水溶性槲皮素的研究

水溶性槲皮素的研究佘戟１）莫丽儿康铁邦梁念慈（广东医学院化学教研室、医用生物化学研究所，湛江５２４０２３）槲皮素（ｑｕｅｒｃｅｔｉｎ；３，３′，４′，５，７－ｐｅｎｔａｈｙｄｒｏｘｙｆｌａｖｏｎｅ）存在于许多植物体内，它的提取和纯化已有大量的报道．国...     

硫酸特布他林干粉吸入剂制备工艺的优化研究

目的:优化硫酸特布他林干粉吸入剂的制备工艺。方法:采用喷雾干燥技术制备硫酸特布他林干粉吸入剂,采用双层液体碰撞器测定其体外肺沉积率,扫描电镜观察干粉的表观形貌,热重分析仪测定干粉的水分含量,激光粒度测定仪测定粒径大小,以产品收率、水分含量、粉末的空气动力学粒径及体外肺沉积率为考察指标,通过正交设计结合多指标综合评价法优化最佳制备工艺。结果:通过正交试验-多指标综合评价,最佳制备工艺为:喷雾压力190 kPa,干燥风速0.7 m3.min-1,供液速度7.0 mL.min-1,入口温度120℃。结论:按最佳制备工艺制得的干粉收率为50.54%,水分含量为0.467%,空气动力学粒径为1.80μm,体外肺沉积率为55.19%。正交试验结合多指标综合评价法用于硫酸特布他林干粉吸入剂制备工艺的优化有效可用。     

A fluorescent curcumin-based Zn(II)-complex reactivates mutant (R175H and R273H) p53 in cancer cells

Background

Mutations of the p53 oncosuppressor gene are amongst the most frequent aberration seen in human cancer. Some mutant (mt) p53 proteins are prone to loss of Zn(II) ion that is bound to the wild-type (wt) core, promoting protein aggregation and therefore unfolding. Misfolded p53 protein conformation impairs wtp53-DNA binding and transactivation activities, favouring tumor growth and resistance to antitumor therapies. Screening studies, devoted to identify small molecules that reactivate mtp53, represent therefore an attractive anti-cancer therapeutic strategy. Here we tested a novel fluorescent curcumin-based Zn(II)-complex (Zn-curc) to evaluate its effect on mtp53 reactivation in cancer cells.

Methods

P53 protein conformation was examined after Zn-curc treatment by immunoprecipitation and immunofluorescence assays, using conformation-specific antibodies. The mtp53 reactivation was evaluated by chromatin-immunoprecipitation (ChIP) and semi-quantitative RT-PCR analyses of wild-type p53 target genes. The intratumoral Zn-curc localization was evaluated by immunofluorescence analysis of glioblastoma tissues of an ortothopic mice model.

Results

The Zn-curc complex induced conformational change in p53-R175H and -R273H mutant proteins, two of the most common p53 mutations. Zn-curc treatment restored wtp53-DNA binding and transactivation functions and induced apoptotic cell death. In vivo studies showed that the Zn-curc complex reached glioblastoma tissues of an ortothopic mice model, highlighting its ability to crossed the blood-tumor barrier.

Conclusions

Our results demonstrate that Zn-curc complex may reactivate specific mtp53 proteins and that may cross the blood-tumor barrier, becoming a promising compound for the development of drugs to halt tumor growth.     

子宫颈鳞癌患者放疗前后血清铜、锌的变化

目的检测子宫颈癌患者放疗前后血清铜、锌的变化，检测子宫颈癌患者与正常人血清铜、锌水平的比较。材料与方法采用三电极直流等离子原子发射光电直读光谱仪法检测。结果子宫颈鳞癌患者血清铜（ＳＣｕ）、铜锌比值（Ｃｕ／Ｚｎ）显著高于正常对照组（Ｐ＜０．００１），Ⅲ期患者又显著高于Ⅱ期患者（Ｐ＜０．００１）；放疗后癌灶完全消退者（ＣＤ）ＳＣｕ、Ｃｕ／Ｚｎ明显下降（Ｐ＜０．０１）；部分消退者（ＰＤ）二值仍处于高水平。结论：ＳＣｕ和Ｃｕ／Ｚｎ可以检测放疗疗效，估计预后，有一定临床意义。     

Protective effects of Terminalia arjuna against Doxorubicin-induced cardiotoxicity

