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1.
Plant Physiol ; 194(3): 1906-1922, 2024 Feb 29.
Article in English | MEDLINE | ID: mdl-37987562

ABSTRACT

Salinity is a severe abiotic stress that limits plant survival, growth, and development. 14-3-3 proteins are phosphopeptide-binding proteins that are involved in numerous signaling pathways, such as metabolism, development, and stress responses. However, their roles in salt tolerance are unclear in woody plants. Here, we characterized an apple (Malus domestica) 14-3-3 gene, GENERAL REGULATORY FACTOR 8 (MdGRF8), the product of which promotes salinity tolerance. MdGRF8 overexpression improved salt tolerance in apple plants, whereas MdGRF8-RNA interference (RNAi) weakened it. Yeast 2-hybrid, bimolecular fluorescence complementation, pull-down, and coimmunoprecipitation assays revealed that MdGRF8 interacts with the transcription factor MdWRKY18. As with MdGRF8, overexpressing MdWRKY18 enhanced salt tolerance in apple plants, whereas silencing MdWRKY18 had the opposite effect. We also determined that MdWRKY18 binds to the promoters of the salt-related genes SALT OVERLY SENSITIVE 2 (MdSOS2) and MdSOS3. Moreover, we showed that the 14-3-3 protein MdGRF8 binds to the phosphorylated form of MdWRKY18, enhancing its stability and transcriptional activation activity. Our findings reveal a regulatory mechanism by the MdGRF8-MdWRKY18 module for promoting the salinity stress response in apple.


Subject(s)
Malus , Salt Tolerance , Salt Tolerance/genetics , Malus/metabolism , 14-3-3 Proteins/genetics , 14-3-3 Proteins/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Stress, Physiological/genetics
2.
Front Plant Sci ; 14: 1065032, 2023.
Article in English | MEDLINE | ID: mdl-36890893

ABSTRACT

Apomixis is the asexual reproduction through seeds that leads to the production of genetically uniform progeny. It has become an important tool in plant breeding because it facilitates the retention of genotypes with desirable traits and allows seeds to be obtained directly from mother plants. Apomixis is rare in most economically important crops, but it occurs in some Malus species. Here, the apomictic characteristics of Malus were examined using four apomictic and two sexually reproducing Malus plants. Results from transcriptome analysis showed that plant hormone signal transduction was the main factor affecting apomictic reproductive development. Four of the apomictic Malus plants examined were triploid, and pollen was either absent or present in very low densities in the stamen. Variation in the presence of pollen was associated with variation in the apomictic percentage; specifically, pollen was absent in the stamens of tea crabapple plants with the highest apomictic percentage. Furthermore, pollen mother cells failed to progress normally into meiosis and pollen mitosis, a trait mostly observed in apomictic Malus plants. The expression levels of meiosis-related genes were upregulated in apomictic plants. Our findings indicate that our simple method of detecting pollen abortion could be used to identify apple plants that are capable of apomictic reproduction.

3.
Pharmaceuticals (Basel) ; 15(5)2022 Apr 23.
Article in English | MEDLINE | ID: mdl-35631346

ABSTRACT

Mitochondria are structurally and functionally unique organelles in male gametes. Apparently, as the only organelles remaining in mature sperm, mitochondria not only produce adeno-sine triphosphate (ATP) through oxidative phosphorylation (OXPHOS) to support sperm mobility, but also play key roles in regulating reactive oxidation species (ROS) signaling, calcium homeostasis, steroid hormone biosynthesis, and apoptosis. Mitochondrial dysfunction is often associated with the aging process. Age-dependent alterations of the epididymis can cause alterations in sperm mitochondrial functioning. The resultant cellular defects in sperm have been implicated in male infertility. Among these, oxidative stress (OS) due to the overproduction of ROS in mitochondria may represent one of the major causes of these disorders. Excessive ROS can trigger DNA damage, disturb calcium homeostasis, impair OXPHOS, disrupt the integrity of the sperm lipid membrane, and induce apoptosis. Given these facts, scavenging ROS by antioxidants hold great potential in terms of finding promising therapeutic strategies to treat male infertility. Here, we summarize the progress made in understanding mitochondrial dysfunction, aging, and male infertility. The clinical potential of antioxidant interventions was also discussed.

