Objective: To prepare astragaloside IV dripping pills (ASDP) and assess their therapeutic effects on mice with doxorubicin hydrochloride-induced dilated cardiomyopathy (DCM). Significance: Astragaloside IV (AS) exhibits pharmacological effects in treating cardiovascular diseases, however, its clinical application is hindered by poor solubility and low bioavailability. The study sheds light on new therapeutic strategy of DCM and development of AS formulations. Methods: The ASDP prepared by solid dispersion technology were optimized and characterized through scanning electron microscopy (SEM), X-ray diffraction (XRD), differential scanning calorimetry (DSC), as well as evaluations of appearance, average weight, hardness, disintegration time, drug content, solubility and dissolution behavior. The therapeutic effects of ASDP on mice with doxorubicin hydrochloride-induced DCM were performed via echocardiography, heart weight index measurements, pathological examination of heart tissues, and determination of serum levels of angiotensin II (Ang II), B-type natriuretic peptide (BNP), and suppressor of tumorigenicity 2 (ST2). Results: ASDP presented as round, white pills with an average weight of 27.61 mg, a short disintegration time (approximately 3 min), a hardness of 4.9 ± 0.2 N and drug content of 64.5 ± 0.12mg/g. Compared to AS, ASDP significantly improved solubility and dissolution rate. In the doxorubicin hydrochloride-induced DCM mouse model, ASDP alleviated cardiac dysfunction and hypertrophy, reduced necrosis, and decreased serum levels of Ang II, BNP and ST2. Conclusion: ASDP, which enhance the solubility and dissolution of AS, demonstrate significant therapeutic efficacy against DCM, suggesting their potential as a promising candidate for DCM treatment.

Licorice is a frequently applied herb with potential edible and medicinal value based on various flavonoids and triterpenes. However, studies on detailed flavonoid and triterpene metabolism and the molecular basis of their biosynthesis in licorice are very limited, especially under drought conditions. In the present study, we carried out transcriptome, proteome, and metabolome experiments. To ultimately combine three omics for analysis, we performed a bioinformatics comparison, integrating transcriptome data and proteome data through a Cloud platform, along with a simplified biosynthesis of primary flavonoids and triterpenoids in the KEGG pathway based on metabolomic results. The biosynthesis pathways of triterpenes and flavonoids are enriched at both gene and protein levels. Key flavonoid-related genes (PAL, 4CL, CHS, CHI, CYP93C, HIDH, HI4OMT, and CYP81E1_7) and representative proteins (HIDH, CYP81E1_7, CYP93C, and VR) were obtained, which all showed high levels after drought treatment. Notably, one R2R3-MYB transcription factor (Glyur000237s00014382.1), a critical regulator of flavonoid biosynthesis, achieved a significant upregulated expression as well. In the biosynthesis of glycyrrhizin, both gene and protein levels of bAS and CYP88D6 have been found with upregulated expression under drought conditions. Most of the differentially expressed genes (DEGs) and proteins (DEPs) showed similar expression patterns and positively related to metabolic profiles of flavonoid and saponin. We believe that suitable drought stress may contribute to the accumulation of bioactive constituents in licorice, and our research provides an insight into the genetic study and quality breeding in this plant.

Methylophiopogonanone B (MOB), one of the homoisoflavonoids isolated from Ophiopogon japonicus, has been demonstrated to possess antioxidative and antitumor activities. The aim of this work was to investigate the metabolism of MOB using liver microsomes and hepatocytes. MOB was individually incubated with rat, monkey, and human hepatocytes to generate the metabolites. To investigate the bioactivation pathways, MOB was incubated with liver microsomes in the presence of glutathione (GSH). All the metabolites were detected and identified using LC with a quadrupole Orbitrap mass spectrometer. Under the current conditions, nine metabolites were identified in hepatocyte incubations. Of these metabolites, M7 derived from hydroxylation was identified as the most abundant metabolite in hepatocyte incubation. MOB was metabolized via demethylation, hydroxylation, and glucuronidation. In liver microsomes, five GSH conjugates were detected and identified. MOB was subjected to bioactivation through demethylation yielding M9, which further formed quinone-methide and ortho-quinone intermediates, followed by GSH conjugation. This work is the first to study the metabolism of MOB, which will help us understand its disposition and efficacy.

