Relationship between rooting ability and endogenous phytohormone changes in successive continuous generation cuttings of Buxus sinica var. parvifolia, an endangered woody species in China

The objectives of this study were to investigate the effects of successive continuous generation （SCG） cuttings of Buxus sinica var. parvifolia on the rejuvenation of ortets at the hormone level, detect levels of indoleacetic acid （IAA）, abscisic acid （ABA）, isopentenyladenosine （iPA）, zeatin riboside （ZR） and gibberellin4 （GA4） during the rooting process of different generations by means of enzyme-linked immunosorbent assay （ELISA） technology and statistically analyze the rooting characteristics of cuttings. The results show that, the root systems of subsequent generations （generation 1998, 2000） developed satisfactorily, only a few initial generation （generation 1990, 1994） cuttings rooted and the root growth was inhibited. Around the period of callus formation and root projection through the epidermis, there was a marked, regular variation in the levels of IAA, ABA and GA4 between the initial generations and subsequent generations.

Buxus Leaves from the Oligocene of Guangxi, China and Their Biogeographical Significance

The present paper reports two new species of Buxus from the Oligocene Ningming Formation of Guangxi, South China, on the basis of several compressed fossil leaves.Buxus ningmingensis sp.nov.is characterized by an elliptic to ovate shape; pinnate venation with marginal secondary, long hairs on the adaxial side of the midvein base; and amphistomatic, anomocytic stomata with a prominent outer ring.All of these characteristics suggest a closest affinity to the living Buxus microphylla subsp.sinica.Another new species identified as Buxus preaustro-yunnanensis sp.nov.is characterized by the diagnostic formation of its admedially branched tertiaries, the trunks of which are often strongly thickened by sclereides.This species is most similar to the extant Buxus austroyunnanensis.The co-occurrence of Buxus ningmingensis sp.nov.and Buxus preaustro-yunnanensis sp.nov.suggests that these two Buxus lived during the Oligocene under a warm, tropical to subtropical climate, similar to the current climate of Ningming.Our findings provide the earliest fossil evidence of Buxus leaves with detailed illustrations of leaf architectural and cuticular features.The occurrence of these two species indicates that Buxus began to diversify in Guangxi no later than the Oligocene.

Triterpenoid alkaloid derivatives from Buxus rugulosa

Four new triterpenoid alkaloid derivatives,buxrugulines A-D(1-4),together with four known ones(5-8),were isolated from the leaves and stems of Buxus rugulosa.The structures of compounds 1-4 were elucidated by NMR and MS spectroscopic analysis.All compounds were assayed for their cytotoxicities against HL-60,SMMC-7721,A549,MCF-7,and SW480 cells lines.

Common Mechanism of Lignification of Compression Wood in Conifers and Buxus

Woody plants develop a specialized secondary xylem known as reaction wood to enable formation of an ideal shape. Reaction wood in coniferous species is known as compression wood, and that of woody angiosperms as tension wood. However, the genus Buxus which is classified as an angiosperm, forms compression-wood-like reaction wood. We investigated the mechanism of lignification in coniferous compression wood and Buxus reaction wood: 1) Several lignin synthesis genes were upregulated in differentiating reaction wood of Buxus microphylla;2) B. microphylla possesses a specific laccase gene that is expressed specifically in differentiating reaction wood (BmLac4);3) laccase activity localization was closely related to lignification of reaction wood, and laccase activity was high in the secondary wall middle layer;4) in reaction wood cell walls, galactan was present in the outer portion of the secondary wall middle layer, and the level of xylan was reduced. These findings suggest that lignification in B. microphylla reaction wood is identical to that of coniferous compression wood. These may represent general mechanisms of increasing lignin content in various reaction woods.

Effects of Total Alkaloids in Buxus microphylla Leaves on Aorta Smooth Muscle of Rats and Their Mechanisms

