Effect of Artemisia annua (Asteraceae) Extracts on Hemolysis in Individuals with G6PD-Deficiency

Individuals with Glucose-6-phosphate dehydrogenase (G6PD) deficiency are susceptible to hemolytic anemia when exposed to pro-oxidant substances. This study investigates the hemolytic impact of Artemisia annua (A. annua) extracts in G6PD-deficient subjects through a mixed experimental approach. In the in vitro phase, red blood cells from G6PD-deficient individuals and rats induced with Dehydroepiandrosterone (DHEA) were exposed to various concentrations of A. annua infusion, with distilled water and physiological saline as positive and negative controls respectively. The in vivo study involved G6PD-deficient Wistar rats divided into three groups receiving A. annua infusion, quinine (positive control), and distilled water (negative control) via gavage. Blood samples were collected for biochemical and hematological analyses. Notably, at a 40% concentration of A. annua infusion, there was a significant increase in the hemolysis rate of G6PD-deficient red blood cells compared to controls (p A. annua exhibited elevated aspartate aminotransferase (129.25 ± 4.55 U/L vs. 80.09 ± 4.03 U/L;p A. annua infusion tested positive for saponins. These findings underscore the risk of hemolysis in G6PD-deficient individuals upon ingesting A. annua.

Resistance of Microbial Community of Artemisia annua L.to Pathogenic Fungi

[Objectives]This paper was to figure out whether the dominant bacterial community has the role and effect of bacterial community and its defense mechanism against potential pathogenic fungi in Artemisia annua,and thus establish a systematic model of bacteria-fungus-plant.[Methods]Fifty-eight strains of bacteria and one strain of pathogenic fungi,Globisporangium ultimatum,were used for the experiments.These 58 bacterial strains were assembled into a bacterial community,and the bacteria with abundance in the top 1%were reassembled into a dominant bacterial community as measured by 16S rDNA.[Results]The growth of A.annua seedlings inoculated with bacterial communities and pathogenic fungi or dominant bacterial communities and pathogenic fungi was significantly better than that of A.annua seedlings inoculated with pathogenic fungi during in vitro confrontation,which was evident in both enzymatic and non-enzymatic antioxidant assays.[Conclusions]The results suggest that the dominant bacterial community has a crucial role as a representative core microbial community of synthetic bacterial community,which can protect plants by interfering with the growth of phytopathogenic fungi mediated by chemical signals,and can be used as the main synthetic community of biocides to achieve the effect of biocontrol.

Study on Soothing and Oil Control Anti-acne Effects of Artemisia Annua Extract

Preliminary exploration of the soothing,oil control,acne removing effects and its mechanisms of Artemisia annua extract.The soothing effect of Artemisia annua extract was tested by hyaluronidase biochemical reaction.The soothing and oil-controlling effects were investigated by cell model.The inhibitory effect on propionibacterium acnes was studied by suspension quantification.The results showed that Artemisia annua extract could effectively inhibit the degradation of hyaluronic acid(P<0.01).Artemisia annua extract significantly inhibited the secretion of inflammatory cytokines TNF-αand IL-6 in RAW264.7 cells(P<0.05).Artemisia annua extract at 0.5%,0.25%,0.125%could significantly inhibit the secretion of oil by SZ95 cells(P<0.05).The minimum inhibitory concentration of Artemisia annua extract against propionibacterium acnes was 0.625%,and the inhibitory rate against propionibacterium acnes increased with the increase of the concentration of Artemisia annua extract.In summary,Artemisia annua extract can achieve acne efficacy through soothing and oil control,and this function may be achieved by reducing hyaluronic acid degradation,inhibiting inflammatory pathways produced by inflammatory factors TNF-αand IL-6,inhibiting oil secretion,and inhibiting the growth of propionibacterium acnes.

