Profiling the Change of Key Chemical Ingredients in Combination of Aconitum carmichaeli Debx. and Bletilla striata (Thunb.) Reichb.f. by UPLC-QTOF/MS with Multivariate Statistical Analysis

In the present study, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) based chemical profiling approach to rapidly evaluate chemical diversity after codecocting of the combination of Aconitum carmichaeli Debx.(wu-tou in Chinese, WT) and Bletilla striata(Thunb.) Reichb.f.(bai-ji in Chinese, BJ) incompatible pair. Two different kinds of decoctions, namely WT-BJ mixed decoction: mixed water extract of each individual herbs, and WT-BJ co-decoction: water extract of mixed two constituent herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-QTOF/MS analysis, the datasets of tR-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis(OPLS-DA) to holistically compare the difference between these two kinds of decoction samples. Once a clear classification trend was found in score plot, extended statistical analysis was performed to generate S-plot, in which the variables(tR-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components that correlate to these ions were regarded as the most changed components during co-decocting of the incompatible pair. The identities of the changed components can be identified by comparing the retention times and mass spectra with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of the known compounds published in the literatures. Using the proposed approach, global chemical difference was found between mixed decoction and co-decoction, and hypaconitine, mesaconitine, deoxyaconitine, aconitine, 10-OH-mesaconitine, 10-OH-aconitine and deoxyhypaconitine were identified as the most changed toxic components of the combination of WT-BJ incompatible pair during co-decocting. It is suggested that this newly established approach could be used to practically reveal the possible toxic components changed/increased of the herbal combination taboos, e.g. the Eighteen Incompatible Medications(Shi Ba Fan), in traditional Chinese medicines.

Comparative study on the effects of hepatic arterial embolization with Bletilla Striata or gelfom in the treatment of primary hepatic carcinoma

AIMS To study the effect and safety of hepatic arte- rial embolization (HAE) with Bletilla Striata powders of traditional Chinese drug in the treatment of primary hep- atic carcinoma (PHC). METHODS From May 1990 to September 1993,106 patients with PHC were treated by HAE with Bletilla Striata powders (n=56) or gelfoam powders (n=50) respectively under the controlled technical conditions. Authors analyzed the effects and complications in two groups. RESULTS Bletilla Striata powders produced exten- sive permanent proximal embolization of hepatic artery and collateral circulation formed. Intermission of treat- ment was as long as 6-12 months and tumor necrosis and shrinking were obvious. The survival rates of 1,2 and 3 years were 81.9%,44.9% and 33.6% respec- tively and the mean survival time was 19.8 months without serious complications. All clinical effects of Bletilla Striata group were better than those of glfoam group. CONCLUSIONS Bletilla Striata powders are superi- or to gelfoam powders as an angioembolus for hepatic carcinoma.

Advances in Tissue Culture Technology of Bletilla striata

The reports about tissue culture technology of Bletilla striata were summa- rized in this paper, and explants for tissue culture, induction differentiation of callus, rooting culture and transplanting, effect of growth regulator and influence of external environment were expounded. This study provides support for further research.

The Rapid Propagation and Domestication of Wild Bletilla striata

The research explored rapid multiplication on Bletilla striata and the re- suits showed that the medium of MS＋2.5 mg/L6-BA＋0.2 mg/L NAA performed the best in induction, with induction rate at 98%; the medium with MS＋IAA2.5mg/L＋ 2.5 mg/L6-BA＋2.0 mg/L NAA was the best in terms of induction and multiplication; the medium with 1/2MS＋0.01 mg/L NAA 1＋0.01 mg/L IAA promoted rooting.The harden- ing temperature was the optimal at 23℃-28 ℃ and relative humidity of 80%-90% the best.

A New Bibenzyl Derivative from Bletilla striata

A new compound was isolated from Bletilla striata. Its structure was elucidated as 5-hydroxy-4-(p-hydroxybenzyl)-3, 3-dimethoxybibenzyl by spectroscopic method.

