Study on Scavenging and Disconnecting Effects of Nitrosation Reaction by Extracts of Tetradium Ruticarpum and Glycyrrhizae

In order to investigate and compare the effects of Tetradium ruticarpum and Glycyrrhizae extracts in terms of scavenging sodium nitrite and inhibiting N-ni- troso compounds formation, the 1_9（3^3） orthogonal test was conducted to extract the active components. In addition, the capacities of scavenging sodium nitrite and of in- hibiting the N-nitroso compounds formation were measured to confirm the optimum condition of extracting. When the scavenging rate was considered as the main index, the maximum capacities of scavenging sodium nitrite by the extracts of Tetradi- urn ruticarpum and Glycyrrhizae were 0.836 5 and 0.558 0, respectively. Further- more, when the disconnection rate was considered as the main index, the maximum capacities of inhibiting the N-nitroso compounds formation by the extracts of Tetradi- urn ruticarpum and G/ycyrrhizae were 0.922 8 and 0.625 6, respectively. As a re- suit, the capacity of inhibiting nitrosation reaction by Tetradium ruticarpum was shown to be much stronger than G/ycyrrhizae, no matter the scavenging rate or the disconnection rate as the main index to be considered.

Intellectual Property Protection and High Quality Development of Radix Glycyrrhizae for Treatment of COVID-19

Radix Glycyrrhizae is the dominant native medicinal material variety in the north and northwestern medicinal materials producing areas.It is a main Chinese medicine of effective TCM drugs and formulas for the treatment of new coronavirus disease(COVID-19).This paper introduces the medicinal value of Radix Glycyrrhizae,involving the labeling,cultural heritage,and creative intellectual property rights of the Chinese medicinal materials,analyzes the poverty-stricken areas that are ecologically suitable for Radix Glycyrrhizae cultivation,and the superior counties and production bases of the Chinese medicinal materials.Besides,mainly from the aspects of perfect intellectual property rights,the establishment of authentic medicinal material production bases,and the construction of quality control systems,etc.,it discusses the rural revitalization strategy and the development strategy of traditional Chinese medicine(TCM),agricultural intellectual property protection and high-quality development strategy of Radix Glycyrrhizae.

A systematic summary of natural compounds in Radix Glycyrrhizae

Objective： To accumulate data from studies on the compounds identified from Gancao （Radix Glycyrrhizae, GC）, andthen systematically summarize and classify these compounds according to their structural characteristics. Methods： Fivedatabases （CNKI, VIP, Wanfang data, CBM, and Pubmed） were used to search for studies on the chemical structure ofcompounds from GC. The retrieval time of the respective databases was from their inception to March 2016. Accordingto the inclusion and exclusion criteria, the papers were carefully screened, and the data were extracted. Results： A totalof 653 compounds from GC were collated from 252 articles, including flavonols （n = 201） belonging to 10 subgroups,terpenes and saponins （n = 167）, coumarins （n = 30） belonging to 3 subgroups, aliphatics （n = 206）, aromatics （n = 35）,and others （n = 14）. The flavones, represented by liquiritin and liquiritigenin, were the most reported compounds isolatedfrom GC, followed by terpenes and saponins such as glycyrrhizic acid and glycyrrhetinic acid. Conclusion： The morethan 600 natural compounds in GC may be responsible for GC’s anti-inflammatory, phlegm-expelling, cough- andpain-relieving activities.

Simultaneous Determination of Saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by High Performance Liquid Chromatography

To establish a method for determining five saponins(notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ammonium glycyrrhizinate) in Glycyrrhizae, Notoginseng and Ginseng, the high performance liquid chromatography with diode array detector(HPLC-DAD) method was applied to an Inertsil ODS-SP column(4.6 mm×250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.05% phosphoric acid in a gradient elution manner. The flow rate was 1.0 mL/min. The column temperature was 30 ℃ and the detection wavelengths were 203 nm and 237 nm, respectively. The linear ranges were 0.700,0—7.000,0 μg for R1(r=1.000,0), 0.751,1— 7.511,4 μg for Rg1(r=1.000,0), 0.677,2—6.771,6 μg for Re(r=1.000,0), 0.733,9—7.339,1 μg for Rb1(r= 1.000,0), and 0.540,0—5.399,8 μg for ammonium glycyrrhizinate(r=0.999,9), respectively. In addition, their average recoveries were 100.28%, 105.83%, 104.09%, 99.36% and 98.54%, respectively. The relative standard deviations(RSDs) of precision, reproducibility and recovery were all less than 1.5%. The results indicate that the method is simple, accurate and reproducible so that it can be used for the simultaneous determination of the five saponins in Chinese patent medicines containing the three kinds of herbs.