Effects of Fungal Elicitors on Cell Growth and Artemisinin Accumulation in Hairy Root Cultures of Artemisia annua

The artemisinin accumulation in the hairy root cultures of Artemisia annua L. was enhanced via a treatment of three fungal elicitors separately ( Verticillium dahliae Kleb., Rhizopus stolonifer (Ehrenb. ex Fr.) Vuill and Colletotrichum dematium (Pers.) Grove). Among these three elicitors, V. dahliae had the highest inducing efficiency, but none of them manifests any noticeable effects on the cell growth of the hairy root cultures. The artemisinin content of the hairy root cultures treated with V. dahliae elicitor was 1.12 mg/g DW, which was 45% higher than the control (0.77 mg/g DW). The results showed that elicitation was dependent on the elicitor concentration, the incubation period and the physiological stage at which the hairy root cultures were treated. In addition, the authors found that for V. dahliae , the optimum concentration was 0.4 mg carbohydrate per millilitre medium, the strongest response of A. annua hairy root cultures to the elicitation was at the late exponential growth stage, and the highest artemisinin content of the hairy root cultures was on the 4th day post treatment.

Elicitation on Artemisinin Biosynthesis in Artemisia annua Hairy Roots by the Oligosaccharide Extract from the Endophytic Colletotrichum sp. B501

The oligosaccharide elicitor from the mycelial wall of an endophytic Colletotrichum sp. B501 promoted the production of artemisinin in Artemisia annua L. hairy root culture. When hairy roots of 22-day-old cultures (later growth phase) were exposed to the elicitor (20 mg/L) for 4 d, the maximum content of artemisinin reached 1.15 mg/g, a 64.29% increment over the control. The electron X-ray microanalysis disclosed the rapid accumulation of Ca 2+ in the elicited cortical cells of hairy root. The electronic microscope observation revealed the high electron density area in vacuole of elicited cells. During the first day of elicitation the peroxidase activity of hairy roots was improved sharply. Some cellular morphological changes including cell shrinkage, condensation of cytoplasm and nuclear fragmentation, coincident with the appearance of DNA ladders, were observed after the third day of elicitation. It was suggested that the oligosaccharide elicitor triggered the programmed cell death, which may provide the substance or chemical signal for artemisinin biosynthesis.

Molecular Cloning, Escherichia coli Expression and Genomic Organization of Squalene Synthase Gene from Artemisia annua

A 1 539 by squalene synthase (AaSQS) cDNA was cloned from a high-yield Artemisia annua L. strain 001 by reverse transcription-polymerise chain reaction (RT-PCR). The amino acid sequence of AaSQS is 70%, 77%, 44% and 39%a identical to that of squalene synthases from Arabidopsis thaliana, tobacco, human and yeast, respectively. The AaSQS genomic DNA has a complex organization containing 14 exons and 13 introns. Full-length or C-terminal truncated cDNA was subcloned into prokaryotic expression vector pET30a and the constructed plasmid was introduced to Escherichia coli strain BL21 (DE3) for induced overexpression. No squalene synthase protein with expected molecular mass was observed in E. cola containing the putative full-length squalene synthase cDNA, however, overexpression in E. coli was achieved by truncating 30 amino acids of hydrophobic region at the carboxy terminus.

Cloning, E. coli Expression and Molecular Analysis of a Novel Sesquiterpene Synthase Gene from Artemisia annua

A 1 886 bp full-length sesquiterpene synthase (AaSES) cDNA was cloned front a high-yield Artemisia annua L. strain 001 by a rapid amplification of cDNA end (RACE) strategy. AaSES is 59% identical to Artemisia cyclase cDNA clone cASC125, 50% identical to epi-cedrol synthase from A. annua , 48% identical to amorpha-4, 11-diene synthase from A. annua, 39% identical to the 5-epi-aristolechene synthase from tabacco, 38 % identical to vetispiradiene synthase front H. muticus, 41 % identical to the, delta-cadinene synthase from cotton. The coding region of the cDNA was inserted into a procaryotic expression vector pET-30a and overexpressed in E. coli BL21 ( DE3). The cyclase proteins extracted front bacterial culture were found largely in an insoluble protein fraction. AaSES expresses in leaves, stems a-rid flowers, not in roots as indicated by Northern blotting analysis.