目的 研究芡茎多糖单糖组成及抗炎活性.方法 水提取总多糖,层析法分离纯化多糖组分,高效凝胶渗透色谱(HPGPC)测定各组分分子量;高效液相色谱-蒸发光散射法(HPLC-ELSD)测定多糖组分的单糖组成;通过二甲苯致小鼠耳廓肿胀模型,考察芡茎多糖(EFPP)抗炎作用.结果 从中纯化出EFPP1、EFPP2、EFPP3 3个组分多糖,其中EFPP1、EFPP2含有量较高;EFPP1组分由鼠李糖、阿拉伯糖、甘露糖、葡萄糖4种单糖组成,其物质的量之比为12.06∶9.544∶3.967∶9.131;EFPP2组分由鼠李糖、阿拉伯糖、甘露糖、葡萄糖4种单糖组成,其物质的量之比13.69∶11.64∶18.14∶6.616;芡茎多糖(EFPP)能降低小鼠耳廓肿胀度、降低致炎小鼠血清中TNF-α,IL-1β和IL-6含有量.结论 芡茎多糖是为非均一多糖,具有抗炎作用.

目的:比较益母草传统加工饮片与一体化加工饮片在化学成分含量与药理作用方面的差异,阐述益母草一体化加工工艺的合理性.方法:选择盐酸水苏碱、盐酸益母草碱、芦丁、金丝桃苷、异槲皮苷的含量为指标,比较2种加工工艺益母草饮片中化学成分含量的差异;以二甲苯致小鼠耳廓肿胀模型,考察2种加工工艺益母草饮片抗炎作用的差异;采用大鼠急性血瘀模型,考察2种加工工艺益母草饮片对全血黏度和凝血功能指标的影响.结果:益母草一体化工艺饮片中盐酸水苏碱、盐酸益母草碱、芦丁、金丝桃苷、异槲皮苷的质量分数分别为1.558％,0.168％,0.137％,0.113％,0.078％;传统工艺饮片中则分别为1.482％,0.134％,0.125％,0.082％,0.071％.2种工艺饮片均能降低致炎小鼠耳肿胀度、降低致炎小鼠血清中肿瘤坏死因子-α(TNF-α),白细胞介素(IL)-1β和IL-6的含量,产生抗炎作用;同剂量组间比较,一体化工艺组整体优于传统工艺组.2种加工饮片均能降低急性血瘀模型大鼠的全血黏度,延长凝血酶时间(TT),凝血酶原时间(PT)和活化部分凝血活酶时间(APTT),降低血浆纤维蛋白原(FIB)的质量浓度;同剂量组间比较,一体化工艺组整体优于传统工艺组.结论:与传统加工工艺相比,益母草一体化加工工艺在保证饮片质量的同时,降低了生产成本,且所得饮片化学成分与药理作用均略高于传统加工饮片,表明该药材选择一体化加工具有一定的合理性.

In this study, a fast, simple, precise, and sensitive hydrophilic interaction liquid chromatography (HILIC) method was established for simultaneous determination of free amino acids in three different varieties of duckweed including Spirodela polyrhiza (L.) Schleid., Landoltia punctata (G. Mey.) Les & D. J. Crawford, and Lemna aequinoctialis Welwitsch by ultrahigh performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Method validation was processed in terms of linearity, precision, stability, repeatability, and accuracy as well as limits of detection and quantification. The developed method was applied for quantification of 59 batches of samples. Then chemometric analysis was used to evaluate different duckweeds by principle component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). The results demonstrated that there was no significant difference in FAAs' profile among three varieties of duckweed.