Terminalia arjuna has been marked as a potential cardioprotective agent since vedic period. The present study was aimed to investigate the effects of butanolic fraction of Terminalia arjuna bark (TA-05) on Doxorubicin (Dox)-induced cardiotoxicity. Male wistar rats were used as in vivo model for the study. TA-05 was administered orally to Wistar rats at different doses (0.42 mg/kg, 0.85 mg/kg, 1.7 mg/kg, 3.4 mg/kg and 6.8 mg/kg) for 6 days/week for 4 weeks. Thereafter, all the animals except saline and TA-05-treated controls were administered 20 mg/kg Dox intraperitonially. There was a significant decrease in myocardial superoxide dismutase (38.94%) and reduced glutathione (23.84%) in animals treated with Dox. Concurrently marked increase in serum creatine kinase-MB (CKMB) activity (48.11%) as well as increase in extent of lipid peroxidation (2.55-fold) was reported. Co-treatment of TA-05 and Dox resulted in an increase in the cardiac antioxidant enzymes, decrease in serum CKMB levels and reduction in lipid peroxidation as compared to Dox-treated animals. Electron microscopic studies in Dox-treated animals revealed mitochondrial swelling, Z-band disarray, focal dilatation of smooth endoplasmic reticulum (SER) and lipid inclusions, whereas the concurrent administration of TA-05 led to a lesser degree of Dox-induced histological alterations. These findings suggest that butanolic fraction of Terminalia arjuna bark has protective effects against Dox-induced cardiotoxicity and may have potential as a cardioprotective agent.     

藻酸双酯钠治疗缺血性脑血管病102例效果分析

应用藻酸双酯钠对102例缺血性脑血管病患者进行治疗。观察用药前后症状与体征及血流动力学的变化。结果;显效率为70.32％,有效率为29.68％,总有效率达100％。     

新基因ZNFD的克隆及其多克隆抗体的制备

目的:PCR扩增得到新基因ZNFD(Znf domain containing protein),构建pET32a-ZNFD原核表达载体,诱导蛋白表达及制备多克隆抗体。方法:通过PCR的方法克隆新基因ZNFD,选取部分抗原免疫原性较高的部分ORF序列,克隆到原核表达载体pET32a上,利用大肠杆菌Rosetta-gami(DE3)表达系统表达ZNFD融合蛋白。纯化目的蛋白免疫家兔,制备多克隆抗体。通过琼脂糖双向扩散实验和Western blot鉴定其效价和特异性。结果:经测序鉴定已成功克隆ZNFD基因。通过构建原核表达载体pET32a-ZNFD,在大肠杆菌Rosetta-gami(DE3)中使用IPTG诱导表达,得到相对分子质量约为45 kD的融合蛋白。纯化蛋白免疫家兔,制备的多克隆抗体具有较强的免疫特异性。结论:得到纯化的ZNFD蛋白,制备的多克隆抗体达到了一定的效价,且具有高特异性,为进一步研究ZNFD的功能奠定了实验基础。     

双波长分光光度法连续测定血清中铜和锌

以锌试剂为显色剂,在普通分光光度计上用双波长分光光度法连续测定血清中铜和锌.测得健康人血清中铜浓度为15.42μmol/L,锌浓度为16.83μmol/L,回收率分别为96%～102%和93%～104%,标准偏差分别为0.04和0.05,变异系数分别为3.8%和4.9%.提示该方法具有较好的准确度和精密度.     

沐舒坦和硫酸镁佐治毛细支气管炎临床观察

目的探讨沐舒坦(氨溴索)和硫酸铁对毛细支气管炎的疗效及安全性。方法2004年11-2005年2月住院毛细支气管炎患儿40列为对照组,2005年-2006年1月住院毛细支气管炎患儿48例为治疗组。两组均采用常规治疗,治疗组加用盐酸沐舒坦和硫酸镁静脉滴注。结果治疗组肺部哮鸣音消失时间及住院日的缩短优于对照组。结论沐舒坦和硫酸镁联合应用是佐治毛细支气管炎有效的药物。     

锌影响镉对人未成熟树突状细胞毒性作用

目的探讨锌对镉诱导的人未成熟树突状细胞(iDC)毒性的影响。方法分离人周围血单核细胞(PBMC)并诱导为iDC后,分组给予生理氯化钠溶液(对照组)、氯化镉(CdCl2组)、硫酸锌(ZnSO4组)、锌预处理后加氯化镉(Zn+Cd组)、锌离子特异性螯合剂(TPEN)预处理后加氯化镉(TPEN+Cd组)处理。噻唑蓝(MTT)颜色反应法检测细胞活力;单细胞凝胶电泳(SCGE)检测DNA链断裂损伤;吖啶橙(AO)与碘化丙啶(PI)核双染观察细胞凋亡并通过流式细胞仪检测凋亡细胞中染色体断裂的亚二倍体(SubG1)。结果 CdCl2组iDC活力抑制率随CdCl2呈剂量依赖性显著增高(P<0.05);与ZnSO4组或CdCl2组相比,Zn+Cd组和TPEN+Cd组细胞活力抑制率明显升高(P<0.05);相同CdCl2剂量(20μmol/L)时,Zn+Cd组和TPEN+Cd组细胞的彗星尾长和尾矩明显增大(P<0.05);各处理组出现凋亡细胞核染色和SubG1阳性细胞比例明显增加。结论镉化合物暴露导致iDC活力降低,其诱导的细胞毒性明显高于锌;细胞内锌稳态干预可增加镉介导的iDC内DNA损伤相关的毒作用敏感性。