The prevalence of Toxoplasma gondii infection in free-range chickens in Jilin province, northeastern China was investigated. A total of 1095 serum samples were collected from nine administrative regions from July to October, 2012, and antibodies to T. gondii were examined by an in-house enzyme-linked immunosorbent assay (ELISA) using Toxoplasma lysate antigen (TLA). The detection results were confirmed by Western blot. The overall seroprevalence of T. gondii in free-range chickens was 17.6% (95% confidence interval [CI], 15.4–19.4%), ranging from 13.3% (95% CI, 6.3–20.4%) in Siping to 23.6% (95% CI, 15.7–31.6%) in Liaoyuan. There was no significant difference in T. gondii infection among different regions in Jilin province (P> 0.05). The widespread presence of T. gondii infection in freerange chickens of Jilin province implies the wide contamination with T. gondii oocysts in the living environment of people, and free-range chickens might be an important source of infection for humans.

OBJECTIVETo study the effects of fertilizers with the different proportional of nitrogen, phosphorus and potassium on growth and active ingredient of Astragalus membranaceus.

METHODField experiment was conducted based on the D-saturation optimal design with three factors of nitrogen, phosphorus and potassium. The effects on growth and active ingredient of A. membranaceus were analyzed.

RESULTFertilization promoted the seedling growth and provided abundant supply of nutrition for growth of root, yield and accumulation of active ingredient at the later growth stage, and increased the accumulation of dry matter of stem-leaf and root system. The effect of nitrogen, phosphorus and potassium application on the total dry matter accumulation of A. membranaceus was as following: nitrogen > potassium > phosphorus; the effect on the stem-leaf dry matter accumulation was as following: nitrogen > phosphorus > potassium; the effect on the root dry matter accumulation was as following: nitrogen > potassium > phosphorus. Nitrogen, phosphorus and potassium fertilizer increased the root yield of A. membranaceus. Obviously, the effect on the root yield was as following: nitrogen > potassium > phosphorus. The application of different proportional with nitrogen, phosphorus and potassium increased the content of polysaccharide and astragaloside, but had no distinct effect on the content of total flavonoids. The effect on the content of polysaccharide was as following: potassium > phosphorus > nitrogen, but the effect on the conten,t of astragaloside was as following: nitrogen > potassium > phosphorus.

CONCLUSIONNitrogen and potassium fertilizer application had more important effect on growth, yield and the contents of polysaccharide and astragaloside in A. membranaceus. During medicinal plants cultivation process, it should pay attention to the application of nitrogen fertilizer and potassium fertilizer and make balance application of nitrogen, phosphorus and potassium fertilizer.

Astragalus membranaceus ; drug effects ; growth & development ; metabolism ; Fertilizers ; Nitrogen ; pharmacology ; Phosphorus ; pharmacology ; Potassium ; pharmacology

Besides UCP1-dependent thermogenesis pathways, UCP1-independent thermogenesis pathways also could increase heat production in adipose tissue to combat obesity. N-Acyl amino acids (NAAs) have been suggested as novel endogenous uncouplers to induce mitochondria UCP1-independent thermogenesis in adipose tissue. Here, we use mouse skeletal muscle C2C12 cells which lack of UCP1 as UCP1 negative cell models. Comparing with its corresponding common fatty acid—oleate, one of the NAAs—N-Oleoylglycine (NOGly), which is highly expressed in the plasma of HFD mice, is selected to study their effects and mechanisms on mitochondrial thermogenesis. We found that 60 μmol / L oleate could induce mitochondrial oxidative phosphorylation protein levels, as well as increase mitochondria thermogenesis-related genes (COX8b, DIO2, UCP3) expression (P < 0. 05) . However, 60 μmol / L NOGly damaged the production and oxidative phosphorylation of mitochondria, significantly down-regulated expression of thermogenic genes (PGC1a, COX8b, COX2, DIO2, UQCRFS1and UCP3) (P< 0. 01), induced the production of reactive oxygen species (ROS) in the mitochondria, and enhanced the oxidative stress in cells. Our study found that oleate can induce UCP1-independent thermogenesis under 60 μmol / L addition dose, whereas NOGly does not due to the induction of oxidative stress in cells.

