Background: Semiliquidambar cathayensis Chang is an extremely valuable and endangered medicinal plant. To investigate the exploitation and rational utilization of S. cathayensis, this study conducted metabolomics analysis of the leaves and bark of artificially cultivated S. cathayensis at different developmental stages. Methods: These metabolites were detected and identified by Ultra-Performance Liquid Chromatography–tandem Mass Spectrometry (UPLC-MS/MS) technology, and then univariate statistical analyses, multivariate pattern analyses, and pathway analyses were carried out. Results: As a result, a total of 801 metabolites were detected in S. cathayensis; differential metabolites in leaves at different developmental stages were mainly enriched in pathways related to flavonoids, whereas differential metabolites in bark at different developmental stages were mainly aromatic compounds, amino acids, and flavonoids, among others. This study revealed that young leaves are ideal for use in treating rheumatism, regulating blood pressure, and lowering blood glucose, while old leaves are better suited for skincare products and extracting materials to prevent neurodegenerative diseases and support women’s ovarian health. As for bark, four-year-old S. cathayensis bark is optimal for extracting myricetin. If the pharmaceutical, chemical, food, and industrial fields require extensive extraction of L-phenylalanine, trans-3-hydroxycinnamate, and 4-hydroxyphenylacetate, and if the medical field needs to extract anti-allergy, liver protection, and anti-coagulant ingredients, the two-year-old S. cathayensis bark is the best choice. Conclusions: Thus, this study established a solid theoretical framework for the rational, effective, and sustainable utilization of S. cathayensis leaves and bark.

Summary A novel metabolomics analysis technique, termed matrix‐assisted laser desorption/ionization mass spectrometry imaging‐based plant tissue microarray (MALDI‐MSI‐PTMA), was successfully developed for high‐throughput metabolite detection and imaging from plant tissues. This technique completely overcomes the disadvantage that metabolites cannot be accessible on an intact plant tissue due to the limitations of the special structures of plant cells (e.g. epicuticular wax, cuticle and cell wall) through homogenization of plant tissues, preparation of PTMA moulds and matrix spraying of PTMA sections. Our study shows several properties of MALDI‐MSI‐PTMA, including no need of sample separation and enrichment, high‐throughput metabolite detection and imaging (>1000 samples per day), high‐stability mass spectrometry data acquisition and imaging reconstruction and high reproducibility of data. This novel technique was successfully used to quickly evaluate the effects of two plant growth regulator treatments ( i.e. 6‐benzylaminopurine and N‐phenyl‐N′‐1,2,3‐thiadiazol‐5‐ylurea) on endogenous metabolite expression in plant tissue culture specimens of Dracocephalum rupestre Hance ( D. rupestre ). Intra‐day and inter‐day evaluations indicated that the metabolite data detected on PTMA sections had good reproducibility and stability. A total of 312 metabolite ion signals in leaves tissues of D. rupestre were detected, of which 228 metabolite ion signals were identified, they were composed of 122 primary metabolites, 90 secondary metabolites and 16 identified metabolites of unknown classification. The results demonstrated the advantages of MALDI‐MSI‐PTMA technique for enhancing the overall detection ability of metabolites in plant tissues, indicating that MALDI‐MSI‐PTMA has the potential to become a powerful routine practice for high‐throughput metabolite study in plant science.