Effects of Ag-carrying Zirconium Phosphate on the Kinetics of Growth of the Roots of Culture Artemisia annua

在植物组织和细胞培养过程中 ,尤其是在生物反应器培养中的染菌问题 ,一直是制约植物细胞培养工业化的难题。通过比较各种防腐剂的抑菌效果 ,确定银型磷酸锆盐抗菌粉为青蒿根培养的最佳防腐剂。银型磷酸锆盐抗菌粉在浓度为 30mg/L时 ,既能降低培养液的染菌几率 ,又不明显抑制青蒿根的生长及青蒿素的生物合成。在添加 30mg/L抗菌粉的培养液中进行的青蒿根生长、pH值变化以及残糖、铵离子和硝酸根离子消耗的动力学研究表明 ,在 4 0d内青蒿根在培养液中生长良好 ,营养成分的消耗和对照呈相似的趋势。

Target-site Mechanisms Involved in Annual Bluegrass(Poa annua L.) Tolerance to Fenoxaprop-P-ethyl

Annual bluegrass （Poa annua L.） was found to be tolerant to fenoxaprop- P-ethyl as well as quizalofop-P-ethyl, haloxyfop-R-methyl, clodinafop-propargyl, fluaz- ifop-P-butyl, cyhalofop-butyl, sethoxydim and tralkoxydim, whereas it was sensitive to clethodim and tepraloxydim. The acetyI-CoA carboxylase （ACCase） IC50 values of five P. annua biotypes were 10.46 to 11.98-fold higher than the susceptible Japanese foxtail （Alopecurus japonicus Steud.）. The presence of the polymorphic lie and Leu at 1 781, which the presence of Leu at 1 781 had been reported to be in- volved in the resistance of grass weeds to ACCase inhibitors, was subsequently i- dentified in the ACCase of P. annua. Furthermore, the expression level of gene that encoding P. annua ACCase was found to be approximately 4.67 to 7.37-fold higher than A. japonicus, possibly explaining the P. annua target site tolerance to fenoxaprop-P-ethyl.

黄花蒿(Artemisia annua L.)提取物对两种病原真菌的生物活性

以石油醚Ⅰ(30℃～60℃)、石油醚Ⅱ(60℃～90℃)、乙醇和丙酮等4种溶剂对黄花蒿的根、茎、叶进行初步提取,采用生长速率法测定不同提取物对玉米小斑病菌、棉花枯萎病菌的抑菌活性。结果表明,黄花蒿提取物对玉米小斑病菌的生物活性好于棉花枯萎病菌;黄花蒿叶的提取物抑菌效果最好,根的抑菌效果最差;叶的石油醚(60℃～90℃)提取物对玉米小斑病菌EC50为156.32mg/L;而叶的丙酮提取物对玉米小斑病菌的EC50为82.37mg/L。

Studies on Acaricidal Bioactivities of Artemisia annua L.Extracts Against Tetranychus cinnabarinus Bois.(Acari:Tetranychidae)

The aim of this study was to determine the best extraction technique, the most suitable solvent, the optimal plant parts, and the acaricidal activities of Artemisia annua L. The petroleum ether （30-60℃）, petroleum ether （60-90℃）, ethanol, acetone, and water parallel and sequenced extracts were obtained from the leaves, stems and roots of different period of A. annua L. in April, May, June, July and September respectively. And then the acaricidal bioactivities against Tetranychus cinnabarinus of all extracts were determined by the slide-capillary method in the laboratory. The results indicated that the acaricidal bioactivities elevated as the development of A. annua plant at the concentration of 5 mg mL-L The general tendency exhibited the sequence of July 〉 June 〉 May 〉 April, but September decreased comparing to July. However, the most effective extracts in five months were all acetone parallel extract of A. annua leaf, and the corrected mortalities treated after 48 h ranged from 74 to 100%. The median lethal concentrations （LC50） against T. cinnabarinus of acetone parallel extracts ofA. annua leaves in September, July, June, May and April were 0.5986, 0.4341, 0.8376, 0.9443 and 1.3817 mg mL^-1, respectively, treated after 48 h. The 13 groups were isolated from acetone extracts ofA. annua leaves in July by column chromatography, both the 1 lth and 12th groups exhibited strong bioactivities. The median lethal concentrations of the 1 lth and 12th groups against T. cinnabarinus were 0.3683 and 0.1586 mg mL^-1, respectively. The acetone parallel extract ofA. annua leaf in July was the most toxic to T. cinnabarinus and the corrected mortality was 100% after 48 h. The acetone parallel extract of the 1 lth and 12th groupswere the most active components, acted as the emphases in further study.

