[Research on distribution and quality suitability division of Desmodium styracifolium].

The research on distribution and quality suitability division of Desmodium styracifolium were formulated by Maxent and ArcGIS model based on the content of schaftoside and polysaccharide of D. styracifolium and its field research in the south and southwest areas of China (Guangdong, Guangxi, Hainan and Yunnan), and the most suitable habitats of distribution suitability and quality suitability were screened. The distribution suitability results indicated that average air temperature in April,mean temperature of coldest quarter, soil type, coldness index were found as the four dominant factors contributing to the plant distribution. The quality suitability results indicated that: ①Polysaccharide content and precipitation in April show significant positive correlation;Schaftoside content and mean temperature of April, mean temperature of coldest quarter show significant negative correlation. Schaftoside content shows significant negative correlation with the precipitation in October and November and the sunshine duration in April and May, while there is a significant positive correlation between schaftoside content and precipitation in April and temperature seasonality standard deviation, and a highly significant positive correlation was found between schaftoside content and precipitation in February and March. ②The quality zoning map was drawn depend on general content of polysaccharide and schaftoside as the index of quality. And this research provides scientific location basis for the production regionalization, cultivation bases selection and directive breeding of D. styracifolium.

[Effects of storage time on quality of Desmodium styracifolium seeds].

The dynamic changes of germination percentage, germination potential, thousand-seed weight, antioxidase activity in Desmodium styracifolium seeds with different storage time were tested, and electrical conductivity, contents of soluble sugar, soluble protein, starch in seed leach liquor were also determined in order to reveal the mechanism of seed deterioration. The results as the following. (1) The germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds declined, while the seed coat color darkened with the extension of storage time. (2) The activities of superoxide dismutase (SOD) and peroxidase (POD) decreased with the prolongation of storage period. The SOD activity declined fastest in 1,095-1,185 d of storage, while the POD activity declined significantly in 365-395 d of storage. (3) The electrical conductivity and the contents of soluble sugar, starch in seed leach liquor increased, while the content of soluble protein declined with the extension of storage time. (4) Correlation analysis indicated that the germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds have a significantly positive correlation with SOD and POD activity, while have a significantly negative correlation with the electrical conductivity, contents of soluble sugar and starch. It can be concluded that during the storage of D. styracifolium seeds, physiological and biochemical changes including decrease in antioxidase activity, rise in electrical conductivity, degradation effluent of soluble sugar and starch, degradation of soluble protein were the main factors leading to the seed deterioration.

[De novo sequencing and analysis of root transcriptome to reveal regulation of gene expression by moderate drought stress in Glycyrrhiza uralensis].

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as ß-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.

Insulin upregulates the expression of epithelial sodium channel in vitro and in a mouse model of acute lung injury: role of mTORC2/SGK1 pathway.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are characterized by proteinaceous pulmonary edema and severe arterial hypoxemia with a mortality of approximately 40%. Stimulation of epithelial sodium channel (ENaC) promotes Na(+) transport, a rate-limiting step for pulmonary edema reabsorption. Insulin is known to participate in the ion transport; however, its role in pulmonary edema clearance and the regulatory mechanism involved have not been fully elucidated. In the current study, in a lipopolysaccharide-based mouse model of ALI, we found that insulin alleviated pulmonary edema by promoting ENaC-mediated alveolar fluid clearance through serum and glucocorticoid induced kinase-1 (SGK1). In alveolar epithelial cells, insulin increased the expression of α-, ß-, and γ-ENaC, which was blocked by the mammalian target of rapamycin complex 2 (mTORC2) inhibitor or knockdown of Rictor (a necessary component of mTORC2), and SGK1 inhibitor, respectively. In addition, an immunoprecipitation study demonstrated that SGK1(Ser422) phosphorylation, the key step for complete SGK1 activation by insulin, was conducted through PI3K/mTORC2 pathway. Finally, we testified the role of mTORC2 in vivo by demonstrating that PP242 prevented insulin-stimulated SGK1 activation and ENaC increase during ALI. The data revealed that during ALI, insulin stimulates alveolar fluid clearance by upregulating the expression of α-, ß-, and γ-ENaC at the cell surface, which was, at least, partially through activating mTROC2/SGK1 signaling pathway.