Transcription factor is a key trans-acting factor to mediate stress response by regulating gene expression. Plants have developed a series of mechanisms to modulate development, stress response, signaling and disease resistance at transcription level. DNA binding with one finger (DOF), containing one C₂-C₂ zinc finger domain, is a special plant transcription factor. Specifically, the conserved domain at N-terminus of DOF has multiple functions, including interacting with DNA and protein, which could be involved in plant development and stress response. Although many DOF family genes are characterized in plant stress response, it is not clear if DOF genes have functions in cereal plants. In the present paper, the role of DOF family genes on cereal plants were discussed based on a comprehensive phylogenetic relationship analysis, expression profiles in different tissues and various environmental conditions. The results obtained here will provide an important reference for further understanding the mechanism of gramineous crops in stress resistance.
DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Plant ; Phylogeny ; Plant Proteins ; metabolism ; Plants ; genetics ; Transcription Factors ; metabolism ; Zinc Fingers

We aimed to obtain the recombinant aminopeptidase encoded by Listeria monocytogenes (L. monocytogenes) gene lmo1711, and characterized the enzyme. First, the amino acid sequences of Lmo1711 from L. monocytogenes EGD-e and its homologues in other microbial species were aligned and the putative active sites were analyzed. The putative model of Lmo1711 was constructed through the SWISS-MODEL Workspace. Then, the plasmid pET30a-Lmo1711 was constructed and transformed into E. coli for expression of the recombinant Lmo1711. The his-tagged soluble protein was purified using the nickel-chelated affinity column chromatography. With the amino acid-p-nitroaniline as the substrate, Lmo1711 hydrolyzed the substrate to free p-nitroaniline monomers, whose absorbance measured at 405 nm reflected the aminopeptidase activity. The specificity of Lmo1711 to substrates was then examined by changing various substrates, and the effect of metal ions on the catalytic efficiency of this enzyme was further determined. Based on the bioinformatics data, Lmo1711 is a member of the M29 family aminopeptidases, containing a highly conserved catalytic motif (Glu-Glu-His-Tyr-His-Asp) with typical structure arrangements of the peptidase family. The recombinant Lmo1711 with a size of about 49.3 kDa exhibited aminopeptidase activity and had a selectivity to the substrates, with the highest degree of affinity for leucine-p-nitroaniline. Interestingly, the enzymatic activity of Lmo1711 can be activated by Cd²⁺, Zn²⁺, and is strongly stimulated by Co²⁺. We here, for the first time demonstrate that L. monocytogenes lmo1711 encodes a cobalt-activated aminopeptidase of M29 family.

INTRODUCTIONThis study aims to assess the early tumour outcome and morbidity associated with radiation therapy (RT) in tumours of the central nervous system (CNS).

MATERIALS AND METHODSPatients receiving RT with radical intent were entered on a prospective database. Tumour types were categorised into glioma, base of skull, pituitary, germ cell or primitive neuroectodermal tumour (PNET) and other malignant CNS tumours. Study endpoints were overall survival and progression free survival. Acute and late toxicity endpoints included Common Terminology Criteria version 3.0 (CTC) grade 3 or 4 events, need for admission during RT and change in performance status at 12 months.

RESULTSOne hundred and fifty-two patients with CNS tumours were managed with radical intent over the 4-year period. The median age was 49 years and 68.4% were Eastern Co-operative Group (ECOG) 0-1 performance status. The major pathology groups were glioma (59.9%) and base of skull tumours (17.1%). Gross total resection was performed in 28.3% only and RT was delayed after diagnosis until time of progression in 19.7%. For the 91 patients with glioma, the median survival and 2-year survival rate was 19.1 months and 44.1%, respectively. The 2-year survival rates for the subgroups of WHO Grade I or II, III and IV were 100%, 52% and 35%, respectively. For the non-glioma tumour groups, the relapse varied with pathology. Toxicity was minimal with only 3 acute and 3 late CTC grade 3 or 4 events occurring. Overall, 47 or 31% of patients required some inpatient hospitalisation during RT, although this was determined to have some causative relationship to RT in only 12 or 8% of patients. In the 12 months post-RT, performance status was stable or improved in 76.2% of patients, and most deterioration was associated with tumour relapse.