Objective To investigate the effects of total alkaloids in Buxus microphylla leaves(ABML)on isolated rats thoracic aorta rings,and then to explore the possible mechanisms underlying the effects.Methods Thoracic aortas of Wistar rats were isolated,removed,and mounted onto an organ bath.The effects of ABML at different concentration on the contraction of isolated thoracic aorta rings(with and without endothelium)precontracted with KCl or PE were observed with organ bath technique.Dose-effect curves of CaCl2 were recorded by organ bath technique.The concentration of intracellular Ca 2+ ([Ca 2+ ]i)increased by PE,KCI,and caffeine in the presence of ABML was determined using Ca 2+ sensitive fluorescence indicator Fura-2/AM loaded thoracic aorta vascular smooth muscle (VSM)cells of rats.Results In aorta rings precontracted with PE and KCl,ABML produced concentration- dependent relaxation in both intact and denuded endothelium ring groups.There was no difference in the inhibition of contraction between the intact and denuded endothelium ring groups at the same concentration.Exposure of isolated thoracic aorta rings to ABML led to a significant reduction in the contracting response induced by CaCl2,and shifted the cumulative concentration-response curves to right.ABML could significantly inhibit the extracellular Ca 2+ influx induced by PE and KCl under[Ca 2+ ]0 of 1.5 mmol/L,with inhibitory ratios of 40.2%and 49.9%,respectively.In the case of Ca 2+ -free,ABML could significantly inhibit the intracellular Ca 2+ release induced by PE,with inhibitory ratio of 72.4%.Conclusion ABML relaxes thoracic aorta VSM cells by suppressing influx of extracellular Ca 2+ via voltage-dependent Ca 2+ channel and receptor-operated Ca 2+ channel.

Lignans with(N,N-diethyl)methyl amino group from Buxus rugulosa

Buxrugulosides A–E,four lignan glycosides(1–4)and a protocatechuate derivative(5)featuring a rare(N,Ndiethyl)methyl amino group at aromatic rings,were obtained from the aerial parts of Buxus rugulosa,which is famous for treating coronary heart disease.Their structures including absolute configurations were elucidated by HRMS,1 D and 2 D NMR,and by comparing their CD data with previous reports.Compound 1 was a rare sesquilignan,and all of these compounds were the first example of lignans with(N,N-diethyl)methyl amino group.

Buxu Tongyu Granule Alleviates Myocardial Ischemia by Activating Vascular Smooth Muscle Cell Soluble Guanylate Cyclase to Inhibit Abnormal Vasomotion

Myocardial ischemia is a serious threat to human health,and vascular dysfunction is its main cause.Buxu Tongyu(BXTY)Granule is an effective traditional Chinese medicine(TCM)for treating myocardial ischemia.However,the underlying mechanism of BXTY is still unclear.In this study,we demonstrate that BXTY ameliorates myocardial ischemia by activating the soluble guanylate cyclase(sGC)-30,50-cyclic guanosine monophosphate(cGMP)-protein kinase G(PKG)signaling pathway in vascular smooth muscle cells(VSMCs)to dilate the arteries.BXTY was given by gavage for ten consecutive days before establishing an animal model of acute myocardial ischemia in mice via the intraperitoneal injection of pituitrin.The results showed that BXTY alleviated the symptoms of myocardial ischemia induced by pituitrin in mice,including electrocardiogram abnormalities and changes in plasma enzymes.In addition,BXTY dilated pre-constricted blood vessels and inhibited the vasoconstriction of the superior mesenteric artery in a dose-dependent but endothelial-independent manner.These effects were eliminated by preincubating vascular rings with the sGC inhibitors NS 2028 or ODQ,or with the PKG inhibitor KT 5823.Moreover,BXTY increased the protein expression of sGC-b1 and the intracellular second messenger cGMP level in mouse aortic vascular smooth muscle cells(MOVAs).NS 2028 or ODQ reversed these effects of BXTY.The expression level of the cGMP downstream effector protein PKG-1 increased after treating MOVAs with BXTY.NS 2028,ODQ,or KT 5823 also reversed this effect of BXTY.In conclusion,BXTY can improve the symptoms of acute myocardial ischemia in mice,and activating the sGC-cGMP-PKG pathway in VSMCs to induce vasodilation is its key pharmacodynamic mechanism.

宜昌黄杨对夏季淹水的生理生态学响应

宜昌黄杨(Buxus ichangensis)主要分布于长江三峡地区的消落带,对该地区的水土保持和岸坡地稳定起着十分重要的作用。作者通过模拟夏季淹水研究了宜昌黄杨的形态特征、光合作用和根系活力的变化特征,揭示了宜昌黄杨对淹水的生态学响应规律。结果表明宜昌黄杨在淹水过程中产生了皮孔、不定根等形态适应特征,经过60d的淹水处理,植株仍全部存活,表现出较强的适应能力。但是植株的生长仍然受到淹水的显著影响:淹水条件下宜昌黄杨的最大光合速率、气孔导度和根系活力与对照相比都显著下降,60d后分别只有对照的19.24%、24.04%和4.78%。在不同处理阶段这些指标下降的幅度不同:初期的下降幅度最大,后期下降幅度减少,其中根系活力在后期还有所回升。叶绿素A的含量随淹水延长持续下降,后期有所回升,而叶绿素B和类胡萝卜素的变化却不大。最后,作者对该物种在水电工程库区退化消落带植被生态恢复中的应用提出了一些建议。