Identification of potential anti-pneumonia pharmacological components of Glycyrrhizae Radix et Rhizoma after the treatment with Gan An He Ji oral liquid

Glycyrrhizae Radix et Rhizoma,a traditional Chinese medicine also known as Gan Cao(GC),is frequently included in clinical prescriptions for the treatment of pneumonia.However,the pharmacological components of GC for pneumonia treatment are rarely explored.Gan An He Ji oral liquid(GAHJ)has a simple composition and contains GC liquid extracts and paregoric,and has been used clinically for many years.Therefore,GAHJ was selected as a compound preparation for the study of GC in the treatment of pneumonia.We conducted an in vivo study of patients with pneumonia undergoing GAHJ treatments for three days.Using the intelligent mass spectrometry data-processing technologies to analyze the metabolism of GC in vivo,we obtained 168 related components of GC in humans,consisting of 24 prototype components and 144 metabolites,with 135 compounds screened in plasma and 82 in urine.After analysis of the metabolic transformation relationship and relative exposure,six components(liquiritin,liquiritigenin,glycyrrhizin,glycyrrhetinic acid,daidzin,and formononetin)were selected as potential effective components.The experimental results based on two animal pneumonia models and the inflammatory cell model showed that the mixture of these six components was effective in the treatment of pneumonia and lung injury and could effectively downregulate the level of inducible nitric oxide synthase(iNOS).Interestingly,glycyrrhetinic acid exhibited the strongest inhibition on iNOS and the highest exposure in vivo.The following molecular dynamic simulations indicated a strong bond between glycyrrhetinic acid and iNOS.Thus,the current study provides a pharmaceutical basis for GC and reveals the possible corresponding mechanisms in pneumonia treatment.

Screening for cyclooxygenase 2 inhibitors from natural compounds of Radix Glycyrrhizae using computer simulation

Objective： To explore the molecular basis of the effects of Gancao （Radix Glycyrrhizae, GC） on inflammation throughthe inhibition of cyclooxygenase 2 （COX-2）. Methods： The Discovery Studio 4.5 System was used to predict thephysicochemical properties of GC molecular compounds. The Ligand Profiler was used to screen for natural GCcomponents that could combine with the COX-2 pharmacophores. The AutoDock Vina 1.1.2 software was used for themolecular docking of the natural GC components with the COX-2 protein. Results： The aromatics were the closest to thenon-steroidal anti-inflammatory drugs in terms of the three properties, namely molecular weight, molecular surface area,and molecular solubility, followed by the flavonoids; whereas the terpenoids/saponins differed most from thenon-steroidal anti-inflammatory drugs in terms of the three properties; and the aliphatics were inconsistent. One hundredand eighteen small molecules were obtained through the pharmacophore screening using GC. The molecular bindingenergy （MBE） results demonstrated that the MBE value of the flavonoids/aromatics, obtained from their binding with theCOX-2 protein, was lower than that obtained from their binding with the substrate, metabolism of arachidonic acid,whereas the MBE value of the aliphatics/terpenoids, obtained from their binding with the COX-2 protein, was higherthan that obtained from their binding with the substrate, arachidonic acid. Finally, further filtration, based on thephysicochemical properties and the molecular binding energies of the small molecules, was carried out. Forty-twonatural GC components, including 35 flavonoid and 7 aromatic constituents, with low binding energies and potentialinhibitory effects on COX-2, were screened. Conclusion： Using the three-step program, pharmacophore screening,molecular docking, and physicochemical properties analysis, we screened out 35 flavonoid molecules and 7 aromaticmolecules, which may be potential COX-2 inhibitors, from GC. Two of the 35 flavonoid molecules （licochalcone A andglabridin） have been confirmed in the laboratory to have inhibitory effects on COX-2. Our findings provide a materialbasis for the development of non-steroidal GC drugs.

