Isolation and Expression Analysis of Two Genes Encoding Cinnamate 4-Hydroxylase from Cotton(Gossypium hirsutum)

Two genes （GhC4H1 and GhC4H2） that encode putative cotton cinnamate 4-hydroxylases that catalyze the second step in the phenylpropanoid pathway were isolated from developing cotton fibers. GhC4H1 and GhC4H2 each contain open reading frames of 1 518 base pairs （bp） in length and both encode proteins consisting of 505 amino acid residues. They are 90.89% identical to each other at the amino acid sequence level and belong to class I of plant C4Hs. GhC4H1 and GhC4H2 genomic DNA are 2 247 and 2 161 bp long, respectively, and contain two introns located at conserved positions relative to the coding sequence. GhC4HI and GhC4H2 promoters were isolated and found to contain many cis-elements （boxes P, L and AC-1 element） previously identified in the promoters of other phenylpropanoid pathway genes. Histochemical staining showed GUS expression driven by the GhC4H1 and GhC4H2 promoters in ovules and fibers tissues. GhC4H1 and GhC4H2 were also widely expressed in other cotton tissues. GhC4H2 expression reached its highest level during the elongation stage of fiber development, whereas GhC4H1 expression increased during the secondary wall development period in cotton fibers. Our results contribute to a better understanding of the biochemical role of GhC4H1 and GhC4H2 in cotton fiber development.

New Process for the Synthesis of A Sun-screening Agent Isooctyl P-methoxy Cinnamate

A sun-screening agent isooctyl p-methoxy cinnamate was prepared by reaction of p-methoxyl benzaldehyde with isooctyl acetate using titanium tetrachloride as catalyst. The optimum conditions are as follows： molar ratio of p-methoxyl benzaldehyde to isooctyl acetate and titanium tetrachloride is 1 ： 1.1 ： 0.5; reaction temperature is 30℃; and reaction time is 6 h. The yield of isooctyl p-methoxy cinnamate can reach 98%. The method of the synthesis of isooctyl p-methoxy cinnamate is efficient and economical.

HPLC Determination of Harpagoside and Cinnamic Acid in Radix Scrophulariae

A gradient HPLC method was established for the determination of harpagoside and cinnamic acid in Radix Scrophulariae (Xuanshen) and a proposition was put forward for the lowest content of the characteristic and active constituent (harpagoside) for qualified Radix Scrophulariae. The experimental conditions were as follows: Ultrasphere ODS column (250 mm 4.6 mm, 5 mm), mobile phase: acetonitrile-water (containing 1.0% acetic acid) (20:8050:50) (20 min), flow-rate 1 mLmin-1, room temperature, detection wavelength 278 nm. Twenty-eight samples of Xuan-shen (Radix Scrophulariae) from different districts of China were analyzed and the contents of harpagoside and cinnamic acid in Xuan-shen were 0.041~0.244% and 0.012~0.068% respectively. The recoveries (RSD)% were 97.13(0.80)% for harpagoside and 99.38(0.51)% for cinnamic acid. The method is simple and accurate. It can be used for the quality control of Radix Scrophulariae. We propose that the content of harpagoside in qualified Radix Scrophularia should be no less than 0.05%.

Determination of Plasma Concentration of Cinnamic Acid by High-Performance Liquid Chromatography and Its Pharmacokinetics in Rats after Oral Administration of Zi-Shen Pill

Aim To develop a simple and sensitive high-performance liquid chromatographicmethod for determination of plasma concentration of cinnamic acid and pharmacokinetic study in ratsafter a single oral dose of traditional Chinese medicinal preparation Zi-Shen pill. Method Plasmasamples were acidified with hydrochloric acid and extracted with ethyl acetate . Cinnamic acid wasdetermined by HPLC using a G_(18) column. A mobile phase ofmethanbl-acetonitrile-water-triethyl-amine (7:22:73 = 0.2, V/V), with the pH adjusted to 4.0 withphosphoric acid, and with a UV detector set at 340 nm. Results The standard curve was linear overthe range of 1.92- 192.0 μg·mL^(-1). The LLOQ was 1.92 μg·mL^(-1) . The RSDs of within-day andbetween-day precision were < 8%. The mean recovery was 82.0% . Conclusion After validation, themethpd has been used to investigate the pharmacokinetic profiles of the traditional Chinesemedicinal preparation Zi-Shen pill.