Comparative efficacy and safety of Chinese patent medicines of iron deficiency anemia during pregnancy:A network meta-analysis

BACKGROUND Iron deficiency anemia(IDA)is a prevalent nutritional disorder during pregnancy.Clinical studies indicate that incorporating Chinese patent medicines(CPMs)with oral iron(OI)in treating IDA in pregnancy can reduce adverse effects and improve clinical outcomes.Nonetheless,the comparative efficacy of different CPMs remains unclear.AIM To assess the safety and effectiveness of different CPMs for treating IDA during pregnancy using network meta-analysis.METHODS We conducted a search for randomized controlled trials(RCTs)that combined CPM and OI for IDA treatment in pregnancy,spanning from 2013 to the present.Data analysis was performed using Rev Man 5.3 and Stata 14.0 on literature that satisfied the quality criteria.RESULTS The analysis included 45 RCTs,encompassing 4422 pregnant patients with IDA.Six CPMs were examined,including Shengxuebao Mixture,Shengxuening Tablets(SXN),Yiqi Weixue CPMs(YQWX),Jianpi Shengxue CPMs(JPSX),Yiqi Buxue Tablets,and Compound Hongyi Buxue Oral Liquid(FFHY).Findings indicated that FFHY+OI significantly improved the clinical effective rate.SXN+OI was most effective in boosting red blood cells counts and hemoglobin levels.YQWX+OI showed superior results in improving serum ferritin,and SXN+OI was most effective in increasing serum iron levels.JPSX+OI was optimal in reducing adverse pregnancy outcomes,while YQBX+OI effectively minimized adverse events.A cluster analysis suggested that SXN+OI could be the potentially optimal therapeutic regimen for IDA in pregnancy.CONCLUSION This study demonstrates that the combination of OI with CPMs offers better outcomes than OI alone.Based on clinical efficacy and other measured outcomes,SXN+OI emerges as the most effective treatment modality for improving the health of pregnant patients with IDA.

珍珠黄杨DNA的提取方法比较及ISSR反应体系的优化

为从珍珠黄杨幼叶中提取高质量的总DNA,比较了1%CTAB、2%CTAB、4%CTAB和1.5%SDS4种DNA提取方法。结果表明:2%CTAB提取的DNAA260/A280值最好,ISSR-PCR扩增效果最佳,是有效提取珍珠黄杨基因组DNA的方法。并在此基础上,以(CT)8G为引物,采用单因素实验法,确立了适合珍珠黄杨的ISSR-PCR反应体系:在总体积20μL的反应体系中,含30ngDNA模板,2.0mmol/LMg2+,0.30μmol/L引物,0.20mmol/LdNTPs,1UTaq酶。

黄杨属植物化学成分的研究进展

综述了近20年来黄杨属植物中生物碱、黄酮、甾醇、香豆素、木脂素和酸类化合物等7大类化学成分的研究结果,并介绍了黄杨的抑制酶活性、抗HIV病毒、抗菌及抗肿瘤等新的药理作用,为我国丰富的黄杨资源的开发利用提供参考。

瓜子黄杨扦插育苗技术

从苗圃地选择与整地作床、插穗采集和处理、扦插、插后管理、移植、苗期管理、苗木出圃等方面总结了瓜子黄杨(Buxus sinia)扦插培育技术,以期为瓜子黄杨的推广种植提供参考。

Protective Effects of Danggui Buxue Tang on a Human Umbilical Vein Endothelialcell Damage Induced by Advanced Glycation End Products

Advanced glycation end products (AGEs) have been regarded as a pivotal inducer in diabetes and kinds of diabetic nephropathy. The present studies explored the effects of Danggui Buxue Tang (DBT) that is a Chinese medicinal de- coction on negative charge to Human Umbilical Vein Endothelial Cell (HUVEC) and the related mechanism. Alcian blue staining was established to evaluate the intensity of negative charge on HUVEC. Proteoglycan expressions of AGP and avidin were determined by SDS-PAGE. We observed that DBT can significantly increase negative charge on HU-VEC and up-regulated AGP and avidin expressions and ameliorate AGEs-induced HUVEC apoptosis. Therefore, all results showed DBT had prevention effects against the progression of AGEs-induced damage, and this decoction might be promising agent against proteinuria in diabetic nephropathy.

