Bishonokiol A Induces Multiple Cell Death in Human Breast Cancer MCF-7 Cells

Objective: A dimeric neolignan, bishonokiol A (BHNKA) isolated from Magnolia grandiflora, significantly inhibits the proliferation of human breast cancer cells. However, the exact mechanism of BHNKA induced breast cancer cell death is unknown. In this study, we investigated the pharmacological mechanism underlying BHNKA induced MCF-7 cell death. Methods: Cell viability measurement was performed by the MTT assay. Flow cytometry with PI staining, DAPI staining, and electron microscopy were used to analyze cellular death modes. In addition, western blotting, siRNA transfection, ATP assay, and fluorescence microscopy were used to determine the mechanism of BHNKA induced MCF-7 cell death. Results: BHNKA induced cell death by apoptosis, necroptosis and autophagy at the same concentration and time in MCF-7 cells, and electron microscopy confirmed these results. The mechanism of BHNKA triggered apoptosis and autophagy in MCF-7 cells was primarily due to an increase in the Bax/Bcl-2 ratio and simultaneous up-regulation of LC3-II protein expression, respectively. BHNKA induced necroptosis by activation of the RIP1-RIP3-MLKL necroptosis cascade, up-regulation of cyclophilin D (CypD) protein expression to stimulate ROS generation. We further demonstrated that siRNA-mediated down-regulation of CypD protected against BHNKA induced cell death. Conclusions: These results suggest that BHNKA may be a potential lead compound for development as an anti-breast cancer agent for induction of multiple cell death pathways.     

药用植物金荞麦中黄酮类化合物积累的时空变化规律

目的 探究药用植物金荞麦不同生育期各器官中黄酮类化合物积累的时空分布规律,为金荞麦合理采收、资源化利用和高价值开发提供科学依据。方法 以根茎繁殖不同生长年限的金荞麦为研究对象,在不同生育时期采集根、茎、叶、花器官,采用高效液相色谱法测定各器官样品中儿茶素、表儿茶素、芦丁和总黄酮含量。结果 随着金荞麦生育期推进,植株不同器官的黄酮类化合物积累增加,以秋季地上部分枯萎后的含量最高;不同生育时期叶片中的儿茶素、表儿茶素、芦丁和总黄酮含量均显著高于根茎和茎秆（P<0.05）。不同器官间儿茶素、表儿茶素、芦丁和总黄酮含量差异有统计学意义（P<0.05）,花中儿茶素、芦丁和总黄酮含量最高;表儿茶素含量在茎秆中最低。黄酮类物质含量在根茎不同部位差异有统计学意义（P<0.05）,表儿茶素和总黄酮含量在根茎不同组织部位表现为须根>韧皮部>根茎（不含须根）>木质部（P<0.05）,儿茶素和芦丁分别在须根和韧皮部含量最高。1年生与2年生金荞麦植株不同器官的黄酮类化合物含量差异无统计学意义。结论 在不考虑产量的前提下,根茎繁殖金荞麦种植1年即可采收,适宜采收期为11月上旬。     

Dark Tea: A popular beverage with possible medicinal application

Tea is a famous beverage that is produced from leaves of Camellia sinensis. Amongst the six major tea categories in China, dark tea is the only one that involves microbial fermentation in the manufacturing process, which contributes unique flavors and functions for the tea. In the recent decade, the reports about the biofunctions of dark teas have increased rapidly. Therefore it may be the proper time to consider dark tea as one potential homology of medicine and food. In this viewpoint, our current understanding of the chemical constituents, biological activities and possible health beneficial effects of dark teas were introduced. Some future directions and challenges to the development perspectives of dark teas were also discussed.     

