蜂胶黄酮Pinobanksin-3-acetate对结肠癌HCT-116细胞增殖及凋亡的影响

为探讨蜂胶黄酮(Pinobanksin-3-acetate,PB3A)对结肠癌HCT-116细胞增殖及细胞凋亡的影响,采用四甲基偶氮唑盐(MTT)比色法,检测不同浓度、不同时间PB3A、5-氟尿嘧啶(5-FU)及两种药物联合作用对HCT-116细胞生长所产生的影响,倒置显微镜观察细胞的形态学变化特点,Annexin V-FITC/PI双染色、流式细胞仪检测药物作用24 h后的细胞凋亡率。结果显示,PB3A对HCT-116细胞增殖有明显的抑制作用,并呈浓度和时间依赖性。其抑制活性与5-FU几乎没有显著性差异,联合用药具有协同效应;在一定剂量范围内,可见凋亡细胞明显增多。流式细胞仪分析结果表明,细胞凋亡率明显上升,呈剂量依赖性。PB3A对HCT-116细胞具有明显的增殖抑制和诱导细胞凋亡作用。

Theoretical Studies on the Aminolysis of Ester: Effects of the Catalysis and Substituent to the Reaction of Methyl Indole-3-acetate with Ammonia

A comprehensive exploration of the aminolysis mechanism for methyl indole-3-acetate with ammonia is carried out by employing the B3 LYP/6-311++G(d,p), M06-2 X/6-311++G(d,p) and MP2/6-311++G(d,p)//M06-2 X/6-311++G(d,p) levels. Two alterative reaction channels of the concerted and addition/elimination stepwise processes including the uncatalyzed, base-catalyzed reactions are taken into consideration. Subsequently, the substituent effects and solvent effects in methanol are also evaluated at the M06-2 X/6-311++G(d,p) level. The calculated results indicate that the calculated values of M06-2 X level are quite close to those of MP2, the stepwise pathway has more advantages to the concerted one for all of the reaction processes and the catalyst facilitates the proton migration and decreases the energy barriers as well. It is shown that the most preferred mechanism is the based-catalyzed stepwise process, the substituent of NH2 group slightly accelerates all the aminolysis reaction processes, and the solvent effect does not remarkably change the mechanism of the reaction.

Lateral root elongation in maize is related to auxin synthesis and transportation mediated by N metabolism under a mixed NO_(3)^(–) and NH_(4)^(+) supply

A mixed nitrate (NO_(3)^(–)) and ammonium (NH_(4)^(+)) supply can promote root growth in maize (Zea mays),however,the changes in root morphology and the related physiological mechanism under different N forms are still unclear.Here,maize seedlings were grown hydroponically with three N supplied in three different forms (NO_(3)^(–)only,75/25 NO_(3)^(–)/NH_(4)^(+)and NH_(4)^(+)only).Compared with sole NO_(3)^(–)or NH_(4)^(+),the mixed N supply increased the total root length of maize but did not affect the number of axial roots.The main reason was the increased total lateral root length,while the average lateral root (LR) length in each axle was only slightly increased.In addition,the average LR density of 2nd whorl crown root under mixed N was also increased.Compared with sole nitrate,mixed N could improve the N metabolism of roots (such as the N influx rate,nitrate reductase (NR) and glutamine synthase (GS)enzyme activities and total amino content of the roots).Experiments with exogenously added NR and GS inhibitors suggested that the increase in the average LR length under mixed N was related to the process of N assimilation,and whether the NR mediated NO synthesis participates in this process needs further exploration.Meanwhile,an investigation of the changes in root-shoot ratio and carbon (C) concentration showed that C transportation from shoots to roots may not be the key factor in mediating lateral root elongation,and the changes in the sugar concentration in roots further proved this conclusion.Furthermore,the synthesis and transportation of auxin in axial roots may play a key role in lateral root elongation,in which the expression of ZmPIN1B and ZmPIN9 may be involved in this pathway.This study preliminarily clarified the changes in root morphology and explored the possible physiological mechanism under a mixed N supply in maize,which may provide some theoretical basis for the cultivation of crop varieties with high N efficiency.