Preliminary pharmacological evaluation of Martynia annua Linn leaves for wound healing

Objective:To evaluate the wound healing potential of fractions from ethanol extract of Martynin annua(M.annua) Linn leaves.Methods:Ethanol extract of Af.annua Linn leaves was fractionate into three different fractions(MAF-A,MAF-B and MAF-C) which were screened for wound healing potential using two models:excision and incision on rats.The thin layer chromatography(TLC) profile of all fractions were analysed and TLC of luteolin was also done.The PovidoneIodine Ointment was used as reference for comparision.Excision and incision wounds were created on dorsal portion of rats for study.Wound contraction,biochemical parameters(protein level and hydroxyproline level) and histopathological study were performed in excision wound model whereas incision model was used for determination of tensile strength.Results:The wound contraction and tensile strength of skin tissues were observed significantly greater in MAF-C fraction treated group than other two fractions(P<0.01).In excision wound method(on day 18) protein content and hydroxyproline were found significantly higher in MAF-C group than control group(P<0.01).Histopathological study also showed better angiogenesis,matured collagen fibres and fibroblast cells as compared with the control group.Conclusions:In conclusion, our findings suggest that fiaction MAF-C from ethanol extract of M.annua leaves is found most effective in wound healing.

Trichome-Specific Expression of Amorpha-4,11-Diene Synthase, a Key Enzyme of Artemisinin Biosynthesis in <i>Artemisia annua</i>L., as Reported by a Promoter-GUS Fusion

Artemisia annua L. produces small amounts of the sesquiterpenoid artemisinin, which is used for treatment of malaria. A worldwide shortage of the drug has led to intense research to increase the yield of artemisinin in the plant. In order to study the regulation of expression of a key enzyme of artemisinin biosynthesis, the promoter region of the key enzyme amorpha-4,11-diene synthase (ADS) was cloned and fused with the β-glucuronidase (GUS) reporter gene. Transgenic plants of A. annua expressing this fusion were generated and studied. Transgenic plants expressing the GUS gene were used to establish the activity of the cloned promoter by a GUS activity staining procedure. GUS under the control of the ADS promoter showed specific expression in glandular trichomes. The activity of the ADS promoter varies temporally and in old tissues essentially no GUS staining could be observed. The expression pattern of GUS and ADS in aerial parts of the transgenic plant was essentially the same indicating that the cis-elements controlling glandular trichome specific expression are included in the cloned promoter. However, some cis-element(s) that control expression in root and old leaf appears to be missing in the cloned promoter. Furthermore, qPCR was used to compare the activity of the wild-type ADS promoter with that of the cloned ADS promoter. The latter promoter showed a considerably lower activity than the wild-type promoter as judged from the levels of GUS and ADS transcripts, respectively, which may be due to the removal of an enhancing cis-element from the ADS promoter. The ADS gene is specifically expressed in stalk and secretory cells of glandular trichomes of A. annua.