Leonurus japonicas is a common clinical medicine, with the effect of activating blood circulation and regulating menstruation, inducing diuresis to alleviate edema, and clearing heat and detoxification. It has been used to treat menoxenia, dysmenorrhea, amenorrhea, lochia, edema of the body, oliguresis, sores, ulcerations, and other diseases in human. Recently, more than 120 compounds, including alkaloids, diterpenes, flavonoids, phenylethanoid glycosides, phenylpropanoids, coumarins, triterpene, organic acid, volatile oils and so on, have been isolated from L. japonicas. L. japonicas and its active ingredients have pharmacological effects in uterus, cardiovascular system, blood, and kidney. In this review, we summarized domestic and foreign literatures about chemical composition and pharmacological activity of L. japonicas in recent 20 years, in the hope of providing reference for further studies of L. japonicas.

A gas chromatography-mass spectrometry(GC-MS)method was established for the analysis of volatile components in Mentha haplocalyx, and seven principal components were quantified by gas chromatography(GC). Based on these analyses, the differences of volatile components in M. haplocalyx from Jiangsu, Anhui and other regions were compared. The results showed that the volatile oil of M. haplocalyx was divided into four chemical types：menthol-menthone type, pulegone-menthone type, piperitone-menthol type, piperitone epoxide type, and menthol-menthone type was the principal type. Menthol was the highest and pulegone was the lowest. The differences of M. haplocalyx from Anhui and other regions were obvious. The major volatile components and the differences of M. haplocalyx from different regions were confirmed and a quantitative method was established for the determination of volatile components, which provided the basis for improving the quality standard of M. haplocalyx.

Objective:The study is aimed to establish an HPLC method for simultaneous determination of tanshinol,protocatechuic acid,protocatechualdehyde,salvianolic acid B,salvianolic acid A,dihydrotanshinone I,tanshinone I,cryptotanshinone and tanshinone IIA in commercial products of Salviae Miltiorrhizae Radix et Rhizoma (CPSMR),and evaluate the qualities of 20 samples from different specifications of CPSMR.Methods:20 batches of S.miltiorrhiza were collected from Zhejiang,Shandong,Jiangsu,Hebei,Henan,Sichuan and other habitats.The determination was performed on a Waters xBridgeTM C18 column(250 mm×4.6 mm,5 μm)with a gradient mobile phase consisting of methanol(A)-0.03% phosphoric acid solution(B)at a flow rate of 1.0 mL·min-1.The column temperature was 25 ℃ and the detection wavelength was 270 nm.Cluster analysis was conducted with DPS software for comprehensive quality evaluation of 20 batches of CPSMR.Results:All the 9 effective constituents showed good linearity.The RSD of the precision,repeatability and stability tests were less than 3%.The average recoveries were in the range of 95.13%-101.35%.In view of habitats,CPSMR from Sichuan province were better than those from Hebei,Jiangsu and Zhejiang province.CPSMR from Henan and Shandong have the samples purchased from pharmacies.Conclusion:The contents of 9 effective constituents are different among Salviae Miltiorrhizae Radix et Rhizoma of differet specifications in markets from various habitats.SMRR products from Shandong and Henan have higher contents of tanshinones and salvianolic acids.HPLC can be used for the simultaneous determination of 9 effective constituents in SMRR products,which could be applied for the comprehensive quality evaluation of SMRR products.