Anti-tumor Activities of Novel Estrogen Compound 17aα-D-Homo-Ethynylestradiol-3-Acetate

Objective： To study the anti-tumor activities of novel estrogen compound 17a α-D-homo-ethynylestradiol-3-acetate in vitro and in vivo. Methods： In vitro anti-tumor activity was assayed in adenoma cells A549 and human liver cancer cells Bel-7402 using MTT method, and half-inhibitory concentration （IC50） were observed. In vivo the pulmonary adenoma LA795 cells was selected and the conventional assay method of anti-tumor activity was employed. 5, 7.5, 10 mg/kg of 17a α-D-homo-ethynylestradiol-3-acetate was administered by i.p., and tumor-inhibitory rate, thymus and spleen indexes, bone marrow cells （BMC） were observed. Results： IC50 of 17a α-D-homo-ethynylestradiol-3-acetate in vitro for A549 and Bel-7402 cells were 12.28 μg/ml and 17.79 μg/ml, respectively. In vivo the highest tumor-inhibitory rates for LA795 was 60.0% （P〈0.01）. The drug had hardly any side-effect in spleen indexes, thymus indexes, and BMC compared with control mice. Nevertheless, compared with the positive control drug cyclophosphamide （CY）, thymus and spleen indexes, BMC showed obvious differences （P〈0.01）. Conclusion： 17a α-D-homo-ethynylestradiol-3-acetate has obvious anti-tumor activities in vitro and in vivo with low side-effect, thus worth further investigation.

Amperometric Determination of lndole-3-acetic Acid Based on Platinum Nanowires and Carbon Nanotubes

Platinum nanowire （PtNW） can be grown by electrodeposition in polycarbonate membrane, with the average diameter of the nanowires about 250 nm. The PtNW and multiwalled carbon nanotubes （CNT） are then dispersed into chitosan （CHIT） solution. The resulting PtNW-CNT-CHIT material brings new capabilities for electrochemical devices by using the synergistic action of the electrocatalytic activity of PtNW and CNT. By dropping the PtNW-CNT-CHIT film onto the glassy carbon （GO） electrode surface, and after evaporation an amperometric sensor for the determination of indole-3-acetic acid （IAA） was developed. The oxidation current of IAA increased significantly at the PtNW-CNT-CHIT film coated GC electrode, in contrast to that at the CNT-CHIT modified GC. The linear response of the sensor is from 50 ng/ml to 50 μg/ml with a detection limit of 25 ng/mL.

Synthesis,Spectroscopic Characterization,Thermogravimetric and Biological Activity Evaluation of Te(Ⅳ),Se(Ⅳ),V(Ⅲ),Nb(Ⅴ),Ta(Ⅴ)Complexes With Indole-3-Acetic Acid Plant Hormone Ligand

Te(Ⅳ),Se(Ⅳ),V(Ⅲ),Nb(Ⅴ)and Ta(Ⅴ)complexes of indole-3-acetic acid(IAAH)ligand were synthesized,characterized by elemental analysis and various spectroscopic techniques like,IR,1H-NMR,X-ray powder diffraction,UV-Visible,thermogravimetry analysis,magnetic measurements,molar conductance and surface morphology using SEM.All the synthesized complexes of IAAH ligand have 1∶2 stoichiometry of the types[Te(IAA)2(NH 3)2]·2Cl(Ⅰ),[Se(IAA)2(NH 3)2]·2Cl(Ⅱ),[V(IAA)2(NH 3)(Cl)](Ⅲ),[Nb(IAA)2(Cl)3](Ⅳ),and[Ta(IAA)2(Cl)3](Ⅴ).Spectral analysis indicates octahedral geometry for the Te(Ⅳ),Se(Ⅳ)and V(Ⅲ)complexes,whereas both Nb(Ⅴ)and Ta(Ⅴ)have a seven-coordination.The bonding sites are the oxygen atoms of carboxylate group for the deprotonated indole-3-acetic acid(IAA)ligand.The thermogravimetry analysis studies gave evidence for the presence of other coordinated molecules(Cl or NH 3)in the composition of IAA complexes,which were further supported by IR and micro analytical measurements.The higher molar conductance data of tellurium and selenium(Ⅳ)complexes reveal that these chelates are electrolytes,while low conductivity values for the vanadium(Ⅲ),niobium and tantalum(Ⅴ)chelates indicated a non-electrolytes.To test the antibacterial property of the five complexes in this study,four bacterial strains Klebsiella(G-),Escherichia coli(G-),Staphylococcus aureus(G+)and Staphylococcus epidermidis(G+)were used in the investigation.The effects of the five complexes in the cytotoxicity of Caco-2 and Mcf-7 human cancer cell lines were studied Neutral red uptake assay for the estimation of cell viability/cytotoxicity protocol.