Tierexperimentelle Untersuchungen zur Effektivittder transarteriellen Chemoembolisation (TACE) unter Einsatz von Bletilla striata

Zusammenfassung： Evaluierung der Effektivitat yon Bletilla striata bei transerteriellen Chemoembolisation （TACE） im Kleintiermodell. Nach subkapsularer Implantation （Tag 0） eines soliden Morris Hepatoms （2 mm^3） bei ACI-Ratten （n=30） wurden am 13. Tag die Tumorvolumina （V1） magnettomographisch bestimmt. Im Anschlu erfolgten nach Laparatomie die retrograde Katheterisierung der Arteria gastroduodenalis und die Anwendung folgender Therapieprotokolle.. （A） TACE （0,1 mg Mitomycin C ＋ 0,1 mL Lipiodol） ＋ Bletilla striata （1,0 mg） （n=10）; （B） TACE （ Bletilla striata （1,0mg） ＋ Ligatur （n=10）;（C） TACE allein （Kontrollgruppe, n=10）. Zur Effektivitatsbeurteilung der unterschiedlichen Therapieprotokolle erfolgte am 28. Tag eine erneute magnettomographische Bestimmung der Tumorvolumina （V2）. Das durchschnittliche Tumorvolumen der Gruppen A, B und C vor und nach interventioneller Therapie betrug 0,0355 cm^3 und 0, 2248 cm^3 , 0,0374 cm^3 und 0,0573 cm^3 sowie 0,0380 cm^3 und 0,3674 cm^3. Das errechnete durchschnittliche Verhaltnis （V2/V1） betrug 6,28, 1,53 sowie 9,14. Im Vergleich zur Kontrollgruppe C konnte somit fur Therapiegruppe B eine signifikant （P〈0.01） geringere Tumorvolumenzunahme im Beobachtungszeitraum ermitteh werden, wkhrend zwischen Gruppe A und Gruppe C keine statistisch signifikanten （P〉0.05） Unterschiede im Tumorwachstum festgestelh werden konnten. Das Wachstum der induzierten Lebertumoren konnte unter Behandlung mittels TACE ＋ Bletilla striata ＋Ligatur im Tiermodell statistisch signifikant im Vergleich zur Kontrollgruppe gehemmt werden. Schlüsselworter： Lebertumor; TACE; Bletilla striata

Experimentelle Embolisation unterschiedlicher hepatischer Gefaβe beim Hund mit Bletilla striata

Diese vergleichende Untersuchung uber Embolisation differenter hepatischer GefaBe bei Hunden mit Bletilla striata und Gelfoam ergab,d aB letilla striala als Embolisationsmaterial bessere Erfolge zeigt als Gelfoam.Die Mechanismen der Embolisation mit Bletilla striata sind folgende :1)Bletilla striata wird nicht leicht von Geweben absorbiert;2)Bletilla striata bewirkt mechanische Blockierung;3)Bletilla striata beeinflt das Gerinnungssystem und dasAntikoagulationssystem;4)Bletilla siriata kann die Geafwandschadigen und dadurch sekundare Blockierung auslosen.

Research Progress on Chemical Constituents and Biological Activity of Kummerowia striata(Thunb.) Schindl.

The purpose of this paper is to review the research progress on the chemical composition and pharmacological action of Kummerowia striata(Thunb.)Schindl.,so as to provide reference for the chemical research of natural medicine and the discovery of new drugs.K.striata(Thunb.)Schindl.contains a variety of active constituents,such as flavonoid glycosides,phenols and amino acids,and has a wide range of pharmacological activities,including anti-allergy,analgesia and anti-inflammation,anti-oxidation,anti-tumor and so on.It is mainly used in the folk for the treatment of cold and fever,summer-heat and dampness,vomiting and diarrhea,dysentery and so on.