Bryophyllum pinnatum (L. f.) Oken is a kind of herbaceous plant of Crassulaceae, although studies have shown that Bryophyllum pinnatum can treat gastritis, the main active compounds and molecular mechanisms are still unclear. In this study, ultra performance liquid chromatography/mass spectrometry (UPLC-MS) analyses and a strategy of network pharmacology were used to elucidate the potential molecular mechanisms of Bryophyllum pinnatum against gastritis. 85 compounds in Bryophyllum pinnatum were identified through UPLC-MS analysis, 7 main active compounds (quercetin, eicosapentaenoic acid (EPA), isorhamnetin, beta-sitosterol, cubebininolide, kaempferol, artemetin) were obtained based on online database. Kyoto encyclopedia of genes and genomes (KEGG) analysis demonstrated that 93 common targets were mainly enriched in pathways in cancer, PI3K-Akt signaling pathway, FoxO signaling pathway, TNF signaling pathway. AKT1, SRC, EGFR, MAPK1 and MAPK8 targets were the core targets. The molecular mechanisms of Bryophyllum pinnatum to exert anti-gastritis effect were mainly related to the control of cell physiological activities and reduction of stomach acids and gastric proteases. In summary, our study reveals that Bryophyllum pinnatum has the characteristics of multitarget, multi-compound and multi-pathway to exert anti-gastritis effect, which provides the direction and reference for the subsequent in-depth verificationof the mechanisms of anti-gastritis effect of Bryophyllum pinnatum.

为了建立简单、高效的狭苞橐吾组织培养再生体系,本研究以狭苞橐吾的子叶作为外植体,探究不同种类、不同浓度的植物生长调节剂组合对狭苞橐吾离体组织培养的影响.研究结果显示,不同浓度的6-BA与2,4-D组合均能诱导狭苞橐吾子叶产生愈伤组织.当2,4-D浓度为3.0 mg/L、6-BA浓度为1.0、2.0、4.0 mg/L时,愈伤组织诱导率较高,诱导率均在90％ 以上.适宜诱导不定芽的培养基为MS+6-BA 2.0 mg/L+NAA 1.0 mg/L,诱导率为48.00％.在MS+TDZ 1.0 mg/L+NAA 2.0 mg/L培养基中,丛芽的诱导效果较好,诱导率为98.67％.适宜的生根培养基为1/2MS+IAA 0.5 mg/L,其生根率为95.56％,平均生根条数为9.36条,平均长度约为10.48 cm.

Aim To study the mechanism of the ethanol extract from Leucopaxillus giganteus（LGEE） in treating breast cancer based on network pharmacology and molecular experimental validation. Methods Some chromatographic methods were used to isolate the chemical constituents of LGEE,and their structures were elucidated based on spectral data. The antitumor activities of LGEE were determined by MTT assay. The predicted targets of LGEE were selected by TCMSP and Pharmmaper, and Genecards database was used to screen the targets. GO and KEGG analysis of target genes were performed. Molecular docking was used to test the binding of active components to core targets. Western blotting was used to validate the regulating function of chlorogenic acid on CHEK2 and CASP3 targets of MDA-MB-231 cells. Results Thirteen compounds were identified including clitocine, chlorogenic acid and so on. LGEE displayed anticancer activities against MDA-MB-231 with the inhibition percent（87.35±1.55）%,at the concentration of 200 mg·L -1 . A total of 203 drug targets were obtained from the 13 active compounds, 78 targets intersected with disease targets for breast cancer were screened out, and 21 core targets including CHEK2 and CASP3 were obtained. The results of KEGG analysis showed that it might treat breast cancer through signaling pathways, such as p53 signaling pathway, FOXO signaling pathway and so on. Chlorogenic acid from LGEE was validated to regulate CHEK2 and CASP3 targets, which proved the reliability of results of pharmacology. Conclusions Based on the network pharmacology, the possible targets and signaling pathways for treating breast cancer by LGEE can be preliminary explored, which may provide a reference for the study of the mechanism of the LEGG.

