生物制品热原实验用家兔胃肠道疾病的原因探讨和相应对策

目的 做好生物制品热原实验用家兔的实验期管理工作,保障热原实验数据的准确性.方法 将《中国药典》2010年版对实验动物的要求与我国现阶段实验兔产业的现状和普通级家兔生长发育规律结合起来,分析生物制品热原实验用家兔胃肠道疾病多发的原因,探讨相应的管理对策.结果与结论 实验管理部门必须加强对动物供应商的技术指导和监督,双方密切合作,才能在实际工作中取得良好效果.     

家兔热原实验中预检温对实验结果的影响

本文从预检温的筛选方式、筛选标准、稳定时间和不同基础体温的家兔对热原的敏感性等方面探讨了做好家兔热原实验值得注意的一些问题,并就美国药典和欧洲药典的要求进行了介绍和比较.建议对预检温筛选家兔采用至少与家兔热原实验一样的方式;为减少复试率和不合格率,应尽量不使用基础体温较低家兔进行实验.     

SPF家兔和普通家兔在热原实验中的比较研究

目的:将不同等级(普通级与SPF级)的家兔用于生物制品、细菌内毒素国家标准品的热原检测,研究普通级与SPF级家兔适应期对环境的适应能力、体温的均一性及对热原质的敏感性上是否存在差异,最终选择出更适合于热原检测的等级家兔。方法:按照中国药典2005版三部附录ⅫD热原检查法,分别对SPF级与普通级家兔进行适应期与筛选实验的比较、对细菌内毒素(G-)的敏感性比较以及对生物制品热原检测准确性的比较。结果:适应期、筛选合格率、筛选实验体温分布情况及筛选实验体温波动情况比较存在显著性差异;对内毒素敏感性和同一制品热原检查无显著性差异。结论:SPF级家兔在适应期中的适应性优于普通级家兔;SPF级家兔筛选合格率优于普通级家兔;SPF级家兔与普通级家兔对内毒素敏感性无明显差异;SPF级家兔与普通级家兔对同一制品热原检查准确性无明显差异。     

热原检查实验次数对家兔致热灵敏度的影响

目的 针对热原检查实验次数是否改变家兔对热原物质灵敏度进行研究.方法 于实验第1d、第3d、第10d和第24d给家兔静脉注射不同浓度内毒素溶液各4次.记录各组家兔体温变化并进行统计学处理.结果 5EU·mL-1·kg-1组家兔在第1次和第2次给药后判定为不合格,第3次和第4次给药后判定为合格;而10EU·mL-1·kg-1组家兔4次给药后均判定为不合格.结论 热原检查使用9～10次的家兔对细菌内毒素的灵敏度未发生明显变化,但家兔短期内多次接触≥临界内毒素剂量可产生耐受性,减少其对热原物质的灵敏度.     

基于关联规则算法分析中/长链脂肪乳注射液热原检查影响因素研究

目的 探索中/长链脂肪乳注射液热原检查中的不同影响因素对结果的影响，提高中/长链脂肪乳注射液热原检查结果的准确性和可靠性。方法 选取苏州市药品检验检测研究中心近5年对中/长链脂肪乳注射液热原检查的检验数据进行统计，采用Apriori关联规则分析热原实验中家兔升温幅度与家兔体质量、性别、基础体温、环境温度、家兔使用次数、实验样品与前次实验是否相同、实验间隔时间之间的关联关系。结果 家兔升温幅度≥0.4℃与家兔基础体温38.0~38.7℃、环境温度波动1.6~3.0℃、家兔第1~3次使用、实验间隔天数为2~5 d和前次使用相同样品呈现强关联。结论 在中/长链脂肪乳注射液热原检查过程中，应尽量选用基础体温和使用次数不同的家兔，同时尽量避免使用前次样品为中/长链脂肪乳注射液的家兔，实验间隔天数适当延长，并且保持较小环境温度波动，以保证中/长链脂肪乳注射液热原检查的准确性和可靠性。     

家兔热原试验中体温变化分析

家兔热原试验中体温变化分析沈琴华（江苏常州市药品检验所２１３００３）《中国药典》１９９０年版对家兔的饲养条件、实验条件等作了严格规定，但许多基层单位动物房仍没达到要求，影响了用家兔法检测热原的结果。现将我所１９９０～１９９４年间热原检测时所用１３６７...     

