耗氧量与质量呈正相关(P0.05),窒息点随质量增大而增大(P0.05)。蚂蟥耗氧率与质量呈负相关,螺蛳耗氧率随质量增大呈先增大后减小趋势。投喂方式对蚂蟥终末质量、SGR,WGR和死亡率影响不显著,对进食比例影响显著(P0.05)。蚂蟥最适投喂频率为3 d投喂1次,应根据蚂蟥和螺蛳各阶段耗氧量及窒息点,结合养殖密度、水体面积以及换水频率,科学合理计算出螺蛳的投喂量。     

Purification and characterization of a novel fibrinolytic enzyme from Whitmania pigra Whitman

A fibrinolytic enzyme was purified from the dry body of Whitmania pigra Whitman. The fibrinolytic enzyme was purified to homogeneity with a yield of 0.003% and a purification of 630.7 fold. The molecular weight of the enzyme was estimated to be 26.7 kDa by reduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was tested by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and it showed that the enzyme was a novel fibrinolytic enzyme. The optimal pH and temperature of the enzyme were 8.5 and 55°C, respectively. Enzyme activity was enhanced by Na⁺, Mg^2+, and K⁺. On the contrary, the proteolytic activity was significantly inhibited by Mn²⁺, Fe²⁺, Fe³⁺, ethylenediaminetetraacetic acid (EDTA), and ethylenebis(oxyethylenenitrilo)tetraacetic acid (EGTA). Fibrinolytic and fibrinogenolytic assays showed that the enzyme preferentially hydrolyzed fibrinogen Aα-chains, followed by Bβ- and γ-chains. The α-, β-, and γ-γ-chains of fibrin were also degraded by the enzyme.