The Difference of Sensitivity between BXPC-3 and K562 Cells by Treatments with Combination of Indole-3-acetic Acid and Horseradish Peroxidase

The difference of sensitivity to indole- 3-acetic acid （ IAA ） combined with horseradish peroxidase （HRP） in K562 and BXPC- 3 cells was investigated. The cell proliferation was determined by MTF assay. The cell cycle and apoptosis of K562 and BXPC-3 cells were examined by a fluorescence flow cytometer （FCM） and terminal deoxynacleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） respectively. The experimental results show that IAA and HRP could inhibit BXPC- 3 cell proliferation greatly compared with K562 cell during the first 48 h . The cell cycle was arrested predominantly at G2/ M phase in K562 and BXPC- 3 cells. The cell apoptosis of K562 and BXPC- 3 was induced by IAA/ HRP. There was a significant difference between the two cell lines since BXPC-3 cells were more sensitive than K562 cells by treatments with combination of IAA and HRP.

THE APPLICATION OF SEROTONIN AND 5-HVDROXYINDOLE-3-ACETIC ACID IN THE DIAGNOSIS OF APUDOMA

In our report, the values of whole blood serotonin (5-HT) in 87 apudoma patients (diagnosed by operation and pathology) were summarized. Among them, the levels of urinary 5-hydroxyindole-3-acetic acid (5-HIAA) of 44 patients were also tested. The results showed that both parametres of apudoma patients were higher than those of non-apudoma, post-operative patients of apudoma as well as the normal. The increasing extent of the levels of whole blood 5-HT and urinary 5-HIAA in small intestinal carcihoid was the most obvious one but that of rectum was not. The referable diagnostic values suggested were: 5-HT>130 ng ml, 5-HIAA>30 mg/ 24 hours.

The auxin transporter OsAUX1 regulates tillering in rice(Oryza sativa)

Tillering is an important agronomic trait of rice(Oryza sativa)that affects the number of effective panicles,thereby affecting yields.The phytohormone auxin plays a key role in tillering.Here we identified the high tillering and semi-dwarf 1(htsd1)mutant with auxin-deficiency root characteristics,such as shortened lateral roots,reduced lateral root density,and enlarged root angles.htsd1 showed reduced sensitivity to auxin,but the external application of indole-3-acetic acid(IAA)inhibited its tillering.We identified the mutated gene in htsd1 as AUXIN1(OsAUX1,LOC_Os01g63770),which encodes an auxin influx transporter.The promoter sequence of OsAUX1 contains many SQUAMOSA PROMOTER BINDING PROTEIN-LIKE(SPL)binding sites,and we demonstrated that SPL7 binds to the OsAUX1 promoter.TEOSINTE BRANCHED1(OsTB1),a key gene that negatively regulates tillering,was significantly downregulated in htsd1.Tillering was enhanced in the OsTB1 knockout mutant,and the external application of IAA inhibited tiller elongation in this mutant.Overexpressing OsTB1 restored the multi-tiller phenotype of htsd1.These results suggest that SPL7 directly binds to the OsAUX1 promoter and regulates tillering in rice by altering OsTB1 expression to modulate auxin signaling.

Effects of Zinc Oxide Particles with Different Sizes on Root Development in Oryza sativa

Given the consistent release of zinc oxide(ZnO)nanoparticles into the environment,it is urgent to study their impact on plants in depth.In this study,grains of rice were treated with two different concentrations of ZnO nanoparticles(NP-ZnO,10 and 100 mg/L),and their bulk counterpart(B-ZnO)were used to evaluate whether ZnO action could depend on particle size.To test this hypothesis,root growth and development assessment,oxidative stress parameters,indole-3-acetic acid(IAA)content and molecules/enzymes involved in IAA metabolism were analyzed.In situ localization of Zn in control and treated roots was also performed.Though Zn was visible inside root cells only following nanoparticle treatment,both materials(NP-ZnO and B-ZnO)were able to affect seedling growth and root morphology,with alteration in the concentration/pattern of localization of oxidative stress markers and with a different action depending on particle size.In addition,only ZnO supplied as bulk material induced a significant increase in both IAA concentration and lateral root density,supporting our hypothesis that bulk particles might enhance lateral root development through the rise of IAA concentration.Apparently,IAA concentration was influenced more by the activity of the catabolic peroxidases than by the protective action of phenols.

