刺果甘草全基因组Survey及叶绿体基因组特征分析＊

目的 基于基因组Survey分析对刺果甘草Glycyrrhiza pallidiflora Maxim.基因组大小和杂合率进行估计，并通过叶绿体基因组序列特征对其在甘草属Glycyrrhiza L.中的系统发育位置进行研究。方法 使用二代测序技术对刺果甘草进行测序，采用K-mer方法对测序reads进行分析，估算刺果甘草基因组大小和杂合率，使用生物信息学方法进行叶绿体基因组组装、注释和系统发育分析。结果 Survey分析结果显示其基因组大小约为577.82 Mb，杂合度约为0.31%，重复序列比例约为53.72%。叶绿体基因组长度为127，267 bp，不具有典型的四分体结构，总GC含量为34.32%，包含110个基因，其中76个蛋白质编码基因，30个tRNA基因和4个rRNA基因。系统发育分析表明，刺果甘草与圆果甘草G. squamulosa Franch.亲缘较接近。结论 刺果甘草存在低杂合和重复序列较多的特点，为了更好地对全基因组进行序列拼接和组装，可尝试采用三代测序结合二代测序的分析策略进行基因组组装；刺果甘草叶绿体全基因组比对和系统发育分析，为后续开展甘草属遗传多样性研究和分子鉴定标记筛选提供了重要依据。     

甘草种子与种苗质量分级标准研究

目的:研究甘草种子、种苗质量分级标准,为甘草药材规范化生产奠定基础。方法:以全国主产区所流通的甘草种子、种苗为样本,通过收集种子的发芽率、净度、含水量、千粒质量,种苗的根长、芦头直径和百株质量等指标信息,采用K-均值聚类分析法、标准差法、平均值法并结合生产实践对甘草种子、种苗进行分级,并开展田间栽培比较试验,对等级划分的合理性进行验证。结果:甘草种子质量等级划分为3个等级,各级甘草种子的千粒质量不低于8 g,含水量不低于10%,一级种子:发芽率≥90%,净度≥90%;二级种子:90%>发芽率≥85%,90%>净度≥85%;三级种子:85%>发芽率≥80%,85%>净度≥80%。甘草种苗质量等级划分为3个等级,一级种苗:根长≥45 cm,芦头直径≥0.8 cm,1.3 kg≤百株质量<0.8 kg;二级种苗:45 cm>根长≥35 cm,0.8 cm>芦头直径≥0.6 cm,0.8 kg≤百株质量<0.4 kg;三级种苗:35 cm>根长≥25 cm,0.6 cm>芦头直径≥0.4 cm,0.4 kg≤百株质量≤0.2 kg。结论:经验证,甘草种子、种苗质量等级划分合理,便于生产实践应用。     

中国产8种甘草的理论分类及亲缘关系的理论判别

目的:根据广义生物遗传与变异信息理论方程推导的2个生物相似度常数P_(g1)=69%,P_(g2)=61%及生物亲缘关系判别函数P_g=1/lnN_d对中国产8种甘草进行理论分类及亲缘关系判别,并提出了生物的理论品种概念。方法:基于8种甘草所含有的59种代表性黄酮类化合物,计算各种甘草之间的共有成分率及变异成分率,采用2个常数及判别函数确定各自的特征序列,根据特征序列进行分类。结果:8种甘草可以分为3类,且8种甘草之间具有紧密的亲缘关系。根据59种特征黄酮类化学成分,可以给出比经典分类结果更精细确定的分类结果,可将严格的理论分类定义为理论品种。结论:广义生物遗传与变异信息理论方程结合植物的多种化学成分,可以对生物进行绝对的理论分类及亲缘关系判别,实现基于数理原理,而不是经验知识的准确科学生物分类,提出了判别生物亲缘关系的理论标准,为确定具有相似功效的中药材提供严谨的科学依据。     

The Flavonoid 7,4′‐Dihydroxyflavone Prevents Dexamethasone Paradoxical Adverse Effect on Eotaxin Production by Human Fibroblasts