4.
Biomed Res Int ; 2019: 2692970, 2019.
Article in English | MEDLINE | ID: mdl-30800665

ABSTRACT

In previous studies, Gentianella acuta (Michx.) Hulten was reported to contain xanthones, iridoids, terpenoids, and sterols and is mainly used to cure hepatitis, jaundice, fever, headache, and angina pectoris. In this study, we used bioassay guided fractionation to identify compounds from G. acuta and investigated their activity against hydrogen peroxide (H2O2)-induced apoptosis of H9c2 cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and glutamate-cysteine ligase catalytic (GCLC) expression were assessed using quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was evaluated using western blot. The results showed that all four compounds had protective effects on H9c2 cells. The transcription levels of HO-1 and GCLC significantly increased in H9c2 cells pretreated with norswertianolin (1), swetrianolin (2), demethylbellidifolin (3), and bellidifolin (4). However, compared to the model group, the transcription levels of Nrf2 were not enhanced by pretreatment with compounds 1, 2, and 4. The protein expression levels of HO-1 and GCLC in H9c2 cells were greater than that in the H2O2-treated group, and the expression of Nrf2 was not significantly changed except by swetrianolin treatment; inhibitors can reverse the protective effect by ZnPP (15 µM), BSO (10 µM), and brusatol (10 µM). The results indicated that the four compounds isolated from G. acuta inhibited the oxidative injury induced by H2O2 by activating the Nrf2/ARE pathway in H9c2 cells and provide evidence that G. acuta may be a potential therapeutic agent for the treatment of cardiovascular diseases.


Subject(s)
Gentianella/chemistry , Hydrogen Peroxide/pharmacology , Plant Extracts/pharmacology , Protective Agents/pharmacology , Animals , Apoptosis/drug effects , Cell Line , Glutamate-Cysteine Ligase/metabolism , Heme Oxygenase-1/metabolism , NF-E2-Related Factor 2/metabolism , Oxidative Stress/drug effects , Rats , Transcription, Genetic/drug effects , Xanthenes/pharmacology , Xanthones/pharmacology
5.
Plant Physiol ; 179(1): 88-106, 2019 01.
Article in English | MEDLINE | ID: mdl-30333149

ABSTRACT

SIZ1 (a SIZ/PIAS-type SUMO E3 ligase)-mediated small ubiquitin-like modifier (SUMO) modification of target proteins is important for various biological processes related to abiotic stress resistance in plants; however, little is known about its role in resistance toward iron (Fe) deficiency. Here, the SUMO E3 ligase MdSIZ1 was shown to be involved in the plasma membrane (PM) H+-ATPase-mediated response to Fe deficiency. Subsequently, a basic helix-loop-helix transcription factor, MdbHLH104 (a homolog of Arabidopsis bHLH104 in apple), which acts as a key component in regulating PM H+-ATPase-mediated rhizosphere acidification and Fe uptake in apples (Malus domestica), was identified as a direct target of MdSIZ1. MdSIZ1 directly sumoylated MdbHLH104 both in vitro and in vivo, especially under conditions of Fe deficiency, and this sumoylation was required for MdbHLH104 protein stability. Double substitution of K139R and K153R in MdbHLH104 blocked MdSIZ1-mediated sumoylation in vitro and in vivo, indicating that the K139 and K153 residues were the principal sites of SUMO conjugation. Moreover, the transcript level of the MdSIZ1 gene was substantially induced following Fe deficiency. MdSIZ1 overexpression exerted a positive influence on PM H+-ATPase-mediated rhizosphere acidification and Fe uptake. Our findings reveal an important role for sumoylation in the regulation of PM H+-ATPase-mediated rhizosphere acidification and Fe uptake during Fe deficiency in plants.


Subject(s)
Iron/metabolism , Malus/enzymology , Proton-Translocating ATPases/metabolism , Ubiquitins/physiology , Cell Membrane/metabolism , Malus/metabolism , RNA, Messenger/metabolism , Rhizosphere , Sumoylation , Ubiquitins/genetics , Ubiquitins/metabolism
6.
J Plant Physiol ; 232: 216-225, 2019 Jan.
Article in English | MEDLINE | ID: mdl-30537609