血液制品热原试验动物最佳体温的探讨

应用ＣＪ－ＲＺＹ型热源自动测试仪，按生物制品规程，预检５２０只正常家兔体温，对检温合格的３１１只家兔体温进行统计学处理，获得实验家兔的最佳体温范围为３８．３２℃￣３８．９４℃进一步将家兔法热原试验由于动物本身体温的影响而造成的假阳性率降至最低水平。同时也为今后再版生物制品规程制定家兔法热原试验使用的家兔体温的范围提供依据。     

不同时间家兔体温变化规律的测定

<正> 热原检查是保证注射剂质量和安全用药的重要项目之一。家兔法检查热原是世界各国药典共同采用的方法。影响热原检查的因素很多,但是家兔本身的体温变化情况是一个很重要的因素。为了探讨家兔体温变化的情况,并从中总结它的规律性以提高实验的成功率和可靠性,我们应用国产热原测温仪,对88只家兔体温进行了测定,并进行了总结分析。     

影响家兔法检查热原的诸因素

家兔法检查热原是各国药典采用的方法。由于天然热原分子的不均一性,被试家兔的个体差异和实验条件较难完全一致,因此有时初试阳性的制品,经复试又可符合规定。资料认为,错判的可能性在7～17%之间。对如何增加家兔热原试验的精确度和可靠性,引起了药学工作者的密切关注。现将影响家兔法检查热原的因素作一介绍,俾使引起实际工作者的注意。     

热原试验中影响家兔体温下降的几个因素

有关家兔的热原试验报道很多,而热原试验中家兔体温下降的报道却少见,普遍认为热原试验中家兔体温升高是正常现象,体温下降是异常现象.为此本文就二年度6种输液制剂的家兔1694只次家兔热原试验情况进行考查分析,以使在热原测试过程中,减少降温现象的出现.     

TNF alpha measurement in rat and human whole blood as an in vitro method to assay pyrogens and its inhibition by dexamethasone and erythromycin

To ensure the safety of potential drugs, pyrogen tests are traditionally performed in rabbits. New methods have been developed as alternatives to the test to reduce the use of experimental animals. Among these methods there are the Limulus amoebocyte lysate test and the determination of cytokine production by human leukocytes and whole blood. When exposed to a range of concentrations of endotoxins, human and rat whole blood release TNFalpha at amounts that are detectable by a commercially available enzyme-linked immunosorbent assay (ELISA). Our results show that the sensitivity of human and rat blood to endotoxins from Salmonella abortus equi and Escherichia coli is similar. In rat blood, TNFalpha was detected after contact with the pyrogens only in fresh blood, collected on the same day of incubation with the pyrogenic substances. The measurement of TNFalpha production would be a reliable alternative to the rabbit pyrogen test. However, given that the addition of erythromycin and dexamethasone inhibited the production of this cytokine, this method is limited when parenteral formulations contain these two drugs. Similar inhibition has been observed in the rabbit test. Additional experiments will be necessary to demonstrate that the rat whole blood test system is useful and reliable for the pyrogens evaluation.     

塞克硝唑注射液中细菌内毒素的定量检测研究

目的建立塞克硝唑注射液中细菌内毒素定量分析的方法.方法采用动态比浊定量法考察塞克硝唑注射液对鲎试剂的干扰作用,并与热原检查法进行对比试验分析.结果塞克硝唑注射液的2倍稀释液用动态浊度法定量检测时已无干扰,内毒素回收率在50%～200%范围内,样品检查结果与家兔法一致.结论可用动态浊度法对塞克硝唑注射液中的细菌内毒素进行质量控制.     

Assay of pyrogens by interleukin-6 release from monocytic cell lines.

A novel in-vitro system has been developed for the detection and quantification of pyrogen in pharmaceutical products. The measured variable was evoked secretion of the pyrogenic cytokine interleukin-6 from MONO MAC 6 monocytic cells incubated with the product. The interleukin-6 was detected using a specific and sensitive ELISA developed for this purpose. The test system detected pyrogenic contamination in 3 batches of therapeutic human serum albumin which had caused adverse reactions in recipients. The contamination was not detected in conventional tests: the rabbit pyrogen test and the limulus amoebocyte lysate test.     