Objective: Asa medicinal plant, the resource of Rhodiola dumulosa is deficient along with the large collection. For the protection and utilization of R. dumulosa, the influence of plant growth regulators (PGRs) on callus induction and adventitious shoots differentiation, polysaccharide production and the antioxidant activity were tested. Methods: Internodes of R. dumulosa were used as explants and cultured on MS medium plus different plant growth regulators (PGRs). The anti-oxidative activities of polysaccharides were evaluated using radical scavenging assays. Results: By response surface plot, 0.85 mg/L N6-benzyladenine (BA), 0.34 mg/L naphthaleneacetic acid (NAA) and 0.33 mg/L 2,4-dicholorophenoxyacetic acid (2,4-D) were the optimal factors for callus induction (90.03%) from internodes explants on MS medium. The fresh weight of green callus increased 47.26 fold, when callus was inoculated on MS + thidiazuron (TDZ) 0.5 mg/L + NAA 2.0 mg/L. Adventitious buds regenerated from callus on the media of MS were fortified with BA 1.0 mg/L plus NAA 0.5 mg/L, and the induction rate was 40.00%. MS plus indole-3-butyric acid (IBA) 1.0 mg/L produced the highest rooting rate with 10 to 15 roots in a length of 2-3 cm per shoot. The content of total polysaccharides in callus developed on MS + TDZ 0.5 mg/L + NAA 2.0 mg/L and MS + BA 1.0 mg/L + NAA 0.5 mg/L was as high as 1.72%2.15%. At the dose of 0.5 mg/mL polysaccharides extracted from different callus induced on MS + NAA 2.0 mg/L + TDZ 0.5 mg/L or MS + BA 1.0 mg/L + NAA 0.5 mg/L or MS + BA 0.5 mg/L + 2,4-D 0.5 mg/L, the ABTS radical eliminating percentages were 82.78%, 80.18% and 68.59%, respectively, much higher than that of wild plant. Conclusion: A rapid micropropagation system for R. dumulosa has been developed. The combination of TDZ and NAA or BA and NAA can increase the yield of the total polysaccharides. The polysaccharides isolated from callus and whole wild plants had stronger free radicals scavenging activities, indicating that polysaccharides from R. dumulosa are the potential pharmaceutical supplements. (c) 2022 Tianjin Press of Chinese Herbal Medicines. Published by ELSEVIER B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Bryophyllum pinnatum (Lam.) Oken is an ornamental and ethno-medicine plant, which can grow a circle of adventitious bud around the leaf margin. The dynamic change of metabolites during the development of B. pinnatum remains poorly understood. Here, leaves from B. pinnatum at four developmental stages were sampled based on morphological characteristics. A non-targeted metabolomics approach was used to evaluate the changes of endogenous metabolites during adventitious bud formation in B. pinnatum. The results showed that differential metabolites were mainly enriched in sphingolipid metabolism, flavone and flavonol biosynthesis, phenylalanine metabolism, and tricarboxylic acid cycle pathway. The metabolites assigned to amino acids, flavonoids, sphingolipids, and the plant hormone jasmonic acid decreased from period Ⅰ to Ⅱ, and then increased from period Ⅲ to Ⅳ with the emergence of adventitious bud (period Ⅲ). While the metabolites related to the tricarboxylic acid cycle showed a trend of first increasing and then decreasing during the four observation periods. Depending on the metabolite changes, leaves may provide conditions similar to in vitro culture for adventitious bud to occur, thus enabling adventitious bud to grow at the leaf edge. Our results provide a basis for illustrating the regulatory mechanisms of adventitious bud in B. pinnatum.

The aim of this study was to analyze the bioactive components of Syringa reticulata subsp. amurensis (SRA) seeds, and the potential mechanism of SRA seed against chronic bronchitis is elucidated using network pharmacology. In total, 235 constituents were detected from SRA seed by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS), and 28 active ingredients were identified through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform database (TCMSP), which was classified into 10 different categories including flavonoids, diterpenoids, coumarins, alkaloids, isoflavones, phenylpropanoids, sesquiterpene, anthraquinones, phenols, and lignans. Among them, 6 components vestitol(V1), wogonin(W2), cheilanthifoline(C1), miltirone(M2), glabridin(G3), and myricanone (M3) were selected as potential key constituents of SRA seed against chronic bronchitis through network pharmacology analysis. Then the 7 key targets (PTGS2, TP53, TNF, VEGFA, IL1 beta, IL6, and JUN) were obtained based on the target prediction and protein-protein interaction (PPI). In conclusion, we use UPLC-ESI-MS analysis and network pharmacology to reveal the chronic bronchitis mechanisms of SRA seed, which provides new theoretical research ideas for the sustainable development of endangered multifunctional plants.