Research on the effect of Bletilla striata and the mechanism of the treatment of bronchoplumonary inflammation based on network pharmacology

Objective:To explore the effect of active compounds of Bletilla Striata,and to predict its mechanism of the treatment of bronchoplumonary inflammation.Methods:Retrieved active compounds of Bletilla Striata from relevant databases include TCMSP,while retrieved action targets of active compounds and bronchoplumonary inflammation.Received corresponding gene symbols of targets by querying UniProt database.Then drew the compound-target network of Bletilla Striata and action targets network between active compounds and bronchoplumonary inflammation using Cytoscape 3.2.1.Subsequently performed GO functional and KEGG enrichment analysis of key targets through relevent databases including Metascape,meanwhile,analysed Protein-protein interaction using STRING.Results:Bletilla Striata contained 29 active components and 632 corresponding targets.The more important targets included AURKA,BRAF,RAF1,MMP1 and ESR2,213 in total.GO functional enrichment analysis resulted in 2,398 items(P<0.05),including 2096 biological process(BP)items,133 cellular component(CC)items,169 molecular function(MF)items.KEGG enrichment analysis resulted in 160 signal pathways items(P<0.05).There were 33 common targets between 335 action targets of bronchoplumonary inflammation and 95 more important active components of Bletilla Striata.GO functional enrichment analysis resulted in 547 items(P<0.05),including 499 BP items,19 CC items,29 MF items.KEGG enrichment analysis resulted in 46 signal pathways items(P<0.05),mainly involving the IL-17 signaling pathways,platelet activation,endocrine resistance,Fc epsilon RI signaling pathway,Th17 cell differentiation,etc.6 core targets including MAPK14,MMP9,HSP90AA1,ROCK2,PRKCB and PTGS2 were obtained by PPI analysis.They were involved in IL-17 signaling pathway,platelet activation,leukocyte migration,VEGF signaling pathway and Th17 cell differentiation respectively.Conclusion:The study explains"multi-component-multi-target-multi-pathway-multi-disease"is the characteristic of Bletilla Striata using network pharmacology,and predict the active compounds of Bletilla Striata can be used for treatment of bronchoplumonary inflammation with the effect of regulating to anti-inflammation and tissue repair and remoding.This study theoretically verified the functional utility of Bletilla Striata and provided scientific basis and reference direction for further experimental research.

Compound Planting of Bletilla striata under Forest in the Three Gorges Dam Area

[Objectives] The aim was to study the artificial cultivation of Bletilla striata to realize large scale and standardization planting. [Methods] The comparison tests were conducted on B. striata with different canopy densities of the Magnolia officinalis forests,different compound planting densities and different tending measures and management. [Results] When the stand canopy density was 0. 4-0. 6,the per unit yield of B. striata was 5. 4%,6. 8% higher than that at the canopy density of less than 0. 4 and more than 0. 6,respectively. When the planting density was 30 cm × 30 cm,the per unit yield increased by 16. 1%,12. 0%,13. 1% respectively compared with the planting density of 20 cm × 20 cm,25 cm × 25 cm,35 cm × 35 cm. When B. striata was planted from October to November,the per unit yield was 5. 6% higher than that planted from December to January of the following year,and 21. 3% higher than that from February to March of the second year. When farmyard manure was applied during the cultivation,the per unit yield was 31. 7% and 18. 4% higher than the application of chemical fertilizer and compound fertilizer. When weeding 4 times per year,the per unit yield increased by 240. 1%,137. 0% and 43. 9% respectively from that weeding 1 times,2 times,3 times per year. [Conclusions]When planting B. striata,the stand canopy density of 0. 4-0. 6 could make it receive absolutely shelters and the lighting conditions required for the growth,thereby bringing in high emergence rate,good growth potential and high yield. The best planting effect of B. striata could achieve by planting from October to November with the planting density of 30 cm × 30 cm,which can play the maximum benefit of individual plants. Moreover,weeding 4 times per year combined with the use of farmyard manure can promote the development and growth of tubers,which can greatly improve the yield of B. striata.

共载黄芪甲苷-黄连素/白及多糖微球的制备与性能研究

利用白及多糖的生物黏附性制备共载黄芪甲苷-黄连素/白及多糖微球(AS-IV-BBR/BSP-Ms),并对其性能进行评价。结果表明,采用乳化交联法制备得到共载AS-IV-BBR/BSP-Ms,微球粒径在30~40μm,Zeta电位为-21.4 mV,且30 min内溶胀率达54.3%;红外光谱分析、X-射线衍射分析表明,黄芪甲苷和黄连素被包裹于白及多糖微球内部;DSC分析表明,在t=121℃时,出现尖锐吸热峰,热稳定性较好;体外抗氧化活性实验表明,该微球DPPH自由基的半数清除浓度(IC_(50))为8.23 mg/mL。该制备工艺稳定可行,微球性能评价达到预期目标,可为下一步药效学评价提供实验依据。

