Fruits extracts of Hovenia dulcis Thunb.suppresses lipopolysaccharide—stimulated inflammatory responses through nuclear factor—kappaB pathway in Raw 264.7 cells

Objective:To investigate the anti-inflammatory effects and the action mechanism of the fruits of Horenia dulcis(H.dulcis) in lipopolysaccharide(LPS)-induced mouse macrophage Raw 264.7cells.Methods:The extract of H.dulcis fruits(EHDF) were extracted with 70%ethanol.Mouse macrophages were treated with different concentrations of EHDF in the presence and absence of LPS(1 μg/mL).To demonstrate the inflammatory mediators including nitric oxide,inducible nitric oxide synthase and cyclooxygenase(COX)-2 expression levels were analyzed by usingin vitro assay systems.COX-derived pro-inflammatory cytokines including interleukin-1 β.tumor necrosis factor- α and prostaglandin F_2 were determined using ELISA kits.Cell viability,heme oxygenase-1 expression,nuclear factor-kappaB and nuclear factor F.2-related factors 2 translocation were also investigated.Results:EHDF potently inhibited the LPS-stimulated nitric oxide,inducible nitric oxide synthase.COX-2,interleukin-1 β and tumor necrosis factor- α expression in a dose-dependent manner.EHDF suppressed the phosphorylation of inhibited kappaB-alpha and p65 nuclear translocation.Treatment of macrophage cells with EHDF alone induced the heme oxygenase-1 and nuclear translocation of nuclear factor E2-reIated factor 2.Conclusions:These results suggest that the ethanol extract of H.dulcis fruit exerts its anti-inflammatory effects by inhibiting inhibited kappaBalpha phorylation and nuclear translocation of nuclear factor-kappaB.

Hepatoprotective effects of Hovenia dulcis seeds against alcoholic liver injury and related mechanisms investigated via network pharmacology

BACKGROUND Alcoholic liver disease(ALD)is a worldwide health problem,and natural products have been shown to improve ALD due to their antioxidant activities.Some parts of Hovenia dulcis(H.dulcis),such as roots,peduncles,and stems,provide health benefits.Nevertheless,the effects and mechanisms of H.dulcis seeds on ALD have not yet been fully elucidated.AIM To determine H.dulcis antioxidant activity,evaluate its effects against ALD,and investigate the related mechanisms via network pharmacology.METHODS The antioxidant activity of H.dulcis seed was determined by both ferric-reducing antioxidant power and trolox equivalent antioxidant capacity assays.The total phenolic and flavonoid contents were determined by Folin–Ciocalteu method and aluminum chloride colorimetry,respectively,and polysaccharide was determined by phenol-sulfuric acid method.The effects of H.dulcis seeds against alcoholic liver injury were investigated in mice with water extract pretreatment for 7 days followed by alcohol administration.Moreover,the mechanisms of action were explored with network pharmacology.RESULTS The results showed that H.dulcis seeds possessed strong antioxidant activity(245.11±10.17μmol Fe2+/g by ferric-reducing antioxidant power and 284.35±23.57μmol TE/g by trolox equivalent antioxidant capacity)and contained remarkable phenols and flavonoids,as well as a few polysaccharides.H.dulcis seeds attenuated alcohol-induced oxidative liver injury,showing reduced serum alanine and aspartate aminotransferases,alkaline phosphatase,and triglyceride,elevated hepatic glutathione,increased activities of superoxide dismutase and catalase,and reduced malondialdehyde and hepatic triglyceride.The results of network pharmacology analysis indicated that kaempferol,stigmasterol,and naringenin were the main bioactive compounds in H.dulcis seeds and that modulation of oxidative stress,inflammation,gut-derived products,and apoptosis were underlying mechanisms of the protective effects of H.dulcis seeds on ALD.CONCLUSION The results of this study demonstrate that H.dulcis seeds could be a good natural antioxidant source with protective effects on oxidative diseases such as ALD.