[Health risk assessment of heavy metals in atmospheric dust of Qingdao City].

Based on the 89 atmospheric dust samples and soil samples that were collected around Qingdao, we tested and analyzed the contents of Cd, Cr, Cu, Hg, Ni, Pb, Zn. Based on these analysis results, the risk of heavy metals in atmospheric dusts to human health were assessed by using the US EPA Health Risk Assessment Model. Analysis showed that the average contents of Cd, Cr, Cu, Hg, Pb, Zn in the atmospheric dust of Shinan, Shibei and Laoshan districts were the highest. Therefore, the air pollution of these districts was more serious than the districts of Licang, Chengyang and Huangdao. Comparing the average contents of heavy metals in atmospheric dust with those in soil, we found that only the content of Hg in atmospheric dust collected from the districts of Shinan, Shibei and Laoshan was lower than that in the corresponding soil. All the contents of other heavy metals in atmospheric dust were higher than those in corresponding soil. As a whole, the heavy metals in atmospheric dust of Qingdao City showed slight difference and were less harmful to human health. However, it was harmful in some samples to human health if the contents of Cr and Pb in atmospheric dusts of Shinan, Laoshan and Chengyang districts were always kept at such high densities. Besides, the accumulation of heavy metals in atmospheric dust through various approaches and categories may obviously increase the risk of damaging human health.

[Analysis of quality variation and genetic diversity of Desmodium styracifolium from different provenances].

OBJECTIVE: To explore the quality variation and genetic diversity of Desmodium styracifolium from different provenances, and lay a foundation for rational exploitation on germplasm resources and fine variety breeding of D. styracifolium. METHOD: Amplified fragment length polymorphism (AFLP) markers were developed to analyze genetic diversity in D. styracifolium from 18 resources. NTSYSpc-2. 11F software was used to analyze the similarity among the D. styracifolium germplasms and construct the genetic phylogenetic tree. The schaftoside content in D. styracifolium from different provenances was determined by HPLC. RESULT: A total of 844 fragments were amplified with 8 primers, in which 717 were polymorphic bands, accounting for 84. 27% of the total detected variation. All the specimens from 18 resources could be grouped into 3 clusters by cluster analysis. The schaftoside contents of D. styracifolium germplasms differed significantly, with the highest content in the germplasm from Sanya, Hainan. CONCLUSION: Significant quality variation and genetic diversity can be observed among D. styracifolium germplasms. The diverse germplasm resources should be explored and the fine variety should be selected to breed.

[Roles of glucosinolates in the interrelationships between Brassicaceae plants and insects: a review].

Glucosinolates (GS) are the important secondary metabolites of Brassicaceae plants, playing an important role in regulating the interrelationships between Brassicaceae plants and insects. GS can protect Brassicaceae plants against euryphagous herbivorous pests because of the toxicity of GS and their breakdown products. However, oligophagous pests which have evolved manifold metabolic pathways to cope with the defensive compounds depended fully on GS and their volatile breakdown products for host-plant recognition and orientation. The GS ingested by herbivores are also toxic to carnivores, and can directly deter predators. On the other hand, predators and parasitoids are attracted by the volatile breakdown products of GS from the Brassicaceae plants damaged by herbivores. Based on the recent findings, this paper reviewed the defensive function of GS against herbivores, host selection of oligophagous pests, GS metabolic pathways of herbivores, induction of GS by herbivores, and effects of GS on the third tropic level. Future directions and techniques in this research field were also suggested.

[Cloning and expression regulation of 1-deoxy-D-xylulose-5-phosphate reductoisomerase cDNA from Alpinia officinarum].

