胰岛素抵抗细胞模型的胰岛素降解酶表达

目的 了解胰岛素降解酶（ＩＤＥ）基因、酶蛋白表达水平与胰岛素敏感性的关系,并初步探讨胰岛素降解抑制剂氯喹改善胰岛素敏感性的分子机制。方法 以的代大鼠肝细胞胰岛抵抗（ＩＲ）模型为研究对象,分别采用免疫印迹技术和逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）技术检测ＩＲ细胞模型的ＩＤＥ酶蛋白表达（ＥＩＰ）及基因表达水平（ＥＩＧ）,同时检测ＩＤＥ活性和反映细胞胰岛素敏感性的＾１４Ｃ－２－脱氧葡萄糖掺入率及＾１４Ｃ     

简化的差示PCR技术在伯氏疟原虫氯喹抗性基因分离中的应用

目的　为探察伯氏疟原虫氯喹抗性株 (RC株 )和敏感株 (N株 )的遗传学差异。方法　先维持对 RC株连续大剂量氯喹治疗 (5 0 m g/ (kg.d) ,再通过简化逆转录差示多聚酶链反应技术 (s DDRT- PCR)寻找 RC株和 N株疟原虫 c DNA差异片段并对所产生的可疑差异片段进行反复的 DNA回收和 PCR鉴定 ,然后选其肯定的差异片段 (N2 5 和 R2 5 )作克隆测序和序列分析及同源性检索。结果　尽管差异片段 N2 5 和 R2 5 的克隆分子 N2 5 2 和 R2 5 1 的 DNA序列相同性为 99.8% ,但由于多点突变导致它们的编码氨基酸及其模拟二级结构出现很大差异。BL AST(2 .0 6 )检索未能在基因库中发现与 R2 5 1 和 N2 5 2 相似的基因序列 ,不过它们的部分核苷酸序列 (从 12 8nt到 189nt)与褐鼠磷脂酶 Bm RNA部分序列 (从 10 5 3nt到 1114nt)具有高度同源性 :在 R2 5 1 和N2 5 2 分别为 93.5 %和 91.9%。结论　简化的差示 PCR技术结合差异片段的反复验证和序列分析可用于氯喹抗性相关基因的分离。     

氯喹对食管癌细胞系放射增敏的实验研究

目的 探讨氯喹对食管癌TE-1细胞系的放射增敏作用及其主要机制.方法 采用MTT法检测不同浓度氯喹对TE-1细胞的生长抑制作用.分别用单纯照射或照射前联合氯喹、照射后联合氯喹作用于TE-1细胞,作用6 h后用Western blot测定自噬相关蛋白LC3和Beclin1的表达,用Lyso-Tracker Red DND-99/Hochest 33258进行荧光染色,并用荧光显微镜观察细胞内酸性囊泡(AVOs)的变化.克隆形成实验分析细胞增殖的改变,拟合剂量-生存曲线并计算放射增敏参数.结果 氯喹对TE-1细胞的生长抑制作用呈浓度依赖性.辐射显著诱导TE-1细胞LC3和Beclin1的表达,并促进LC3-Ⅰ转换为LC3-Ⅱ.与单纯照射及照射前加药相比,照射后加药显著降低TE-1细胞内AVOs的荧光强度(F=16.44, P<0.05)和克隆形成率,照射前加药和照射后加药的SER_D0分别为1.037和1.439(t=8.30, P<0.05).结论 氯喹能增加食管鳞癌TE-1细胞的放射敏感性,其机制可能与抑制自噬作用相关.     

氯喹抑制肾癌细胞活性促进舒尼替尼诱导的细胞凋亡

目的：研究氯喹（经常用于抑制自噬体与溶酶体的融合,是一种晚期自噬的抑制剂）是否能抑制肾癌细胞的增殖以及对舒尼替尼(sunitinib, ST)诱导的细胞凋亡的影响。方法：以肾癌细胞系786-O和ACHN为模型,利用3-(4,5-二甲基)-5-(3 羧甲基苯环)-2-(4-硫基苯)-2H-四唑盐复合物检测法［3-(4,5-dimethylthiazol-2-yl) 5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt, MTS］观察氯喹对细胞活性的影响;透射电子显微镜、免疫杂交等手段检测氯喹影响ST诱导的凋亡、自噬的情况。应用早期自噬抑制剂3-甲基腺嘌呤(3-methyladenine, 3-MA)、RNA干扰技术敲降自噬相关蛋白Ulk1 (unc-51-like kinase 1)和微管相关蛋白1轻链3融合蛋白(microtubule associated protein 1 light chain 3 fusion protein, LC3)确定早期自噬对ST引起的细胞凋亡的影响。结果：氯喹和ST均可以抑制786-O和ACHN细胞的增殖,氯喹可促进ST诱导的细胞凋亡。与氯喹不同,抑制早期自噬减少了ST诱导的细胞凋亡和活性丢失。结论：舒尼替尼既可以引起细胞自噬,也可以促进细胞的凋亡;早期、晚期自噬对ST诱导的细胞凋亡作用不同,氯喹可以作为一种潜在的抗肾癌药物进行相关的临床研究。     