Cytokinin induced shoot regeneration and flowering of Scoparia dulcis L.(Scrophulariaceae)-an ethnomedicinal herb

Objective:To develop an improved protocol for micropropagation of ethnomedicinally important Scoparia dulcis(S.dulcis) L.Methods:Explants were inoculated on MS basal medium supplemented with kinelin and 6-benzylaminopurine for shoot bud induction.To enhance the shoot induction,various auxins like 3-indoleacetic acid or 3-indolebutyric acid or a-naphthylacetic acid were tested along with 2.32 M KI and 4.44 μ M BAP.The regenerated shoots were rooted in half strength MS medium supplemented with various concentrations of IAA,IBA or NAA.After roots were developed,the plantlets were transplanted to pots filled with vermiculate and sand and kept in growth chamber with 70%-80%humidity under 16 h photoperiod.After acclimatization,the plantlets were transferred to the garden and survival percentage was calculated.Data were statistically analyzed and means were compared using Duncan's multiple range test(P<0.05).Results:An in vitro method was developed to induce high frequency shoots regeneration from stem,mature leaf and young leaf explants of S.dulcis.Shoot induction on young leaf explants was most successful in MS medium supplemented with combination of two cytokinins(2.32 μ M KI and 4.44 μ M BAP) 2.85 μ M IAA,10%CM and 1 483.79 μM adenine sulfate.A single young leaf explant was capable of producing 59 shoots after 13 days of culture. Flower was induced in medium supplemented with combination of KI and BAP.Conclusions: Cytokinins are the key factor to induce the direct shoot regeneration and flowering of S.dulcis.

A comparative study of the antioxidant,antimicrobial,cytotoxic and thrombolytic potential of the fruits and leaves of Spondias dulcis

Objective:To investigate the antioxidant,antimicrobial,cytotoxic and thrombolytic property of the fruits and leaves of Spondias dulcis（S.dulcis）.Methods:Methanolic extracts of fruits and leaves of S.dulcis were partitioned with chloroform and dichloromethane.The antioxidant potential of the crude extract and partitioned fractions were evaluated in terms of total phenolic content,total flavonoid content,DPPH radical scavenging potential,reducing potential and total antioxidant capacity by specific standard procedures.The antimicrobial activity was evaluated using disc diffusion method.The cytotoxicity was evaluated by using brine shrimp lethality bioassay and compared with vincristine sulfate.The thrombolytic activity was compared with streptokinase.Results:The methanolic fruit extract exhibited the highest phenolic content,flavonoid content and antioxidant capacity,among the other extracts,with the highest DPPH radical scavenging activity at a concentration of 10μg/mL(IC50:1.91μg/mL)and maximum reducing power at a concentration of 100μg/ml.(EC50:3.58μg/mL).Though all extract showed moderate antimicrobial activity against the bacterial strains,weak or no activity against fungus.The range of LC50value of all extracts was 1.335-14.057μg/mL which was far lower than the cut off index for cytotoxicity.All extracts exhibited statistically significant（P【0.001）thrombolytic activity.Conclusions:Our study suggested that 5.dulcis exhibits antimicrobial activities against a wide variety of strains while it possesses significant antioxidant,cytotoxic and thrombolytic aclivily.

Effects of Phosphate Fertilizer on Yield and Quality of Eleocharis dulcis

The effects of different application levels and species of phosphatic fertilizer on Eleocharis dulcis were studied. The results showed that the growth stage of Eleocharis dulcis was not significantly influenced by different levels of phosphatic fertilizer application, with phosphorus level in 195-375 kg/hm^2. With increasing of the phosphorus level, starch and total soluble sugar accumulated, but the yield was not improved significantly. The analysis on the economic benefits showed that the best phosphorus level was 195 kg/hm^2. The growth stage and yield of Eleocharis dulcis were not considerably influenced by different proportions of calcium-magnesia phosphate fertilizer and ammonium dihydrogen phosphate. With increasing of ammonium dihydrogen phosphate, big-fruit rate of Eleocharis dulcis kept growing, but the accumulations of starch and total soluble sugar were decreasing.