The rhizome of Alpinia officinarum is a widely used Chinese herbal medicine. The essential oil in A. officinarum rhizome is mainly composed of 1, 8-cineole and other monoterpenes, as the major bioactive ingredients. In plants, monoterpenes are synthesized through the methylerythritol phosphate (MEP) pathway in the plastids, and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) is an enzyme catalyzing a committed step of the MEP pathway. In the present study, the full-length cDNA encoding DXR was cloned from the rhizome of A. officinarum, using homology-based RT-PCR and rapid amplification of cDNA ends (RACE) techniques. The new cDNA was designated as AoDXR and submitted to GenBank to be assigned with an accession number HQ874658. The full-length cDNA of AoDXR was 1 670 bp containing a 1 419 bp open reading frame encoding a polypeptide of 472 amino acids with a calculated molecular mass of 51.48 kDa and an isoelectric point of 6.15. Bioinformatic analyses revealed that AoDXR showed extensive homology with DXRs from other plant species and contained a conserved plastids transit peptide, a Pro-rich region and two highly conserved NADPH-binding motifs in its N-terminal region characterized by all plant DXRs. The phylogenetic analysis revealed that AoDXR belonged to angiosperm DXRs. The structural modeling of AoDXR showed that AoDXR had the typical V-shaped structure of DXR proteins. The tissue expression pattern analysis indicated that AoDXR expressed strongly in leaves, weak in rhizomes of A. officinarum. Exogenous methyl jasmonate (MeJA) could enhance the expression of AoDXR and the production of 1, 8-cineole in A. officinarum rhizomes. The cloning and characterization of AoDXR will be helpful to reveal the molecular regulation mechanism of monoterpene biosynthesis in A. officinarum and provides a candidate gene for metabolic engineering in improving the medicinal quality of A. officinarum rhizome.

Cloning and expression of 1-aminocyclopropane-1-carboxylate oxidase cDNA induced by thidiazuron during somatic embryogenesis of alfalfa (Medicago sativa).

Embryogenic callus (EC) induced from petioles of alfalfa (Medicago sativa L. cv. Jinnan) on B5h medium turned green, compact and non-embryogenic when the kinetin (KN) in the medium was replaced partially or completely by thidiazuron (TDZ). The application of CoCl2, which is an inhibitor of 1-aminocyclopropane-1-carboxylate oxidase (ACO), counteracted the effect of TDZ. Ethylene has been shown to be involved in the modulation of TDZ-induced morphogenesis responses. However, very little is known about the genes involved in ethylene formation during somatic embryogenesis (SE). To investigate whether ethylene mediated by ACO is involved in the effect of TDZ on inhibition of embryogenic competence of the alfalfa callus. In this study we cloned full-length ACO cDNA from the alfalfa callus, named MsACO, and observed changes in this gene expression during callus formation and induction of SE under treatment with TDZ or TDZ plus CoCl2. RNA blot analysis showed that during the EC subcultural period, the expression level of MsACO in EC was significantly increased on the 2nd day, rose to the highest level on the 8th day and remained at this high level until the 21st day. However, the ACO expression in the TDZ (0.93 µM)-treated callus was higher than in the EC especially on the 8th day. Moreover the ACO expression level increased with increasing TDZ concentration during the subcultural/maintenance period of the callus. It is worth noting that comparing the treatment with TDZ alone, the treatment with 0.93 µM TDZ plus 50 µM CoCl2 reduced both of the ACO gene expressions and ACO activity in the treated callus. These results indicate that the effect of TDZ could be counteracted by CoCl2 either on the ACO gene expression level or ACO activity. Thus, a TDZ inhibitory effect on embryogenic competence of alfalfa callus could be mediated by ACO gene expression.

QTL mapping for test weight by using F(2:3) population in maize.

Test weight is an important trait in maize breeding. Understanding the genetic mechanism of test weight is important for effective selection of maize test weight improvement. In this study, quantitative trait loci (QTL) for maize test weight were identified. In the years 2007 and 2008, a F(2:3) population along with the parents Chang7-2 and Zheng58 were planted in Zhengzhou, People's Republic of China. Significant genotypic variation for maize test weight was observed in both years. Based on the genetic map containing 180 polymorphic SSR markers with an average linkage distance of 11.0 cM, QTL for maize test weight were analysed by mixed-model composite interval mapping. Five QTL, including four QTL with only additive effects, were identified on chromosomes 1, 2, 3, 4 and 5, and together explained 25.2% of the phenotypic variation. Seven pairs of epistatic interactions were also detected, involving 11 loci distributed on chromosomes 1, 2, 3, 4, 5 and 7, respectively, which totally contributed 18.2% of the phenotypic variation. However, no significant QTL x environment (QxE) interaction and epistasis x environment interaction effects were detected. The results showed that besides the additive QTL, epistatic interactions also formed an important genetic basis for test weight in maize.