Wound healing effect of flavonoid rich fraction and luteolin isolated from Martynia annua Linn,on streptozotocin induced diabetic rats

Objective:To evaluate wound healing potential of flavonoid fractions of Martynia annua (M.annua) Linn,leaves in diabetic rats on the basis of folkloric information and preliminary study.Methods:The flavonoid compound luteolin and apigenin were isolated from dried leaves of plant by column chromatography.The two concenlrations(0.2%and 0.5%w/w) of luteolin and flavonoid fraction were selected for topically applied as ointment on diabetic wound. The Povidone Iodine Ointment USP was used as a reference.On 18th days,protein content, hydroxyproline and antioxidants(SOD,CAT and GSH) level in granuloma tissues were determined. Results:The results showed that,percent wound contraction were observed significantly(P【0.01) greater in MAF fraction and 0.5%w/w of luteolin treatment groups.Presence of matured collagen fibres and fibroblasts with better angiogenesis were observed in histopathological studies. Conclusions:In conclusion,our findings suggest that flavonoid fraction(MAF) and luteolin(0.5% w/w) may have potential benefit in enhancing wound healing in diabetic condition,possibly due to Gee-radical scavenging activity of plant.

Efficient Somatic Embryogenesis and Organogenesis of Self-Pollination <i>Artemisia annua</i>Progeny and Artemisinin Formation in Regenerated Plants

To enhance the understanding of artemisinin biosynthesis, we have successfully bred self-pollination Artemisia annua plants. Here, we report efficient somatic embryogenesis and organogenesis of self-pollination plants and artemisinin formation in regenerated plants. The first through sixth nodal leaves of seedlings are used as explants. On agar-solidified MS basal medium supplemented with TDZ (0.6 mg/l) and IBA (0.1 mg/l), all explants after inoculation of less than 3 weeks start to form embryogenic calli, which further produce globular, torpedo, heart and early cotyledon embryos. In all six positional leaves, explants from the sixth leaf show the rapidest responses to induction of embryogenic calli and somatic embryos. On this medium, somatic embryos continuously develop into adventitious buds, which can form adventitious roots on a rooting medium containing NAA (0.5 mg/l). Meanwhile, on agar-solidified MS basal medium supplemented with BAP (1 mg/l) and NAA (0.05 mg/l), approximately 100% of explants from leaves #3-6 form calli in less than 3 weeks of inoculation and adventitious buds via organogenesis in 3-4 weeks. In all six positional leaves, explants from the sixth leaf exhibit the rapidest response to induction of calli and adventitious buds. Nearly 100% adventitious buds can form adventitious roots on the rooting medium. Regenerated plants from both somatic embryogenesis and organogenesis complete self-pollination to produce seeds in 80-90 days of growth in growth chamber. LC-ESI-MS analysis demonstrates that regenerated plants biosynthesize artemisinin. These results show the highly efficient regeneration capacity of self-pollination A. annua plants that can form a new platform to enhance the understanding of artemisinin biosynthesis and metabolic engineering.

Effects of traffic pollution on the genetic structure of Poa annua L.populations

The genetic composition of Poa annua L. populations with a series of traffic pollution was studied by starch electrophoresis. Five enzyme systems were stained. The results showed that: (1) Traffic pollution can dramatically change genotypic frequencies at some loci of P. annua populations. Significant deviations from Hardy Weinberg equilibrium were observed on loci Fe 1 and Me due to the excess of heterozygotes in some populations. (2) The effective number of alleles per locus and the observed and expected heterozygosity were higher in the pollution series than in the clear control site(Botanic Park population), but the increase was not related with the pollution extent. (3) Most genetic variation was found within populations, and only 6 21% was among populations of the polluted series. Slightly higher differentiation( F ST =7 98%) was observed when the control population was included. (4) The calculated gene flow(Nm) is 2 8841 per generation. The mean of genetic identity is 0 9864 and the genetic distance average to 0 0138

Comparative Study on Population Ecological Distribution and Extracellular Enzyme Activities of Endophytic Fungi in <i>Artemisia annua</i>

The endophytic fungi in different tissues of Artemisia annua was isolated and purified to explore their ecological distribution and tissue preference, and the extracellular enzyme activities of dominant endophytic fungi were determined to characterize the metabolic function of endophytic fungi. The results showed that a total of 67 endophytic fungi were obtained from Artemisia annua tissues. The number and species of endophytic fungi in different tissues were significantly different. The number, colonization rate (CR) and isolation rate (IR) of endophytic fungi in root were significantly higher than those of stem and leaf. The dominant endophytic fungi, diversity and similarity coefficient of endophytic fungi also showed significant difference among tissues. The extracellular enzyme activities of endophytic fungi in different tissues are significantly different. The enzyme activities of endophytic fungi isolated from root are significantly higher than those isolated from stem and leaf. The research results showed that the endophytic fungi in Artemisia annua had significant tissue preference, and the metabolic function of endophytic fungi showed significant difference among tissues. This will lay a foundation for further research, development and utilization of endophytic fungi, and also provide a theoretical basis for screening functional endophytic fungi in Artemisia annua.

