Targeting colorectal cancer with Herba Patriniae and Coix seed:Network pharmacology,molecular docking,and in vitro validation

BACKGROUND Herba Patriniae and Coix seed(HC)constitute a widely utilized drug combination in the clinical management of colorectal cancer(CRC)that is known for its diuretic,anti-inflammatory,and swelling-reducing properties.Although its efficacy has been demonstrated in a clinical setting,the active compounds and their mechanisms of action in CRC treatment remain to be fully elucidated.AIM To identify the active,CRC-targeting components of HC and to elucidate the mechanisms of action involved.METHODS Active HC components were identified and screened using databases.Targets for each component were predicted.CRC-related targets were obtained from human gene databases.Interaction targets between HC and CRC were identified.A“drug-ingredient-target”network was created to identify the core components and targets involved.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were conducted to elucidate the key pathways involved.Molecular docking between core targets and key components was executed.In vitro experiments validated core monomers.RESULTS Nineteen active components of HC were identified,with acacetin as the primary active compound.The predictive analysis identified 454 targets of the active compounds in HC.Intersection mapping with 2685 CRC-related targets yielded 171 intervention targets,including 30 core targets.GO and KEGG analyses indicated that HC may influence the phosphoinositide 3-kinase(PI3K)/Akt signaling pathway.Molecular docking showed that acacetin exhibited an optimal interaction with AKT1,identifying PI3K,AKT,and P53 as key genes likely targeted by HC during CRC treatment.Acacetin inhibited HT-29 cell proliferation and migration,as well as promoted apoptosis,in vitro.Western blotting analysis revealed increased p53 and cleaved caspase-3 expression and decreased levels of p-PI3K,p-Akt,and survivin,which likely contributed to CRC apoptosis.CONCLUSION Acacetin,the principal active compound in the HC pair,inhibited the proliferation and migration of HT-29 cells and promoted apoptosis through the PI3K/Akt/p53 signaling pathway.

The Role of Coix lacryma-jobi Linn. in Wastewater Treatment System of Constructed Wetland

The dynamic of growth and nutrient （N and P） absorption of Coix lacryma-jobi Linn.in constructed wetland （CW）,as well as the effect of constructed wetland harvest on N,P absorption in eutrophic water were studied.The results revealed that,in Summer and Autumn,especially in August,C.lacryma-jobi could obtain higher biomass and N,P accumulations,thereby achieving the most efficient purification; from the breeding value point of view,N,P contents could be improved by the appropriate increase of harvest times of C.lacryma-jobi,while too many harvest times was inappropriate to the wastewater purification by C.lacryma-jobi and the nutrients accumulation.So,C.lacryma-jobi should be harvested completely in August during its growth period,and twice-3 times of harvests was appropriate.

Research Progress of Biology in Coix

Coix has a long cultivation history as a minor crop in China, but many problems including unclear origin, chaos of species, loss and inaccurate use of germplasm resources exist in its research and production due to the lack of systematic study. In recent years, the importance of Coix has been realized with the exploration of its effects in treating and preventing cancer. The systematic study of Coix has met a superb development opportunity with the rapid development of biotechnology especially the deep-sequencing technique. Therefore, this paper presented here the current status of origin and evolution, classification, cytology and molecular biology progress of Coix, analyzed the ploidy of Coix and the genetic relationships with Zea, Sorghum and so on from the point of cytology, and then based on the molecular biology, analyzed the important significance of DNA molecular marker, genetic mapping and gene library in the research on identification of genetic relationship between species and genetic breeding, which could provide useful information for further Coix research.