Uridine-induced hyperthermia in the rabbit

Uridine injection in 0.6% saline elevated rabbit temperatures (mean = 0.9 °C) in the USP XX pyrogen test. Hyperthermia was delayed in onset and peaking 3–4 h post injection, but the injection was negative in the limulus amoebocyte lysate (LAL) assay. Uridine from five lots of different sources exceeded USP XX limits in the rabbit pyrogen test and proved negative in the LAL assay. Because the dose of uridine was high, several procedures were used to determine if an impurity was the cause of temperature elevation. Uridine remained pyrogenic in spite of ultrafiltration (10 000 nominal mol. wt), recrystallization and preparative scale HPLC. Sterile filtration and autoclaving also did not affect the response. Hyperthermia, therefore, appears to be an inherent property of uridine. Uridine was also found to release endogenous pyrogen in-vitro from human mononuclear cells. Uridine has been reported to induce fever in man, thus the USP rabbit pyrogen test predicted for the clinical response.     

Comparative evaluation of the human whole blood and human peripheral blood monocyte tests for pyrogens

Two different in vitro tests for pyrogens, using human peripheral blood monocytes (PBMNC) and diluted whole blood (WBC), respectively, were applied to different classes of parenteral medicinal products. Many of these products did not have a specified endotoxin limit concentration that was established as the maximum valid dilution to comply with the test. The results of the in vitro tests for pyrogens were compared with the results from the Limulus amoebocyte lysate (LAL) and rabbit pyrogen tests. The Second International Standard for endotoxin was used to calibrate all of the assays and the International Standard for IL-6 was used to calibrate the IL-6 ELISA which provided the readout for the in vitro tests for pyrogens. Preparatory tests were conducted to ensure that the "criteria for validity and precision of the standard curve" were satisfied and that the drugs being tested did not interfere in the tests. The PBMNC/IL-6 test had a detection limit of 0.06 EU/ml and spike recoveries were 62-165%. The whole blood/IL-6 test also had a detection limit of 0.06 EU/ml and spike recoveries were 58-132%. The application to the detection of non-endotoxin pyrogens needs to be evaluated in more detail, but the two in vitro tests for pyrogens showed good agreement overall, both with each other and with the LAL test and the rabbit pyrogen test for the detection of endotoxins.     

灯盏花素注射液细菌内毒素检查法的研究

目的:建立灯盏花素注射液的细菌内毒素检查方法。方法:按《中国药典》2005年版二部附录ⅪE进行实验和结果判断。用所建立的细菌内毒素检查法与家兔法同时对148批样品进行比较试验。结果:灯盏花素注射液的浓度最大不干扰浓度为0.3 mg.mL-1;家兔法与细菌内毒素检查法的结果具有一致性;灯盏花素注射液对细菌内毒素的致热活性有一定的抑制作用。结论:用细菌内毒素检查法替代家兔法检查热原是可行的,其限度拟订为1 EU.mg-1。     

医疗机构仓储管理系统信息化平台应用初步研究

本研究以上海市血液中心仓储管理为研究模型，建立适用于医疗机构（包括采供血机构）仓储管理的系统信息化平台，从申购、购入、确认、入库、标识、贮存、监控、发放（或退货）到使用整条流程线的信息化管理进行阐述，对医疗机构的仓储管理情况进行有效控制并跟踪仓储业务的物流和成本管理全过程，实现了较完善的仓储信息化管理。     

鲎试剂法检测平衡液中细菌内毒素

目的 建立平衡液的细菌内毒素检查方法。方法 参照中国药典2000年版细菌内毒素检查法及热原检查法。结 果 干扰试验表明,供试品对细菌内毒素检查法不存在干扰作用。结论 细菌内毒素检查法可代替家兔法进行平衡液热原 检查。     

参麦注射液热原检查的适用性研究

目的对热原检查标准制订时开展适用性研究的必要性进行探讨。方法选用参麦注射液按《中国药典》2010年版一部附录ⅩⅧB＂中药注射剂安全性检查法应用指导原则＂进行热原检查适用性研究。结果参麦注射液热原检查不干扰给药剂量为3mL/kg;参麦注射液给药剂量为5mL/kg时,对细菌内毒素致家兔体温升高有干扰。结论中药注射剂的热原检查有必要进行适用性研究。     

重组人红细胞生成素注射液中细菌内毒素凝胶检查法的研究

目的本文介绍了一种重组人红细胞生成素注射液的细菌内毒素检查法。方法应用鲎法检查重组人红细胞生成素注射液中的细菌内毒素,考察重组人红细胞生成素注射液对细菌内毒素检查法的干扰。结果重组人红细胞生成素注射液稀释150倍对细菌内毒素检查法无干扰。结论使用凝胶法检查重组人红细胞生成素注射液中的细菌内毒素是可行的,可用细菌内毒素凝胶法代替家兔热原检查法。