ABSTRACT

In plants, SIZ1 regulates abiotic and biotic stress responses by promoting the SUMOylation of proteins. The apple MdSIZ1 protein has conserved domains similar to those of Arabidopsis AtSIZ1. Real-time fluorescent quantitative analysis showed that MdSIZ1 gene expression was induced by phosphate-deficient conditions. In addition, the level of SUMOylation was also significantly increased under these conditions. The MYB transcription factor MdPHR1 might be a target for the SUMO protein, which is a phosphorus starvation-dependent protein. Subsequently, an MdSIZ1 expression vector was constructed and transformed in Arabidopsis mutant siz1-2 and apple callus. The MdSIZ1 transgenic Arabidopsis partially complemented the defect phenotype of siz1-2 under phosphate-deficient conditions. The survival rate, length of primary root, and number or density of lateral roots were similar between the transgenic lines and wild type (WT). Under phosphate-deficient conditions, the SUMO conjugate and fresh weight of the MdSIZ1 transgenic apple callus were improved compared with WT. The MdSIZ1 transgenic apple callus grew under phosphate-deficient conditions, whereas the MdSIZ1 sense apple callus did not. Therefore, MdSIZ1 is involved in the regulation of the phosphate-deficiency response in apple.


Subject(s)
Ligases/physiology , Malus/physiology , Phosphates/deficiency , Plant Proteins/physiology , Arabidopsis , Gene Expression Regulation, Plant , Ligases/metabolism , Malus/enzymology , Malus/metabolism , Plant Proteins/metabolism , Plants, Genetically Modified , Sumoylation
7.
Plant Cell Environ ; 40(10): 2068-2080, 2017 Oct.
Article in English | MEDLINE | ID: mdl-28440563

ABSTRACT

MdMYB1 acts as a crucial component of the MYB-bHLH-WD40 complex to regulate anthocyanin biosynthesis in red-skinned apples (Malus domestica), but little is known about its post-translational regulation. Here, a small ubiquitin-like modifier E3 ligase MdSIZ1 was screened out as an MdMYB1-interacting protein with a yeast two-hybridization approach. The interaction between MdSIZ1 and MdMYB1 was further verified with pull-down and CoIP assays. Furthermore, it was found that MdSIZ1 directly sumoylated MdMYB1 proteins in vivo and in vitro, especially under moderately low temperature (17 °C) conditions, and that this sumoylation was required for MdMYB1 protein stability. Moreover, the transcription level of MdSIZ1 gene was remarkably induced by low temperature and phosphorus deficiency, and MdSIZ1 overexpression exerted a large positive influence on anthocyanin accumulation and red fruit coloration, suggesting its important role in the regulation of anthocyanin biosynthesis under stress conditions. Our findings reveal an important role for a small ubiquitin-like modifier modification of MYB transcription factors in regulation of anthocyanin biosynthesis in plants.


Subject(s)
Anthocyanins/metabolism , Cold Temperature , Malus/metabolism , Plant Proteins/metabolism , Sumoylation , Ubiquitin-Protein Ligases/metabolism , Anthocyanins/biosynthesis , Biosynthetic Pathways , Fruit/metabolism , Models, Biological , Proteasome Endopeptidase Complex/metabolism , Protein Binding , Protein Stability , Proteolysis
8.
J Plant Physiol ; 198: 69-80, 2016 Jul 01.
Article in English | MEDLINE | ID: mdl-27152458

ABSTRACT

SUMOylation, the conjugation of target proteins with SUMO (small ubiquitin-related modifier), is a type of post-translational modification in eukaryotes and involves the sequential action of activation (E1), conjugation (E2) and ligation (E3) enzymes. In Arabidopsis, the AtSIZ1 protein is a SUMO E3 ligase that promotes the conjugation of SUMO proteins to target substrates. Here, we isolated and identified a SUMO E3 ligase, MdSIZ1, in apple, which was similar to AtSIZ1. SUMOylation analysis showed that MdSIZ1 had SUMO E3 ligase activity in vitro and in vivo. SUMO conjugation was increased by high temperatures, low temperatures, and abscisic acid (ABA). The ectopic expression of MdSIZ1 in Arabidopsis siz1-2 mutant plants partially complemented the morphological mutant phenotype and enhanced the levels of SUMO conjugation. Taken together, these results suggest that MdSIZ1-mediated SUMO conjugation of target proteins is an important process that regulates the adaptation of apple plants to various environmental stresses.