Simultaneous Determination of Four Amino Acids in Fufang Ejiao Buxue Granule by HPLC

[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a Waters Sunfire C 18 column(4.6 mm×250 mm,5μm).A binary mobile solvent was used:mobile solvent A was acetonitrile-0.1 mol/L sodium acetate solution(adjusting pH to 6.5 with 36%acetic acid)(7∶93)and mobile solvent B was acetonitrile-H 2O(4∶1).The mobile phase was delivered at a flow rate of 1.0 mL/min with a gradient elution profile(0-13 min,100%A→93%A;13-17.9 min,93%A→88%A;17.9-29.0 min,88%A→85%A;29-39 min,85%A→66%A;39-45 min,66%A→0%A).The column temperature was at 43℃.The detection wavelength was 254 nm.[Results]The injection volume of L-hydroxyproline,glycine,alanine,and L-proline showed a good linear relationship with the chromatographic peak area in the range of 0.012 to 0.117,0.022 to 0.218,0.010 to 0.097,0.016 to 0.160μg,separately.The average recovery rate(n=6)was 96.4%,97.3%,97.1%,and 99.4%,respectively;the relative standard deviations were 1.2%,1.9%,1.7%,and 0.9%,respectively.[Conclusions]This method is simple in operation and good in reproducibility,and provides a reliable method for controlling the quality of Fufang Ejiao Buxue Granules.

Danggui Buxue decoction promotes proliferation and differentiation of Caco-2 cells and antagonizes obesity by stimulating apolipoprotein-IV expression

Background:Chinese medicine has been proposed as a novel approach to the prevention of metabolic disorders such as obesity.Danggui Buxue decoction,a decoction prepared from Huangqi(Astragali Radix)and Danggui(Angelicae Sinensis Radix),has been used to nourish vitality and enhance blood circulation in traditional Chinese medicine.However,the effect of Danggui Buxue decoction on obesity is still primarily unknown.Methods:Cell proliferation,differentiation,and apolipoprotein-IV transcription were investigated to explore the function of Danggui Buxue decoction by methyl thiazolyl tetrazolium assay,alkaline phosphatase assay,and luciferase assay,respectively.Results:Danggui Buxue decoction promoted cell growth by up to 15%(P=0.034)and induced cell differentiation by up to 38%(P=0.006)at 0.3 mg/mL.Moreover,Danggui Buxue decoction enhanced the transcription of apolipoprotein-IV by 2.4 times(P=0.027),activating its promoter by 28.9%(P=0.031)at 0.3 mg/mL.In addition,Danggui Buxue decoction functioned in a dose-dependent manner.Conclusion:These results suggest that Danggui Buxue decoction promotes cell differentiation by enhancing apolipoprotein-IV transcription and alkaline phosphatase activity,and it may also have a potential anti-obesity effect because apolipoprotein-IV transcription is closely related to the reduction of food intake.

Dissecting the underlying pharmaceutical mechanism of Danggui Buxue decoction acting on idiopathic pulmonary fibrosis with network pharmacology

Backgroud:Danggui Buxue decoction(DBD),a classical prescription in traditional Chinese medicine,has been found to have protective effect on bleomycin-induced pulmonary fibrosis in rats by reducing alveolar inflammation and fibrosis.However,the biological activity of individual chemical components and mechanism of action of whole formula are not clear.Methods:Potential targets of active ingredients of DBD were collected through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and SymMap database.Target genes related to idiopathic pulmonary fibrosis were obtained from the Online Mendelian Inheritance in Man database,Therapeutic Targets Database and Gkb database.Then,the common targets were obtained by overlapping the potential targets of active ingredients in DBD and diseases related targets.The selected targets were subjected to Kyoto Encyclopedia of Genes and Genomes signaling pathway and Gene Ontology analysis,and the network map of active component-target-pathway was established using Cytoscape 3.7.1 software.The active components of DBD with most targets were selected for fibrosis-related marker verification.The mRNA and protein expression of fibrosis markers,α-smooth muscle actin,collagen 1 and fibronectin,were detected in TGF-β1-induced fibroblast cell line after treatment with the active components.Results:The 14 active ingredients,such as quercetin and kaempferol,were screened from DBD.It acts on 26 targets like estrogen receptor 2 and prostaglandin-endoperoxide synthase 2,and mainly involves 38 signaling pathways such as cell inflammation and autophagy.Kaempferol and quercetin are the two compounds with the highest network regulation,which can inhibit the transformation of fibroblasts into myofibroblasts and reduce the expression of fibrosis markersα-smooth muscle actin,collagen 1 and fibronectin.Conclusion:The integration mode of multi-component,multi-target,multi-channel and mechanism of DBD in the treatment of idiopathic pulmonary fibrosis are predicted by means of network pharmacology.Our study could indicate the direction of further anti-fibrotic mechanism research.