白芨外泌体的分离及对体外人永生化角质形成细胞(Hacat)的凋亡保护研究

探索鲜白芨(Bletilla striata)外泌体对体外人永生化角质形成细胞(Hacat)的生物活性,为其在护肤品上的开发提供数据。聚乙二醇(PEG)沉淀法提取白芨外泌体,透射电镜鉴定,BCA法测蛋白浓度;细胞计数试剂(Cell Counting Kit-8,CCK8)法检测Hacat细胞存活率,流式双染检测凋亡,蛋白印迹法检测凋亡相关通路蛋白。白芨外泌体呈茶杯状具双层膜,平均粒径69.63 nm,每克鲜白芨含外泌体浓度为5.24E+8 Particles,内含蛋白19.53μg;白芨外泌体(≥5μg/mL)可显著提高体外Hacat细胞存活率(P<0.05);10μg/mL白芨外泌体处理48h,可显著降低H_(2)O_(2)氧化造模引起的Hacat细胞早凋亡、晚凋亡和总凋亡的细胞比例(P<0.01),同时,模型组caspase-9和caspase-3表达显著上升,外泌体处理能显著降低其表达,而Bcl-2表达在组间无差异(P<0.05)。PEG沉淀法分离的白芨外泌体,可促进体外Hacat生长,抑制H_(2)O_(2)引起的细胞凋亡且与蛋白caspase-9和caspase-3相关。

星点设计-效应面法优化白及多糖pH-离子敏感原位凝胶的处方

目的优化白及多糖pH-离子敏感原位凝胶的处方。方法以去乙酰结冷胶用量、海藻酸钠用量、氯化钙用量、枸橼酸钠与氯化钙的质量比为考察变量,采用单因素实验初步确定各变量的范围。以25℃凝胶前体溶液黏度和37℃胶凝黏度为考察指标,采用星点设计-效应面法优选白及多糖pH-离子敏感原位凝胶的处方工艺,并进行预测分析。结果白及多糖pH敏感原位凝胶的最优处方为:去乙酰结冷胶用量为0.50%,海藻酸钠用量为1.10%,氯化钙用量为0.07%,枸橼酸钠与氯化钙的质量比为4∶1。该处方条件下,25℃前体溶液黏度为(704.00±3.00)mPa·s,37℃胶凝黏度为(12544.70±47.33)mPa·s。结论白及多糖pH-离子敏感原位凝胶稳定性好,前体溶液在25℃流动性良好,凝胶强度良好,可为基于白及多糖原位凝胶的后续研究提供实验基础。

中药白及在治疗慢性创面修复中的应用进展

白及Bletilla striata(Thunb.)Reichb.f.是一种传统中药,具有收敛止血、消肿生肌等功效,常用于外伤出血、疮疡肿毒等。古籍记载及现代临床发现白及具有治疗慢性难愈性创面的功效,为深入研究白及治疗慢性难愈性创面的活性物质基础、药理作用及机制,因此对近年来国内外有关白及的报道进行了综述。在创面修复过程中,白及具有止血、抗炎、抗病原微生物、抗氧化、免疫调节、促进表皮生长因子增殖、分泌胶原蛋白等药理作用,可以通过调节伤口的微环境来促进慢性难愈性伤口的愈合。同时提出了白及在临床应用中存在的问题和解决办法,展望了白及在慢性创面修复中的应用前景,为合理开发白及资源用于临床伤口护理提供理论依据。