花生根际多功能高效促生菌的筛选鉴定及其效应研究

为了获得具有固氮、解磷解钾及合成生长素的多功能花生根际促生菌(PGPR),提高和改善河南花生种植区的花生产量和品质。从华北地区花生根际砂质潮土中筛选出5株多功能促生菌HS4、HS7、HS9、HS10、HS11,通过对其各功能指标的对比挑选出菌株HS9,其固氮酶活性、解磷解钾以及产IAA能力分别达到15.53 nmol C2H4/h·m L、279.23 mg/L、22.5 mg/L和40.96 mg/L。经形态观察、生理生化指标以及16S r DNA保守序列鉴定,确定该菌株为弯曲芽胞杆菌(Bacillus flexus)。最后通过花生盆栽实验验证其促生能力。盆栽实验表明:接种HS9能显著提高土壤中速效磷、钾含量,极显著地提高了土壤中IAA的含量;植株的根总长、根表面积、根体积和根尖数分别增加了109.60%、84.30%、76.08%和386.24%,促进植株对养分的吸收和利用,提高植株生物量以及养分含量;植株鲜重、株高、叶绿素值(Soil and plant analyzer development,SPAD)及氮、磷、钾含量分别提高了70.05%、28.35%、16.06%、23.11%、83.04%和23.95%。该多功能菌株对花生具有良好的促生作用,具有广阔的农业应用前景。

Changes in endogenous hormone levels and redox status during enhanced adventitious rooting by rare earth element neodymium of Dendrobium densiflorum shoot cuttings

The effects of neodymium nitrate （Nd^3＋） on the adventitious rooting of Dendrobium densiflorum shoot cuttings were studied. The addition of Nd^3＋ （5 μmol/L） to culture medium significantly increased rooting frequency. Histological investigation showed that Nd^3＋ did not change the process of root initiation. Nd^3＋ did not influence total endogenous cytokinin levels, but significantly increased the level of endogenous indole-3-acetic acid （IAA） in the base of shoot cuttings. Compared to the control, the ratio of IAA/cytokinins was very high in the Nd^3＋ treatment. These results suggested that the enhanced rooting frequency may be related to the increase in endogenous IAA level in Nd^3＋ treatment. Analysis of enzyme activities showed that the enhanced accumulation of the endogenous IAA by Nd^3＋ should not be attributed to inhibition of IAA decomposition by IAA oxidase or promotion of cytokinin decomposition by cytokinin oxidase. Besides, Nd^3＋ increased the ratio of reduced glutathione （GSH） and oxidized glutathione （GSSG） in the process of adventitious rooting while the ratio of ascorbate （ASC） to dehydroascorbate （DHA） was not affected.

Enhancement of Growth and Grain Yield of Rice in Nutrient Deficient Soils by Rice Probiotic Bacteria

Plant associated bacteria are promising alternatives to chemical fertilizers for plant growth and yield improvement in an eco-friendly manner.In this study,rice associated bacteria were isolated and assessed for mineral phosphate solubilization and indole-3-acetic acid(IAA) production activity in vitro.Six promising strains,which were tentatively identified as phylotaxon Pseudochrobactrum sp.(BRRh-1),Burkholderia sp.(BRRh-2),Burkholderia sp.(BRRh-3),Burkholderia sp.(BRRh-4),Pseudomonas aeruginosa(BRRh-5 and BRRh-6) based on their 16 S rRNA gene phylogeny,exhibited significant phosphate solubilizing activity in National Botanical Research Institute phosphate growth medium,and BRRh-4 displayed the highest phosphate solubilizing activity,followed by BRRh-5.The p H of the culture broth declined,resulting in increase of growth rate of bacteria at p H 7,which might be due to organic acid secretion by the strains.In presence of L-tryptophan,five isolates synthesized IAA and the maximum IAA was produced by BRRh-2,followed by BRRh-1.Application of two most efficient phosphate solubilizing isolates BRRh-4 and BRRh-5 by root dipping(colonization) of seedling and spraying at the flowering stage significantly enhanced the growth and grain yield of rice variety BRRI dhan-29.Interestingly,application of both strains with 50% of recommended nitrogen,phosphorus and potassium fertilizers produced equivalent or higher grain yield of rice compared to the control grown with full recommended fertilizer doses,which suggests that these strains may have the potential to be used as bioinoculants for sustainable rice production.