Anticancer Activities of Substituted Cinnamic Acid Phenethyl Esters on Human Cancer Cell Lines

Caffeic acid phenethyl ester (CAPE) and sixteen substituted cinnamic acid phenethyl esters were prepared via conventional procedures in order to test their in vitro anticancer activities by either MTT assay or SRB assay on six different human cancer cell lines. The results indicated that in the concentration of 10 μmol·L -1 the lead compound CAPE possessed anticancer activities against human HL 60, Bel 7402, and Hela cell lines, and two other compounds possessed potent anticancer activities against Bel 7402 and Hela cell lines.

Effects of Cinnamic Acid on Bacterial Community Diversity in Rhizosphere Soil of Cucumber Seedlings Under Salt Stress

To investigate the effects of a plant autotoxin, cinnamic acid, on bacterial communities in the rhizosphere soil of cucumber seedlings under salt stress, we used cucumber as the experimental material, cinnamic acid as the autotoxin, and NaCl to apply salt stress. Bacterial communities in the rhizosphere soil were analyzed using polymerase chain reaction （PCR）, denaturing gradient gel electrophoresis （DGGE）, and clone sequencing. Salt stress decreased the diversity of bacterial species in rhizosphere soil of cucumber seedlings at several growth stages. Cinnamic acid exacerbated the effects of salt stress at high concentrations, but alleviated its effects at low concentrations. Cloning and sequencing results indicated that DGGE bands amplified from soil samples showed high homology to uncultured bacterial species. Cinnamic acid at 50 mg kg^-1 soil improved cucumber growth and was the most effective treatment to alleviate the effects of salt stress on bacterial communities.

Synthesis and Crystal Structure of (E)-4-(Benzyloxy)-2-(cinnamoyloxy)-N,N,N-trimethyl-4-oxobutan-1-aminium Chloride as a Double-prodrug

A novel hydrochloride quaternary ammonium salt （E）-4-（benzyloxy）-2-（cinnamo- yloxy）-N,N,N-trimethyl-4-oxobutan-l-aminium chloride （Cz3HzgNO4Cl2, Mr = 454.37） has been synthesized via the sequence of acetylation and esterification by using L-carnitine （L-4-N-trimethy- lammonium-3-hydroxybutyric acid, LC） and cinnamic acid as the starting materials, and its crystal structure was determined by single-crystal X-ray diffraction method. The crystal belongs to monoclinic, space group P212121 with a = 10.1670（4）, b = 10.4488（4）, c = 22.9795（11）A, V = 2441.18（18）A3, Z = 4, Dc = 1.236 g/cm3,μ（MoKa） = 0.293 mm-1, F（000） = 960, Flack factor = -0.01（11）, the final R = 0.0489 and wR = 0.1550 for 3350 observed reflections （I 〉 2σ（/）） and R = 0.0953 for all 5648 unique reflections. The crystal structure involves a conjugated system which shows a reverse olefin structure.

RP-HPLC Determination and Pharmacokinetic Comparison of Cinnamic Acid in Rat Plasma After Administration of Di-Gu-Pi Decoction and Pure Cinnamic Acid

A sensitive, simple, and accurate method was developed for the determination and pharmacokinetic comparison of cinnamic acid in rat plasma after the administration of a Traditional Chinese Medicinal preparation, Di-Gu-Pi decoction, and pure cinnamic acid using RP-HPLC. Di-Gu-Pi was extracted with 5% aqueous sodium bicarbonate, which was followed by purification with ion exchange column chromatography. The plasma samples taken from rats were deproteinized with methanol. The reversed-phase （HPLC） system with a Diamonsil C18 column and methanol-acetonitfile-water （8： 32： 60, volume ratio） （adjusted to pH = 3.0 with glacial acetic acid） as the mobile phase was employed for the separation of cinnamic acid in the plasma samples. The detection was set at 272 nm and 3-（p-fluoro-phenyl）-2-propenoic acid was chosen as the internal standard. The calibration curve was linear in a range from 0. 10 to 25.0μg/mL （R2 = 0. 9988, n = 9）. The precision was 3.42%-10. 10%; the between-day precision was 2. 84%-8.91% ; the accuracy was - 1.51%-1.26% ; the mean recovery was 99. 9%. The method was found to be sensitive, simple, accurate and appropriate for the determination of cinnamic acid.