Subject(s)
Malus/enzymology , Plant Proteins/metabolism , Small Ubiquitin-Related Modifier Proteins/metabolism , Ubiquitin-Protein Ligases/metabolism , Abscisic Acid/pharmacology , Amino Acid Sequence , Arabidopsis/anatomy & histology , Arabidopsis/genetics , Cloning, Molecular , Cold Temperature , Gene Expression Regulation, Plant/drug effects , Genes, Plant , Genetic Complementation Test , Germination/drug effects , Heat-Shock Response/genetics , Malus/drug effects , Malus/genetics , Mutation/genetics , Phylogeny , Plant Leaves/anatomy & histology , Plant Leaves/drug effects , Plant Proteins/chemistry , Plants, Genetically Modified , Sequence Analysis, Protein , Species Specificity , Stress, Physiological/genetics
9.
Biomed Environ Sci ; 28(1): 80-4, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25566866

ABSTRACT

To study the microbiological contamination of kitchen dishcloths in Chinese housholds, 1010 'in-use' kitchen dishcloths were collected from residential premises in Beijing and Shanghai, and they were sent to the laboratory for microbiological quality analysis. The aerobic plate counts for dishcloths were 10-109 cfu/cm2 in the range of 150 cfu/cm2 to 1.776×109 cfu/cm2 (Beijing) and 62.5 cfu/cm2 to 8.75×108 cfu/cm2 (Shanghai). Nineteen species of bacteria were detected in the dishcloths, most of which were conditional pathogenic bacteria. This study found a significant difference in the aerobic plate counts of dishcloths with regard to type, number of the days used, activities used for, and some family factors. The findings of the study highlight the potential for contamination of kitchen dishcloths within homes.


Subject(s)
Hospitals , Mass Screening , Squamous Intraepithelial Lesions of the Cervix/diagnosis , Uterine Cervical Dysplasia/diagnosis , Female , Humans
10.
Molecules ; 19(9): 13432-47, 2014 Aug 29.
Article in English | MEDLINE | ID: mdl-25178064

ABSTRACT

Momordica charantia Linn. is used as an edible and medicinal vegetable in sub-tropical areas. Until now, studies on its composition and related activities have been confined to compounds of low molecular mass, and no data have been reported concerning the plant's polysaccharides. In this work, a crude polysaccharide of M. charantia (MCP) fruit was isolated by hot water extraction and then purified using DEAE-52 cellulose anion-exchange chromatography to produce two main fractions MCP1 and MCP2. The immunomodulatory effects and physicochemical characteristics of these fractions were investigated in vitro and in vivo. The results showed that intragastric administration of 150 or 300 mg·kg-·d⁻¹ of MCP significantly increased the carbolic particle clearance index, serum haemolysin production, spleen index, thymus index and NK cell cytotoxicity to normal control levels in cyclophosphamide (Cy)-induced immunosuppressed mice. Both MCP1 and MCP2 effectively stimulated normal and concanavalin A-induced splenic lymphocyte proliferation in vitro at various doses. The average molecular weights of MCP1 and MCP2, which were measured using high-performance gel permeation chromatography, were 8.55×104 Da and 4.41×105 Da, respectively. Both fractions exhibited characteristic polysaccharide bands in their Fourier transform infrared spectrum. MCP1 is mainly composed of glucose and galactose, and MCP2 is mainly composed of glucose, mannose and galactose. The results indicate that MCP and its fractions have good potential as immunotherapeutic adjuvants.


Subject(s)
Immunologic Factors/pharmacology , Momordica charantia/chemistry , Plant Extracts/pharmacology , Polysaccharides/pharmacology , Animals , Cell Line, Tumor , Cell Proliferation , Cytotoxicity, Immunologic/drug effects , Fruit/chemistry , Hemolysin Proteins/biosynthesis , Immunologic Factors/isolation & purification , Inhibitory Concentration 50 , Killer Cells, Natural/drug effects , Killer Cells, Natural/metabolism , Macrophages/drug effects , Macrophages/metabolism , Male , Mice , Phagocytosis/drug effects , Plant Extracts/isolation & purification , Polysaccharides/isolation & purification , Spectroscopy, Fourier Transform Infrared , Spleen/cytology
11.
Molecules ; 18(9): 11601-13, 2013 Sep 18.
Article in English | MEDLINE | ID: mdl-24051475