Mobilization of Peripheral Blood Stem Cells Using Regimen Combining Docetaxel with Granulocyte Colony-stimulating Factor in Breast Cancer Patients.

OBJECTIVE: To evaluate the effectiveness and safety of the mobilization of peripheral blood hematopoietic stem cells by combining docetaxel with granulocyte colony-stimulating factor (G-CSF) in breast cancer patients. METHODS: A total of 57 breast cancer patients were treated with docetaxel 120 mg/m(2). When the white blood cell (WBC) count decreased to 1.0×10(9)/L, patients were given G-CSF 5 µg/kg daily by subcutaneous injection until the end of apheresis. Peripheral blood mononuclear cells (MNC) were isolated by Cobe Spectra Apheresis System. The percentage of CD34(+) cell was assayed by flow cytometry. RESULTS: At a median 6 of days (range 3-8) after the administration of docetaxel, the median WBC count decreased to 1.08×10(9)/L (range 0.20-2.31). The median duration of G-CSF mobilization was 3 days (range 2-7). The MNC collection was conducted 8-12 days (median 10 days) after docetaxel treatment. The median MNC was 5.35×10(8)/kg (range 0.59-14.07), the median CD34(+) cell count was 2.43×10(6)/kg (range 0.16-16.69). The CD34(+) cell count was higher than 1.00×10(6)/kg in 47 of 57 cases (82.46%) and higher than 2.00×10(6)/kg in 36 cases (63.16%). The CD34(+) cell count was higher than 2.00×10(6)/kg in 27 collections (23.68%). The MNC count and the CD34(+) cell count were correlated with the bottom of WBC after docetaxel chemotherapy (r=0.364, 0.502, P=0.005, 0.000). The CD34(+) cell count was correlated with the MNC count (r=0.597, P=0.000). The mobilization and apheresis were well tolerated in all patients. Mild perioral numbness and numbness of hand or feet were observed in 3 cases. No serious adverse events were reported. CONCLUSION: Mobilization of peripheral blood hematopoietic stem cell by combining docetaxel with G-CSF was effective and safety in breast cancer patients.

[Community structure and time-space distribution characteristic of airborne fungi in a municipal landfill site].

In order to determine the community structure and time-space distribution characteristics of airborne fungi in waste landfill sites, supervisory points were set in sanitary landfill area, leachate treatment area and living area in a Waste Sanitary Landfill Site in Beijing. Airborne fungi were collected by Andersen six stages sampler, and the study on the community structure and distribution of airborne fungi was carried out systematically. The results show that fifteen genera of culturable fungi are identified (exclusion Non-sporing), Cladosporium, Aspergillus, Penicillium and Non-sporing are the dominant genera. The concentration of airborne fungi in sanitary landfill area and leachate treatment area are about 1750 CFU x m(-3), which is higher than that in living area (p < 0.05). Monthly change curve of the airborne fungi concentration shows double peak type, the peak value appears in May and September-October, which can reach at 5 000 CFU x m(-3). It indicates a trend that the concentration is higher at 09:00-11:00 than that at 15:00-16:00 from April to July, but it performs reserved trend from August to the next January. 75% of the airborne fungi distributes in stage III - V. The counted median diameters (CMD) of airborne fungi in three functional areas are 2.9 microm, 2.9 microm and 2.8 microm respectively, there are no statistical difference (p > 0.05).

[Reaction of atrazine with protein and its application to detection of protein].