Simultaneous determination of five major components of Glycyrrhizae Radix et Rhizoma in Xiaoer Zhike Tangjiang with standardized reference extract

In this study, we developed a novel and simple HPLC-DAD method for simultaneous qualitative and quantitative determination of five major components of Glycyrrhizae Radix et Rhizoma （GRR） in Xiaoer Zhike Tangjiang （XEZKTJ） with standardized reference extract （SRE）. The five analytes （liquiritin apioside, liquiritin, isoliquiritin apioside, liquirigenin and glycyrrhizic acid） were well separated with good linearity, precision, stability and repeatability. The recovery rates ranged from 95.69% to 100.80%. The content of the five compounds in 34 batches of commercial XEZKTJ products was determined using standardized GRR extract （SRE method） and individual chemical reference standards （CRS method）. Highly similar results were obtained from the two methods, demonstrating the feasibility of the proposed SRE method. Taken together, we proposed an efficient and low-cost way to perform multi-component quality control of XEZKTJ in this study.

Variations in the Composition of Fructus Evodiae after Processing with Radix Glycyrrhizae Extract

Objective： To evaluate the changes in the concentrations of five components in Fructus Evodiae used in Chinese medicine, including evodiamine and glycyrrhizic acid, during processing of Fructus Evodiae with Radix Glycyrrhizae extract by using high performance liquid chromatography （HPLC） and to provide a scientific basis for different clinical uses of processed and unprocessed Fructus Evodiae. Methods： The concentrations of the Fructus Evodiae components in processed or unprocessed Fructus Evodiae were evaluated by HPLC using a YMC J＇sphere ODS-H80 column （4.6 mm × 250 ram, 5 μm） with acetonitrile-water-tetrehydrofuran-acetic acid （41：59：1：0.2, v/v/v/v） as the mobile phase. The detection wavelength was 225 nm, the column temperature was 35 %, the flow rate was 1.0 mL/min, and the injection volume was 10 μL. The concentrations of the Radix Glycyrrhizae components were determined by HPLC with a KromasiI-C18 column （4.6 mm × 250 mm, 4 μm） and a gradient elution of acetonitrile （A） and 0.05% aqueous phosphoric acid （B） as the mobile phase. The detection wavelength was 237 nm, the column temperature was 35 ℃, the flow rate was 1.0 mlEmin, and the injection volume was 10μL. Results： The calibration curves of evodia lactone, evodiamine, rutaecarpine, liquiritin, and glycyrrhizin showed good linear relationships （r〉0.99）. The recoveries of evodia lactone, evodiamine, rutaecarpine, liquiritin, and glycyrrhizin were 96.59%, 104.18%, 101.91%, 97.75%, and 97.95%, respectively. The concentrations of the components in processed Fructus Evodiae were obviously different to those in unprocessed Fructus Evodiae. Conclusions： The developed method is rapid and accurate. The results provide a reference for processed Fructus Evodiae and the changes that could be expected in its effects compared to unprocessed Fructus Evodiae.

HPLC法测定不同产地甘草中甘草苷的含量

[目的]建立甘草中甘草苷含量测定的HPLC法,测定不同产地甘草中甘草苷的含量,以控制其质量。[方法]HPLC色谱条件为：分析柱为Kromasil C18柱（4.6 mm×25 cm,5μm）;流动相为乙腈-0.5%醋酸溶液（V/,V 1∶4）;流速为1.0 ml/min;柱温为25℃;紫外检测波长为270 nm;采用外标法定量测定。[结果]甘草苷在0.1~1.2μg范围内呈良好的线形关系（r=0.999 9）,其平均加样回收率和RSD值分别为97.86%和2.04%（n=6）。[结论]该方法操作简单、准确、快速、重现性好,可作为甘草苷的定量分析方法。

甘草种植产业发展现状及对策研究

在分析甘草种植产业发展现状、存在的问题的基础上,提出了以提高药农的经济效益、满足国内外市场对甘草及甘草制品需求为目标的的甘草种植产业发展对策,为甘草的可持续发展提供依据。

大孔吸附树脂纯化甘草黄酮类化合物的研究

[目的]探讨大孔吸附树脂对甘草(RADIXRHIZOMAGLYCYRRHIZAE)黄酮的纯化条件。[方法]采用5种大孔吸附树脂ADS-7、D101、HPD-300、HPD-910和AB-8吸附甘草黄酮类化合物,并考察了样品浓度,洗脱剂乙醇浓度,洗脱剂乙醇体积等影响吸附性能的因素。[结果]ADS-7型大孔吸附树脂对甘草黄酮类化合物有较大的吸附率和洗脱率,其用于纯化甘草黄酮的最佳上样浓度是1.2 mg/ml,洗脱剂为5BV体积分数为0.70的乙醇,纯化后的黄酮粉黄酮含量47.1%,精制率211.2%。[结论]ADS-7树脂对黄酮类化合物有较好的吸附纯化作用,可为甘草黄酮的纯化提供参考依据。