八大品种菊花中不同成分的含量比较

目的 研究八大品种菊花中不同成分的含量。方法 以绿原酸、木犀草素-7-O-β-D-葡萄糖苷及芹菜素-7-O-β-D-葡萄糖苷为指标，采用高效液相色谱法对不同品种的菊花进行了含量测定。结果 绿原酸含量以济菊和祁菊最高，川菊最低；木犀草素-7-O-β-D-葡萄糖苷含量以济菊最高，而毫菊最低；芹菜素-7-O-β-D-葡萄糖苷含量以济菊最高，而最低为贡菊、毫菊和怀菊。结论 绿原酸和黄酮类成分均高者才是优良品种。     

Spray freeze drying for dry powder inhalation of nanoparticles

Formulating nanoparticles for delivery to the deep lung is complex and many techniques fail in terms of nanoparticle stability. Spray freeze drying (SFD) is suggested here for the production of inhalable nanocomposite microcarriers (NCM). Different nanostructures were prepared and characterized including polymeric and lipid nanoparticles. Nanoparticle suspensions were co-sprayed with a suitable cryoprotectant into a cooled, stainless steel spray tower, followed by freeze drying to form a dry powder while equivalent compositions were spray dried (SD) as controls. SFD-NCM possess larger specific surface areas (67–77 m²/g) and lower densities (0.02 g/cm³) than their corresponding SD-NCM. With the exception of NCM of lipid based nanocarriers, SFD produced NCM with a mass median aerodynamic diameter (MMAD) of 3.0 ± 0.5 μm and fine particle fraction (FPF ⩽ 5.2 μm) of 45 ± 1.6% with aerodynamic performances similar to SD-NCM. However, SFD was superior to SD in terms of maintaining the particle size of all the investigated polymeric and lipid nanocarriers following reconstitution (S_f/S_i ratio for SFD ≈ 1 versus >1.5 for SD). The SFD into cooled air proved to be an efficient technique to prepare NCM for pulmonary delivery while maintaining the stability of the nanoparticles.     

Ginsenoside Rg1 ameliorates blood–brain barrier disruption and traumatic brain injury via attenuating macrophages derived exosomes miR-21 release

During the traumatic brain injury (TBI), improved expression of circulatory miR-21 serves as a diagnostic feature. Low levels of exosome-miR-21 in the brain can effectively improve neuroinflammation and blood–brain barrier (BBB) permeability, reduce nerve apoptosis, restore neural function and ameliorate TBI. We evaluated the role of macrophage derived exosomes-miR-21 (M-Exos-miR-21) in disrupting BBB, deteriorating TBI, and Rg1 interventions. IL-1β-induced macrophages (IIM)-Exos-miR-21 can activate NF-κB signaling pathway and induce the expressions of MMP-1, -3 and -9 and downregulate the levels of tight junction proteins (TJPs) deteriorating the BBB. Rg1 reduced miR-21-5p content in IIM-Exos (RIIM-Exos). The interaction of NMIIA–HSP90 controlled the release of Exos-miR-21, this interaction was restricted by Rg1. Rg1 could inhibit the Exos-miR-21 release in peripheral blood flow to brain, enhancing TIMP3 protein expression, MMPs proteolysis, and restricting TJPs degradation thus protected the BBB integrity. Conclusively, Rg1 can improve the cerebrovascular endothelial injury and hold the therapeutic potential against TBI disease.     

基于网络药理学研究丹蛭降糖胶囊治疗糖尿病的分子机制

目的 采用网络药理学方法研究丹蛭降糖胶囊治疗糖尿病的潜在分子机制。方法 借助中药系统药理学平台和Swiss Target Prediction数据库检索丹蛭降糖胶囊的化学成分、对应作用靶标及靶标基因。通过GeneCards获取糖尿病相关靶点,将复方活性成分靶点和糖尿病靶点构建韦恩图,以寻找交集靶点,获得丹蛭降糖胶囊治疗糖尿病的预测靶标。利用Cytoscape 3.7.2软件构建药材-化合物-靶点（基因）网络,筛选关键化合物。利用STRING网站建立交集靶点蛋白相互作用网络,选出关键靶点基因。借助R软件对共同靶点进行GO分析和KEGG通路富集分析。结果 丹蛭降糖胶囊作用于糖尿病的活性成分有25个,相关靶基因76个。GO和KEGG分析结果显示,丹蛭降糖胶囊治疗糖尿病的机制共涉及1 702条生物学过程和149条信号通路。结论 丹蛭降糖胶囊治疗糖尿病的关键成分槲皮素、木犀草素、山奈酚等可能通过对AGE-RAGE、IL-17、TNF、HIF-1等信号通路的调控,最终达到治疗糖尿病的效果。其作用机制可能与抗炎,降低氧化应激水平,参与细胞的凋亡与损伤有关。     