Electron Momentum Spectroscopy of Valence Orbitals of n-Propyl Iodide： Spin-Orbit Coupling Effect and Intramolecular Orbital Interaction

The binding energy spectrum and electron momentum distributions for the outer valence orbitals of n-propyl iodide molecule have been measured using the electron momentum spectrometer employing non-coplanar asymmetric geometry at impact energy of 2.5 keV plus binding energy. The ionization bands have been assigned in detail via the high accuracy SACCI general-R method calculation and the experimental momentum profiles are compared with the theoretical ones calculated by Hartree-Fock and B3LYP/aug-cc-pVTZ（C,H）6-311G？？（I）. The spin-orbit coupling effect and intramolecular orbital interaction have been analyzed for the outermost two bands, which are assigned to the iodine 5p lone pairs, using NBO method and non-relativistic as well as relativistic calculations. It is found that both of the interactions will lead to the observed differences in electron momentum distributions. The experimental results agree with the relativistic theoretical momentum profiles, indicating that the spin-orbit coupling effect dominates in n-propyl iodide molecule.

Effects of Exogenous Cinnamic Acids on the Growth and Physiological Characteristics of Cucumber Seedlings

In order to study the effects of exogenous cinnamic acids on plant growth, contents of photosynthetic pigment, root activities and ATPase activities of root membrane at cucumber seedling stage, the seedlings of Shandong Mici cucumber were tested. The results showed that seedlings growth, contents of photosynthetic pigment, root activities and ATPase activities of root membrane were inhibited by cinnamic acids. The growth and root activities of seedlings were significantly （P〈0.05） lower in the soil amended with 100 mg kg^-1 cinnamic acids compared to the control. The contents of chlorophyll a, chlorophyll a＋b and carotenoid of seedlings significantly （P〈0.05） decreased in the soil amended with 200 mg kg^-1 cinnamic acids, whereas ATPase activities exhibited a higher sensitivity and greatly decreased in the soil amended with 50 mg kg^-1 cinnamic acids. These results suggested that cinnamic acids could induce a stress condition, and the stress intensities increased with enhanced cinnamic acid concentration.

Synthesis and Characterization of Rare Earth Tesnary Complexes with Cinnamic Acid and O-Phenanthroline

Thirteen solid rare earth tesnary complexes with cinnamic acid C 6H 5CH=CHCOOH, HL and o phenanthroline (phen), RE(phen)L 3 were synthesized. Their IR absorption spectra, molar conductance, TG DTA, X ray power diffraction analysis and fluorescence spectra were studied. The results show that all coordination compounds are oxidized and decomposed around 400 ℃ to 540 ℃. They are all crystalline material. The fluorescence spectra indicate that the fluorescence emission intensity of Eu(phen)L 3 is the highest among them.

Isobaric Vapor-Liquid Equilibrium of Binary Systems： p-Xylene ＋ （Acetic Acid, Methyl Acetate and n-Propyl Acetate） and Methyl Acetate ＋ n-Propyl Acetate in an Acetic Acid Dehydration Process

The vapor-liquid equilibrium data of four binary systems (acetic acid +p-xylene, methyl acetate +n-propyl acetate, n-propyl acetate +p-xylene and methyl acetate +p-xylene) are measured at 101.33 kPa with Ellis equilibrium still, and then both the NRTL and UNIQUAC models are used in combination with the HOC model for correlating and estimating the vapor-liquid equilibrium of these four binary systems. The estimated binary VLE results using correlated parameters agree well with the measured data except the methyl acetate +p-xylene system which easily causes bumping and liquid rushing out of the sampling tap due to their dramatically different boiling points. The correlation results by NRTL and UNIQUAC models have little difference on the average absolute deviations of temperature and composition of vapor phase, and the results by NRTL model are slightly better than those by UNIQUAC model except for the methyl acetate +n-propyl acetate system, for which the latter gives more accurate correlations.