Cardiac hypertrophy (CH), as one of the major causes of morbidity and mortality in the world, has become an independent and predictive risk factor for adverse cardiovascular events. However, progress in treatment remains sluggish in recent years. Therefore, compounds derived from non-toxic nature plants are urgently needed. Stachydrine (STA), which is isolated from Leonurus, has various activities, including resistance to cardiovascular disease, but little is known about its effect on CH or the mechanisms. We herein investigated the effect of STA on isoproterenol-induced CH and the underlying mechanisms. Treatment with STA significantly increased the ratios of heart weight/body weight, left ventricle weight/body weight and the cross-sectional areas of cardiomyocytes. In addition, STA significantly decreased the mRNA levels of atrial natriuretic peptide, B-type natriuretic peptide and β-myosin heavy chain. Furthermore, isoproterenol-induced fibrosis in rats receiving STA was significant attenuated, as evidenced by decreased ratio of fibrotic area/total area and decreased mRNA levels of collagens I and III. Given down-regulation of interleukin-6, tumor necrosis factor-α, interferon-γ (IFN-γ) and IFN-1β, treatment with STA significantly reversed the expressions of pro-inflammatory induced by isoproterenol. Moreover, STA attenuated the oxidative stress level in serum of isoproterenol-induced CH rats, as shown by increased activity of superoxide dismutase and decreased malondialdehyde level. STA inhibited the expressions of phosphorylated IκBα, NF-κB p65, JAK2 and STAT3 in vivo. Thus, both NF-κB and JAK/STAT signalings played essential roles in mediating the anti-CH effect of STA. Collectively, STA has a potent protective effect on isoproterenol-induced CH, with therapeutic implication for CH.

The present study investigated the potential hypoglycemic and antioxidant effects of polysaccharides extracted from the petioles and pedicels of Euryale ferox Salisb. (EFPP) on alloxan-induced hyperglycemic mice. The EFPP had a total carbohydrate of 65.72 ± 2.81%, uronic acid of 4.56 ± 0.62% and protein of 0.58 ± 0.12%, with an average molecular weight from 1.02 kDa to 11.45 kDa and monosaccharide composition of Man, GlcA, Rha, Glc, Gal and Ara at a molar ratio of 0.12 : 0.01 : 9.57 : 0.41 : 1.00 : 0.24. Administration with EFPP, especially high dose EFPP, was beneficial to reverse body weight loss, reduce blood glucose levels, enhance serum insulin levels, improve oral glucose tolerance, increase hepatic glycogen content and GCK activity, and modulate the mRNA expression of GCK in the liver. Meanwhile, EFPP had protective effects against alloxan-induced oxidative injury in mice, via increasing the activities of SOD, CAT and GSH-Px and decreasing the MDA contents in the liver and kidney of the mice. EFPP ameliorated the damage in pancreas, kidney and liver tissues, which was confirmed by histopathological observation. The results suggested that EFPP possess hypoglycemic and antioxidant activities, and could be a potential source of natural hypoglycemic and antioxidant agents for functional foods or complementary medicines.

A new isocoumarin derivative 8,5'-dihydroxy-6'-methoxy-4-phenyl-5,2'-oxidoisocoumarin (1) and a new stilbenoid derivative methyl 5-hydroxy-2-(2-hydroxyphenyl)benzofuran-4-carboxylate (2) together with nine known compounds (3-11) were isolated from the tubers of Sparganium stoloniferum Buch.-Ham.. Another stilbenoid derivative (3) and a xanthone (4) were identified as new natural products and compounds 5-10 were obtained for the first time from the genus Sparganium. All their structures were elucidated by comprehensive spectroscopic analysis and comparison with available literature information.

A high performance liquid chromatograph (HPLC) method was established for the simultaneous determination of five components (Harpagide,Verbascoside,Angoroside-C,Harpagoside,Cinnamic acid) in Scrophulariae Rhizome and Scrophulariae Radix,in order to investigate the impact of Scrophulariae Rhizome on the overall quality of Scrophulariae Radix.The determination was performed on a Waters 2695 HPLC with a 2998 DAD detector under 210,280 nm and 330 nm.In addition,the DPPH free radical scavenging rates of Scrophulariae Radix were determined at 517 nm by Ultraviolet Spectrophotometry.Consequently,the five components reached the baseline separation and there were no significant differences in the DPPH free radical scavenging rates between the extracts of Scrophulariae Rhizome and Scrophulariae Radix.Each batch of Scrophulariae Radix generally contained Scrophulariae Rhizome,and the total contents of Harpagide and Harpagoside in Scrophulariae Rhizome was similar to those in Scrophulariae Radix.Therefore,the existence of Scrophulariae Rhizome had little impact on the overall quality of Scrophulariae Radix.