Genome-wide identification and expression analysis of Aux/IAA gene family in strawberry(Fragaria vesca)

Auxin signaling and its components(Auxin/Indole-3-Acetic Acid(Aux/IAA))are critical for plant growth and development.Here,we performed a genome-wide annotation and identified twenty-one Aux/IAA genes in strawberry(Fragaria vesca).Most FveIAAs were located on chromosomes 1,2,4,5,and 6,while no FveIAAs were found in chromosomes 3 and 7.Phylogenetic analysis divided these genes into nine subfamilies.Most FveIAAs contained the DNA-binding and Aux/IAA domains,as well as motifs I-IV.There were 2-6 exons in the FveIAA genes based on the gene structure analysis.Also,we found that four pairs of FveIAA genes underwent segment duplications.Moreover,four pairs of orthologous genes were observed between strawberry and Arabidopsis.Cis-element analysis in the promoter region indicated that FveIAAs may be involved in light,phytohormones,stress responses,and growth processes.Prediction of protein-protein interaction revealed that 17 of 21 FveIAA proteins were involved in the auxin-related signaling pathways.Additionally,FveIAAs showed tissue-specific expression and responded to IAA treatment.Thus,this systematic annotation of the FveIAA family would provide a fundamental basis for further functional and evolutionary analysis and to understanding the role of FveIAAs in strawberry growth and development.

Sulfur nutrient availability regulates root elongation by affecting root indole-3-acetic acid levels and the stem cell niche

Sulfur is an essential macronutrient for plants with numerous biological functions. However, the influence of sulfur nutrient availability on the regulation of root development remains largely unknown. Here, we report the response of Arabidopsis thaliana L. root development and growth to different levels of sulfate, demonstrating that low sulfate levels promote the primary root elongation. By using various reporter lines, we examined in vivo IAA level and distribution, cell division,and root meristem in response to different sulfate levels.Meanwhile the dynamic changes of in vivo cysteine, glutathione,and IAA levels were measured. Root cysteine, glutathione, and IAA levels are positively correlated with external sulfate levels in the physiological range, which eventually affect root system architecture. Low sulfate levels also downregulate the genes involved in auxin biosynthesis and transport, and elevate the accumulation of PLT1 and PLT2. This study suggests that sulfate level affects the primary root elongation by regulating the endogenous auxin level and root stem cell niche maintenance.

The Effect of Exogenous Phytohormones on Resistance of Wheat Calluses to <i>Tilletia caries</i>(D.C.) Tul. &C. Tul.

The influence of exogenic hormones (indole-3-acetic acid (IAA), abscisic acid (ABA) and kinetin) on defense reaction of wheat (Triticum aestivum L.) calli to the bunt agent Tilletia caries (D.C.) Tul. & C. Tul. was studied. ABA and kinetin induced the oxalate oxidase activity, increased the Н2О2 level, decreased germination of fungi teliospores and induced on calli the occurrence of dense sites non-infected by pathogen. On the contrary, IAA led to the decrease of oxalate oxidase activity, loosening of calli and increase germination of bunt agent teliospores and growth of fungi mycelium, besides stimulated rhizoids formation of wheat calli. Probably, the accumulation of Н2О2 in wheat calli under the influence of kinetin and ABA connected with activity of oxalate oxidase is one of the factors increasing defense reaction of wheat to bunt agent.

