Physiological Effect and Quality Influences of Hedyotis diffusae Extract on Tobacco Infected by TMV

The extract from Hedyotis diffusae was used to treat tobacco plants infected by TMV before and after incidence. The effect of the changes of some physiological and biochemical indices related with the virus-resistance in tobacco leaves on tobacco quality ware determined. The resuits showed that H. diffusae extract could remarkably increase the total phenol content in tobacco leaves ,strength the activity of phenylalanine ammonialyase （ PAL） , improve the membrane permeability and tobacco quality. Prevention treatment was better than therapy treatment. In conclusion, H. diffusae extract could induce plants to produce disease resistance and strength tha antiviral capability of tobacco against TMV.

Anti-tumor research progress of Herba Hedyotidis Diffusae

Cancer,as one of the major public health problems in the world,dramatically harms human health.In recent years,studies have found that Herba Hedyotidis Diffusae not only plays a huge role in anti-infection but also has a strong effect on anti-tumor.Several studies have shown that the components contained in Herba Hedyotidis Diffusae have different levels of antitumor results in multiple pathways.This article summarizes the antitumor elements in Herba Hedyotidis Diffusae,and describes the antitumor mechanism of Herba Hedyotidis Diffusae.Provide new ideas for further clinical use.

Anti-tumor Effect and Protective Effect on Chemotherapeutic Damage of Water Soluble Extracts from Hedyotis diffusa

Bai-Hua-She-She-Cao Hedyotis diffusa Willd. (Ru-biaceae) is a medicinal herbwidely distributed in northeast Asian countries. In traditional Chinese medicine, it has the effectof 'clearing away heat and toxic material, promoting blood circulation and removing blood stasis'.It is a well known Chinese folk-medicine used for the treatment of appendicitis, sore throat, mumps,acne, sebo-rheic dermatitis and various kinds of tumors, such as tumors of digestive tract,carcinoma of liver. It was reported that the MeOH extract of H. diffusa demonstrated a significantantitumor activity and ursolic acid succeeded in being isolated from the MeOH extract as an activecomponent . Shan BN, et al suggested that the direct aqueous extract of H. diffusa hadimmuno-modulating activity and antitumor activity in vitro through stimulating the immune system tokill or engulf tumor cells. But regarding anti-tumor activity in vivo of water soluble extracts fromH. diffusa, no detail was reported. Therefore, we prepared water soluble extracts (H_1 and H_2)from H. diffusa and evaluated their anti-tumor property in vivo experiments as well as protectiveeffect on chemo-therapeutic damage.

大豆叶部病害研究的简易离体叶片培养技术

为寻找简易有效的大豆叶部病害研究方法,利用大豆离体叶片,进行了相关试验。试验结果表明,大豆离体复叶培养30 d,新根长达16.8～41.2 cm,平均根长国>24 cm,移至营养杯30 d后,叶色及叶片仍保持完好。经离体叶片法和整株法接种大豆锈病病原菌后,不同品种和不同菌株表达出不同的病斑反应型;接种大豆白粉病病原菌,也同样表达出在不同品种和不同接种浓度条件下,大豆白粉病发生程度不同;两种接种方试验结果一致。本方法试验条件可控、易控,省时,省空间,因此可考虑应用于大豆叶部病害研究的各种试验。

微波辅助酶法提取白花蛇舌草多糖的工艺优化

为优化微波辅助酶法提取白花蛇舌草(Hedyotis diffusa Willd.)多糖工艺。在单因素试验基础上,应用中心组合设计,采用二次多项式逐步回归分析对提取工艺条件进行优化,确定微波辅助酶法提取白花蛇舌草多糖工艺最佳条件为酶解时间2.9 h,酶解温度60℃,酶用量0.9%,p H 6.0;按此工艺条件,提取3次,白花蛇舌草多糖平均含量可达31.02%。

微波辅助酶法提取白花蛇舌草总黄酮的工艺研究

为优化微波辅助酶法提取白花蛇舌草(Hedyotis diffusa)总黄酮工艺,在单因素试验基础上,应用中心组合设计,采用二次多项式逐步回归分析方法对提取工艺条件进行了优化,确定微波辅助酶法提取白花蛇舌草总黄酮工艺最佳条件为酶解时间1.5 h、酶解温度45℃、酶用量1.4%、p H 5.5,按此工艺条件进行验证,白花蛇舌草总黄酮提取率可达4.42%。