Objective A method was established for separation, purification, enrichment and enumeration of peltate glandular trichomes from Schizonepetae Spica which combined with qualitative and quantitative analysis of major constituents of its inclusions. Methods Frozen brush method and sedimentation of water concentration connected to technology of materials affinity were applied to separate and concentrate peltate glandular trichomes respectively, and extracts of inclusions was qualified by GCMS. Microscope enumeration was considered used to quantitatively analyze three monoterperes in single peltate glandular trichome and the calyx of Schizonepetae Spica. Results 29 chemical compounds were identified from peltate glandular trichomes’ inclusions of Schizonepetae Spica,constituting 97. 45% of the total peak area. 35 characteristic common peaks exceeding 97% of the total peak area was both present in peltate glandular trichomes’ inclusions and essential oils of Schizonepetae Spica. The difference contents of d-limonene, dl-menthone, pulegone, E-carveol acetate, et al between peltate glandular trichomes’ inclusions and essential oils of Schizonepetae Spica were large. The content of d-limonene,dl-menthone, pulgone in a single peltate glandular trichome was 27. 660,24. 334 and 396. 390 ng respectively through the method of microscope enumeration. Conclusion The research provides a method of separation, purification enrichment and enumeration of peltate glandular trichomes. The chemical constituent of essential oil which is extracted by water vapor is different from peltate glandular trichomes’ inclusions and could not reflect the ratio and content, while prediction model which based on microscope enumeration is precision.

Jujube (Ziziphus jujuba) fruit is widely consumed as food and traditional Chinese medicine in Asian countries due to its potential effects for human health. To facilitate selection of the maturity stage providing optimum health benefits, jujube fruits were analysed at six stages of growth (S1-6) for triterpenic acids, nucleosides, nucleobases, and sugars by UHPLC-MS/MS or HPLC-ELSD methods. The content levels of most triterpenic acids and sugars increased with ripening, and reached the highest at S5 and S6, respectively. The accumulation of the cyclic nucleotides (cAMP and cGMP) was mainly in the later stage of ripening (S5-6). Therefore, if taking triterpenic acids as the major quality indicator, S5 should be the ideal time to harvest jujube fruit, and the full ripen stage (S6) maybe the best choice when taking sugars and cyclic nucleotides as the most important components.

目的建立超高效液相色谱三重四级杆串联质谱法（UPLC-MS/MS）同时测定浮萍中9种核苷类成分的方法,分析比较不同产地样品中9种成分的差异。方法采用Waters XbridgeTM Amide色谱柱（150 mm×4.6 mm,3.5μm）;以0.1%甲酸的水（A）-0.1%甲酸的乙腈（B）溶液为流动相,梯度洗脱,体积流量为0.5 m L/min;柱温30℃,采用电喷雾离子源,正离子检测方式,得到相应的提取离子流图,以峰面积进行定量,用主成分分析和聚类分析方法进行综合评价。结果 9种核苷类成分具有良好的线性关系,r值均大于0.998 9,方法精密度、重复性和稳定性的RSD值均小于3.5%,加样回收率在94.4%~101.9%,RSD在1.73%~3.58%。不同产地浮萍药材中9种核苷在组成和量上均有差异,其中2′-脱氧尿苷、2′-脱氧肌苷在13个产地样品中量均较低,以胞嘧啶最低;尿苷、肌苷、黄嘌呤量较高;13个产地中安徽黄山浮萍中9种核苷总量最高。结论该方法操作简便、准确、重复性好,可为浮萍药材的质量标准制定提供依据。