A 3D Homochiral Mn(Ⅱ) Coordination Polymer Based on the Ligand 2,2'-(Ethane-1,2-diylbis(oxy)) Dicinnamic Acid

A 3D homochiral coordination polymer [Mn（edca）]n（1） was synthesized from a flexible coupled cinnamic acid ligand, 2,2？-（ethane-1,2-diylbis（oxy）） dicinnamic acid（H_2 edca） and MnCl_2×4 H_2 O under solvothermal conditions. The complex crystallizes in orthorhombic, space group P2_12_12_1 with a = 5.5785（8）, b = 13.0393（18）, c = 23.133（3） A, V = 1682.7（4） A^3, M_r = 407.27, D_c = 1.608 Mg/cm^3, F（000） = 836, Z = 4, the final R = 0.0240 and wR = 0.0590 for 2983 observed reflections（I 〉 2σ（I））. In 1, the edca2＋ anions alternately bridge the Mn（Ⅱ） cations to form one-dimensional（1 D） infinite helical chains of [Mn（edca）]n, which are further connected by the ligands generating a three-dimensional（3D） homochiral network. The photoluminescence of 1 was also investigated in solid state at ambient temperature.

Indirect determination of cefradine with n-propyl alcohol-ammonium sulfate-water system by extraction-flotation of cuprous thiocyanate

A new method was developed for the determination of cefradine by extraction-flotation of CuSCN. The experiment indicated that in the presence of 0.20 mol/L NaOH the degradation of cefradine took place in water bath at 100 ℃. The thiol group （-SH） of the degradation product could reduce Cu（Ⅱ） to Cu（Ⅰ） for the formation of the emulsion CuSCN in the presence of NH4SCN at pH 4.0. By determining the residual amount of Cu（Ⅱ） in the solution and calculating the flotation yield of Cu（Ⅱ）, the indirect determination of cefradine can be obtained. This method has been applied to determine cefradine in capsules, human serum and urine samples, respectively.

TWO NEW CINNAMIC ACID DERIVATIVES FROM MAGNOLIA BIONDII PAMP

Two new cinnamic acid derivatives named biondinin C and biondinin D were isolated from the flower buds of Magnolia biondii pamp. The structures were elucidated by the spectral data including ~1H, ~1H COSY, ~1H-^(13)C COSY spectra.

A Novel Chiral Cd-organic Network Based on Flexible Coupled Cinnamic Acid

A new metal-organic network, [Cd（CH_3OH）_2（epa）]_n（1）, was synthesized from an achiral coupled cinnamic acid, 3,3？-（（ethane-1,2-diylbis（oxy））bis（2,1-phenylene））diacrylic acid（H_2epa）, and Cd（NO3）2·4H2O under solvothermal conditions. The complex crystallizes in orthorhombic, space group P2_12_12 with a = 9.8286（8）, b = 21.0004（16）, c = 5.5169（4） A, V = 1138.71（15） A3, M_r = 528.81, D_c = 1.542 Mg/cm3, F（000） = 536, Z = 2, the final R = 0.0345 and wR = 0.1101 for 2006 observed reflections（I 〉 2σ（I））. In 1 the epa^（2＋） anions alternately bridge the Cd（II） cations to form a one-dimensional（1D） infinite homochiral helical chain of [Cd（epa）]n. The chiral homohelical chains via hydrogen bonds formed a two-dimensional（2D） network. The photoluminescence of 1 was also investigated in solid state at ambient temperature.

Cinnamic acid regulates the intestinal microbiome and short-chain fatty acids to treat slow transit constipation

