人ERRα基因小分子干扰RNA慢病毒载体构建及鉴定

目的构建并鉴定靶向人ERRα基因的小分子干扰RNA的慢病毒载体。方法针对ERRαmRNA设计了4条si RNA,并构建pGCSIL-GFP-siERRα慢病毒质粒,PCR扩增阳性克隆并测序鉴定。用pGCSIL-GFP-siERRα、pHelp-er1.0和pHelper2.0质粒共转染293T细胞包装产生慢病毒,测定病毒滴度。将慢病毒干扰RNA及含有ERRα过表达载体共转染293T细胞,Western-blot检测ERRα表达,观察蛋白表达抑制效果。结果 PCR和测序结果与设计的干扰序列一致,病毒滴度达2×109TU/ml。转染细胞中ERRα蛋白表达显著降低。结论成功构建高表达、高效率的人ERRα基因小分子干扰RNA慢病毒载体,为进一步研究ERRα在细胞核内转导中的作用机制和靶向ERRα治疗奠定基础。     

Multi-walled carbon nanotube-induced inhalation toxicity: Recognizing nano bis-demethoxy curcumin analog as an ameliorating candidate

Human beings and ecosystems are being possibly exposed to CNTs, as there is a rise in global production rate of carbon nanotubes (CNTs). This may affect the health of humans and increases the environmental risk. We have already reported the pulmonary toxicity due to the inhalation of MWCNTs. We claim that a compound with anti-inflammatory and antioxidant activity may ameliorate the CNT-induced toxic effect. With this view, we have investigated the ameliorative effect of intravenously-administered nano bis-demethoxy curcumin analog (NBDMCA) against MWCNTs-induced inhalation toxicity by examining the lung histopathology for inflammatory cell dynamics, pulmonary remodeling and estimating the inflammatory biomarkers in the broncho-alveolar lavage fluid. We observed that NBDMCA could ameliorate the injury as evidenced by the decline in the levels of markers of inflammation, cell damage, and the histopathological changes induced by MWCNTs. We conclude that NBDMCA may be used to reduce the risk of MWCNTs-induced inhalation toxicity.     

石菖蒲抗癫痫作用及其机制的研究进展

癫痫是最常见的神经系统疾病之一,发病率逐年增加,目前其主要治疗手段是药物治疗,但是效果有限。石菖蒲具有开心窍、益心智、安心神等功效,是祖国医学治疗癫痫症的常用味药;石菖蒲的多种提取物及其有效成分α-细辛醚具有抗癫痫作用,在癫痫的多种发病机制中发挥了降低神经兴奋性、保护神经元等作用。同时,石菖蒲对神经系统还具有兴奋性和抑制性双向调节作用。本文主要综述了目前癫痫发病机制的研究进展,在此基础上探讨中药石菖蒲的抗癫痫作用及其可能的作用靶点,论述其发展为辅助抗癫痫药物的可能性。     

X射线照射对RAW264·7细胞TNF-α和NF-κB表达的影响

目的探讨X射线照射诱发小鼠巨噬细胞分泌TNF-α和NF-κB的规律。方法以8Gy X射线单次照射小鼠巨噬细胞系RAW264.7细胞,并于照后不同时间收集细胞培养上清液,采用ELISA法检测RAW264.7细胞TNF-α分泌水平;以8Gy X射线单次照射小鼠巨噬细胞系RAW264.7细胞,并于照后不同时间收集细胞并进行裂解,采用Western blot实验检测NF-кB（p65）的表达。结果 X射线照射能使小鼠巨噬细胞分泌TNF-α增加,在照射后34h达到一次分泌峰值;细胞核中NF-κB（p65）分布增多,并且随着时间的延长其核移位明显增多,照射后4h达到一次最高值。结论 X射线照射能诱发小鼠巨噬细胞TNF-α和NF-κB表达增强。     

密蒙花总黄酮对去势雄鼠干眼症模型角膜和泪腺组织中TNF-α,IL-1β表达的影响

目的:探讨密蒙花总黄酮对去势雄鼠干眼症动物模型角膜及泪腺组织炎症因子表达的影响与组织损伤的关系。方法:选取150只健康1月龄Wistar雄性大鼠随机分为5组(正常组、假手术组、手术对照组、雄激素治疗组、密蒙花总黄酮治疗组,分别用A,B,C,D,E组表示),每组30只。将C,D,E三组实验鼠切除双侧睾丸及附睾;B组只切开阴囊,不切除睾丸,作为假手术对照组,A组不作处理。自造模后1wk,待伤口基本愈合开始用药。A,B,C组以生理盐水灌胃,1次/d;D组用丙酸睾酮注射液大腿肌肉注射,3d1次;E组以密蒙花提取物生理盐水混悬液灌胃,1次/d。各组实验动物分别于用药1,3,5mo后随机取10只,行SchirmerⅠ试验、泪膜破裂时间(break-up time,BUT)及角膜荧光素染色检查并处死各组动物,取双眼泪腺、角膜组织采用免疫组织化学方法观察各组角膜和泪腺组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),白细胞介素1β(interleukin 1β,IL-1β)蛋白的表达。结果:A与B组泪腺、角膜上皮细胞中TNF-α,IL-1β阳性表达均不明显;C组泪腺、角膜上皮细胞中TNF-α,IL-1β阳性表达的细胞数明显高于D,E组(P<0.05);D,E各组间差异不显著(P>0.05)。结论:去势雄鼠干眼症角膜和泪腺组织局部炎症反应可能是角膜损伤和泪腺分泌功能丧失的原因之一,TNF-α,IL-1β表达增加与其炎症反应有关。密蒙花总黄酮能够下调去势雄鼠角膜和泪腺局部TNF-α,IL-1β蛋白的表达,可能是其治疗干眼症的关键机制之一。     