乙醇-磷酸氢二钾双水相体系萃取白花蛇舌草总黄酮的工艺研究

利用乙醇-磷酸氢二钾双水相体系对白花蛇舌草(Hedyotis diffusa)总黄酮进行萃取分离。通过单因素实验分别考察了m(K2HPO4)、超声时间及超声温度对白花蛇舌草总黄酮萃取效果的影响,并用响应面法优化了萃取工艺。经优化后,白花蛇舌草总黄酮的最佳萃取条件为:m(K2HPO4)2.0g,超声时间37min,超声温度57℃。在此条件下,白花蛇舌草总黄酮的萃取率可高达99.45%。本方法精密度良好,准确度较高,可为白花蛇舌草总黄酮的高效萃取提供参考。

A novel polyherbal formulation containing thymoquinone attenuates carbon tetrachloride-induced hepatorenal injury in a rat model

Objective:To evaluate a novel polyherbal formulation(BSVT)containing the standardized extracts from the leaves of Boerhavia diffusa,Solidago virgaurea,Vitex negundo,and thymoquinone in CCl4 induced hepatorenal toxicity in rats.Methods:A total of 36 rats were divided into six groups including normal control,CCl4(2 mL/kg,i.p.),CCl4(2 mL/kg,i.p.)+Cystone?(750 mg/kg p.o.),CCl4(2 mL/kg,i.p.)+BSVT(25 mg/kg,p.o.),CCl4(2 mL/kg,i.p.)+BSVT(50 mg/kg,p.o.),and CCl4(2 mL/kg,i.p.)+BSVT(100 mg/kg,p.o.).All treatments were given for four weeks.Serum levels of aspartate transaminase,alanine transaminase,alkaline phosphatase,cholesterol,total protein,serum urea,blood urea nitrogen and creatinine were assessed.Superoxide dismutase,malondialdehyde,and glutathione peroxidase were evaluated in tissue homogenate.The histopathological study of liver and kidney tissues was also done.Results:Aspartate transaminase,alanine transaminase,alkaline phosphatase,cholesterol,serum urea,blood urea nitrogen and creatinine were significantly elevated(P<0.001)while total protein was considerably reduced in the CCl4 group as compared to the normal control(P<0.001),which indicated hepatorenal toxicity.In addition,superoxide dismutase and glutathione peroxidase activities were significantly decreased(P<0.001)while malondialdehyde levels were increased markedly(P<0.001).Treatment with BSVT formulation recovered these parameters towards a normal level in a dose-dependent manner.Conclusions:BSVT formulation ameliorates the hepatorenal toxicity in a dose-dependent manner.Furthermore,clinical studies are required to confirm its efficacy.

白花蛇舌草总黄酮提取工艺及其抗氧化活性研究

研究白花蛇舌草(Hedyotis diffusa Willd)总黄酮的提取工艺及其体外抗氧化活性。以白花蛇舌草总黄酮提取率为评价参数,通过单因素试验、正交试验优化提取工艺;测定了总还原力及Fe2+螯合力、羟自由基和超氧阴离子清除能力研究白花蛇舌草总黄酮的抗氧化活性。结果表明,用95%乙醇与白花蛇舌草粉末按40∶1(m L∶g),在90℃下提取180 min,此时提取率最高,达(2.22±0.01)%。体外抗氧化活性结果显示白花蛇舌草总黄酮具有较强的总还原力,Fe2+螯合率最高为90%,羟自由基和超氧阴离子的清除率分别达79%、77%。白花蛇舌草总黄酮具有显著的体外抗氧化活性,具有进一步开发利用的价值。

Phytochemical screening and in vitro bioactivities of the extracts of aerial part of Boerhavia diffusa Linn.