Potential Mechanism of Danggui Buxue Decoction in Treating Iron Deficiency Anemia Based on Network Pharmacology and Molecular Docking Technology

[Objectives]To explore the potential mechanism of Danggui Buxue Decoction in treating the iron deficiency anemia(IDA)based on network pharmacology and molecular docking technology.[Methods]The active components and target proteins of Danggui Buxue Decoction were searched in databases such as TCMSP,OMIM,GeneCards,Drugbank,String,Metascape,etc.,and the target proteins shared with IDA were screened out,and the information about the signal pathways and biological functions of these target proteins was obtained.[Results]17 active components of Danggui Buxue Decoction and 24 potential targets for the treatment of IDA were obtained.With the aid of String database and Cytoscape software,the protein interaction network was obtained and the network topology analysis was performed.Four potential core targets with higher scores were obtained,namely F2,NOS2,NOS3,and PPARG.Using the Metascape database,GO function enrichment analysis and KEGG pathway enrichment analysis were performed on the potential targets of Danggui Buxue Decoction in the treatment of IDA,and the important biological processes,cell composition,molecular functions and signal pathways related to the target were screened through the R language.The results show that biological processes are related to positive regulation of growth,cell composition is related to membrane microdomain,and molecular functions are related to oxidoreductase activity.The signal pathways involved are mainly AGE-RAGE signal pathway,TNF signal pathway and IL-17 signal pathway.Finally,the molecular docking results confirmed that the active components of Danggui Buxue Decoction have a good binding ability with the target.[Conclusions]Danggui Buxue Decoction treats the IDA through multiple components,multiple targets,multiple signal pathways,and multiple biological functions.

Study on the Molecular Mechanism of Danggui Buxue Decoction in Intervention of Perimenopausal Syndrome Based on Network Pharmacology and Molecular Docking

[Objectives]This study was conducted to explore the intervention mechanism of Danggui Buxue Decoction in perimenopausal syndrome based on network pharmacology and molecular docking.[Methods]The chemical components and targets of Danggui Buxue Decoction were acquired through the TCMSP database,and the main targets of perimenopausal syndrome were obtained through the GeneCards database.The component targets and disease targets were intersected,and combining with active components and Chinese herbs in the decoction,a traditional Chinese medicine-component-target network was constructed using Cytoscape 3.7.1 software.The STRING platform was employed for protein-protein interaction analysis.The DAVID analysis platform was used to conduct target GO and KEGG enrichment analysis,so as to predict the action mechanism Danggui Buxue Decoction.Finally,an active component-disease target-signal pathway network diagram was constructed.[Results]Twenty two components in Danggui Buxue Decoction related to perimenopausal syndrome and 120 corresponding targets were obtained,including active components such as 1,7-dihydroxy-3,9-dimquercetin and kaempferol,and key targets such as TNF,ESR1 and PPARG.The results of GO analysis and KEEG analysis indicated that Danggui Buxue Decoction might regulate the transcription of RNA polymerase II promoter,DNA templating,gene expression,signal transduction,hypoxia response and other biological processes by regulating multiple signal pathways such as chemical carcinogenesis-receptor activation,cancer pathways,lipid and atherosclerosis,tryptophan metabolism,malaria,steroid hormone biosynthesis and chemical carcinogenesis-DNA adduct.[Conclusions]Danggui Buxue Decoction intervenes in perimenopausal syndrome through multiple components,targets and pathways,providing a basis for elucidating the intervention mechanism of Danggui Buxue Decoction and expanding its clinical application.

Exploring the mechanism of Danggui Buxue Decoction in treating osteoporosis from vitamin D-axis

Osteoporosis is a common disease in the middle-aged and elderly population and is a global health problem that needs to be solved urgently.Vitamin D deficiency is closely related to osteoporosis,and it is of practical significance to prevent and treat kidney and strengthen bone therapy.A large number of literature have confirmed that qi and blood to represent Danggui Buxue Decoction has a reliable therapeutic effect on osteoporosis,but the mechanism is still unclear.Based on the previous work of the research group,it is believed that the vitamin D axis may be a potential target for TCM Bushen Recipe.Syndrome and vitamin D deficiency have similar pathological changes,so it is suggested that Danggui Buxue Decoction may regulate osteoporosis by regulating the vitamin d axis.

Immunotherapy with activation of NK cells

Given the role of CAR-T cell in the fight against cancer cells,as well as the role of NK cell as an innate immune cell with anti-tumor function too,we hypothesize that Insert the CD16 antibody-producing gene or activator protein CD16,CD16 is one of the most important markers of NK cell activation for ADCC,into the CAR-T cell panel.So with this hypothesis in addition to the role of CAR-T cells to fight cancer,the role of NK cells in the fight against cancer too.