Anti-tumor effect of coix seed based on the theory of medicinal and food homology

Coix seed is a dry and mature seed of Coix lacryma-jobi L.var.ma-yuen(Roman.)Stapf in the Gramineae family.Coix seed has a sweet,light taste,and a cool nature.Coix seed enters the spleen,stomach,and lung meridians.It has the effects of promoting diuresis and dampness,strengthening the spleen to prevent diarrhea,removing arthralgia,expelling pus,and detoxifying and dispersing nodules.It is used for the treatment of edema,athlete's foot,poor urination,spleen deficiency and diarrhea,dampness and obstruction,lung carbuncle,intestinal carbuncle,verruca,and cancer.The medicinal and health value is high,and it has been included in the list of medicinal and food sources in China,which has a large development and application space.This article reviews the current research achievements in the processing methods and anti-tumor activities of Coix seed and provides examples of its clinical application in ancient and modern times,aiming to provide reference for further research on Coix seed and contribute to its clinical application and development.Through the analysis of the traditional Chinese patent medicines,and simple preparations and related health food of Coix seed queried by Yaozhi.com,the source,function,and dosage form of Coix seed were comprehensively analyzed,with a view of providing a reference for the development of Coix seed medicine and food.

Screening and Analysis of Germplasm Resources of Wild Coix lacryma-jobi L. with High Content of Coixin

[Objectives] To screen and analyze germplasm resources of wild Coix lacryma-jobi L. with high content of coixin. [Methods] From the 150 kinds of collected wild C. lacryma-jobi L. germplasm resources, 24 kinds of excellent resources were screened and detected. Morphological statistics were carried out, and the content of coixin in the four organ of roots, stems, leaves and seeds were measured, and the approved cultivar white husk Liugu of C. lacryma-jobi L. was taking as the control. [Results] Among the organs of the plant of C. lacryma-jobi L., the roots had the highest content of coixin, so the roots of C. lacryma-jobi L. can be used as the medicinal parts of C. lacryma-jobi L. [Conclusions] The wild C. lacryma-jobi L.Y229, Y164, Y166, Y83, and Y84 stains had excellent yield, and their root coixin content was significantly higher than that of the control variety, so they can be used for cultivation as excellent germplasm, so as to obtain new variety(line) of C. lacryma-jobi L.with higher content of coixin.

薏苡属(CoixL.)2种1变种的遗传多样性及亲缘关系的初步分析

利用垂直板型聚丙烯酰胺凝胶电泳技术,检测了薏苡属(CoixL.)2种1变种:薏苡(CoixlacrymajobiL.)6个居群、念珠薏苡(C.lacrymajobivar.maximaMakino)3个居群和薏米(C.chinensisTod.)2个居群共11个居群5个酶系统6个位点上9个等位基因的分布情况,结果表明:薏苡、念珠薏苡和薏米的遗传多样性水平普遍较低,每个位点等位基因平均数(A)分别为12,11和12;多态位点百分数(P)分别为167,113和167;平均期望杂合度(He)分别为006,003和007。聚类分析不能从遗传上将以上的2种1变种区分开。

Effect of Planting Density,Fertilizer Level,and Number of Seedlings per Hole on Yield of Coix Lachryrma-jobi L.

In order to study high yield cultivation technology for Coix Lachryrma-jobi L.,and discuss the effect of planting density,fertilizer level,and number of seedlings per hole on yield of Coix Lachryrma-jobi L.,this paper took domesticated variety Xingren Small White Shell of Coix Lachryrma-jobi L. as an example. Using L9( 34) orthogonal design,it selected optimum combination of planting density,fertilizer level,and number of seedlings per hole to guide planting of Coix Lachryrma-jobi L.. Experiment indicated that the optimum planting density( spacing in the rows and spacing between rows) of Coix Lachryrma-jobi L. is 40 cm x 80 cm; the optimum fertilizer level is pure nitrogen 15 kg,pure phosphorus 15 kg and pure potassium 15 kg per mu; the optimum number of seedlings per hole is 5 seedlings per hole. Through variance analysis,it indicated that planting density exerts significance effect on the yield of Coix Lachryrma-jobi L.