目的 基于UHPLC-Q-Exactive-Orbitrap-MS技术阐明树舌灵芝主要成分,结合网络药理学探讨其防治新型冠状病毒肺炎的作用机制.方法 采用UHPLC-Q-Exactive-Or-bitrap-MS技术鉴定树舌灵芝的主要成分;采用Swiss Target Prediction数据平台对鉴定成分进行靶点预测,对核心靶基因进行GO和KEGG富集分析,运用Cytoscape 3.8.0绘制关联网络图,同时通过分子对接评价树舌灵芝核心成分与血管紧张素转化酶Ⅱ(ACE2)及3种新型冠状病毒(SARS-CoV-2)蛋白的结合作用.结果 该研究从树舌灵芝中鉴定化合物62个;30个活性化合物主要作用在IL-6、PTGS2和MAPK1等32个核心靶点,通过PI3K/Akt、TNF及肺结核等多条与肺损伤保护相关通路,发挥防治COVID-19的作用;分子对接显示小白菊内酯、1,4-二羟基-2-萘甲酸和瑞香素等核心成分与ACE2及3种SARS-CoV-2蛋白的亲和作用较好.结论 该研究初步阐明树舌灵芝的化学成分及潜在作用机制,为筛选树舌灵芝防治COVID-19的药效成分及深入阐明作用机制提供了科学的理论依据.

Eucommia ulmoides Oliver is a precious economic crop, which can be used as raw industrial materials and traditional Chinese medicine. Secondary metabolites may vary in type, content, and proportion due to different origins and cultivars, and are closely related to the intrinsic quality of the medicinal plant. In this study, a metabolomics method was applied to investigate the effects of regions and cultivars on the changes of endogenous metabolites in samaras of ‘Qinzhong’ and ‘Huazhong’, two cultivars of E. ulmoides. The differential metabolites mainly enriched in pathways related to flavonoids when the same cultivar was cultivated in different regions. The differential metabolites between the two cultivars were primarily concentrated in flavonoids, sphingolipids, and glutathione. Therefore, it is speculated that flavonoids play an essential role in the adaption of the environment and formation of different cultivars. The research provides a direction for the rational utilization of E. ulmoides samaras.

Anaphalis hancockii Maxim is often used as an aromatic plant in China. To effectively protect and utilize the resources of A. hancockii, a completely rapid micropropagation system has been developed. Additionally, ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS) technology, coupled with multivariate statistical analysis and pathway enrichment analysis, was used to assess the regulatory influences of 6-benzylaminopurine (6-BA) and kinetin (KT) applied in A. hancockii. The results showed that when MS medium was supplemented with 4.0 mg/L 2,4-D and 2.0 mg/L KT, the effect of inducing callus from leaf, petiole, and root explants was the best. At this time, the leaf callus induction rate was 94.67

本文采用网络药理学-分子对接手段探索二陈丸治疗2型糖尿病的物质基础及作用机制.从TCMSP中药数据库获取二陈丸的活性成分及靶点,结合GeneCards数据库获取2型糖尿病的靶点信息,利用venny映射得到治疗2型糖尿病的潜在靶点,通过String数据库对潜在靶点进行蛋白相互作用(PPI)网络预测,再通过DAVID数据库对关键靶点进行GO和KEGG分析,借助Cytoscape 3.8.0软件构建"活性成分-疾病靶点-通路"的复杂网络通路,最后使用Autodock vina软件对二陈丸主要活性成分和作用靶点进行分子对接,进一步揭示二陈丸治疗2型糖尿病的作用机制.结果表明,二陈丸治疗2型糖尿病的关键成分有柚皮素、黄芩素和刺芒柄花素等17个,这些成分调控显著富集于TNF signaling pathway等信号通路的MMP9、NOS3和CAT等21个核心靶点,分子对接结果显示刺芒柄花素、柚皮素、黄芩素与MMP9、CAT和NOS3具有较强的亲和力.本研究通过网络药理学分析二陈丸治疗糖尿病的物质基础及作用机制,为其进一步研究奠定基础.