The resonance light scattering (RLS) technique and UV-Vis absorption spectra were applied to the investigation of the interaction between atrazine and bovine serum albumin(BSA). Under acidic conditions, the formation of atrazine-BSA supermolecule by Van der Waals force and N/O--H...pi hydrogen bonds leads to a red shift of absorption band and strong RLS enhancement of atrazine. The characteristics and intensity of RLS were related to the pH, the concentration of atrazine, and temperature. Under the optimum conditions, the enhanced RLS intensities are in proportion to the concentration of BSA in the range of 0.05-100 microg x mL(-1). Based on the enhancement of the RLS, a simple and sensitive method for the determination of BSA was established. The detection limit (3sigma) is 12 ng x mL(-1). Synthesis samples were determined with satisfactory results.

Influence of CaNa2 EDTA on topical 5-aminolaevulinic acid photodynamic therapy.

BACKGROUND: We assessed whether the CaNa2 EDTA could improve the accumulation of protoporphyrin IX (PpIX) and photosensitisation in HEp-2 cells as well as the depth of treatment of skin cancers on the topical 5-Aminolaevulinic acid (5-ALA) PDT. METHODS: HEp-2 cells were incubated with 5-ALA (0-1 mmol/L) and CaNa2EDTA (0-1 mmol/L) for 4 hours, intracellular protoporphyrin IX content was quantified by extraction, and cell viability was assessed by use of the methyl-tetrazolium (MTT) assay four hours after exposure to light. In comparison with the pictures before and after treatment, depth of treatment could be determined using a Acuson Sequioa 512 phase-array system in paired experiments. RESULTS: PpIX accumulation increased with increasing extracellular concentrations of ALA (0-1 mmol/L). Adding 1 mmol/L of CaNa2EDTA increased 30% PpIX accumulation over the same period of incubation in the concentration of 1 mmol/L ALA. Significant difference was observed between the 5-ALA alone group and 5-ALA combined CaNa2 EDTA group in the PpIX accumulation (P < 0.01). Cell viability after exposure to light decreased with adding CaNa2 EDTA, a statistical difference in a same fluence above 1.2 J/cm2 between two groups was demonstrated (P < 0.05, P < 0.01 respectively). Depth of treatment of skin cancers were increased in CaNa2 EDTA-treated group. CONCLUSION: CaNa2 EDTA could improve the PpIX accumulation and photosensitisation in HEp-2 cells. Clinically, CaNa2 EDTA could increase the depth of treatment in the cutaneous cancers.

[Determination of deoxyribonuceic acids by resonance light scattering technique with benzidine in the presence of the sensitizing sodium dodecyl sulfate].

The resonance light scattering (RLS) spectra of benzidine with deoxyribonucleic acid (ctDNA) in the presence of Sodium Dodecyl Sulfate had been studied. RLS intensity of SDS-benzidine system was enhanced by trace ctDNA and was proportional to the concentrationof ctDNA. Under optima conditions, the linear ranges of the calibration were 0.3-4 mg x L(-1), the linear regression equation was I = 37.17c (ctDNA, mg x L(-1)) + 208.9, r = 0.9992. The limit of detection were 0.17 mg x L(-1) (3delta), RSD% = 3.5%. The method had been applied to the determination of DNA in synthetic samples and gene tobacco sample, and the results were satisfactory.

[Study on the resonance scattering light spectra of atrazine-RNA and its analytical application].

The Resonance Light Scattering (RLS) spectra of atrazine with yeast ribonucleic acid (yRNA) had been studied. At pH 1.50, the enhanced RLS of atrazine-yRNA system is characterized by the maximum scattering peaks at 320 nm and the RLS intensity is proportional to the concentration of yRNA. Under optimal conditions, the linear ranges of the calibration were 0.6-5.0 microg x mL(-1), the linear regression equation was I = 25.88 + 140.0 c(yRNA, microg x mL(-1)), r = 0.9975, with the limit of detection 20.7 ng x mL(-1) (3delta). The method was applied to the determination of yRNA in synthetic samples and RNA in salt cress (Thellungiella halophila) sample satisfactorily. Mechanism studies show that there are two interaction modes between atrazine and RNA: electrostatic mode and intercalative mode.