Effects of Drought Stress on Morphological and Physiological Indices of New Coix Varieties in Yunnan

In order to study the effects of drought stress on the morphological and physiological indexes of Yunnan Coix at different growth stages, two new varieties of Yunnan Coix Wenyi 4 and Wenyi 5 were used as experimental materials, potted and compared with CK by normal irrigation. Morphological indices such as plant height, stem diameter and physiological indices such as protein, soluble sugar, chlorophyll content and relative conductivity (REC) of the two varieties under drought stress at seedling stage, tillering stage and jointing stage were determined, and data statistics and variance analysis were performed for each index. The results showed that the morphological indices, plant height of Wenyi 4 and Wenyi 5 were significantly affected by drought stress at seedling stage, while the stem diameter was significantly affected by drought stress at seedling and jointing stage. In terms of physiological indices, the relative conductivity, protein and soluble sugar contents of Wenyi 5 increased at a higher degree, whereas chlorophyll content decreased at a higher degree, indicating that Wenyi 5 was greatly affected by drought stress. Through the analysis of drought resistance of the tested materials at different stages by polar ordination method, the results were as follows: jointing stage of Wenyi 5>jointing stage of Wenyi 4>seedling stage of Wenyi 4>seedling stage of Wenyi 5>tillering Stage of Wenyi 4>Tillering Stage of Wenyi 5. For the growth period, the drought resistance was as follows: jointing stage>seedling stage>tillering stage. In conclusion, Wenyi 4 has strong drought resistance and is suitable for droughtresistant cultivation.

薏苡(Coix.L)正常幼苗与白化苗的同工酶分析

比较薏苡正常幼苗与其白化苗的过氧化物酶及酯酶的同工酶的结果表明,正常绿苗的过氧化物酶酶带有8条,白化苗有10条(多出A_5、A_7);正常绿苗的酯酶同工酶带有4条,白化苗有3条,白化苗缺少B_2带,并且B_(1-1)与B_1带相近似。结果表明白化与过氧化物酶及酯酶同工酶的表达有关系。

Anti-lipid-oxidation effects and edible safety evaluation of the oil extracted by a supercritical CO_(2) process from coix seed fermented by Monascus purpureus

The physicochemical properties and composition of coix seed oil produced by Monascus purpureus fermentation and supercritical CO_(2)extraction were determined.Anti-lipid-oxidation and edible safety were evaluated using a cholesterol-fish oil model,acute oral toxicity assay,and genetic toxicity assay in vitro and in vivo,respectively.The results show that the extraction oil from fermented coix seed(FCS-O)had good physicochemical quality and abundant active components with physiological function.In particular,γ-tocotrienol,γ-oryzanol,coixenolide and oleic acid concentrations reached 72.83μg/g,745.96μg/g,9.65 mg/g and 316.58 mg/100 g DW,respectively.The FCS-O exhibited higher antioxidant capability in inhibiting lipid oxidation and peroxidation.Compared to the blank control,the concentrations of 7-ketocholestreol and peroxide only were 8.42μg/mL and 16.16 mmol/kg at 168 h of oxidation(P<0.01).In addition,the FCS-O has been confirmed to be a very safe edible oil,with no acute toxicity(LD50>10 g/kg bw,considered actually non-toxic)and no induced mutagenicity,cytotoxicity or genotoxicity.These results serve as a good safety reference for future application of the oil from fermented coix seed.The development and utilization of this kind of oil will be beneficial as a food,food ingredient,nutritional supplement,or natural food antioxidant to promote good health function.

Extraction of Flavonoids from Leaves of Coix lacryma-jobi L.

[Objectives] To extract the flavonoids from leaves of Coix lacryma-jobi L. [Methods] Ethanol extraction method was adopted,spectrophotometry was used,and single factor experiment and orthogonal experiment were carried out to study the effects of ethanol percentage,extraction temperature,solid-to-liquid ratio and extraction time on the extraction of total flavonoids from leaves of C. lacryma-jobi L.[Results] The order of 4 factors influencing the extraction of flavonoids from leaves of C. lacryma-jobi L. was: solid-to-liquid ratio > extraction time > ethanol percentage > extraction temperature. When the extraction temperature was 70℃,the extraction time was 1. 5 h and the solid-liquid ratio was 1: 10,the ethanol percentage was 60%,the extraction effect was the best,extraction of flavonoids was 0. 107 5 mg/m L.[Conclusions] This study is expected to provide a theoretical basis for further development and utilization of C. lacryma-jobi L.