In vitro propagation technology with plant growth regulators (PGRs) is generally applied in the cultivation of Scabiosa tschiliensis, which can solve collection difficulties and limited resources of S. tschiliensis. Nevertheless, comprehensive metabolomic evaluation on S. tschiliensis with PGR effects is still lacking. In this work, a non-targeted metabolomics approach, coupled with statistical and pathway enrichment analysis, was used to assess the regulatory influences of 6-benzylaminopurine (6-BA) and kinetin (KT) applied in S. tschiliensis. The results showed that the PGRs affect metabolism differentially, and the addition of 6-BA and KT can increase different secondary metabolites. In the two PGR groups, some primary metabolites such as L-phenylalanine, L-tyrosine, L-arginine, L-asparagine, and D-proline were significantly reduced. We suspect that under the action of PGRs, these decreased amino acids are derived into secondary metabolites such as umbelliferone, chlorogenic acid, and glutathione. Additionally, some of those secondary metabolites have a biological activity and can also promote the plant growth. Our results provide a basis for the targeted cultivation and utilization of S. tschiliensis, especially the expression of metabolites related to PGR application.

为有效利用藏橐吾Ligularia rumicifolia(Drumm.)S.W.Liu资源,以其子叶为外植体进行离体培养研究.结果发现,诱导愈伤组织的适宜培养基为MS+TDZ 1.0 mg/L+NAA 1.0 mg/L,诱导愈伤组织产生不定芽的适宜培养基为MS+6-BA 2.0 mg/L+NAA 0.5 mg/L,诱导率为69.56％.诱导子叶产生不定芽的适宜培养基为MS+6-BA 3.0 mg/L+NAA 0.8 mg/L,诱导率为72.33％.不定芽在1/2 MS+IAA 0.5 mg/L的培养基中生根率为70.00％.

本文通过调查发现,安国药市交易的常用中草药共有540种,隶属于125科368属植物.其中豆科、菊科、唇形科、蔷薇科、百合科、芸香科等为优势科,柑橘属、莲属、大戟属、贝母属、鹅绒藤属、蒿属等为优势属.安国地产道地中草药主要包括祁紫菀、祁花粉、祁芥穗、祁山药、祁沙参、祁白芷、祁菊花和祁薏米等.

目的:基于DNA条形码技术及高分辨率熔解曲线(HRM)方法鉴定肉苁蓉药材的基原.方法:通过聚合酶链式反应(PCR)对肉苁蓉样品的ITS2序列进行扩增;将待鉴定肉苁蓉样品的ITS2序列与Genbank数据库进行BLAST比对;利用MEGA 6.06软件通过建树法进行NJ树聚类分析;通过HRM方法快速鉴定肉苁蓉药材基原.结果:21份待鉴定肉苁蓉样品ITS2序列的BLAST比对结果中,11份样品与沙苁蓉高度一致,其余10份样品与《中华人民共和国药典》2015年版一部收载品种肉苁蓉(Cistanche deserticola)或管花肉苁蓉(C istanchetubulosa)高度一致;通过ITS2序列的NJ聚类分析,将21份待鉴定肉苁蓉样品分别聚类到沙苁蓉、管花肉苁蓉、盐生肉苁蓉、肉苁蓉4个种;通过HRM方法鉴定肉苁蓉药材样品,鉴定结果与NJ聚类分析结果一致.结论:通过DNA条形码和HRM技术均能有效鉴定不同种的肉苁蓉药材样品,且结果一致,起到互相验证的作用,为肉苁蓉及其他中药民族药品种的快速鉴定提供了参考依据.