响应面优化硒化白及多糖的制备工艺及其体外抗氧化活性研究

通过优化硒化白及多糖的制备工艺,探究是否获得较高抗氧化活性的硒化白及多糖。以白及多糖(Bletilla striata polysaccharide,BSP)为原料,亚硒酸钠(Na_(2)SeO_(3))为硒化剂,冰乙酸为催化剂对白及多糖进行硒化修饰制备硒化白及多糖(Selenized Bletilla striata polysaccharide,BSP-Se)。以硒含量为指标,利用单因素实验和响应面法对制备工艺进行优化。通过傅里叶红外光谱(Fourier transform infrared spectroscopy,FT-IR)确定白及多糖是否硒化成功,并通过测定DPPH自由基、ABTS^(+)自由基以及羟基自由基的清除能力综合评价硒化白及多糖的体外抗氧化活性。结果显示,在最佳工艺条件为Na_(2)SeO_(3)与BSP的质量比1.5、冰乙酸与BSP用量比3、反应时间6 h、反应温度80℃的条件下制备的BSP-Se硒含量最高,可达到7.831 mg/g。FT-IR分析显示硒与多糖通过Se-O-C和O-Se-O结合,表明白及多糖已被成功硒化;自由基清除能力分析显示BSP-Se对DPPH、ABTS^(+)和·OH自由基的IC_(50)值分别为2.875、4.431和0.314 mg/mL,显著低于BSP的IC_(50)值5.828、8.475和0.833 mg/mL(P<0.05),表明硒化修饰提高了BSP的抗氧化能力。因此,通过硒化修饰可以提高BSP的抗氧化能力,为进一步研究硒多糖作为新型抗氧化剂在食药领域、保健品领域、日化领域的应用提供一定的理论基础。

直播条件下水分胁迫对白及种子萌发的影响

为解决白及直播种子萌发不稳定的问题,本研究以椰糠为基质,设置4个含水量梯度,研究直播条件下水分胁迫对白及种子萌发过程的影响。结果表明:种子直播后主要经历吸水膨胀成球状体(a)、球状体转绿(b)、球状体一端长出第一片叶(c)、长出丛毛状假根(d)、长第二片叶(e)和长根(f)6个阶段。不同含水量基质上,最高萌发率为68.3%,最低萌发率为0,总体趋势为随着基质含水量增加萌发率提高,从不同阶段转化率来看,水分条件影响的关键阶段为b和e阶段;从萌发时间看,最终完成长根需要82.3~86.7 d,总体趋势随着基质含水量的增加萌发时间减少,其中从播种到c阶段转化时间随基质含水量增加而减少,e到f阶段转化时间随基质含水量增加而增加。基于以上结果,在白及种子直播育苗生产中,可以在萌发前期增加基质含水量,最后阶段适当降低基质含水量来提高萌发率,缩短萌发时间。

白及锈病病原菌的鉴定及抗锈病资源筛选

为鉴定引起广西种植区白及锈病的病原菌种类且筛选抗锈病的白及资源,该研究对白及锈病病原菌进行分离,并采用形态学和分子生物学的方法对病原菌进行鉴定,同时通过人工接种病原菌法对23份白及进行锈病抗性评价以及筛选抗锈病的白及资源。结果表明:(1)从白及感病叶片中分离的锈病病原菌X2夏孢子呈金黄色,卵圆形或椭圆形,大小为(21.43~30.95)μm×(13.10~19.05)μm;冬孢子呈橘红色、红褐色,倒卵形或棍棒状,大小为(17.25~30)μm×(5.5~6.65)μm。(2)把菌株X2全长689 bp的ITS序列(OQ826009)与GenBank已登陆的序列进行相似性分析发现,菌株X2与Coleosporium sp.(KY783686.1)匹配度最高,序列一致性为95.86%,但系统发育树表明,X2与Coleosporium bletiae(MN108161.1,OP363680.1)聚为一类群;结合形态学和分子生物学的方法,鉴定菌株X2为Coleosporium bletiae。(3)人工接种菌株X214 d后,23份白及的病情指数范围在0~70.7之间,并根据病情指数将23份白及划分为6个抗性等级,即表现为免疫的白及1份,病情指数为0;表现为高抗的白及4份,病情指数为1.7~4.7;表现为抗病的白及6份,病情指数为5.6~9.4;表现为中抗的白及5份,病情指数为12.7~18.3;表现为感病的白及5份,病情指数为32.0~49.1;表现为高感的白及2份,病情指数为62.2~70.7。综上认为,表现为免疫和高抗的5份白及(分别来自云南红河、广西恭城、广西百色、贵州遵义、湖北宜昌)病情指数低、抗锈病能力强,可推广应用或作为培育抗锈病优良种质的亲本材料。该研究结果为后续开展白及锈病的有效防治与致病机理研究提供了支撑。