Effects of Different Herbicides on Weed Control,Agronomic Characters and Grain Quality of Coix lacryma-jobi L.

[Objectives]The paper was to study the effects of different herbicides on weed control,agronomic characters and grain quality of Coix lacryma-jobi L.[Methods]Six commonly used herbicides were selected for the trial,including atrazine SC,butachlor SC,monosulfuron WP,MCPA-starane EC,clopyralid AS and tribenuron-methyl WP.[Results](i)Atrazine,butachlor and monosulfuron had no chemical injury to C.lacryma-jobi L.,while clopyralid caused slight chemical injury.(ii)The fresh weight control effects of atrazine,butachlor and monosulfuron on weeds increased first and then decreased.The initial control effect of MCPA-starane and clopyralid reached more than 90%;the control effect of monosulfuron increased gradually in the late stage,reaching 97%at 60 d post spraying,and the overall control effect was optimal.(iii)Compared with manual weeding,herbicides had no significant effect on plant height of C.lacryma-jobi L.However,compared with no weeding,other herbicides except tribenuron-methyl inhibited plant growth to a certain extent.Herbicides had different effects on main stem diameter,tiller number per plant and grain number per plant of C.lacryma-jobi L.(iv)Different herbicides had no significant effect on 100-grain weight,but had significant effect on yield.(v)Compared with manual weeding,atrazine,butachlor and clopyralid decreased the protein content of C.lacryma-jobi L.,and the use of atrazine,butachlor,monosulfuron,MCPA-starane and tribenuron-methyl decreased the fat content of C.lacryma-jobi L.;butachlor and MCPA-starane decreased the glycerol trioleate content of C.lacryma-jobi L.Compared with no weeding,chemical herbicides had no significant effect on the protein content,while atrazine decreased the fat content,and butachlor and MCPA-starane decreased the glycerol trioleate content of C.lacryma-jobi L.[Conclusions]The use of herbicides significantly increases crop yield compared with no weeding,but has an impact on agronomic characters and grain quality.

Effects of Thallium Stress on Photosynthesis, Chlorophyll Fluorescence Parameters and Antioxidant Enzymes Activities of Coix Lacryma-jobi

Levels of T1 （Thallium） in soil from 0 （control） to 50 μg/L through 0.2, 0.5, 1 and 2.5μg/L were directly and positively correlated to levels of T1 in plant tissue, the accumulation being maximum in roots, intermediate in leaves and minimum in stems. Thallium, especially at higher concentrations, adversely affected photosynthesis （as judged based on chlorophyll fluorescence parameters）, suggesting inhibition of photo-activation of PSII （Photosystems II）, and also decreased the rate of photosynthesis, the rate of transpiration and stomatal conductivity drastically. Exposure to TI also increased the activity of CAT （Catalase） （except at 1 μg/L） and POD （Peroxidase） （except at 0.2 μg/L）, suggesting that the antioxidant systems in Coix lacryma-jobi were the main contributors of CAT and SOD （Superoxide Dismutase） and that the tolerance of C. lacryma-jobi to T1 is mainly due to this induced antioxidant machinery.

Advances in research on anti-tumor of Coix seed

Coix seed is a dry mature seed of Gramineae plant Coix. It has been confirmed that a plurality of components thereof have pharmacological effects. The mechanism of Coix seed against tumor includes the following aspects:(1) Inhibit the proliferation of tumor cells.(2) Inhibit the invasion and metastasis of tumor cells.(3) Inhibit the formation of tumor blood vessels.(4) Induce apoptosis of tumor cells.(5) The synergistic and attenuating effects on chemotherapy.(6) Enhance the sensitivity of tumor cells to radiation.(7) Adjust the body's immune function.(8) Relieve the pain of advanced tumors. In this paper, we summarized the research adcances of Coix seed foucusing on its anti-tumor effects.