以华北八宝Hylotelephium tatarinowii (Maxim.) H.Ohba愈伤组织为材料,采用不同质量分数的聚乙二醇(PEG)6 000进行胁迫处理,分别测定愈伤组织中过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性变化,通过非变性聚丙烯凝胶电泳分析POD、SOD和酯酶(EST)的同工酶表达特征.研究结果表明,在PEG 6 000质量分数为0.5％～5.0％时,随着质量分数的增加和胁迫时间的延长,POD、SOD和CAT的活性逐渐增强,且愈伤组织生长良好;当PEG 6 000质量分数为8.0％时,随着胁迫时间的延长,POD、SOD和CAT酶活性急剧下降.

目的:通过特异性引物的聚合酶链式反应(PCR)法结合微芯片电泳检测的方法,特异性鉴别宽体金线蛭、日本医蛭、菲牛蛭3种水蛭品种.方法:针对宽体金线蛭、日本医蛭、菲牛蛭3个品种的线粒体同源序列,设计区分正伪品水蛭以及分别鉴定3个品种水蛭样品的特异性引物;通过PCR和微芯片电泳检测的方法验证所设计引物的特异性;将验证得到的特异性引物进行整合,建立3个品种的特异性引物用于三重PCR的方法.结果:确定了用于区分宽体金线蛭、日本医蛭、菲牛蛭3个品种中正伪品的特异性引物KA;确定了宽体金线蛭特异性引物KT,日本医蛭特异性引物RI,菲牛蛭特异性引物FN;建立了3种特异性引物进行三重PCR鉴别3个品种水蛭药材的方法.结论:通过特异性引物结合三重PCR的方法,可以同步鉴定宽体金线蛭、日本医蛭、菲牛蛭的人为预混样品,鉴定结果准确,且用于三重PCR的特异性引物不存在引物之间相互结合或非特异性扩增的现象.

目的:基于DNA条形码技术鉴定制草乌疑似伪品的基原.方法:通过聚合酶链式反应(PCR)对建库样品及制草乌药材的ITS2序列进行扩增;对建库样品进行N-J聚类和变异位点分析,通过与GenBank数据库进行Blast比对,以及通过与建库样品进行比对推断制草乌药材疑似伪品的基原.结果:制草乌药材疑似伪品经Blast比对,结果显示能比对到多个种,且一致率均达到100％;建库样品ITS2序列进行N-J聚类和变异位点分析,结果均显示乌头与北乌头不能区分,其余建库样品均可区分;通过将5.8S与28S区间位点准确的56条制草乌药材疑似伪品的ITS2序列与建库样品的ITS2序列进行变异位点分析,其中2条序列与康定乌头完全一致,18条序列与准噶尔乌头完全一致,6条序列与甘青乌头完全一致,剩余30条序列与乌头、北乌头序列一致.结论:鉴定结果显示,市场制草乌药材存在掺伪及全伪现象,混伪品有康定乌头、甘青乌头、准噶尔乌头,其中准噶尔乌头为主要混伪品,此研究为DNA条形码技术鉴别制草乌正伪品开辟了一条新途径.

目的:基于DNA条形码技术鉴定重楼药材疑似伪品的基原.方法:通过聚合酶链式反应(PCR)对重楼的内部转录间隔区2(ITS2)序列进行扩增;通过与GenBank数据库进行Blast比对,以及利用建树法推断重楼药材疑似伪品的基原.结果:重楼药材疑似伪品经Blast比对,结果显示为狭叶重楼(变种)和黑籽重楼(原变种),一致率均达到99％;通过与前期研究的重楼ITS2序列进行NJ聚类分析,可以判定待鉴定样品的基原与西藏延龄草较为接近;通过将重楼药材疑似伪品与收集的吉林延龄草标本、西藏延龄草标本、宽瓣重楼标本、华重楼标本的ITS2序列进行NJ聚类分析,推断重楼药材疑似伪品的基原为吉林延龄草.结论:确定重楼药材疑似伪品为非2015年版《中华人民共和国药典》收载物种,并推断其基原为吉林延龄草,通过DNA条形码技术为鉴别药材正伪品开辟了一条新途径.