Assessment of Genetic Diversity of Moroccan Cultivated Almond(Prunus dulcis Mill.DA Webb)in Its Area of Extreme Diffusion,Using Nuclear Microsatellites

Assessment of genetic diversity of Moroccan cultivated almond (Prunus dulcis Mill.) grown from seed and cultivated at four eco-geographical regions was performed using 16 nuclear SSRs. 238 alleles were detected with an average of 14.88 alleles per locus, ranging from 4 (locus BPPCT027) to 24 (locus CPSCT018). The size of alleles ranged from 84 bp (locus UDP96-003) to 253 bp (locus UDP96-018). A high genetic diversity of the local almonds is apparent and structured into three major clusters (Oasis cluster, High and Anti Atlas cluster, and Middle Atlas cluster). Compared to the Mediterranean genetic pools, from the East to West, the genetic diversity tends to be limited in Morocco which is the area of its extreme diffusion.

Antioxidant mediated response of Scoparia dulcis in noise-induced redox imbalance and immunohistochemical changes in rat brain

Noise has been regarded as an environmental/occupational stressor that causes damages to both auditory and non- auditory organs. Prolonged exposure to these mediators of stress has often resulted in detrimental effect, where oxidative/nitrosative stress plays a major role. Hence, it would be appropriate to examine the possible role of free radicals in brain discrete regions and the ＂antioxidants＂ mediated response of S. dulcis. Animals were subjected to noise stress for 15 days （100 dB/4 hours/day） and estimation of endogenous free radical and antioxidant activity were carried out on brain discrete regions （the cerebral cortex, cerebellum, brainstem, striatum, hippocampus and hypothalamus）. The result showed that exposure to noise could alleviate endogenous free radical generation and altered anfioxidant status in brain discrete regions when compared to that of the control groups. This alleviated free radical generation （H202 and NO） is well supported by an upregulated protein expression on immunohistochemistry of both iNOS and nNOS in the cerebral cortex on exposure to noise stress. These findings suggest that increased free radical generation and altered anti-oxidative status can cause redox imbalance in the brain discrete regions~ However, free radical scavenging activity of the plant was evident as the noise exposed group treated with S. dulcis[200 mg/（kg · b. w）] displayed a therapeutic effect by decreasing the free radical level and regulate the anti-oxidative status to that of control animals. Hence, it can be concluded that the efficacy of S. dulcis could be attributed to its free radical scavenging activity and anti-oxidative property.

Development of a marketable rice based herbal porridge suitable for diabetics from Scoparia dulcis

Consumption of herbal leafy porridges is a reputed dietary remedy among Sri Lankans in treating diabetes. The aim of the study was to develop a rice based herbal porridge [rice: fresh leaves: scraped coconut kernel 13 - 15: 25 - 30: 10 - 13 (w/w/w)] commercially with Scopariadulcis leaves which has proven antidiabetic effects and low GI (fresh porridge) which benefits diabetics. Two porridges with the same ingredient ratio were produced with different particle sizes. Porridge produced with 100% fine particles (extruded rice, leaves and scraped coconut mixed with rice powder) elicited a high GI for normal (92 ± 22) and diabetic (97 ± 20) subjects. The second porridge prepared with the minimum amount of extruded rice with other ingredients mixed with boiled and dried intact rice grains had a medium GI (normal 58 ± 11, diabetics 61 ± 11). A significant reduction (p < 0.05) in the peak blood glucose was observed in diabetic subjects for porridge 2 (12.4%) compared to porridge 1 (0.7%) when compared to glucose. The reason could be the significantly high (p0.5 mm) (63%) and minimum amount of extruded fine particles in porridge 2 compared to porridge 1 (1.3%).