卤虫卵油脂肪酸组成分析及α-亚麻酸含量测定

目的采用毛细管气相色谱法分析卤虫卵油的脂肪酸主要组成并建立测定卤虫卵油中α-亚麻酸含量的方法。方法采用DB-WAX毛细管柱;程序升温:起始温度150℃,升温速率3℃.min-1,结束温度240℃,保持10 min;氢焰离子化检测器温度250℃。结果卤虫卵油中不饱和脂肪酸占总脂肪酸质量分数超过70%,其中油酸、α-亚麻酸质量分数分别为26.9%、27.7%。α-亚麻酸甲酯质量在0.719～3.60μg内线性关系良好(r=0.999 8,n=5),平均回收率为101.6%,RSD=2.3%。结论毛细管气相色谱法可作为卤虫卵油α-亚麻酸的含量测定方法。     

遗传易感因素与慢性乙型肝炎干扰素治疗反应的相关性

IFN-α以抗病毒及免疫调节双莺作用用于慢性乙型肝炎的治疗,对治疗反应的早期预测有利于治疗的持续.随着人类基因组学和蛋白质组学的研究进展,国内外对HBV感染发病及治疗反应遗传易感因素有较多报道,主要集中于抗病毒蛋白和免疫调节相关细胞因子的基因多态性.此文就遗传易感因素对IFN-α治疗慢性乙型肝炎疗效的影响作了综述.     

异种移植免疫学及其潜在风险的研究进展

同种器官移植成功率的提高导致了供体器官的严重短缺.采用解剖学上与人类相近的动物,比如猪的器官可以解决这种危机.但从猪到人的器官移植需要克服很多障碍,包括免疫学,生理学及其伦理道德问题.超急性排斥反应是猪到人异种器官移植的首要免疫学障碍,目前主要通过敲除半乳糖α1,3半乳糖(galactose-α1,3-galactose,Gal)抗原来克服超急性排斥反应.除此之外,仍有其它的非-Gal抗原可能引起猪到人的移植物的失功,例如N-羟乙酰神经氨酸等.除了免疫学障碍,猪器官携带的病毒及可能引起的异种移植的潜在风险也不容忽视.虽然现在还没有明显的实验数据显示猪到人的病原体的感染,但当猪到人的免疫学障碍被克服后,感染将成为又一研究热点.     

昆丹颗粒对代谢综合征大鼠模型的影响及机制研究

目的观察昆丹颗粒对代谢综合征大鼠模型的影响及探讨其可能机制。方法 Wistar雄性大鼠随机分组,分为空白对照组和造模组,空白对照组灌胃蒸馏水,造模组灌胃高脂高糖高盐乳剂。造模6周后,将造模成功的大鼠按体重随机分为5组：模型组、昆丹高剂量组、昆丹中剂量组、昆丹低剂量组、罗格列酮组,药物干预6周,在用药期间继续灌胃高脂高糖高盐乳剂。试验结束后,检测体重、空腹血糖、血清胰岛素、计算胰岛素敏感指数（ISI）、血脂、肿瘤坏死因子-α（TNF-α）、游离脂肪酸（FFA）、瘦素（lept）。结果昆丹高剂量组可明显降低代谢综合征大鼠的空腹血糖、血清胰岛素、尾动脉收缩压、TNF-α,升高胰岛素敏感指数;昆丹高、中剂量组可显著降低体重、FFA、瘦素。结论昆丹颗粒具有明显的减重、降糖、调脂、降压作用,其机制可能与降低FFA、TNF-α、Lept的水平,改善胰岛素敏感性有关。     

Herbal medicines for the prevention of alcoholic liver disease: A review

Ethnopharmacological relevance

Long-term excess alcohol exposure leads to alcoholic liver disease (ALD)—a global health problem without effective therapeutic approach. ALD is increasingly considered as a complex and multifaceted pathological process, involving oxidative stress, inflammation and excessive fatty acid synthesis. Over the past decade, herbal medicines have attracted much attention as potential therapeutic agents in the prevention and treatment of ALD, due to their multiple targets and less toxic side effects. Several herbs, such as Cnidium monnieri (L.) Cusson (Apiaceae), Curcuma longa L. (Zingiberaceae) and Pueraria lobata (Willd.) Ohwi (Leguminosae), etc., have been shown to be quite effective and are being widely used in China today for the treatment of ALD when used alone or in combination.

Aim of the review

To review current available knowledge on herbal medicines used to prevent or treat ALD and their underlying mechanisms.

Materials and methods

We used the pre-set searching syntax and inclusion criteria to retrieve available published literature from PUBMED and Web of Science databases, all herbal medicines and their active compounds tested on ALD induced by both acute and chronic alcohol ingestion were included.

Results

A total of 40 experimental studies involving 34 herbal medicines and (or) active compounds were retrieved and reviewed. We found that all reported extracts and individual compounds from herbal medicines/natural plants could be beneficial to ALD, which might be attributed to regulate multiple critical targets involved in the pathways of oxidation, inflammation and lipid metabolism.

Conclusions

Screening chemical candidate from herbal medicine might be a promising approach to drug discovery for the prevention or treatment of ALD. However, further studies remain to be done on the systematic assessment of herbal medicines against ALD and the underlying mechanisms, as well as their quality control studies.