Objective:To investigate the bioactivities of crude n-hexane,ethyl acetate and methanol extracts of aerial part of Boerhavia diffusa Linn.(B.diffusa) and its phytochemical analysis.Methods: the identification of phyloconstituents and assay of antioxidant,thrombolytic,cytotoxic, antimicrobial activities were conducted using specific standard in vitro procedures.Results:The results showed that the plant extracts were a rich source of phytoconstituents.Methanol extract showed higher antioxidant,thrombolytic activity and less cytotoxic activity than those of n-hexane and ethyl acetate extracts of A diffusa.Among the bioactivities,antioxidant activity was the most notable compared to the positive control and thus could be a potential rich source of natural antioxidant.In case of antimicrobial screening,crude extracts of the plant showed remarkable antibacterial activity against tested microorganisms.All the extracts showed significant inhibitory activity against Candida albicuns,at a concentration of 1000 μg/disc.Conclusions:The present findings suggest that,the plant widely available in Bangladesh,could be a prominent source of medicinally important natural compounds.

Antitumor effects and compatibility mechanism of Hedyotis Diffusa-Sculellaria Barbata herb pair

OBJECTIVE Hedyotis diffusa Willd and Scutellaria barbata D Don are most commonly used herb pair for cancer treatment in traditional Chinese medicine.This study aimed to evaluated the in vitro and in vivo antitumor effects and the compatibility mechanisms of Hedyotis diffusa-Sculellaria barbata herb pair.METHODS Hedyotic diffusa extract,Scutellariae Barbatae extract,and herb pair extract were prepared and the component were detected by UPLC-Q-TOF-MS.The effects of different concentrations of Hedyotic diffusa extract,Scutellariae Barbatae extract,and herb pair extract were detected using MTS assay.The in vivo antitumor effects were evaluated in Panc28 pancreatic cells bearing nude mice model.The compatibility mechanism of Hedyotis diffusa-Sculellaria barbata herb pair were evaluated based on the network pharmacology,using TCMSP,Cytoscape 3.6.1 software,and DAVID 6.8.The anticancer mechanism were further validated in vitro.RESULTS Both the MTS and in vivo results showed that herb pair extract showed more obvious inhibitory effects on cancer cells compared to each individual.A total of 37 active components were selected from Hedyotis diffusa and Sculellaria barbata,33 kinds of active ingredients are involved in their anti-tumor effects.58 cancer-related targets and 66 KEGG pathways were identified.The potential targets for the herb pair might be prostaglandin G/H synthase 2(PTGS2),HSP90,EGFR,72 ku typeⅣcollagenase,PPAR-γ,et al.In vitro validation result showed that compatibility mech⁃anisms was related with HSP90,EGFR related pathways.CONCLUSION The result of the study preliminarily verified the basic anti-tumor pharmacological effects of Hedyotis diffusa-Sculellaria barbata herb pair,and lays a solid foundation for further studies on the anti-tumor mechanism of the herbal pair.

Introduction and Cultivation of Viola diffusa Ging and Its Gardening Application in Hubei

Distribution of Viola diffusa Ging in Hubei was introduced fi rst,then its biological and ecological characters,introduction and cultivation,and reproduction techniques elaborated. In addition,its gardening application in Wuhan was explored.

Effect of Polysaccharides from Hedyotis diffusa on Immune Cells of Small Intestinal Mucosa of Chicks

[Objectives]This study aimed to investigate the effect of polysaccharides from Hedyotis diffusa on the immune cells of the intestinal mucosa of chicks.[Methods]Total 200 seven-day-old AA chicks were evenly and randomly divided into four groups:control group and three treatment groups.The chicks in the control group(group I)were fed basal diet,and those in the treatment groups were fed with basal diet supplemented with 0.4(group II),0.8(group III)and 1.2(group IV)g/kg H.diffusa polysaccharides(OPS),respectively.At the age of 14,28 and 42 d,10 chicks from each of the groups were sacrificed,respectively for counting of intraepithelial lymphocytes(IELs)and lamina propria secretory IgA(SIgA)positive cells in small intestinal mucosa.[Results]At the age of 14 d,the duodenal and jejunal IEL and SIgA positive cell counts in groups II,III and IV were significantly greater than those in group I(P<0.05).At the age of 28 d,the jejunal IEL count of group IV was significantly lower than those of groups I,II and III(P<0.05);and there was no significant difference in IEL or SIgA positive cell count between group I and the other groups(P>0.05).At the age of 42 d,the duodenal and jejunal IEL counts in group II were significantly greater than those in groups I,III and IV(P<0.05),while the differences in duodenal and jejunal IEL count between groups III,IV and I were insignificant(P>0.05).The duodenal SIgA positive cell counts in the treatment groups were insignificantly different from that in group I(P>0.05).The jejunal SIgA positive cell count in group II was significantly greater than those in groups I,III and IV(P<0.05).There was no significant difference in jejunal SIgA positive cell count between groups III and IV and group I(P>0.05).[Conclusions]OPS at 0.4 g/kg can promote the proliferation of immune cells in the small intestine mucosa of 14-42-day-old AA chicks,and it is recommended to be added to the feed.

