肾康注射液细菌内毒素检查法的研究

目的建立肾康注射液细菌内毒素检查方法,以期能够替代家兔热原法控制产品的热原,达到快速尧高效尧可靠尧简便的 目的遥方法按叶中国药典2015 年版四部曳通则1143 细菌内毒素检查法进行试验遥采用两个不同厂家的的鲎试剂,对7 个不同 批次的样品开展干扰试验,确定最小不干扰稀释倍数,并对样品进行了细菌内毒素检查遥结果供试品4倍稀释液无干扰作用, 细菌内毒素限值定为1.0 Eu/ml遥结论用细菌内毒素检查法代替热原法是可行的遥     

双黄连注射液细菌内毒素检查法的应用

目的:建立双黄连注射液的细菌内毒素检查方法.方法:按照《中国药典》2005年版附录细菌内毒素检查法进行试验.结果:双黄连注射液对鲎试剂与内毒素反应无干扰作用.结论:双黄连注射液采用细菌内毒素检查方法可靠,简单可行,可以考虑代替热原检查.     

两种生物材料细菌内毒素检查和热原检查方法比较的研究

目的本试验对样品管腔支架(镍钛)和支架输送系统进行热原检测.方法采用细菌内毒素检查和家兔检查法.结果细菌内毒素试验表明,两样品不合格;热原试验表明,两样品合格.结论导致鲎试剂凝集不一定就能引起生物体的热原反应.出现这种不相一致结果的原因很多.所以在评价生物材料和医疗器械时,建议做热原试验.     

家兔法与内毒素法检查生物材料热原的适用性研究

药品与生物材料在生物安全性评价方法上有着很大的区别。临床上广泛运用内毒素法检查药品热原,然而,运用细菌内毒素法进行部分组成成分较为复杂的生物材料的热原试验是否适当有待明确。本研究在2005版药典的基础上,分别运用内毒素法和家兔法对两种组织工程支架材料进行热原试验的比较研究,实验结果表明运用内毒素法得到的试验结果为阴性,运用家兔法得到的试验结果为阳性。这两种方法分别测定每种材料所得到的热原试验结果不相符合,表明对组成成分复杂的生物材料,含热原的因素较为复杂,用家兔法进行试验检测热原可能更加灵敏。     

丹参川芎嗪注射液细菌内毒素检查法研究

目的 建立丹参川芎嗪注射液细菌内毒素检查法。方法 依据《中国药典》2015年版四部（通则1143）细菌内毒素检查法,通过预干扰预试验和干扰试验,确定丹参川芎嗪注射液最大无干扰的浓度,并进行方法学验证。结果 将丹参川芎嗪注射液稀释20倍,对细菌内毒素检测法无干扰作用。结论 丹参川芎嗪注射液内毒素检查法可用于检查丹参川芎嗪注射液。     

甲硝唑葡萄糖注射液细菌内毒素检查热原方法研究

本文通过研究甲硝唑葡萄糖注射液对细菌内毒素试验的干扰情况,并与热原检查法(简称 PT)做对比试验,以期建立用细菌内毒素法检查代替热原检查的方法。1 材料鲎试剂(TAL)批号960930,0.5Eu·ml~(-1),0.1ml·支~(1-),970116,0.5Eu·ml~(-1),0.1ml·支~(-1),福州东方鲎试剂厂产,961005,0.5Eu·ml~(-1)。,0.1ml·支~(-1),厦门鲎试剂厂产;细菌内     

两种方法测定灯盏细辛注射液中细菌内毒素含量

目的：建立灯盏细辛注射液细菌内毒素的定性和定量检测方法。方法：按照《中国药典》2010年版一部附录细菌内毒素检查法项下凝胶法和动态浊度法对灯盏细辛注射液进行试验。结果：灯盏细辛注射液注射液稀释25倍（凝胶法）和200倍（动态浊度法）后对鲎试剂及内毒素反应无干扰作用。结论：用凝胶法和动态浊度法检测灯盏细辛注射液中细菌内毒素的含量是可行的。     

家兔法与内毒素法检查生物材料热原的适用性研究

药品与生物材料在生物安全性评价方法上有着很大的区别.临床上广泛运用内毒素法检查药品热原,然而,运用细菌内毒素法进行部分组成成分较为复杂的生物材料的热原试验是否适当有待明确.本研究在2005版药典的基础上,分别运用内毒素法和家兔法对两种组织工程支架材料进行热原试验的比较研究,实验结果表明运用内毒素法得到的试验结果为阴性,...     