四种杀菌剂在白及初加工过程中的残留变化

为明确白及初加工过程中多菌灵、三唑酮、戊唑醇和苯醚甲环唑的残留变化规律和加工因子,在实验室条件下,模拟白及的清洗、水煮和烘干3个初加工过程,采用超高效液相色谱串联质谱法(ultra high performance liquid chromatography-tandem mass spectrometry,UHPLC-MS/MS)检测白及初加工各阶段的农药残留浓度,并计算加工因子。结果表明,白及不同加工过程对4种杀菌剂残留浓度的影响差异较大。清洗过程使农药残留浓度降低4.6%~57.8%,加工因子为0.42~0.95,在清水中分别添加质量分数2%淀粉、2%食盐和2%小苏打对农药的去除效果可提升1.4~7.3倍;水煮过程使农药残留浓度降低38.6%~67.8%,加工因子为0.32~0.61;烘干过程由于浓缩效应使农药残留浓度升高2.1~3.2倍,加工因子为2.08~3.23。白及依次经2%食盐水清洗、水煮4 min、60℃烘干的初加工全过程对4种杀菌剂的降解作用大于因脱水造成的浓缩作用,可降低4种杀菌剂在干白及产品中的残留风险,全过程加工因子为0.34~0.67。因此,以鲜白及中4种杀菌剂残留评价经此初加工后干白及中4种杀菌剂残留的安全性是可行的,研究结果可为白及产品的安全风险评估提供基础数据参考。

白及根腐病病原菌的鉴定及抑制效应研究

为明确鉴定白及块茎腐烂病(根腐病)的病原菌,并筛选能够抑制病原菌的中药材提取物。该研究利用常规组织分离法对病原菌进行分离,通过形态学和分子生物学技术对致病菌株进行鉴定,同时观察了7种中药材提取物对病原菌的抑菌效果。结果表明:(1)从感病叶片、叶鞘及块茎中共分离得到14株真菌和4株细菌,病原菌室内和室外回接表明菌株GF-1致病症状与田间一致,致病率均达到100%。(2)经形态学鉴定,菌株GF-1为附球菌属(Epicoccum)病原菌,菌落白色绒絮状,圆形;菌丝匍匐向外、向上生长,气生,无色,有隔膜,有分枝,具有分生孢子和厚垣孢子。(3)菌株GF-1的ITS序列(全长522 bp)与GenBank中已登录的甘蔗的高粱附球菌(E.sorghinum,MN493119.1)序列一致性最高,达99.62%,与已报道的白及叶斑病致病菌高粱附球菌(E.sorghinum,MF948994.1)的一致性为98.88%。(4)培养基中含有0.1~0.2 g·mL^(-1)的青钱柳等7种中药材提取物,能够完全抑制GF-1菌落的生长;当培养基中含有0.05 g·mL^(-1)的提取物时,肉桂和丁香提取物仍然能够完全抑制菌落的生长。综上认为,引起白及根腐病的致病菌为附球菌属高粱附球菌(E.sorghinum),培养基中分别含有0.1~0.2 g·mL^(-1)的青钱柳、白及、厚朴、八角、肉桂、蛇床子和丁香7种中药材提取物且均能完全抑制致病菌的生长。该研究结果为白及根腐病的防治提供了理论指导。

白及多糖的提取、纯化技术及结构表征研究进展

中药白及Bletilla striata(Thunb.)Reichb.f.是临床上常用的中药材之一,广泛用于各种出血、疮痈肿毒、皮肤皲裂等,具有很高的药用价值,白及多糖是中药白及的主要有效成分。大量研究发现,白及多糖具有止血、抗炎、抗氧化、黏膜保护和抗肿瘤等多种药理活性。同时,白及多糖还具备优良的生物相容性、生物可降解性、生物粘附性等特性,在生物材料及工业领域被广泛应用。近年来,白及多糖的提取、分离、纯化及结构表征相关研究文献较多。然而,各提取工艺及纯化方法对白及多糖的提取率和结构特征差异较大。综述了白及多糖的提取方法、纯化方法和结构特征研究进展,以期为白及多糖的进一步研究和开发提供帮助。