Effects of Oldenlandia diffusa Polysaccharides on the Structures of Chick Small Intestine Tissues

In order to explore the application effect of Oldenlandia polysaccharides(OPS)in the production of AA broilers,2007-day-old AA broilers were randomly divided into 4 groups,namely groups Ⅰ,Ⅱ,Ⅲ,and Ⅳ,each including 50 chicks.The control group(group Ⅰ)was fed with basal diet;and the 3 experimental groups were fed based on the basal diet with the addition of OPS at 0.4 g/kg(group Ⅱ),0.8 g/kg(group Ⅲ),and 1.2 g/kg(group Ⅳ)feed for 1 week,respectively,and then fed the basal diet for 4 weeks.At the ages of 14,28,and 42 d,10 chickens were put into death in the control group and the experimental groups,respectively,and the duodenum and jejunum tissue samples were collected for the analysis of the effects of OPS on the structures of chick intestines.The results showed that at the age of 14 d,the villus lengths of duodenum and jejunum in groups Ⅱ,Ⅲ,and Ⅳ were significantly greater than those in group Ⅰ(P<0.05),and the ratios of duodenal or jejunal villus length to crypt depth in groups Ⅱ and Ⅲ were significantly greater than that in group Ⅰ(P<0.05);at the age of 28 d,the villus length of duodenum in group Ⅱ was significantly greater than that in group Ⅰ(P<0.05);and at the age of 42 d,the ratios of jejunual villus length to crypt depth in groups Ⅱ,Ⅲ and Ⅳ were significantly larger than those of group I(P<0.05).It showed that OPS could promote the small intestine development of AA broilers aged 14 to 42 d.

Cytotoxic, Antimicrobial and Antioxidant Properties of <i>Commelina diffusa</i>Burm. F.

Commelina diffusa Burm. F. is a herbaceous tropical plant with different traditional medicinal uses. Present study is aimed to isolate the bioactive compounds from DCM-Methanol extract of the powdered whole plant of Commelina diffusa and to investigate the cytotoxic, antimicrobial and antioxidant activities of the crude extract and its twelve vacuum liquid chromatographic fraction (CD1-12). Vacuum Liquid Chromatography (VLC) was employed to isolate bioactive metabolites. The physicochemical properties of the isolated compound were examined and the molecular structure was elucidated by NMR spectroscopy.?Cytotoxic, antibacterial, antioxidant activities were evaluated following brine shrimp lethality bioassay, disc diffusion method and DPPH free radical scavenging assay respectively. The isolated compound was identified as steroid (stigmasterol) which had significant cytotoxic effect on vero cell line. The crude extract and its fractions (CD8-CD12) exhibited strong cytotoxicity in brine shrimp lethality bioassay having LC50 values 3.79, 9.19, 29.49, 16.60, 19.36, 44.58 μg/mL respectively. The crude extract showed mild to strong antibacterial activity. Fractions (CD5-CD12) showed mild to strong antibacterial activity against Escherichia coli in comparison with Kanamycin (standard). Strong antioxidant activities were found in crude extract (IC50 value 30.52 μg/ml), CD11 (IC50 value 39.27 μg/ml) and CD12 (IC50 value 19.50 μg/ml). The study suggests Commelina diffusa plant extract to have strong antioxidant and cytotoxic activity which is indicative of presence of compounds with broad spectrum of curative applications. One compound namely stigmasterol has been isolated from the plant.