Eotaxin/CCL‐11 is a major chemoattractant that contributes to eosinophilic inflammation in asthma. Glucocorticoids inhibit inflammation, but long‐time exposure may cause paradoxical adverse effects by augmenting eotaxin/CCL‐11production. The aim of this study was to determine if 7,4′‐dihydroxyflavone (7,4′‐DHF), the eotaxin/CCL11 inhibitor isolated from Glycyrrhiza uralensis, reduces in vitro eotaxin production induced by long‐time dexamethasone (Dex) exposure, and if so, to elucidate the mechanisms of this inhibition. Human lung fibroblast‐1 cells were used to identify the potency of 7,4′‐DHF compared with other compounds from G. uralensis, to compare 7,4′‐DHF with Dex on eotaxin production following 24‐h short‐time culture and 72‐h longer‐time (LT) culture, and to determine the effects of the 7,4′‐DHF on Dex LT culture augmented eotaxin production and molecule mechanisms. 7,4′‐DHF was the most potent eotaxin/CCL‐11 inhibitor among the ten compounds and provided continued suppression. In contrast to short‐time culture, Dex LT culture increased constitutively, and IL‐4/TNF‐α stimulated eotaxin/CCL11 production by human lung fibroblast‐1 cells. This adverse effect was abrogated by 7,4′‐DHF co‐culture. 7,4′‐DHF significantly inhibited Dex LT culture augmentation of p‐STAT6 and impaired HDAC2 expression. This study demonstrated that 7,4′‐DHF has the ability to consistently suppress eotaxin production and prevent Dex‐paradoxical adverse effects on eotaxin production. Copyright © 2017 John Wiley & Sons, Ltd.     

桔梗、甘草及其组成的复方对口腔病原菌生长影响的体外实验

目的 :探讨桔梗、甘草及其组成的复方中药对常见致龋菌和牙周病原菌生长的影响。方法 :用 φ =65 %的乙醇溶液分别对桔梗、甘草及其组成的复方中药进行初步提取 ,观察 3种粗提物对常见口腔致病菌生长的抑制影响 ,并检测其最小抑菌浓度 (minimalinhibitoryconcentration ,MIC)和最小杀菌浓度 (minimalbactericidalconcentration ,MBC)。结果 :桔梗不影响口腔致病菌的生长 ,甘草仅对致龋菌有抑制作用 ,对S .mutansMT814 8和S .sobrinus 6715的MIC均为 3.91mg/ml;而桔梗和甘草组成的复方明显抑制 4种实验菌株的生长 ,对S .mutansMT814 8和S .sobrinus 6715的MIC为 1.96mg/ml ,对P .gingivalis 381的MIC为 3.91mg/ml,B .forsythus 430 37的MIC为 7.81mg/ml。 结论 :桔梗和甘草组成的复方抑菌和杀菌效果均优于单味药     

Hypokaliémie avec pseudo-hyperaldostéronisme : est-ce un syndrome de Lidl® ?

IntroductionIntoxication induced by glycyrrhizin is a common cause of hypokalaemia by pseudo-hyperaldosteronism.ObservationWe hereby present the observation of a 68-year old patient hospitalised following a full hip-prosthesis operation after a deep hypokalaemia at 2.5mM was observed, with ECG signs (flat T waves and appearance of U waves). The kaliuresis was not adapted at 8,4mmol/mmol of creatininuria. We noted a history of axonal and demyelinising polyneuropathy, of psoriasis and chronic ethylism.The evolution after intravenous potassic supplementation and then per os was favourable leading to a normalisation of the blood and urinary potassic concentrations. The blood concentrations of renin and of aldosterone upon admission were lower than the detection threshold and the tests carried out 7 days later were normal with a plasmatic renin of 35.2 pg/mL and a plasmatic aldosterone of 74 pg/mL, therefore indicating a toxic cause. It is the interview of the patient that allowed for the diagnosis, identifying a daily, prolonged and important consumption (around 1L every 2-3 days for several years) of a pastis produced by supermarket brand Lidl®. The composition of the drink mentions ‘liquorice infusion’ without giving any more information as regards to the real concentration; it was later estimated at 170 mg/L by the distributor.DiscussionThe consumption of glycyrrhizin is a well-known aetiology for pseudo-hyperaldosteronism. It is commonly mentioned amongst excessive consumers of liquorice or of non-alcoholic anise drinks. Drinks that are derived from original pastis contain varying levels of glycyrrhizin, which is used as a flavour enhancer and can become toxic in cases of prolonged and important consumption.     