Flavonoid C-glycosides from the seeds of Hovenia dulcis Thunb.

Ten known flavonoid C-glycosides identified as 2′′-O-β-D-glucopyranosyl isovitexin(1),spinosin(2),6′′′-acetyl spinosin(3),6′′′-p-hydroxycinnamoyl-2′′O-β-D-glucopyranosyl isovitexin(4),6′′′-p-hydroxy-cinnamoyl spinosin(5),6′′′-(E)-feruloyl O-β-Dglucopyranosyl isovitexin(6),isospinosin(7),isovitexin(8),swertisin(9),swertiajaponin(10)were purified from the seeds of Hovenia dulcis Thunb.Their structures were confirmed by spectroscopic data analysis and comparison with previous literature.All compounds were obtained from H.dulcis for the first time.

Hydrangeae Dulcis Folium Attenuates Physical Stress by Supressing ACTH-Induced Cortisol in Zebrafish

Objective:To determine the effects of Hydrangeae Dulcis Folium(EHDF)on physical stress,changes in the whole-body cortisol level and behaviour in zebrafish(Danio rerio).Methods:One hundred and seventy-four fish were randomly divided into 4[adrenocorticotropin hormone(ACTH)challenge test:4 fish per group]or 6 groups(behavioural test:10–12 fish per group,whole-body cortisol:4 fish per group).Net handling stress(NHS)was used to induce physical stress.Fish were treated with vehicle or EHDF(5–20 mg/L)for 6 min before they were exposed to stress.And then,fish were sacrificed for collecting body fluid from whole-body or conducted behavioural tests,including novel tank test and open field test,and were evaluated to observe anxiety-like behaviours and locomotion.In addition,to elucidate the mode of action of the anti-stress effects of EHDF,ACTH(0.2 IU/g,i.p.)challenge test was performed.Results:The increased anxiety-like behaviours in novel tank test and open field test under stress were prevented by treatment with EHDF at 5–20 mg/L(P<0.05).Moreover,compared with the unstressed group,which was not treated with NHS,the whole-body cortisol level was significantly increased by treatment with NHS(P<0.05).Compared with the NHS-treated stressed control group,pre-treatment with EHDF at concentrations of 5–20 mg/L for 6 min significantly prevented the NHS-increased whole-body cortisol level(P<0.05).In addition,ACTH challenge test showed that EHDF completely blocked the effects of ACTH on cortisol secretion(P<0.05).Conclusion:EHDF may be a good antistress candidate and its mechanism of action may be related to its positive effects on cortisol release.

Chemical constituents from aerial parts of Scoparia dulcis

Objective: To study the chemical constituents from the aerial parts of Scoparia dulcis.Methods: Various chromatographic techniques were used to separate the constituents and their structures were elucidated using spectroscopic methods and by comparing their data to those reported in the literatures. The α-glucosidase inhibitory activity assay was used to identify potential α-glucosidase inhibitors.Results: Nine compounds were isolated from the aerial parts of S. dulcis. Their structures were identified as Scoparic zolone(1),(2S)-2,7-dihydroxy-2H-1,4-benzoxazin-3(4H)-one(2),(2R)-7-hydroxy-2H-1,4-benzoxazin-3(4H)-one-2-O-β-D-glucopyranoside(3),(2R)-7-methoxy-2H-1,4-benzoxazin-3(4H)-one-2-O-β-D-glucopyranoside(4),(2S)-7-hydroxy-2H-1,4-benzoxazin-3(4H)-one-2-O-β-D-glucopyranoside(5),6-methoxy-benzoxazolin-2(3H)-one(6), 4-acetonyl-3,5-dimethoxy-p-quinol(7), zizyvoside I(8), and3,4-dihydroxy benzeneacetic acid(9). Compound 2 showed the potent α-glucosidase inhibitory activity with an IC50value of(132.8 ± 11.5) μmol/L, which is 28-fold higher than the positive control acarbose.Conclusion: Compound 1 is a new natural product. Compounds 2 and 9 have not been reported in Scoparia before. Compounds 3, 5, 7, 8 are isolated from Scrophulariaceae for the first time.