ABSTRACT

The solution properties of four fractions (LPI-IV) from crude longan pulp polysaccharides (LP3) were analyzed by size-exclusion chromatography combined with laser light scattering, viscometry, complex formation with Congo red, and atomic force microscopy. Their radii of gyration (z)(½) were 43.3, 62.6, 43.2 and 77.3 nm, exponents of (z)(½) = k M(w)(v) were 0.04, 0.50, 0.52 and 0.02, and intrinsic viscosities ([η]) were 9.945, 25.38, 308.2 and 452.1 mL/g, respectively. Moreover, the dependence of [η] on M(w) was established to be [η] = 5.3 × 10⁻²M(w)°·6¹ (mL/g). LPI had both a sphere-like conformation and a triple-helix structure, and LPII-IV existed as flexible chains. LP3, LPI, LPII and LPIII all exhibited direct inhibitory effects on A549, HeLa and HepG2 cells in a positive dose-dependent manner in the range of 50-400 µg/mL. The activities of LPIII, especially the inhibition of HepG2 cell proliferation, were stronger than those of others, which may be partly related to its flexible conformation. The present results support the cancer therapeutic potential of longan polysaccharides.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Fruit/chemistry , Plant Extracts/pharmacology , Polysaccharides/pharmacology , Sapindaceae/chemistry , Antineoplastic Agents, Phytogenic/chemistry , Carbohydrate Conformation , Cell Proliferation/drug effects , Drug Screening Assays, Antitumor , HeLa Cells , Hep G2 Cells , Humans , Molecular Weight , Plant Extracts/chemistry , Polysaccharides/chemistry , Polysaccharides/ultrastructure , Solutions , Viscosity
12.
PLoS One ; 8(7): e69955, 2013.
Article in English | MEDLINE | ID: mdl-23950843

ABSTRACT

The MYB proteins comprise one of the largest families of transcription factors (TFs) in plants. Although several MYB genes have been characterized to play roles in secondary metabolism, the MYB family has not yet been identified in apple. In this study, 229 apple MYB genes were identified through a genome-wide analysis and divided into 45 subgroups. A computational analysis was conducted using the apple genomic database to yield a complete overview of the MYB family, including the intron-exon organizations, the sequence features of the MYB DNA-binding domains, the carboxy-terminal motifs, and the chromosomal locations. Subsequently, the expression of 18 MYB genes, including 12 were chosen from stress-related subgroups, while another 6 ones from other subgroups, in response to various abiotic stresses was examined. It was found that several of these MYB genes, particularly MdoMYB121, were induced by multiple stresses. The MdoMYB121 was then further functionally characterized. Its predicted protein was found to be localized in the nucleus. A transgenic analysis indicated that the overexpression of the MdoMYB121 gene remarkably enhanced the tolerance to high salinity, drought, and cold stresses in transgenic tomato and apple plants. Our results indicate that the MYB genes are highly conserved in plant species and that MdoMYB121 can be used as a target gene in genetic engineering approaches to improve the tolerance of plants to multiple abiotic stresses.


Subject(s)
Genome, Plant , Malus/genetics , Plant Proteins/genetics , Stress, Physiological/genetics , Transcription Factors/genetics , Binding Sites , Chromosomes, Plant , Gene Duplication , Gene Expression Profiling , Gene Expression Regulation, Plant , Solanum lycopersicum/drug effects , Malus/physiology , Multigene Family , Phylogeny , Plant Proteins/pharmacology , Plants, Genetically Modified/physiology , Stress, Physiological/drug effects , Transcription Factors/pharmacology
13.
Plant Physiol Biochem ; 67: 169-77, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23570872

ABSTRACT

Cryptochromes are blue-light photoreceptors involved in regulating many aspects of plant growth and development. Investigations of cryptochromes in plants have largely focused on Arabidopsis (Arabidopsis thaliana), tomato (Solanum lycopersicum), rice (Oryza sativa) and pea (Pisum sativum). Here, we isolated the cryptochrome 1 gene from apple (Malus domestica) (MdCRY1) and analyzed its function in transgenic Arabidopsis. The predicted MdCRY1 protein was most closely homologous to strawberry CRY1. In terms of transcript levels, MdCRY1 expression was up-regulated by light. The function of MdCRY1 was analyzed through heterologous expression in Arabidopsis. Overexpression of MdCRY1 in Arabidopsis is able to rescue the cry1 mutant phenotype, inhibit hypocotyl elongation, promote root growth, and enhance anthocyanin accumulation in wild-type seedlings under blue light. These data provide functional evidence for a role of MdCRY1 in controlling photomorphogenesis under blue light and indicate that CRY1 function is conserved between Arabidopsis and apple. Furthermore, we found that MdCRY1 interacts with AtCOP1 in both yeast and onion cells. This interaction may represent an important regulatory mechanism in blue-light signaling pathway in apple.


Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Cryptochromes/metabolism , Malus/metabolism , Plant Proteins/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Cloning, Molecular , Cryptochromes/genetics , Gene Expression Regulation, Plant , Malus/genetics , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism
14.
Plant Cell Rep ; 32(4): 555-66, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23314496

ABSTRACT

KEY MESSAGE: MdCRY2 was isolated from apple fruit skin, and its function was analyzed in MdCRY2 transgenic Arabidopsis. The interaction between MdCRY2 and AtCOP1 was found by yeast two-hybrid and BiFC assays. Cryptochromes are blue/ultraviolet-A (UV-A) light receptors involved in regulating various aspects of plant growth and development. Investigations of the structure and functions of cryptochromes in plants have largely focused on Arabidopsis (Arabidopsis thaliana), tomato (Solanum lycopersicum), pea (Pisum sativum), and rice (Oryza sativa). However, no data on the function of CRY2 are available in woody plants. In this study, we isolated a cryptochrome gene, MdCRY2, from apple (Malus domestica). The deduced amino acid sequences of MdCRY2 contain the conserved N-terminal photolyase-related domain and the flavin adenine dinucleotide (FAD) binding domain, as well as the C-terminal DQXVP-acidic-STAES (DAS) domain. Relationship analysis indicates that MdCRY2 shows the highest similarity to the strawberry FvCRY protein. The expression of MdCRY2 is induced by blue/UV-A light, which represents a 48-h circadian rhythm. To investigate the function of MdCRY2, we overexpressed the MdCRY2 gene in a cry2 mutant and wild type (WT) Arabidopsis, assessed the phenotypes of the resulting transgenic plants, and found that MdCRY2 functions to regulate hypocotyl elongation, root growth, flower initiation, and anthocyanin accumulation. Furthermore, we examined the interaction between MdCRY2 and AtCOP1 using a yeast two-hybrid assay and a bimolecular fluorescence complementation assay. These data provide functional evidence for a role of blue/UV-A light-induced MdCRY2 in controlling photomorphogenesis in apple.


Subject(s)
Cryptochromes/metabolism , Malus/genetics , Plant Proteins/metabolism , Amino Acid Sequence , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Cloning, Molecular , Cryptochromes/genetics , Genetic Complementation Test , Light , Malus/growth & development , Malus/metabolism , Molecular Sequence Data , Plant Proteins/genetics , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Two-Hybrid System Techniques , Ubiquitin-Protein Ligases/metabolism
15.
Biomed Environ Sci ; 26(1): 40-7, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23294614

ABSTRACT

OBJECTIVE: To assess and compare the Human Papillomavirus (HPV) detection efficiency and the potential clinical utility of PCR sequencing-based technology. METHODS: Four HPV consensus primer sets (GP5+/6+, MGP, MY09/11, and PGMY09/11) were used in order to amplify a broad spectrum of HPV types for HPV infection in 325 cervical samples and the PCR products were sequenced afterwards for the HPV genotyping. RESULTS: The HPV-positive rate was 75.4%, of which 35.5% harbored more than one HPV genotype. A total of 36 different genotypes was found, with HPV 16 (24.1%) being the most prevalent, followed by HPV 58 (13.3%) and HPV 52 (9.6%). There were substantial to almost perfect agreements between different primer sets regarding HPV detection efficiency, with the kappa value varying from 0.751 to 0.925, MGP, and PGMY09/11 were the most effective in detecting multiple infections (P < 0.001). With each of the primer sets, a board range of HPV types could be identified, though there were several differences for a few genotypes. CONCLUSION: The substantial agreement between PCR-sequencing and HC2 for the detection of high-risk HPV (kappa=0.761) indicated that PCR-sequencing is also suitable for routine HPV screening.