BACKGROUND Slow transit constipation(STC)is a disorder with delayed colonic transit.Cinnamic acid(CA)is an organic acid in natural plants,such as Radix Scrophulariae(Xuan Shen),with low toxicity and biological activities to modulate the intestinal microbiome.AIM To explore the potential effects of CA on the intestinal microbiome and the primary endogenous metabolites-short-chain fatty acids(SCFAs)and evaluate the therapeutic effects of CA in STC.METHODS Loperamide was applied to induce STC in mice.The treatment effects of CA on STC mice were assessed from the 24 h defecations,fecal moisture and intestinal transit rate.The enteric neurotransmitters:5-hydroxytryptamine(5-HT)and vasoactive intestinal peptide(VIP)were determined by the enzyme-linked immunosorbent assay.Hematoxylin-eosin and Alcian blue and Periodic acid Schiff staining were used to evaluate intestinal mucosa's histopathological performance and secretory function.16S rDNA was employed to analyze the composition and abundance of the intestinal microbiome.The SCFAs in stool samples were quantitatively detected by gas chromatography-mass spectrometry.RESULTS CA ameliorated the symptoms of STC and treated STC effectively.CA ameliorated the infiltration of neutrophils and lymphocytes,increased the number of goblet cells and acidic mucus secretion of the mucosa.In addition,CA significantly increased the concentration of 5-HT and reduced VIP.CA significantly improved the diversity and abundance of the beneficial microbiome.Furthermore,the production of SCFAs[including acetic acid(AA),butyric acid(BA),propionic acid(PA)and valeric acid(VA)]was significantly promoted by CA.The changed abundance of Firmicutes,Akkermansia,Lachnoclostridium,Monoglobus,UCG.005,Paenalcaligenes,Psychrobacter and Acinetobacter were involved in the production of AA,BA,PA and VA.CONCLUSION CA could treat STC effectively by ameliorating the composition and abundance of the intestinal microbiome to regulate the production of SCFAs.

Determination of Cinnamic Acid in Stephania longa Lour. by HPLC

[Objectives] To establish a HPLC method for the quantitative determination of cinnamic acid in Stephania longa Lour. [Methods]The chromatographic column: Thermo BDS HYPERSIL C_(18)( 4. 6 mm × 250 mm,5 μm); mobile phase: acetonitrile-0. 1% phosphoric acid( 24∶ 76); flow rate: 1 mL/min; column temperature: 30℃; wavelength: 285 nm. [Results]The cinnamic acid was in a good linear relationship in the range of 0. 021 7-0. 076 μg,the sample recovery rate was 100. 46%,the RSD was 2. 20%,and the content was in the range of 0. 004%-0. 022%. [Conclusions] The method is simple,accurate and reproducible,and can be used for the quality control of S. longa Lour.

Crystallization of Curcumin and Cinnamic Acid from Aqueous Solutions of Hydrotropes

Batch crystallization studies of curcumin from hydrotropic solutions of sodium cumenesulphonate (NaCS) and of cinnamic acid from a photosensitive hydrotropic medium of sodium cinnamate (Na-CIN) were carried out, in an agitated reactor for the effect of alternate heating and cooling cycles on crystal morphology. The crystal characterization by Scanning electron microscopy (SEM) and crystal size distribution (CSD) showed formation of spheroidal curcumin crystals while cinnamic acid formed porous aggregates when subjected to thermal cycles. The UV irradiation of cinnamic acid however showed no formation of the aggregates. The type of hydrotrope used and the initial crystal morphologies of curcumin and cinnamic acid are shown to be important factors to result in a different behaviour of the crystal morphology upon thermal cycles. The CSD data were effectively used for estimation of nucleation and growth rate parameters.

Simultaneous Determination of Gallic Acid and Cinnamic Acid Content in Rhubarb by HPLC

[Objective]The paper was to determine the contents of gallic acid and cinnamic acid in Chinese herb rhubarb by HPLC.[Method]High performance liquid chromatography(HPLC)was established with the following conditions:Thermo SCIENTIFIC Hypersil GOLD Dim column(4.6 mm×250 mm,5μm),mobile phase methanol-0.1%phosphoric acid solution,gradient elution,detection wavelength 278 nm,flow rate 1.0 mL/min,column temperature 30℃,injection volume 10 pL.[Result]The content of gallic acid and cinnamic acid in rhubarb was determined by HPLC,and they showed good linear relationship in the linear ranges of 0.125-1.250 pg(E2=0.9998)and 0.0382-0.3820μg(E2=0.9994),respectively.[Conclusion]A HPLC method is established to determine gallic acid and cinnamic acid content in rhubarb.The method will provide a scientific basis for quality control of rhubarb and its traditional Chinese medicine.