Threat of Domestic Power Generators to Terrestrial Ecosystem as Expressed in Some Plant Species

Polluted populations of Euphorbia heterophylla, Chromolaena odorata, Commelina diffusa and Kyllinga pumila were collected fresh from 0.1 m to 0.25 m radii of the exhausted-pipe of power generators used for domestic purposes, whereas, their non-polluted counterparts were collected from distances of 100-110 m away. The length and width of 10 leaves, each of both polluted and non-polluted populations, were measured. Their average leaf areas （LA） were 2.60 ± 0.1 cm2, 14.16 ±0.4 cm2, 5.19 ± 0.8 cm2 and 1.80 ±0.8 cm2 for the polluted populations of each of the species, respectively. Whereas, the LA of their non-polluted counterparts were 4.18±0.7 cm2, 34.39 ±0.2 cm2, 7.51± 0.1 cm2 and 10.76 ± 0.3 cm2, respectively. Damages such as plugged stomatal pores, epidermal cell aberrations and erosion, ruptured stomatal ledges, occasional leaf perforations, irregularly fused cell boundaries and glued leaf surfaces were noticeable in the leaves of the polluted populations. Such were not observed in the non-polluted populations. It was suggested that foliar morphology of these group of plants could serve as phytometer to gauge the threats of power generators used in homes to terrestrial ecosystem.

Establishing a Quality Control System of Hedyotis corymbosa

Objective Hedyotis corymbosa(Shui Xian Cao,水线草,HC)is the main antitumor medicine in China and other southeast Asian countries.A comprehensive quality control system of HC was established in this study,as the current system was ineffective.Methods Microscopy was used to observe the characteristics of the powder and cross-sectional tissue.Thin layer chromatography(TLC)was employed to separate HC from Hedyotis diffusa(Bai Hua She She Cao,白花蛇舌草,HD).Simultaneously,high-performance liquid chromatography(HPLC)was used to establish the characteristic spectrum of HC and determine the concentrations of hedyotiscone A.Regarding the issue of systematicness,concentrations of water,total ash,acid-insoluble ash,heavy metals(including lead,arsenic,mercury,cadmium and copper)and pesticide residue were measured by the methods described in the 2015 edition of Chinese Pharmacopoeia.Results Calcium oxalate needle crystal bundles and oil drops were observed in the cross-sectional tissue.Three characteristic blue fluorescent spots were observed in the HC samples by TLC.There were five distinct characteristic peaks in HC but not in HD,and hedyotiscone A was the sharpest characteristic peak.The generated linear equation of hedyotiscone A was Y=3.15×108X+312.04(r2=1.000)between 0.00005270 and 0.002108 mg/mL,and the average recovery was 100.42%.Precision[relative standard deviation(RSD)0.92%],repeatability(RSD 0.14%),and recovery(RSD 0.85%)met the requirements for quantitative analysis.Hedyotiscone A concentrations ranged from 0.00017%~0.00113%across different habitats.Concentrations of water,ash and acid-insoluble ash were 8.80%~11.2%,7.00%~15.3%and 0.440%~7.70%,respectively.Concentrations of Pb,As,Hg,Cd and Cu were 1.60~4.60,0.650~1.60,0.0300~0.100,0.240~0.690 and 10.1~16.6μg/g,respectively.Pesticide residues were less than 0.00100 ppm.Conclusion The quality control system can effectively distinguish HC from HD,so as to comprehensively control the quality of HC,to provide references for the clinical use of HC.

Flavonoid-Rich Extract of Oldenlandia diffusa(Willd.)Roxb.Inhibits Gastric Cancer by Activation of Caspase-Dependent Mitochondrial Apoptosis