Based on the infrared spectra of Lophatheri Herba and Commelinae Herba, one-dimensional infrared spectra, second derivative spectra and two-dimensional correlated spectra were used to find out the differences between Lophatheri Herba and its imitations, respectively. The common peak ratio and variant peak ratio dual-indexes sequential were calculated and established according to infrared spectra of eleven batches of herbs. Infrared spectral data of Lophatheri Herba cluster analysis was applied to explore the similarity between each sample. The grouping results trend of sequential analysis of dual-indexes and cluster analysis was accordant. The results showed that the differences could be found by multi-level identification, and the source and the quality of the herbs could be effectively distinguished by the two analysis methods. Infrared spectroscopy, used in the present work exhibited some advantages on quick procedures, less sample required, and reliable results, which could provide a new method for the identification of traditional Chinese medicine with the imitations and adulterants, and the control of quality and origin.

目的:建立了芡实中不同构型维生素E的RP-HPLC含量测定方法.方法:采用高效液相色谱分析,WatersXBridge C18柱(4.6mm×150mm,5μm),以A相甲醇和B相水(97∶3)为流动相等度洗脱,流速为1.0mL/min,柱温:30℃,检测波长为295nm.结果:α-生育酚、β+γ-生育酚、δ-生育酚均能够达到基线分离,α-生育酚在0.12～1.2g/L范围内具有良好的线性关系,R21=0.9998;β+γ-生育酚在0.056～0.56g/L范围内具有良好的线性关系,R22=0.9999;δ-生育酚则在0.0096～0.096g/L范围内具有良好的线性关系,R23=0.9998.平均加样回收率分别为100.47％,100.04％,99.10％;RSD分别为0.27％,4.65％,1.92％.安徽安庆芡实中总VE含量最高,可达2.2633mg/g.结论:该方法简便、准确、重现性好,为芡实药材的质量评价提供合理的参考依据.

目的 研究不同干燥方法对何首乌药材中二苯乙烯苷类和蒽醌类活性成分转化的影响,为何首乌药材产地干燥加工适宜方法的建立提供科学依据.方法 以新鲜野生何首乌块根为材料,分别采用阴干、晒干、烘干、冷冻干燥、微波干燥、远红外干燥6种干燥加工方法处理,采用HPLC法同时测定经各种干燥方法制备的样品及不同烘干时间样品中二苯乙烯苷、游离型蒽醌类和结合型蒽醌类化学成分的量.结果 经6种不同干燥方法处理的样品中,二苯乙烯苷的量由高至低依次为晒干＞远红外干燥＞冷冻干燥＞阴干＞烘干＞微波干燥;阴干法和烘干法更适宜于结合型蒽醌类成分转化为游离型蒽醌类成分;采用远红外干燥的样品,其结合型蒽醌的量最高.主成分分析综合评价显示,经远红外干燥法制备的何首乌药材其综合评价指数明显高于其他干燥方式.结论 远红外干燥法为何首乌药材适宜的产地干燥加工方法.

In this study, a sensitive and rapid method for the simultaneous determination of free amino acids without derivatization using hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry (HILIC-MS/MS) was developed. The method was performed on an ultra-high-performance liquid chromatography (UHPLC) separation system coupled with a triple-quadrupole mass spectrometry (TQ-MS) instrument. Sufficient separation of 23 underivatized amino acids was achieved on an Acquity BEH Amide column (2.1 mm × 100 mm, 1.7 μm) in a single run of 12 min. Then the method was applied for the analysis of the free amino acids in 46 batches of Ziziphus jujuba fruits which comprised 39 cultivars from 26 cultivation regions. Multivariate statistical analysis was also used to investigate the differences in free amino acid profiles among the samples. This study showed that HILIC-UHPLC-TQ-MS is an effective technique to analyze underivatized amino acids in the food samples.