扁柏醇联合氯喹对人肺腺癌细胞凋亡的影响

目的 探讨扁柏醇（Hinokitiol）联合氯喹对人肺腺癌（H1975）细胞凋亡的影响。方法 选用人肺腺癌H1975细胞系,应用MTT法检测扁柏醇和（或）氯喹对H1975细胞活性的影响,Westernblot法检测扁柏醇和（或）氯喹对H1975细胞凋亡通路相关蛋白（Cleaved PARP、Cleaved Caspase-3、Bax）表达水平的影响。结果 0、2.5、5、10 μmol/L扁柏醇作用H1975细胞24 h后,呈剂量依赖性地抑制H1975细胞增殖,H1975细胞凋亡相关蛋白（Cleaved PARP、Cleaved Caspase-3、Bax）表达水平升高。当氯喹和扁柏醇联合应用时,与单用扁柏醇相比,显著降低H1975细胞存活率,并提高H1975细胞凋亡率,凋亡相关蛋白Cleaved PARP、Cleaved Caspase-3表达水平显著增加。结论 氯喹增加扁柏醇诱导人肺腺癌H1975细胞凋亡。     

Oxygen therapy for cerebral malaria

Malaria is an important global health issue, killing nearly one million people worldwide each year. There is a disproportionate disease burden, since 89% of cases are of African origin, and 85% of deaths worldwide occur in children under 5 years of age of age.¹ Cerebral malaria (CM) is the most serious complication of infection. Despite prompt anti-malarial treatment, fatalities remain high – mortality rates while undergoing treatment with Artemisinin or quinine-based therapy reach 15% and 22% respectively.² There is, therefore, a need to develop an adjunct therapy to preserve neurological function during the treatment period. Recent experimental research has indicated hyperbaric oxygenation (HBO) to be a rational and effective adjunct therapy.³ This article examines the current understanding of CM, and the possible benefits provided by HBO therapy.     

Inhibition of autophagy and induction of breast cancer cell death by mefloquine,an antimalarial agent

Autophagy has been recognized as a potential target for cancer therapy. The antimalarial drug chloroquine (CQ) is able to inhibit autophagy and therefore is being considered for cancer therapeutics. However, the relatively low potency of CQ prompted us to investigate whether other lysosomotropic agents might be more effective, and thus potentially more useful. We therefore compared the cytotoxic efficacy of CQ, the quinoline analog mefloquine (MQ), and the fluoroquinolones ciprofloxacin and levofloxacin in several human breast cancer cell lines. We found that MQ was the most potent compound tested; it inhibited autophagy, triggered endoplasmic reticulum stress, and caused cell death in T47D and MDA-MB-231. Altogether, our study demonstrates superior potency of MQ over CQ and the ability of MQ to produce anticancer effects in both hormone receptor positive and negative breast cancer cell lines, suggesting its usefulness in treating various types of cancer.     

间日疟原虫氯喹抗性研究进展李

疟疾仍然是一个严重危害人类健康的公共卫生问题, 尤其多见于发展中国家。随着认识的深入, 间日疟也获得 越来越多的重视。有效的药物治疗是控制疟疾甚至消除疟疾的基石, 近年来已有越来越多的间日疟原虫氯喹抗性报道, 间日疟原虫氯喹抗性已成为间日疟防治中一个备受关注的问题。本文就间日疟原虫氯喹抗性的分布现状、 体内外检测方 法和分子检测标志物研究进展进行初步总结。     

Optical coherence tomography in a patient with chloroquine-induced maculopathy

We herein report the optical coherence tomography (OCT) findings in a case of chloroquine-induced macular toxicity, which to our knowledge, has so far not been reported. A 53-year-old lady on chloroquine for treatment of rheumatoid arthritis developed decrease in vision 36 months after initiation of the treatment. Clinical examination revealed evidence of retinal pigment epithelial (RPE) disturbances. Humphrey field analyzer (HFA), fundus fluorescein angiography (FFA) and OCT for retinal thickness and volume measurements at the parafoveal region were done. The HFA revealed bilateral superior paracentral scotomas, FFA demonstrated RPE loss and OCT revealed anatomical evidence of loss of ganglion cell layers, causing marked thinning of the macula and parafoveal region. Parafoveal retinal thickness and volume measurements may be early evidence of chloroquine toxicity, and OCT measurements as a part of chloroquine toxicity screening may be useful in early detection of chloroquine maculopathy.