Objective:To evaluate the apoptosis and cycle arrest effects of Oldenlandia diffusa flavonoids on human gastric cancer cells,determine the action mechanisms in association with the mitochondrial dependent signal transduction pathway that controls production of reactive oxygen species(ROS),and evaluate the pharmacodynamics of a mouse xenotransplantation model to provide a reference for the use of flavonoids in prevention and treatment of gastric cancer.Methods:Flavonoids were extracted by an enzymatic-ultrasonic assisted method and purified with D-101 resin.Bioactive components were characterized by high-performance liquid chromatography.Cell lines MKN-45,AGS,and GES-1 were treated with different concentrations of flavonoids(64,96,128,and 160μg/mL).The effect of flavonoids on cell viability was evaluated by MTT method,and cell nuclear morphology was observed by Hoechst staining.The apoptosis rate and cell cycle phases were measured by flow cytometry,the production of ROS was detected by laser confocal microscope,the mitochondrial membrane potential(MMP)were observed by fluorescence microscope,and the expression of apoptotic proteins related to activation of mitochondrial pathway were measured by immunoblotting.MKN-45cells were transplanted into BALB/c nude mice to establish a xenograft tumor model.Hematoxylin and eosin staining was used to reveal the subcutaneous tumor tissue.The tumor volume and tumor weight were measured,the expression levels of proliferation markers proliferating cell nuclear antigen(PCNA)and Ki-67were detected by immunohistochemistry,and the expression levels of CA72-4 were measured by enzyme linked immunosorbent assay.Results:Oldenlandia diffusa flavonoids inhibited proliferation of MKN-45 and AGS human gastric cancer cells,arrested the cell cycle in G1/S phase,induced accumulation of ROS in the process of apoptosis,and altered MMP.In addition,flavonoids increased Apaf-1,Cleaved-Caspase-3,and Bax,and decreased Cyclin A,Cdk2,Bcl-2,Pro-Caspase-9,and Mitochondrial Cytochrome C(P<0.05).The MKN-45 cell mouse xenotransplantation model further clarified the growth inhibitory effect of flavonoids towards tumors.The expression levels of PCNA and Ki-67 decreased in each flavonoid dose group,the expression level of CA72-4 decreased(P<0.05).Conclusion:Flavonoids derived from Oldenlandia diffusa can inhibit proliferation and induce apoptosis of human gastric cancer cells by activating the mitochondrial controlled signal transduction pathway.

3D Microfluidic System for Evaluating Inhibitory Effect of Chinese Herbal Medicine Oldenlandia diffusa on Human Malignant Glioma Invasion Combined with Network Pharmacology Analysis

Objective:To investigate the anti-invasion efficacy of the ethanol extract of Oldenlandia diffusa Will.(EEOD) on a three-dimensional(3D) human malignant glioma(MG) cell invasion and perfusion model based on microfluidic chip culture and the possible mechanism of action of Oldenlandia diffusa Will.(OD).Methods:The comprehensive pharmacodynamic analysis method in this study was based on microfluidic chip 3D cell perfusion culture technology,and the action mechanism of Chinese medicine(CM) on human MG cells was investigated through network pharmacology analysis.First,the components of EEOD were analyzed by ultraperformance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS).Then,cell viability and apoptosis were assessed to determine the optimum concentration of EEOD for invasion experiments,and two-dimensional(2D) migration and invasion abilities of U87 and U251 MG cells were evaluated using scratch wound and Transwell assays.The possible mechanism underlying the effects of EEOD on glioma was analyzed through a network pharmacology approach.Results:Thirty-five compounds of EEOD were detected by UPLC-Q-TOF/MS.EEOD suppressed the viability of MG cells,promoted their apoptosis,and inhibited their migratory and invasive potentials(all P<0.05).Network pharmacology analysis showed that OD inhibited the invasion of MG cells by directly regulating MAPK and Wnt pathways through MAPK,EGFR,MYC,GSK3B,and other targets.The anti-invasion effect of OD was also found to be related to the indirect regulation of microtubule cytoskeleton organization.Conclusion:EEOD could inhibit the invasion of human MG cells,and the anti-invasion mechanism of OD might be regulating MAPK and Wnt signaling pathways and microtubule cytoskeleton organization.

Anti-tumor activity of Hedyotis diffusa Willd. in mice

The whole plant with root ofHedyotis diffusa Willd. （AE） has been demonstrated to have anti-inflammatory, antioxidant, and anti-bacterial activities. In our study, we aim to examine the anti-tumor effect of alcoholic extract of AE in mice implanted with sarcoma S180 cells （SBT mice）. We also compared the immune system function and the life span of SBT mice with that of control mice. We found that AE displayed a significant inhibitory effect on solid tumor growth, as well as increased the life span. At the dose of 10 mg/kg body weight, both tumor weight and volume were decreased significantly. We also measured several immune fimction markers, inlcuding spleen index （SI）, thymus index （TI） and parameters of hematological. We observed no reduce in these markers, indicatine that AE could inhibit tumor ~rowth without affectino immune function in SBT ,nice.