密蒙花中4种成分的含量测定及其主成分、偏最小二乘判别分析

目的 建立同时测定密蒙花药材中4种成分含量的方法,并进行主成分分析（principal component analysis,PCA）和偏最小二乘判别分析（partial least squares discriminant analysis,PLS-DA）。方法 采用超高效液相色谱法,色谱柱为Waters ACQUITY UPLC BEH C18,流动相为乙腈-0.1％磷酸溶液（梯度洗脱）,流速为0.15 mL/min,检测波长为330 nm,柱温为30 ℃,进样量为2 μL。采用SIMCA 14.1统计软件对含量测定结果进行PCA和PLS-DA研究。结果 4种指标性成分的线性、分离度、稳定性、重复性与精密度良好;24批药材样品有2个主成分,PLS-DA分析确定了2个分类标志物。结论 本研究建立的方法的精密度、稳定性、重复性均较好,可用于密蒙花药材中4种成分含量的同时测定;不同产地的密蒙花药材质量差异较大。     

不同产地甘草水提液和醇提液指纹图谱研究

目的建立不同产地甘草水提液和醇提液的HPLC指纹图谱,以期为不同产地甘草品控及甘草提取加工方法提供借鉴。方法采用HPLC法建立10批不同产地甘草药材的水提液和醇提液的指纹图谱,并对所获得的指纹图谱数据进行相似度评价和层次聚类分析。结果建立了甘草水提液和醇提液的指纹图谱,其中水提液指纹图谱确定了16个共有峰,并指认出6种成分,10批甘草的相似度在0.030～0.999,醇提液指纹图谱确定了60个共有峰,并指认出7种成分,10批甘草的相似度均大于0.85。层次聚类分析结果显示能够很好地将不同地区的甘草水提液和醇提液予以分类,提示产地气候环境、土壤条件等自然因素对甘草的内在品质产生较为显著的影响。结论建立了稳定可靠的甘草水提液和醇提液HPLC指纹图谱评价方法,为甘草质量标准研究及提取加工工艺的优化提供有力的理论依据和指导,为临床用药方法的选择提供科学依据。     

黄蒲通窍胶囊对阿尔茨海默病模型大鼠Wnt/Ca2+信号通路的影响

目的 观察黄蒲通窍胶囊对阿尔茨海默病（Alzheimers disease,AD）模型大鼠学习记忆的影响,探究其潜在作用机制。方法 将30只SD大鼠随机分为空白组、模型组、黄蒲通窍胶囊组,每组10只,腹腔注射D-半乳糖联合双侧海马注射Aβ25-35复制AD大鼠模型;给药28 d,采用Morris水迷宫实验检测大鼠的学习记忆能力,检测后取材,采用苏木精-伊红染色观察海马神经细胞损伤情况,采用Western blot法检测海马组织中Wnt5、Frizzled、PLCβ蛋白表达水平,采用ELISA法检测血清三磷酸肌醇（inositol triphosphate,IP3）表达水平。结果 Morris水迷宫实验结果显示：训练期,各组大鼠的逃避潜伏期随时间的增加呈下降趋势;测试期,与空白组比较,模型组大鼠逃避潜伏期明显延长,目标象限路程与时间百分比明显减少（P＜0.05）;与模型组比较,黄蒲通窍胶囊组大鼠逃避潜伏期明显缩短,目标象限路程百分比和时间百分比明显增加（P＜0.05）。苏木精-伊红染色结果显示：与空白组比较,模型组大鼠海马神经细胞萎缩破裂、层数减少、排列松散;与模型组比较,黄蒲通窍胶囊组大鼠海马神经细胞损伤减轻。Western blot法检测结果显示：与空白组比较,模型大鼠Wnt5、Frizzled、PLCβ表达水平均明显升高（P＜0.05）;与模型组比较,黄蒲通窍胶囊组大鼠Wnt5、Frizzled、PLCβ表达水平均明显降低（P＜0.05）。ELISA法检测结果显示：与空白组比较,模型组大鼠IP3表达水平明显升高（P＜0.05）;与模型组比较,黄蒲通窍胶囊组IP3表达水平明显降低（P＜0.05）。结论 黄蒲通窍胶囊可减少AD模型大鼠海马神经细胞的损伤,改善其学习记忆的能力,其机制可能与抑制Wnt/Ca2+信号通路有关。