Increased artemisinin production by promoting glandular secretory trichome formation and reconstructing the artemisinin biosynthetic pathway in Artemisia annua

Dear Editor,Artemisinin,which has potent antimalarial properties,is a sesquiterpene endoperoxide originally isolated from the traditional Chinese medicinal plant Artemisia annua.However,the artemisinin content in wild-type(WT)A.annua is low(1-10 mg/g dry weight),leading to its erratic supply and price fluctuations[1].

Dried-leaf Artemisia annua: A practical malaria therapeutic for developing countries?

Artemisinin from the plant Artemisia annua （A. annua） L., and used as artemisinin combination therapy （ACT）, is the current best therapeutic for treating malaria, a disease that hits children and adults especially in developing countries. Traditionally, A. annua was used by the Chinese as a tea to treat “fever”. More recently, investiga-tors have shown that tea infusions and oral consumption of the dried leaves of the plant have prophylactic and therapeutic effcacy. The presence of a complex matrix of chemicals within the leaves seems to enhance both the bioavailability and effcacy of artemisinin. Although about 1000-fold less potent than artemisinin in their antiplasmodial activity, these plant chemicals are mainly small molecules that include other artemisinic compounds, terpenes （mainly mono and sesqui）, favonoids, and polyphenolic acids. In addition, polysaccharide constituents of A. an-nua may enhance bioavailability of artemisinin. Rodent pharmacokinetics showed longer T？ and Tmax and greater Cmax and AUC in Plasmodium chabaudi -infected mice treated with A. annua dried leaves than in healthy mice. Pharmacokinetics of deoxyartemisinin, a liver metabolite of artemisinin, was more inhibited in infected than in healthy mice. In healthy mice, artemisinin serum levels were 〉 40-fold greater in dried leaf fed mice than those fed with pure artemisinin. Human trial data showed that when delivered as dried leaves, 40-fold less artemisinin was required to obtain a therapeutic response compared to pure artemisinin. ACTs are still unaffordable for many malaria patients, and cost estimates for A. annua dried leaf tablet production are orders of magnitude less than for ACT, despite improvements in the production capacity. Considering that for 〉 2000 years this plant was used in traditional Chinese medicine for treatment of fever with no apparent appearance of artemisinin drug resistance, the evidence argues for inclusion of affordable A. annua dried leaf tablets into the arsenal of drugs to combat malaria and other artemisinin-susceptible diseases.

Study on the Repairing Effect of Cosmetics Containing <i>Artemisia annua</i>on Sensitive Skin

In view of the increasing sensitivity of consumer skin in recent years, cosmetics containing Artemisia annua extract was tested to evaluate its effectiveness in repairing sensitive skin. Through the experiment of xylene-induced ear swelling in mice, it was found that the inhibition rates of ear swelling in mice induced by xylene in three groups of cosmetics containing Artemisia annua extract reached 60.40%, 73.36% and 74.01%, respectively, close to the positive drug group. Twenty-five sensitive skin volunteers were selected for human clinical trial, and the skin TEWL value, cuticle hydration degree and skin heme (ultra-high concentration) were tested. The results showed that using cosmetics containing Artemisia annua extract for four weeks could effectively increase the hydration degree of cheek cuticle by 63.90% and reduce transepidermal waterloss (TEWL) by 21.51%. The skin heme (ultra-high concentration) decreased by 69.14% and the affected area decreased by 77.47%. The results show that the cosmetics containing Artemisia annua extract can inhibit inflammation, repair skin barrier, improve damaged skin, and reduce redness and other sensitive skin symptoms.