RP—HPLC法测定含保护剂重组人睫状神经营养因子含量

目的:建立含人血白蛋白(HSA)的重组人睫状神经营养因子(rhCNTF)蛋白含量的 RP-HPLC 测定方法。方法:采用Symmetry300反相 C_4色谱柱(150mm×3.9mm,5μm);以0.1%三氟乙酸(TFA)水-0.1%TFA 乙腈分别组成流动相 A 和 B,以初始浓度比70:30进行终浓度比为10:90的31min 线性梯度洗脱,流速为0.6 mL·min~(-1),检测波长为280nm,柱温35℃。rhCNTF 主峰保留时间在27.7～28.4 min,HSA 保留时间在17.0～17.5min。结果:在线性范围为0.061～0.735 mg·mL~(-1)有良好的线性关系(r=0.9995)。结论:本方法简便、快捷,可用于含 HSA 的 rhCNTF 纯品的纯度检测及含量测定。     

动态浊度法测定间苯三酚注射液中细菌内毒素含量

目的探讨间苯三酚注射液的细菌内毒素定量分析法。方法采用2005年版《中国药典（二部）》中的动态浊度法。结果供试品稀释50倍可以排除干扰，回收率在50％～200％范围内，试验结果与家兔升温法的一致。结论用动态文度法测定闯苯三酚注射液的细菌内毒素含量准确有效。     

维拉帕米增强猴肾上皮细胞低密度脂蛋白受体活性

本文用放射配体法观察了维拉帕米(Ver)对恒河猴肾上皮细胞LDL受体活性的影响,发现Ver以浓度和时间依赖的方式明显增强受体介导的结合和内移过程,对降解的影响不显著。Ver作用浓度为44 mmol·L~(-1)效应最明显。     

Changes in antioxidant,inflammatory and metabolic markers during 1 week cultivation of human skin explants

Human skin explant (HSE) seems to be a useful model for dermatological/cosmetic testing. HSE prepared from donor superfluous skin from plastic surgery operations is cheap and easily obtainable compared to reconstructed models. The HSE use, however, may be limited by the degeneration processes during cultivation. The aim was to monitor changes in metabolic activity and selected apoptotic, inflammatory and antioxidant parameters during 7 day cultivation. The significant changes were found in the superoxide dismutase‐2 level from day 5, glutathione S‐reductase level from day 6, metabolic activity and fibulin‐5 level from day 4, cyclooxygenase‐2, interleukin‐6 and interleukin‐10 from day 1 to 2. Other selected markers (lipid peroxidation products and glutathione level, glutathione S‐transferase, catalase, superoxide dismutase and glutathione S‐reductase activity, glutathione peroxidase and glutathione S‐reductase levels) were not modified significantly due to high inter‐individual variability of skin donors. The HSE microstructure as well as cytokeratin‐10 and proliferation marker Ki67 expression was also only minimally affected during cultivation. Collectively, the results demonstrate that HSE represents a good model for short‐term studies focused on the physical and chemical agent toxicity, protective potential of compounds or metabolic biotransformation. However, reduced metabolic activity, increased inflammation and the high inter‐individual variability and sensitivity of donors have to be taken into consideration.     

Therapeutic effects of total alkaloids of Tripterygium wilfordii Hook f. on collagen-induced arthritis in rats

Ethnopharmacological relevance

Tripterygium wilfordii Hook f. is one of Traditional Chinese Medicines which is commonly used to treat rheumatoid arthritis (RA). The total alkaloids were the main constituent part of Tripterygium wilfordii Hook f. It has a great significance to study the effects of the total alkaloids of Tripterygium wilfordii Hook f. (ATW) on RA.

Aim of the study

This paper aims at investigating the therapeutic effect of ATW on RA and its possible mechanism, and providing a theoretical and experimental basis for the clinical use of ATW.

Materials and methods

The model of wistar rats of type II collagen-induced arthritis (CIA) was made, and the rats were perfused a stomach with ATW for 4 weeks continuously. Then the levels of interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)-<alpha>, in the serum of CIA rats were detected by enzyme linked immunosorbent assay (ELISA), and the joint pathological section of CIA rats was observed by hematoxylin and eosin (HE) staining method and the expression of IL-6, IL-8, nuclear factor kappa B (NF-κB), TNF-α were measure by immunohistochemistry staining method.

Results

Compared with model group, ATW could significantly reduce paw swelling and suppresse articular cartilage degenerated. The results also found that there was significant reduction levels of IL-6, IL-8 and TNF-α in serum of CIA rats treated with ATW and ATW inhibited the expression of IL-6, IL-8, NF-κB, TNF-α in synovial tissue.

Conclusion

ATW not only could inhibit the symptom of CIA rats significantly but also could inhibit the production of IL-6, IL-8, TNF-α in serum and the expression of IL-6, IL-8, NF-κB and TNF-α in synovial tissue targeting the inflammatory. ATW would be a drug as a novel botanical drug for the treatment of RA.     

Humoral immunity induced by a live Francisella tularensis vaccine. Complement fixing antibodies determined by an enzyme-linked immunosorbent assay (CF-ELISA)

A complement-fixing enzyme-linked immunosorbent assay (CF-ELISA) with bacterial sonicate as the antigen, human AB serum as the source of complement and alkaline-phosphatase-labelled anti-(human C3e) as the conjugate was developed for detecting antibodies to Francisella tularensis. Humoral responses after inoculation with a live attenuated tularemia vaccine were studied in 13 subjects by this method. Twelve responded with CF antibodies, which in most cases appeared 4 weeks after vaccination, the response time varying from 1 week to 2 months. The highest individual CF titres, of 140–9990, were reached 1–4 months after vaccination (average 2.4 months). The CF titres tended to decrease with time, and at 10 months two of the 12 subjects with an earlier CF positive response were negative again. CF antibodies were nevertheless still detectable in all three subjects studied 1.5 years after vaccination.