Study on the mechanism of Coix seed in the treatment of colon cancer Based on network pharmacology

Objective:Study on the mechanism of Coix seed in the treatment of colon cancer based on network pharmacology.Methods:The main chemical components of Coix seed were identified by TCMP,and the target sites of drug components were screened.The target of the protein was mapped to the corresponding gene target through the Uniprot database.Genecards database was used to find the target gene of human colon cancer disease,and then the disease gene target was combined with the drug gene target to screen the common target of disease-drug composition,and the STRING platform was used to construct the drug component-disease target protein interaction.(PPI)network model.The Bioconductor database was used to analyze the potential targets,related molecular functions and signaling pathways of cocoon in the treatment of colon cancer by GO and KEGG enrichment.The disease-drug component-target-signal pathway network was established by Cytoscape software.Results:It was found that there are 9 pharmacodynamics and 28 targets related to colon cancer treatment,and GO functional enrichment analysis is based on correction.P<0.05 screened 346 molecular functional related items,including 256 biological processes,48 molecular functions,and 42 cell components.KEGG functional enrichment analysis screened 24 related pathways according to the adjusted P<0.05.Conclusion:PTGS2,AR,ESR1,PGR,ADH1C,AKR1B1,LTA4H,NCOA1,NCOA2 and other related targets may be potential targets for the treatment of colon cancer.

The Coix Genome Provides Insights into Panicoideae Evolution and Papery Hull Domestication

Coix is a grass crop domesticated as early as the Neolithic era.It is still widely cultivated for both highly nutritional food and medicinal use.However,the genetic study and breeding of this crop are hindered by the lack of a sequenced genome.Here,we report de novo sequencing and assembly of the 1619-Mb genome of Coix,and annotation of 75.39%repeats and 39629 protein-coding genes.Comparative genomics analysis showed that Coix is more closely related to sorghum than maize,but intriguingly only Coix and maize had a recent genome duplication event,which was not detected in sorghum.We further constructed a genetic map and mapped several important traits,especially the strength of hull.Selection of papery hull(thin:easy dehulling)from the stony hull(thick:difficult dehulling)in wild progenitors was a key step in Coix domestication.The papery hull makes seed easier to process and germinate.Anatomic and global transcriptome analysis revealed that the papery hull is a result of inhibition of cell division and wall biogenesis.We also successfully demonstrated that seed hull pressure resistance is controlled by two major quantitative trait loci(QTLs),which are associated with hull thickness and color,respectively.The two QTLs were further fine mapped within intervals of 250 kb and 146 kb,respectively.These resources provide a platform for evolutionary studies and will facilitate molecular breeding of this important crop.

Evolution and Domestication Footprints Uncovered from the Genomes of Coix

Coix lacryma-jobi,a plant species closely related to Zea and Sorghum,is an important food and medicinal crop in Asia.However,no reference genome of this species has been reported,and its exact phylogeny within the Andropogoneae remains unresolved.Here,we generated a high-quality genome assembly of coix comprising～1.73 Gb with 44485 predicted protein-coding genes.We found coix to be a typical diploid plant with an overall 1-to-1 syntenic relationship with the Sorghum genome,despite its drastic genome expansion(～2.3-fold)due mainly to the activity of transposable elements.Phylogenetic analysis revealed that coix diverged with sorghum～10.41 million years ago,which was～1.49 million years later than the divergence between sorghum and maize.Resequencing of 27 additional coix accessions revealed that they could be unambiguously separated into wild relatives and cultivars,and suggested that coix experienced a strong genetic bottleneck,resulting in the loss of about half of the genetic diversity during domestication,even though many traits have remained undomesticated.Our data not only provide novel comparative genomic and evolutionary insights into the Andropogoneae lineage,but also an important resource that will greatly benefit molecular breeding of this important crop.

Phytochemical screening, antibacterial and anthelmintic activities of leaf and seed extracts of Coix lacryma-jobi L.