CONCLUSIONSRT for CNS tumours using modern techniques was well-tolerated with good tumour outcome and minimal morbidity.

Adolescent ; Adult ; Aged ; Central Nervous System ; physiopathology ; Central Nervous System Neoplasms ; radiotherapy ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Radiotherapy ; methods ; standards ; Singapore ; Survival Analysis

OBJECTIVETo study Astragalus membranaceus absorption characteristic of nitrogen, phosphorus and potassium at different growth stages.

METHODThrough the field experiment and the sampling analysis, the absorbing capacity of nitrogen, phosphorus and potassium as well as the growth of plant at different growth stages in A. membranaceus were analyzed.

RESULTThe dry matter accumulation reached 88.22% of the total accumulation in 100-163 days after seedling emergence, The content of N, P, K in the stem was higher than that in the root system. Nitrogen, phosphorus and potassium absorbing capacity in the stem, the leaf and the root systems was N > K > P. In the whole growth period, nitrogen accumulation reached the highest, and followed with the accumulation of potassium, and the accumulation of phosphorus was the lowest. In the last phase of the exuberant growth period, the accumulation intensity of nitrogen, phosphorus and potassium reduced. During the harvesting time, the accumulation intensity of nitrogen and potassium increased, and the accumulation intensity of phosphorus remained stable.

CONCLUSIONThe dry matter accumulation reached the maximum in 100-163 days after seedling emergence. The absorption strength of nitrogen, phosphorus and potassium in aerial part reached the maximum in 100-132 days after seeding emergence, the maximum absorption comes earlier than that of phosphorus and potassium, at that time the needs of nutrients reach the highest. For producing of 100 kg A. membranaceus 2.32 kg N, 0.323 kg P2O5, 1.62 kg K2O are needed to be absorbed from soil and fertilizer.

Absorption ; Astragalus membranaceus ; growth & development ; metabolism ; Ecosystem ; Fertilizers ; analysis ; Nitrogen ; metabolism ; Phosphorus ; metabolism ; Plant Leaves ; growth & development ; metabolism ; Plant Roots ; growth & development ; metabolism ; Plant Stems ; growth & development ; metabolism ; Plants, Medicinal ; growth & development ; metabolism ; Potassium ; metabolism ; Soil ; analysis ; Time Factors

In order to provide rationale for selection of good germplasm in Rubus chingii, main effective medicinal ingredients of green fruit such as gallic acid, ellagic acid, kaempferol-3-rutinoside, astragalin and tiliroside were measured using UPLC for the samples collected from Chun'an county of Zhejiang province, and such parameters as soluble solid contents of ripe fruit of some samples were also measured to study variation among individuals and correlation. It has been found that there were differences among individuals in the contents of gallic acid, ellagic acid, kaempferol-3-rutinoside, astragalin and tiliroside, which ranged from 0.010 2%-0.027 4%, 0.089 5%-0.291 1%, 0.010 5%-0.114 8%, 0.005 8%-0.041 2% and 0.010 9%-0.086 3%, respectively, with a CV of 18.60%, 27.02%, 44.23%, 44.17% and 47.29%, respectively. Gallic acid was positively correlated with ellagic acid, but negatively with kaempferol-3-rutinoside and astragalin significantly. Significantly positive correlation existed between kaempferol-3-rutinoside, astragalin and linden glycoside as well as between ellagic acid and fruit shape index of ripe fruit and between linden glycoside and the content of soluble solids. 51.35% of the individuals had a content of soluble solids more than 15%. Therefore, abundant variations have been found among individuals in effective medicinal ingredients in R. chingii, which shows great potential for selection, but only do 7.61% of the individuals meet the requirement of Chinese pharmacopoeia in terms of the contents of effective medicinal ingredients. Therefore, selection could be first performed in terms of fruit shape index of ripe red fruit, followed by the contents of ellagic acid and kaempferol-3-rutinoside measured. The individuals, in which the contents of effective medicinal ingredients don't meet the requirement of Chinese pharmacopoeia, could be considered for the selection in terms of edible fresh fruit.
Ellagic Acid ; Fruit ; Glycosides ; Humans ; Plant Extracts ; Rubus