水生蔬菜答农民问(67-2):荸荠新品种选育主要途径有哪些?如何进行良种繁育?

根据荸荠(Eleocharis dulcis)繁殖方式、性状遗传变异特点及育种和栽培历史,认为芽变、实生苗变异、诱变、组织培养后代变异、人工杂交等都是荸荠新品种选育的可能途径。荸荠品种混杂退化的主要原因包括机械混杂、遗传变异、球茎异质性差异、病毒感染等。良种繁育程序从原原种或原种开始时,宜为“原原种→原种→生产用种→大田生产”;从生产大田优选单株开始时,宜为“生产大田→选择单株或球茎→株行圃→株系圃→原种或生产用种→大田生产”;从组培苗开始时,宜从原原种或原种,或提纯复壮后的优选单株或株系球茎上采取顶芽进行初代培养。

水生蔬菜答农民问(67-1):荸荠新品种选育主要途径有哪些?如何进行良种繁育?

根据荸荠(Eleocharis dulcis)繁殖方式、性状遗传变异特点及育种和栽培历史,认为芽变、实生苗变异、诱变、组织培养后代变异、人工杂交等都是荸荠新品种选育的可能途径。荸荠品种混杂退化的主要原因包括机械混杂、遗传变异、球茎异质性差异、病毒感染等。良种繁育程序从原原种或原种开始时,宜为“原原种→原种→生产用种→大田生产”;从生产大田优选单株开始时,宜为“生产大田→选择单株或球茎→株行圃→株系圃→原种或生产用种→大田生产”;从组培苗开始时,宜从原原种或原种,或提纯复壮后的优选单株或株系球茎上采取顶芽进行初代培养。

荸荠种质资源品质性状综合评价

对保存于国家种质武汉水生蔬菜资源圃内90份荸荠[Heleocharis dulcis（Burm.f.）Trin.ex Hensch.]资源感官品质性状（脐部特征、单个球茎质量、甜度、化渣度）和理化品质性状（干物质含量、可溶性糖含量、淀粉含量）进行了测试分析。结果表明,荸荠资源品质性状具有广泛的遗传多样性,数量性状变异系数达18.2%~31.3%,质量性状类型多样,并且存在一定的相关性。采用主成分分析法得到了90份荸荠资源品质的综合排名,排名前20位的均为鲜食型荸荠,该方法可以较好地用于鲜食荸荠品质性状的评价。

荸荠花粉活力测定及贮藏寿命研究

为了解荸荠[Heleocharis dulcis(Burm.f.)Trin.ex Hensch.]花粉生活力和最佳体外萌发条件,分别采用I2-KI、TTC、MTT染色法和离体培养基萌发法对荸荠花粉活力进行了测定,并初步研究了荸荠花粉在常温(25℃)条件和低温(4℃)贮藏条件下活力变化的动态。结果表明,MTT染色法测定结果与离体萌发法最接近,最适宜荸荠花粉活力的快速检测;离体萌发法适宜培养基配方为1%蔗糖+50μg/m L硼酸;荸荠花粉在常温条件下50 min活力降至0.29%的低水平,在低温条件下5 h后完全失去活力。

团风荸荠果汁饮料的研制

以湖北省黄冈市团风县的新鲜荸荠(Eleocharis dulcis)为原料,利用单因素试验和正交试验,以感官评定进行配方优选,得到荸荠果汁饮料的最佳工艺参数为90℃水中烫漂2 min,70℃糊化10 min,果浆含量20%,甜菊糖添加量0.004%,柠檬酸添加量0.10%,黄原胶添加量0.10%。