Objective: To evaluate the possible phytochemical constituents, antibacterial and anthelmintic activity of Coix lacryma-jobi L. (Job's tears) using the chloroform leaves and seed extracts. Methods: The test for antibacterial activity and minimum inhibitory concentration was conducted by the disc diffusion and two-fold dilution method, respectively. In anthelmintic activity test, using Pheretima posthuma model, vermifuge and vermicidal activity were determined by using the chloroform extract at various concentrations. Results: The preliminary phytochemical screening of chloroform extracts of Job's tears leaves indicated the presence of alkaloid, carbohydrate, saponin, glycosides, flavonoids, phenols, tannins and steroids whereas the seeds extract contained glycosides, flavonoids, phenols and steroids, which revealed highest antimicrobial activity against Bacillus cereus and Klebsiella pneumoniae. The lowest minimum inhibitory concentration (12.5–50 mg/mL) was observed against all selected bacteria. On the other hand, it has been observed that chloroform leaves extracts showed shortest time of paralysis (P = 8.17 min) and death (D = 18.23 min) at 100 mg/mL concentration, in comparison with seed extracts (P = 36.83 min and D = 62.33 min) at 100 mg/mL concentration and albendazole (10 mg/mL) used as reference drug (P = 20.17 min and D = 43.67 min), which indicated the plant possessed mild anthelmintic activity. Conclusions: The chloroform extracts (leaves and seeds) showed efficacy for both bacterial infections and parasitic diseases, which ensure the traditional uses of Coix lacryma-jobi L.

Identification of anti-hyperuricemic components from Coix seed

Coix seed is a nutritious food with the effect of reducing uric acid.Few studies have been performed on the relevant active compounds.In this study,uric-acid-lowering active compounds were isolated and screened from Coix seed,and their structures were identified using NMR and mass spectrometry.The study showed that Coix seed effectively reduced serum uric acid and creatinine levels,suppressed liver xanthine oxidase activity,and improved kidney damage in mice due to hyperuricemia.Two major anti-hyperuricemic compounds were identified,namely,1-phenyl-1,3,3-propanetriamine(compound 1)and dihydroferulic acid(compound 2),among which compound 1 was a new compound,and compound 2 was separated from Coix seed for the first time.Two compounds showed potent inhibitory activity in vitro against xanthine oxidase,with IC_(50) values of 230.48 and 373.09μM,respectively.Furthermore,the molecular docking results indicated that the compounds could inhibit enzyme activity by interacting with residues within the active pocket of xanthine oxidase.This study provided a new alternative for the development of uric acid-lowering medications by identifying an anti-hyperuricemic alkaloid from Coix seed.

Shelf life evaluation of formulated cookies from Hanjeli (Coix lacryma-jobi L.) and Moringa leaf flour (Moringa oleifera)

Hanjeli(Coix lacryma-jobi L.)and moringa leaf(Moringa oleifera)remain underutilised despite their high nutritional value for food.In the present research,cookies made from hanjeli(Coix lacryma-jobi L.)with moringa leaf flour(Moringa oleifera)have been developed.Hanjeli cookies were measured for nutritional composition and shelf life assessment.The shelf life evaluation of hanjeli-moringa cookies was calculated using two approaches,which were accelerated shelf life testing(ASLT)methods with critical moisture content and the Arrhenius method.The results of the nutritional composition analysis showed that hanjeli-moringa cookies contain protein(9.14%),fat(24.67%),carbohydrates(61.62%),crude fibre(4.85%),and energy(505.01 Kcal).Using the Arrhenius method,the shelf life of hanjeli-moringa cookies was 95.46 days at room temperature and 97.63 days at 20℃.Meanwhile,the shelf life of cookies using the critical moisture content approach was 170.57 days.Shelf life is influenced by initial moisture content,critical water content of the product,packaging surface area and permeability,saturated vapour pressure,and slope sorption isotherm curve.The formulated cookies have significant amounts of nutrients that are acceptable and safe for consumption,with guaranteed desirable sensory properties.The result of this study shows that hanjeli-moringa cookies can be used as an alternative food for people who need high energy in a practical way of serving or ready to eat.