脱细胞粘膜基质的热原试验研究

目的 考察脱细胞粘膜基质的热原试验方法的可行性。方法 依据《中华人民共和国药典》2010年版二部附录XIE细菌内毒素检查法和ISO10993.12 Biological evaluation of medical devices-Part 12:Sample preparation and reference materials,使用离心法消除样品浸提液的悬浮颗粒,使用动态浊度法验证离心条件是否会影响浸提液中细菌内毒素的检测。依据《中华人民共和国药典》2010年版三部附录XIID热原检查法检测脱细胞粘膜基质的热原试验是否合格。结果 本实验中采用的离心条件不会影响样品浸提液中细菌内毒素的检测,热原试验合格。结论该产品可以采用此离心条件进行热原检测。     

鲎试验测定长链脂肪乳注射液中细菌内毒素的方法学研究

目的：确定长链脂肪乳注射液中细菌内毒素鲎试验检查法的可行性。方法：供试品干扰试验后，采用凝胶鲎试验对样品中细菌内毒素进行检测。结果：三批长链脂肪乳注射液稀释至2倍时对细菌内毒素的检测无干扰作用，所测样品的细菌内毒素含量均小于0．5EU·ml^-1。结论：所建立方法检查长链脂肪注射液中的细菌内毒素可行。     

Analysis of endotoxin fever in rabbits by using a monoclonal antibody to tumor necrosis factor (cachectin).

A murine monoclonal antibody to rabbit tumor necrosis factor (TNF; cachectin) was injected intravenously into an endotoxin-treated rabbit to examine the role of endogenous TNF in fever. Both early and late peaks of biphasic fever evoked by the endotoxin injection were suppressed by the antibody. TNF activity was detected in an endotoxin dose-dependent manner in the blood 1 h after the endotoxin injection, which was coincident with the early-peak fever. Although the late-peak fever responded to the antibody, no significant TNF activity was detected in the blood obtained 1 h before the peak response. The blood was found to contain endogenous pyrogen activity, which was stable after heating at 70 degrees C for 30 min and resistant to in vitro treatment with the antibody. Rabbit TNF injection also elicited biphasic fever in rabbits, the second phase of which was found to be mediated by the similar endogenous pyrogen. These results suggest that endogenous TNF plays an important role in eliciting a febrile response to endotoxin.     

Mechanism of augmentation of endotoxin fever by beta interferon in rabbits: possible participation of tumor necrosis factor (cachectin).

We analyzed the mechanism of the augmentation of endotoxin fever by human beta interferon (IFN) cross-reacting to rabbit cells in rabbits by using a purified rabbit tumor necrosis factor (RaTNF) and a monoclonal anti-RaTNF. The late peak of fever evoked by the injection with both endotoxin and HuIFN was suppressed when the animals were injected previously with anti-RaTNF. IFN also augmented the pyrogenicity of RaTNF in a synergistic manner in rabbits. The blood collected 2 h after the injection of RaTNF plus IFN contained a significant endogenous TNF activity, and the serum was shown to be pyrogenic. The endogenous pyrogen activity in the 2-h blood was heat stable (70 degrees C, 30 min) and was reduced by the in vitro treatment with anti-RaTNF. These results suggest that IFN augments the febrile response of rabbits to endotoxin by stimulating endogenous TNF-mediated TNF production to induce the late peak of fever.     

Rabbit polymorphonuclear leukocytes do not secrete endogenous pyrogens or interleukin 1 when stimulated by endotoxin, polyinosine:polycytosine, or muramyl dipeptide.

Rabbit polymorphonuclear leukocytes were purified from rabbit blood by centrifugation on colloidal silica gradients followed by sedimentation in 4% Ficoll. The purified neutrophils had normal random motility, responded to chemotactic stimuli, phagocytosed zymosan particles, made superoxide, and phagocytosed and killed bacteria. However, they did not secret endogenous pyrogens either spontaneously or in response to stimulation with endotoxin, polyinosine:polycytosine, or muramyl dipeptide. Macrophages isolated on the same gradients secreted some pyrogen spontaneously and secreted considerably more in response to the same three stimuli. This evidence reinforces the idea that macrophages are the only source of endogenous pyrogens, and that pyrogens secreted by cell populations that are rich in neutrophils are to be attributed to the monocytes or macrophages that the cell populations contain.     

Rat blood leucocytes,unlike rabbit leucocytes,do not generate procoagulant activity on exposure to endotoxin.

We have investigated the ability of rat and rabbit leucocytes to generate procoagulant activity (PCA) in response to endotoxin in vitro and in vivo. On prolonged incubation with endotoxin (10 micrograms/ml f.c.) isolated rabbit leucocytes developed strong PCA as measured by clotting and amidolytic assay. In contrast, rat leucocytes failed to produce any PCA even in the presence of huge amounts of endotoxin (200 micrograms/mol f.c.). When rabbits were given two spaced endotoxin injections (25 micrograms/kg consistently showed marked PCA. Again, unlike in the rabbit, rat leucocytes obtained after 2 endotoxin injections (up to 2 mg/kg body wt) showed absolutely no PCA. These findings support the view that leucocytes are involved in endotoxin-induced disseminated intravascular coagulation (DIC) in rabbits. On the other hand the poor response of rat leucocytes to endotoxin might help explain the resistance of rats to DIC and Sanarelli-Shwartzman reaction.     