ICP-MS法测定中国7个主产地的甘草中重金属的含量

[目的]利用ICP-MS法测定7个主产地的甘草中Ni、Cu、Zn、As、Cd、Hg、Pb和Bi共8种重金属的含量,为甘草中重金属的测定提供可靠方法,从而确保甘草入药安全。[方法]在确定ICP-MS工作曲线的基础上,测定7个主产地的甘草中8种重金属的含量,并将测定结果与2010版中国药典一部中重金属标准进行对比。[结果]甘草中符合中国药典2010版(1部)重金属标准的占42.86%,其中As、Hg、Pb 3种重金属是造成甘草中重金属超标的主要因素。[结论]甘草中重金属的含量与土壤、肥料及水分中重金属的含量相关,相关质检部门应深入调查甘草的产地,从而确定甘草重金属超标原因。

Effect of Glycyrrhiza Polysacchride on Growth Performance and Immunity Function of Mice

Aniso-dose glycyrrhiza polysaccharide was injected into abdominal cavity of mice. The effect of glycyrrhiza polysaccharide on growth performance and immune ruction of mice was determined. The results showed that the growth performance and immune function of mice were improved and there were significant differences among the treatment groups and control group.

甘草粗提物对鲫的抗应激及免疫保护功效研究

采用腹腔注射和混饲投喂两种方式研究中药甘草(Glycyrrhiza uralensis Fiseh)粗提物对鲫(Carassius auratus)的抗应激和免疫保护功效。将一部分粗提物按0.5%和2%的质量分数制成药饵投喂实验鲫,分别在第14 d、28 d、35 d、42 d、56 d采集血样;将另一部分粗提物用无菌生理盐水按0.5%和2%的浓度配成针剂腹腔注射鲫,分别在第3 d、7 d、14 d、21 d、28 d采集血样。结果显示:投喂实验中药物组和对照组血清皮质醇水平均呈先升后降趋势,除第14 d外,药物组皮质醇水平都低于同期对照组。在最后一次采样中(56 d),对照组、低剂量组和高剂量组的皮质醇水平分别为147.85 ng/mL、79.41 ng/mL和66.47 ng/mL。腹注实验中对照组皮质醇水平一直呈上升趋势,最后一次采样(28 d)高达274.23 ng/mL,显著高于药物组(P<0.01)。投喂甘草提取物56 d后,用Aeromonas hydrophila对鲫进行人工感染,低剂量组和高剂量组一周后的死亡率分别为45.0%和55.0%,显著低于对照组的70.0%(P<0.05)。腹注甘草提取物后第28 d,鲫受A.hydrophila菌人工感染,对照组、低剂量组和高剂量组的死亡率分别为60.0%、55.0%和35.0%,高剂量组显著低于对照组(P<0.05)。实验表明,投喂和腹注甘草粗提物后,可明显提高鲫的抗应激能力,并能增强鱼体对A.hydrophila的抵抗力。

盐碱条件下接种丛枝菌根真菌对甘草种苗生长的影响

[目的]研究接种丛枝菌根真菌对甘草生长和矿质元素吸收的影响。[方法]测定丛枝菌根侵染率和幼苗干重及N、P、K和Na的含量变化,研究菌根真菌与对甘草生长的作用。[结果]接种丛枝菌根真菌提高了甘草的生物量,提高了对N、P和K的吸收,但降低对Na的吸收。[结论]接种菌根真菌,增强了甘草的生长和抗盐碱能力,其机制可能与对元素的选择性吸收有关。

α-Glucosidase Inhibitors from Glycyrrhiza uralensis Fisch.

Aim To screen for α-glucosidase inhibitor from Glyeyrrhiza uralensis Fisch.. Methods Glycyrrhizic acid, glycyrrhetinic acid, flavonoids of glycyrrhiza, alkaloids of glycyrrhiza, and glycyrrhiza polysaccharides were isolated from the root of Glycyrrhiza uralensis Fisch. respectively. Three compounds were isolated from the flavonoids of glycyrrhiza as guided by the α-glucosidase inhibitory test in vitro. Moreover, the characteristics of inhibitory kinetics of glycyrol and glycyrrhetinic acid were investi- gated. Results The flavonoids of glycyrrhiza and glycyrrhetinic acid had the strongest α-glucosidase inhibitory activity. Glycyrol,β-sitosterol and liquifitin were isolated and identified. Glycyrol was a fast- binding, reversible, noncompetitive α-glucosidase inhibitor, showing IC50 at 0.26 μg·mL^-1 Glycyrrhetinic acid was a fast-binding, irreversible α-glucosidase inhibitor, showing IC50 at 102.4 μg·mL^-1. Conclusion Glycyrol is an effective α-glucosidase inhibitor.