Objective: To optimize the warning threshold values of common communicable diseases in Gansu province, and improve the early warning effect. Method: An early warning model was set up for influenza, scarlet fever, other infectious diarrheal diseases, dysentery, typhoid and paratyphoid, viral hepatitis type E and hand foot and mouth disease (HFMD) respectively in Gansu by using the moving percentile method and cumulative sum method. By calculating the sensitivity, specificity, predictive value of positive test, predictive value of negative test, Youden' index and receiver-operating characteristic curve, the optimum early warning threshold values for communicable diseases in Gansu were selected. Results: The optimum early warning boundary values of influenza, scarlet fever, other infectious diarrheal diseases, dysentery, typhoid and paratyphoid, and viral hepatitis type E were P(90), P(80), P(95), P(90), P(80) and P(90) respectively. The optimum early warning parameters of HFMD were k=1.2, H=5σ. Under the optimum early warning boundary values/parameters, the early warning sensitivities of influenza, scarlet fever, other infectious diarrheal diseases, dysentery, typhoid and paratyphoid, viral hepatitis type E and HFMD were 86.67%, 100.00%, 91.67%, 100.00%, 100.00%, 100.00% and 100.00%, the specificities were 86.49%, 62.22%, 75.00%, 100.00%, 97.92%, 89.13% and 74.47%. The predictive values of positive test were 72.22%, 29.17%, 52.38%, 100.00%, 80.00%, 54.55% and 29.41%, and the predictive values of negative test were 94.12%, 100.00%, 96.77%, 100.00%, 100.00%, 100.00% and 100.00%, and the Youden' indexes were 0.73, 0.62, 0.67, 1.00, 0.98,0.89 and 0.74. Receiver-operating characteristic curve showed that the values/parameters of this warning boundary were the points closest to the upper left of the coordinate diagram. Conclusion: The early warning thresholds of influenza, other infectious diarrheal diseases, dysentery and hepatitis E in Gansu may be raised appropriately and the early warning parameters of HFMD need to be adjusted to improve the effectiveness of early warning.
China ; Communicable Disease Control/methods* ; Communicable Diseases/epidemiology* ; Disease Notification ; Disease Outbreaks/prevention & control* ; Humans ; Models, Theoretical ; Population Surveillance/methods*

To establish a prokaryotic expression and purification protocol for nuclease P1 (NP1), we first obtained a synthetic NP1 by splicing 22 oligonucleotides with overlapping PCR. We constructed and transformed a secretory expression vector pMAL-p4X-NP1 into Escherichia coli host strains T7 Express and Origami B (DE3) separately. Then, the recombinant NP1 was purified by amylose affinity chromatography, and its activity, thermo-stability and metal-ion dependence were investigated systematically. The results indicated that the expressed fusion proteins MBP-NP1 (Maltose binding protein-NP1) existed mainly in soluble form both in host strains T7 Express and Origami B (DE3), but the specific activity of recombinant protein from Origami B(DE3) strain was higher than T7 Express strain (75.48 U/mg : 51.50 U/mg). When the MBP-tag was cleaved by protease Factor Xa, the specific activity both increased up to 258.1 U/mg and 139.2 U/mg. The thermal inactivation experiments demonstrated that the recombinant NP1 was quite stable, and it retained more than 90% of original activity after incubated for 30 min at 80 degrees C. Zn2+ (2.0 mmol/L) could increase enzyme activity (to 119.1%), on the contrary, the enzyme activity was reduced by 2.0 mmol/L Cu2+ (to 63.12%). This research realized the functional expression of NP1 in the prokaryotic system for the first time, and provided an alternative pathway for NP1 preparation.
Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; biosynthesis ; genetics ; metabolism ; Genes, Synthetic ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; Single-Strand Specific DNA and RNA Endonucleases ; biosynthesis ; genetics ; metabolism