超声波法辅助提取荸荠块茎皮中甾醇的工艺研究

采用超声波法提取荸荠[Heleocharis dulcis(Burm.f.)Trin.]块茎皮中的甾醇,对提取溶剂的种类、溶剂体积分数、料液比、提取时间、超声波功率分别进行单因素试验,并在此基础上利用Box-Benhnken中心组合试验和响应面分析法确定了植物甾醇提取的最佳工艺。结果表明,荸荠块茎皮中甾醇的最佳提取工艺条件为60%乙醇、料液比为1∶40(g∶m L)、提取时间为14 min、超声波功率为600 W,实际测得的植物甾醇的紫外吸光度为1.375。

不同磷肥施用水平和磷肥品种搭配对马蹄产量及品质的影响

采用不同磷肥施用水平和不同的磷肥品种搭配,研究其对马蹄[Heleocharis dulcis（Burm.f.）Trin.]生育期、产量和品质等的影响。结果表明,施磷量为13~25 kg/667 m2时,不同的磷肥施用水平对马蹄的生育期无影响;随着磷肥施用量的增加,有利于淀粉和可溶性总糖的积累,但不能显著提高马蹄的产量。从经济效益上看,增施磷肥不能提高马蹄的经济效益,纯收益最好的是施磷水平为13 kg/667 m2。钙镁磷肥和磷酸一铵两种磷肥的不同比例对马蹄生育期和产量均无影响,随着磷肥中磷酸一铵的比例增加,马蹄的大果率增加,淀粉和可溶性总糖的积累随之降低。

Discriminating ability of molecular markers and morphological characterization in the establishment of genetic relationships in cultivated genotypes of almond and related wild species

A total 23 morphological traits, 19 AFLP-primer combinations, 80 RAPD primers and 32 SSR primer pair were used to compare the informativeness and efficiency of random amplified polymorphic DNA （RAPD）, amplified fragment length polymorphism （AFLP） and simple sequence repeat （SSR） markers in establishing genetic relationships among 29 almond cultivars and three related wild species. SSRs presented a high level of polymorphism and greater information content, as assessed by the expected hetrozygosity, compared to AFLPs and RAPDs. The lowest values of expected hetrozygosity were obtained for AFLPs; however AFLPs showed the highest efficiency, owing to their capacity to reveal large numbers of bands per reaction, which led to high values for various types of indices of diversity. All the three techniques discriminated almond genotypes very effectively, except that SSRs failed to discriminate between ＇Monagha＇ and ＇Sefied＇ almond genotypes. The correlation coefficients of similarity were statistically significant for all the three marker systems, but were lower for the SSR data than for RAPDs and AFLPs. For all the markers, high similarity in dendrogram topologies was obtained, although some differences were observed. All the dendrograms, including that obtained by the combined use of all the marker data, reflect relationships for most of cultivars according to their geographic diffusion. AMOVA detected more variation among cultivated and related wild species of almond within each geographic group. Bootstrap analysis revealed that the number of markers used was sufficient for reliable estimation of genetic similarity and for meaningful comparisons of marker types.

Morphological and Physicochemical Properties of Water Chestnut Starches: a Comparative Analysis of Three Varieties

Water chestnut(Eleocharis dulcis) is widely cultivated in many countries for its edible and starchy corms. In this study, starches were separated from Guangxi biqi, Hubei biqi and Anhui biqi, and the morphological and physicochemical characteristics of the starches were systematically investigated. There were significant differences in granule size, total starch and amylose contents among starches from the three water chestnuts which present similar X-ray diffraction patterns and Fourier transform infrared spectroscopy. The starch of Guangxi biqi exhibited higher swelling power and solubility than that of Hubei biqi and Anhui biqi. Thermal and pasting properties of starch were different among the three water chestnuts. When hydrolyzed by porcine pancreatic α-amylase(PPA), starches from Hubei biqi and Anhui biqi had higher hydrolysis degree. The starches from the three water chestnut varieties differed significantly in morphological and physicochemical properties, and these studies may provide useful information for future exploitation and application of water chestnut starch in food and non-food industries.