Evidence of Hydropassive Movement in Stomatal Oscillations of Glycyrrhiza inflata under Desert Conditions

Stomatal conductance was found to change from steady-state to a slate of oscillations during daytime when vapour pressure deficit (VPD) increased to a value of 1 kPa in Glycyrrhiza inflata Batalin grown under the conditions of arid desert in north-west China. The injected metabolic inhibitors (NaN3 or carbonyl cyanide-m-chlorophenyl-hydrazone (CCCP)) slightly reduced the stomatal conductance but did not significantly decrease the intensity of stomatal oscillations (amplitude/average). The oscillation intensity was found to he significantly correlated with VPD and root resistance, but not with the respiration rate. There might exist a minimum threshold of VPD (0.8 kPa) and root resistance (1/4 relative value) that induced stomatal oscillations. These results suggested that stomatal oscillations induced by atmospheric drought stress and root resistance were mainly a type of hydropassive movement.

Effect of Different Concentrations of Mo on Yield and Accumulation of Active Constituents of Glycyrrhiza uralensis Fisch

[Objective] The aim was to explore the effects of different concentrations of molybdenum（Mo）on the growth,yield and the accumulations of active constituents of Glycyrrhiza uralensis.[Method] The seedlings of one-year-old G.uralensis were subjected to four concentrations of Mo（0,0.52,5.20 and 10.40 mg/L Mo）culturing in vermiculite and 0.52 mg/L Mo was the normal concentration in the complete Hoagland nutrition solution.The solution was irrigated to the pots every week.The fresh and dry weights of shoots and roots of the plants were measured by electronic balance.The contents of glycyrrhizic acid and liquiritin of roots were determined by HPLC.[Result] This result showed that both fresh and dry weights of the shoots and roots increased with the increasing of Mo concentrations.During the later period of this experiment,they showed significant differences among different Mo treatments.On the 105 d,both fresh and dry weights of the shoots and roots under 0 mg/L Mo reached the lowest,and the 5.20 and 10.40 mg/L Mo reached the highest.However,the contents of glycyrrhizic acid and liquiritin increased first and then decreased with increasing of Mo concentrations.During the whole experiment,the contents of glycyrrhizic acid and liquiritin under 5.20 mg/L Mo were always higher than that under other Mo treatments.[Conclusion] In conclusion,both the concentrations of 5.20 and 10.40 mg/L Mo could promote the accumulation of the shoots and roots of G.uralensis,but the 5.20 mg/L Mo was more useful for the accumulation of glycyrrhizic acid and liquiritin in the roots.

盐胁迫对胀果甘草种子萌发和幼苗生理特性的影响

[目的]研究盐胁迫对胀果甘草种子萌发和幼苗生理特性的影响。[方法]以胀果甘草种子为材料,分别采用浓度为0、50、100、200和300 mmol/L的NaCl和Na2SO4以及浓度分别为0、25、50、75和100 mmol/L的NaHCO3进行处理,研究盐胁迫对胀果甘草种子萌发和幼苗生理特性的影响。[结果]3种盐胁迫对胀果甘草的种子萌发率、萌发指数和活力指数的影响差异明显,对胀果甘草种子萌发影响的强弱顺序为:NaHCO3>Na2SO4>NaCl。在3种盐胁迫下,幼苗子叶、胚根脯氨酸(Pro)、丙二醛(MDA)和可溶性蛋白含量随盐浓度的升高而升高;幼苗子叶SOD活性随盐浓度的升高一直处于下降趋势;胚根SOD活性随盐浓度升高先上升后下降。[结论]3种盐胁迫对胀果甘草种子萌发和幼苗生理特性均有明显的影响。

超临界流体萃取甘草中角鲨烯的方法研究

采用超临界流体萃取技术提取甘草(Glycyrrhiza uralensis)中的角鲨烯,并用高效液相色谱法测定其含量。结果显示,在丙酮为夹带剂的萃取产物中角鲨烯含量高达0.47%,超临界流体萃取法对甘草角鲨烯的总提取率为0.018%,高于传统索氏提取法及超声辅助提取法所得提取率。