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OBJECTIVE:To establish a method for detecting bacterial endotoxins in methylthioninium chloride injection (MCI) quantitatively. METHODS: Kinetic turbidimetric limulus test was applied to detect bacterial endotoxins in MCI quantitatively, and compared with gel-clot method. RESULTS: MCI was un-interfered with the test for bacterial endotoxins at the concentration of 0.125 mg?mL-1; the content of bacterial endotoxins in all samples tested (10 mg?mL-1) were not more than 0.25 EU?mg -1, which were in accordance with the result of gel-clot method. CONCLUSIONS: Kinetic turbidimetric limulus test provides a new way to detect bacterial endotoxins in MCI quantitatively.
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OBJECTIVE: To investigate the effect of detection index of IL-4 and IgE on rapid examination of drug-induced allergic reaction.METHODS: The guinea pigs were injected with drugs intravenously and intraperitoneously,such as normal saline,horse serum,Qingkailing injection,Xiangdan injection,cerebrolysin injection and Chuankezhi injection once a day for 3 days.Then the content of IL-4 and IgE in serum of guinea pigs was recorded at different time points.Allergic reactions of guinea pigs were observed 8 days after intravenous administration,14 days and 21 days after intraperitoneous administration.RESULTS: As compared with normal saline except for Chuankezhi injection,serum level of IL-4 and total IgE were raised obviously before guinea pigs were challenged either in 8 days after intravenous injection of other drugs or in 14 days and 21 days after intraperitoneous injection.Allergic reaction occurred on all those guinea pigs after they were challenged.CONCLUSION: The detection of IL-4 and IgE provides a new way for rapid test of drug-induced allergic reaction.
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OBJECTIVE:To explore the feasibility of detecting bacterial endoxins in low molecular weight heparins calcium injection LMWHC by tachypleus amebocyte lysate(TAL).METHODS:Bacterial endotoxins in samples were detected qualita?tively and quantitatively by kinetic turbidimetric assay and gel-clot method.RESULTS:Diluted to the concentration of25IU/ml,LMWHC had an obvious interference action to the TAL test,while not at the concentration of12.5IU/ml.The contents of bacterial endotoxins in all samples were less than0.01EU/IU.CONCLUSION:It is feasible to detect bacterial endotoxins in LMWHC by TAL test.
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OBJECTIVE:To study the absorptive effect of activated carbon and desorptive effect of organic solvent on endotoxin.METHOD:The effect of different pretreatment modes,time,temperature,amount of activated carbon as well as treatment time on the absorptive effect of activated carbon were analyzed using limulus reagent test.Different organic solvents and treatment time were used to study the desorptive effect of organic solvent on endotoxin.RESULTS:Activated carbon was treated by high temperature baking or adding 0.5 mol?L-1 NaOH represented same effect in removing endotoxin.It showed that optimal time for absorption of endotoxin was 12 min and the amount of activated carbon was at 1.5%.The absorptive effect was similar under condition of room temperature and 37 ℃.The best effect of desorption achieved by using 75% alcohol as desorption reagent after 8 minutes of shaking.CONCLUSION:The activated carbon could be used to absorb endotoxin effectively under suitable conditions and organic solvent could be used to desorb endotoxin from activated carbon.
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AIM To investigate the feasibility of bacterial endotoxins test for cisplatin injection. METHODS Gel clot and kinetic turbidimetric limulus tests were applied. RESULTS After diluted to 0 14 g?L -1 ,six batches of cisplatin injection didn't interfere with the test for bacterial endotoxins, others except three also didn't interfere with the test at the concentration of 0 28 g?L -1 ; The content of bacterial endotoxins in all samples tested was not more than 1 8 ZU?g -1 . CONCLUSION The kinetic turbidimetric limulus test provides a new way to detect the content of endotoxins in cisplatin injection, samples can be diluted to 0 14 g?L -1 to refute their interfering effects on routine bacterial endotoxins test.
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AIM To examine the role of Cl - channels in endothelin 1 (ET 1) induced proliferation of vascular smooth muscle cells (VSMCs). METHODS Through cell counting and 3H TdR incorporation, together with [Ca 2+ ] i measurement by fura 2/AM fluorescence, we studied the effects of Cl - channel blockers on proliferation of VSMCs induced by ET 1. RESULTS Cl - channel blocker, DIDS, inhibited 10 nmol?L -1 ET 1 induced proliferation of VSMCs in a concentration dependent manner, however,other Cl - channel blockers such as IAA 94, NPPB, SITS, DPC and furosemide, all failed to inhibit ET 1 induced proliferation of VSMCs. DIDS reduced 10 nmol?L -1 ET 1-induced sustained [Ca 2+ ] i elevation but not Ca 2+ release from internal stores; Pretreatment of cells with 1 ?mol?L -1 nifedipine abolished the inhibitory effect of 3 ?mol?L -1 DIDS on proliferation of VSMCs and [Ca 2+ ] i elevation evoked by 10 nmol?L -1 ET 1 , while after cells were pretreated with 10 ?mol?L -1 SK&F96365, the same concentration of DIDS could also inhibit the aforecited effects of ET 1;3 ?mol?L -1 DIDS had no effect on 30 mmol?L -1 KCl induced elevation of [Ca 2+ ] i. CONCLUSION DIDS inhibited Ca 2+ influx and VSMCs proliferation induced by ET 1 through blockade of DIDS sensitive Cl - channels, and these channels may play important roles in modulating Ca 2+ influx and VSMCs proliferation induced by ET 1.