Subject(s)
Alphapapillomavirus/genetics , Alphapapillomavirus/isolation & purification , Papillomavirus Infections/virology , Polymerase Chain Reaction/methods , Adult , Aged , Female , Genotype , Humans , Middle Aged , Young Adult
16.
Molecules ; 17(12): 14954-67, 2012 Dec 17.
Article in English | MEDLINE | ID: mdl-23247368

ABSTRACT

Litchi fruit pericarp (LFP) contains significant amounts of phenolics which have been found to exhibit diverse biological activities. The purpose of this work was to determine the varietal differences in phenolic profiles and antioxidant activity of LFP from nine commercially available cultivars. The total phenolic and flavonoid contents ranged from 9.39 to 30.16 mg gallic acid equivalents/g fresh weight (FW) and from 7.12 to 23.46 mg catechin equivalents/g FW, respectively. The total anthocyanin contents ranged from 1.77 to 20.94 mg cyanidin-3-glucoside equivalents/100 g FW. Three anthocyanins, including cyanidin-3-rutinoside, cyanidin-3-glucoside, malvidin-3-glucoside, were detected, and cyanidin-3-rutinoside was the predominant constituent which contributes from 68.8% to 100% to total anthocyanins, The total procyanidin contents ranged from 4.35 to 11.82 mg epicatechin equivalents/g FW. Procyanidin B2, epicatechin, A-type procyanidin trimer, and procyanidin A2 were detected in all nine litchi varieties. The oxygen radical absorbance capacity activities and DPPH radical-scavenging activities ranged from 430.49 to 1752.30 µmol TE/100 g FW and from 4.70 to 11.82 mg/g (IC50), respectively. These results indicate that there are significant differences in phytochemical profiles and antioxidant activity among the tested varieties. Knowing the phenolic profiles and antioxidant activity of LFP of different varieties gives the insights into its potential application to promote health.


Subject(s)
Antioxidants , Fruit/chemistry , Litchi/chemistry , Phenols , Antioxidants/analysis , Antioxidants/chemistry , Breeding , Catechin/analysis , Catechin/chemistry , Chromatography, High Pressure Liquid , Flavonoids/analysis , Flavonoids/chemistry , Oxidation-Reduction , Phenols/analysis , Phenols/chemistry , Plant Extracts/analysis , Plant Extracts/chemistry
17.
J Agric Food Chem ; 60(44): 11097-103, 2012 Nov 07.
Article in English | MEDLINE | ID: mdl-23075052

ABSTRACT

Penthorum chinense Pursh, widely distributed in eastern Asia, has long been used in China for both food and medicine due to its various bioactivities. The aim of this study was to isolate its active compounds with antioxidant and antihepatocarcinoma properties. P. chinense was extracted with 95% ethanol, 70% ethanol, and water, respectively, and then the 70% ethanol extract was re-extracted, resulting in petroleum ether, ethyl acetate, n-butanol, and water fractions, subsequently. Results showed that the antioxidant and antihepatocarcinoma activities of ethanol extracts were stronger than those of aqueous extract, and the ethyl acetate fraction of 70% ethanol extract showed the highest activities. Four compounds, ß-sitosterol, quercetin, pinocembrin-7-O-[3-O-galloyl-4″,6″-hexahydroxydiphenoyl]-ß-glucose (PGHG), and thonningianins A (Th A), were isolated from the ethyl acetate fraction and identified by UV, MS, and NMR spectroscopic analysis. Th A was isolated from P. chinense for the first time. PGHG and Th A exhibited higher antioxidant and antihepatocarcinoma activities than did other isolated parts of P. chinense . The antihepatocarcinoma activity of Th A was much higher than that of positive control (5-fluorouracil). PGHG and Th A were suggested to be the active chemical compositions responsible for potent antioxidant and antihepatocarcinoma properties of P. chinense , which are worthy of further study.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Antioxidants/pharmacology , Carcinoma, Hepatocellular/pathology , Liver Neoplasms/pathology , Saxifragaceae/chemistry , Cell Line, Tumor , Cell Proliferation/drug effects , Humans , Magnetic Resonance Spectroscopy
18.
Plant Cell Environ ; 35(11): 1884-97, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22519753

ABSTRACT

Low environmental temperatures promote anthocyanin accumulation and fruit colouration by up-regulating the expression of genes involved in anthocyanin biosynthesis and regulation in many fruit trees. However, the molecular mechanism by which fruit trees regulate this process in response to low temperature (LT) remains largely unknown. In this study, the cold-induced bHLH transcription factor gene MdbHLH3 was isolated from an apple tree and was found to interact physically and specifically through two regions (amino acids 1-23 and 186-228) at the N terminus with the MYB partner MdMYB1 (allelic to MdMYB10). Subsequently, MdbHLH3 bound to the promoters of the anthocyanin biosynthesis genes MdDFR and MdUFGT and the regulatory gene MdMYB1 to activate their expression. Furthermore, the MdbHLH3 protein was post-translationally modified, possibly involving phosphorylation following exposure to LTs, which enhanced its promoter-binding capacity and transcription activity. Our results demonstrate the molecular mechanism by which MdbHLH3 regulates LT-induced anthocyanin accumulation and fruit colouration in apple.