Evidence for the involvement of adenosine 3′,5′-cyclic monophosphate in fever genesis

Summary In rabbit cerebrospinal fluid (CSF) during fever induced by endotoxins, myxoviruses, or endogenous pyrogen, concentrations of adenosine 3,5-cyclic monophosphate (cyclic AMP) and prostaglandin E (PGE) are about 2-fold higher in comparison to normal values. In endotoxin treated animals paracetamol reduced the fever reaction and both PGE and cyclic AMP levels. Administration of theophyllin together with endotoxin enhanced the fever reaction and cyclic AMP levels in CSF, but had no influence on stimulation of PGE synthesis. Following injection of PGE₂ into the lateral cerebral ventricles increased cyclic AMP concentrations were found in CSF. The results suggest, that the pyrogenic effect of PGE is mediated by stimulation of cerebral cyclic AMP synthesis.     

Determination of cytokine release after in vivo and in vitro administration of Deodan (a preparation from Lactobacillus bulgaricus "LB51") by the rabbit pyrogen test.

We investigated the in vivo and in vitro cytokine inducing effects of Deodan, an oral preparation from Lactobacillus bulgaricus "LB-51", using the rabbit pyrogen test. In the first experimental approach we administered Deodan, or its chromatographically purified fraction, via the i.m. or i.v. routes. Low doses of Deodan i.m. caused the formation of a single temperature peak, whereas large doses produced a biphasic temperature curve. Intravenous injection of Deodan produced a monophasic fever in all tested doses. Chromatographically purified Deodan injected i.v. to rabbits caused a febrile response with a dose-dependent pattern, strikingly similar to that of lipopolysaccharide. LAL-testing of Deodan, however, showed that the preparation does not contain endotoxin. In in vivo neutralization studies we demonstrated that IL-1, TNF alpha, and IL-6 mediate the rabbit febrile response to Deodan. Interestingly, the effects of Deodan on the production of TNF alpha and IL-6 were more pronounced than its IL-1 inducing activity. In the second approach, we injected supernatants from mononuclear cells incubated with nonpyrogenic doses of Deodan, intravenously to rabbits ("monocyte type" of pyrogen test). Rapid-onset monophasic fevers were observed, typical for the rabbit pyrogen reaction to i.v. administration of exogenous IL-1 and TNF. Finally, we demonstrated the presence of pyrogenic cytokines in the supernatants from macrophages of Deodan-treated mice. Together, these results indicate that Deodan induces the production of cytokines with endogenous pyrogenic activity.     

Interrelationship between the restructuring of liver microvessel endothelium and changes in Kupffer's cells and hepatocytes in the development of gram-negative infection

The morphological study of various types of liver cells revealed early reaction of sinusoid endothelial cells during the development of gram-negative infection (Salmonella typhimurium). The reaction of sinusoid endothelial cells is obviously connected with the direct effect of endotoxin because the first increase of endotoxin level in the blood is detected 60 min after injection of bacteria. The initial signs of Kupffer's cell activation are revealed at the same time. One of the manifestations of endothelial cell reaction is the increase of porosity of the barrier between the blood and Disse's space. It is accompanied by ultrastructural changes in hepatocytes. The second sharp rise of endotoxin level occurs 24 hours after the administration of bacteria and is accompanied by necrobiotic changes in the endothelial cells. It is assumed that the intensity of sinusoidal endothelial cell reaction and Kupffer's cell activation determine the degree of liver damage during the development of infection.     

槲皮素、松萝酸对伴放线放线杆菌产生内毒素的抑制作用研究

目的 研究不同浓度下槲皮素、松萝酸对伴放线放线杆菌产生内毒素的抑制作用.方法 采用显色基质鲎试验法检测不同浓度药物作用下的伴放线放线杆菌菌液中留存的内毒素量.结果 槲皮素组与松萝酸组中伴放线放线杆菌菌液产生内毒素的量明显低于阴性对照组.经检验,t(谢)N =3.890,t松N =6.120,P值均＜0.01,具有显著统计学差异.结论 槲皮素与松萝酸对伴放线放线杆菌产生内毒素均有抑制作用,作用和效果与药物浓度呈正相关性,松萝酸的抑制作用强于槲皮素.     

The Effect of Thorotrast® and Cortisone on Renal Cortical Fibrinolytic Activity in the Rabbit

Previous studies have shown that a single injection of endotoxin inhibits renal cortical fibrinolytic activity in the rabbit. This suggests that the initial injection of endotoxin may prepare for the generalized Shwartzman reaction by depletion of cortical fibrinolytic activity. A fibrin slide technic was used to determine whether Thorotrast® and cortisone prepare for the generalized Schwartzman reaction by a similar mechanism. Renal cortical fibrinolytic activity was inhibited following Thorotrast injection, but no inhibition could be detected following cortisone injection. This suggests that Thorotrast, like endotoxin, prepares for the generalized Shwartzman reaction by depletion of cortical fibrinolytic activity. Failure to demonstrate inhibition of lytic activity following cortisone injection may indicate that cortisone prepares for the generalized Shwartzman reaction by another mechanism, or that the fibrin slide technic was unable to detect quantitative changes in lytic activity.