Differences of Plasma Levels of Tryptophan, Serotonin, 5-Hydroxyindole Acetic Acid, and Kynurenine between Healthy People and Patients of Major Monopolar Depression at Various Age and Gender

Background: It is not well analyzed whether there are differences in plasma levels of tryptophan (TRP) metabolites between healthy control people (HC) and patients of major monopolar depression (MMD). Methods: Ultra high-speed liquid chromatography/mass spectrometry has been used for the simultaneous determination of plasma levels of tryptophan metabolites in depressive patients. Results: There are no significant differences between plasma levels of TRP between HC and MMD. Plasma levels of TRP of HC are higher in young men, young women, old men, and old women in this order. Serotonin (5-HT) levels are higher in MMD than HC. Plasma levels of 5-HIAA of HC are also higher than those of patients of MMD. Plasma levels of kynurenine (KYN) of healthy old men and old women are higher than those of young men and old women. Plasma levels of KYN are higher in old women and young men of MMD than those of HC. Conclusion: Plasma levels of 5-HT are higher in patients of MMD than those of HC, which may suggest that use of drugs inhibiting the 5-HT transportation may increase plasma levels of 5-HT in MMD.

AN AMPEROMETRIC BIOSENSOR BASED ON COVALENT IMMOBILIZATION OF ASCORBATE OXIDASE ON BIOCOMPATIABLE AND LOW-TOXIC POLY(THIOPHENE-3-ACETIC ACID) MATRIX

The biocompatiable and low-toxic poly（thiophene-3-aeetic acid） （PTAA） matrix was successfully electrosynthesized in ionic liquid 1-butyl-3-metbylimidazolium hexafluorophosphate （BmimPF6） in comparison with the electrosynthesis of PTAA matrix in acetonitrile （ACN）. Ascorbate oxidase （AO） was used as a model for the development and application of biosensor. Vitamin C （VC） biosensors were facilely fabricated by the covalent immobilization of AO molecules on PTAA matrices electrosynthesized in ACN containing tetrabutylammonium tetrafluoroborate and BmimPF6, respectively. Electrochemical impedance spectroscopy, scanning electron microscopy and FTIR spectroscopy indicated that AO molecules were covalently immobilized on PTAA matrices. Parameters of the as-obtained biosensors such as working potential, pH and temperature have been optimized. The amperometric biosensor based on PTAA matrix electrosynthesized in BmimPF6 exhibited wider linear range, lower detection limit, higher sensitivity and bioaffinity, and better operational and storage stability than that electrosynthesized in ACN under optimal conditions. The as-obtained biosensor based on PTAA matrix electrosynthesized in BmimPF6 was employed for the detection of VC content in comme：rcial juices, and the result was close to the data given by manufacturers. Excellent results indicate that the PTAA matrix electrosynthesized in ionic liquid is a promising platform for the covalent immobilization of biologically-active species and the development of biosensors.

ELECTROSYNTHESES OF FREE-STANDING POLY(THIOPHENE-3-ACETIC ACID) FILM IN MIXED ELECTROLYTES OF BORON TRIFLUORIDE DIETHYL ETHERATE AND TRIFLUOROACETIC ACID

High quality free-standing poly（thiophene-3-acetic acid） （PTAA）, a water-soluble polythiophene derivative, was uccessfully electrosynthesized in boron trifluoride diethyl etherate （BFEE） ＋ 25% （by volume） trifluoroacetic acid （TFA） at 9wer potential （0.38 V versus Pt）. The carboxyl group makes PTAA highly soluble in water, facilitating its potential pplication as a blue-light-emitting material. PTAA films with conductivity of 7 S cm-1 obtained from this medium showed ,etter redox activity and thermal stability. The structure and morphology of the polymer were studied by UV-Vis, FT-IR, H-NMR spectra and scanning electron microscopy, respectively.

Regeneration Function Analysis of GmESR1 in Transgenic Soybean

ENHANCER OF SHOOT REGENERATION(ESR1)is an important regulator of plant regeneration in vitro,which promotes regeneration of plant.In this study,transgenic positive plants with normal expression of proteins were screened by molecular assay.Through the study of the transgenic plants and the control Dongnong 50,the difference between immature embryo-induced callus and induced shoot bud was observed.The increase in callus weight indicated that GmESR1 gene accelerated the formations of shoot buds.By measuring the changes of hormone in the process of induction callus of transgenic plants,it was found that the contents of indole-3-acetic acid(IAA)and zeatin(ZT)in transgenic lines were significantly increased.It could be concluded that GmESR1 gene promoted the accumulation of hormone and affected regeneration process.In addition,this study also verified the interaction between GmBIM1 gene and GmESR1 gene by bimolecular fluorescence complementation(BiFC).