Subject(s)
Anthocyanins/metabolism , Basic Helix-Loop-Helix Transcription Factors/physiology , Fruit/metabolism , Malus/metabolism , Plant Proteins/physiology , Temperature , Anthocyanins/biosynthesis , Basic Helix-Loop-Helix Transcription Factors/genetics , Basic Helix-Loop-Helix Transcription Factors/metabolism , Climate , Color , Fruit/genetics , Genetic Vectors , Malus/genetics , Phosphorylation , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/metabolism , Protein Processing, Post-Translational , Nicotiana/genetics
19.
Carbohydr Polym ; 87(1): 636-643, 2012 Jan 04.
Article in English | MEDLINE | ID: mdl-34663015

ABSTRACT

An aqueous extract of polysaccharides from longan pulp was chromatographed on a DEAE anion-exchange column to yield four fractions (LPI-IV). Immunomodulatory activities of these polysaccharides were also evaluated in vitro. The purified products, neutral polysaccharide LPI, polysaccharide-protein complex LPII and acidic polysaccharides LPIII and LPIV, exhibited conspicuous differences in their monosaccharide composition, molecular mass and glycosidic linkages. Except for LPI, the other three significantly stimulated lymphocyte proliferation in the dose range of 100-400µg/mL compared with the normal control (P<0.05), and might electively stimulate B cells, but not T cells. Furthermore, their stimulations on normal/lipopolysaccharide-induced proliferation and depressions on concanavalin A-induced proliferation could be ordered as LPIII>LPIV>LPII>LPI. All the fractions had the optimal dose of 100µg/mL on enhancing macrophage phagocytosis. Among them, LPII had the considerable yield and activity for exploiting as a potential immunoadjuvant.

20.
Molecules ; 16(12): 10324-36, 2011 Dec 13.
Article in English | MEDLINE | ID: mdl-22158685

ABSTRACT

The immunomodulatory function of longan pulp polysaccharide-protein complex (LP3) was investigated in immunosuppressed mice models. Compared with the model control, peroral administration of 100 mgkg⁻¹d⁻¹ LP3 could significantly increase/enhance antibody production against chicken red blood cell (CRBC), concanavalin A (ConA)-induced splenocyte proliferation, macrophage phagocytosis, NK cell cytotoxicity against YAC-1 lymphoma cell, and interferon-gamma (INF-γ) and interleukin-2 (IL-2) secretion in serum (P < 0.05). The immunomodulatory effects, except for those on splenocytes and macrophages (P > 0.05), were also observed in mice administered with 50 or 200 mgkg⁻¹d⁻¹ LP3 (P < 0.05). The beneficial effects of 50-200 mgkg⁻¹d⁻¹ LP3 were comparable to those of 50 mgkg⁻¹d⁻¹ ganoderan. The strong immunomodulatory activity of LP3 confirmed its good potential as an immunotherapeutic adjuvant.


Subject(s)
Glycoproteins/chemistry , Glycoproteins/pharmacology , Immunologic Factors/pharmacology , Plant Extracts/pharmacology , Polysaccharides/pharmacology , Sapindaceae/chemistry , Animals , Body Weight/drug effects , Cell Death/drug effects , Cell Proliferation/drug effects , Chickens , Drinking Behavior/drug effects , Feeding Behavior/drug effects , Hemolysin Proteins/blood , Interferon-gamma/blood , Interleukin-2/blood , Killer Cells, Natural/cytology , Killer Cells, Natural/drug effects , Lymphocytes/cytology , Lymphocytes/drug effects , Macrophages/cytology , Macrophages/drug effects , Male , Mice , Phagocytosis/drug effects , Spleen/cytology , Spleen/drug effects , Thymus Gland/drug effects
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