银苓Ⅰ号对铅中毒小鼠非特异免疫功能的影响

目的:探讨中药复方银苓Ⅰ号对铅中毒小鼠非特异免疫功能的影响。方法:小鼠自由饮用醋酸铅溶液造模,分别予银苓Ⅰ号或依地酸钠钙治疗3周;观察外周血弊细胞计数、碳粒廓清指数K及校正指数α、免疫脏器指数等非特异性免疫功能指标的变化。结果:银苓Ⅰ号对外周血白细胞计数、碳粒廓清指数K及校正指数α、免疫脏器指数有明显改善作用,而且碳粒廓清能力优于依地酸钠钙。结论:银苓号可在一定程度上对抗铅中毒所致的非特异免疫损害,效果优于依地酸钠钙。     

锰胁迫对甘草生理和生长特性的影响

目的通过对甘草进行不同质量浓度锰处理的研究,探讨锰胁迫对甘草植株生理和生长特性的影响。方法以1年生甘草移栽苗为实验材料,采用盆栽蛭石的方法,共设置5个锰质量浓度水平,分别为CK(0),1.81,18.1,36.2和54.3mg·L-1,在测定甘草叶片色素含量、光合指标及各抗氧化酶活性的同时,对甘草生长指标和地上、地下生物量进行测定。结果甘草各项生理和生长指标随着锰处理质量浓度的增加均呈现先升高后下降的趋势,且适当质量浓度的锰处理与CK和54.3mg·L-1处理之间比较差异均显著(P<0.05)。在质量浓度18.1mg·L-1处理时,甘草叶片各色素含量和净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)达到最大值,与54.3mg·L-1处理之间比较差异显著(P<0.05)。超氧化物歧化酶(SOD)和过氧化物酶(POD)活性在质量浓度18.1mg·L-1时达到最大值,与54.3mg·L-1处理之间比较差异显著(P<0.05),而过氧化氢酶(CAT)活性在质量浓度36.2mg·L-1处理时达到最大值,随后三者活性均开始下降。同时,甘草株高、地茎和芦头直径在质量浓度36.2mg·L-1处理,地上和地下部分干质量在18.1mg·L-1处理时达最大值后,均开始下降,且与54.3mg·L-1处理之间比较差异显著(P<0.05)。结论适当质量浓度的锰处理能够显著提高甘草的各项生理和生长指标,但过高质量浓度的锰则会对甘草生长产生胁迫,抑制其光合产物的积累。     

甘草废渣中1个新的香豆素类化合物

目的研究甘草Glycyrrhiza uralensis废渣的化学成分。方法采用硅胶吸附柱色谱、Sephadex LH-20柱色谱、高效液相色谱等分离手段进行分离纯化,根据化合物的理化性质和波谱数据鉴定化合物的结构。结果从新疆产甘草废渣75%乙醇提取物中分离得到了5个化合物,分别鉴定为槐香豆素C(1)、3β-O-对羟基-反-肉桂酰-齐墩果酸(2)、3-羰基甘草次酸(3)、3β,18-O-异丙叉-8(14),15-异右松脂烷二烯(4)、3β,18-O-异丙叉-7,15-异右松脂烷二烯(5)。结论化合物1为新的香豆素类化合物,命名为槐香豆素C;化合物2、4、5为首次从该属植物中分离得到。