Adult ; Aged ; Aged, 80 and over ; Coal Mining ; Humans ; Lung Neoplasms ; complications ; microbiology ; Male ; Methicillin Resistance ; Microbial Sensitivity Tests ; Middle Aged ; Pneumoconiosis ; complications ; microbiology ; Staphylococcus aureus ; drug effects ; isolation & purification

The potential of a Sinorhizobium fredii strain producing a copolymer polyhydroxyalkanoate(PHA)from glucose and sodium decanoate substrates was studied in this paper.Using orthogonal design in a flask-shaker culture system,the culture medium,some culture conditions and vital regulation conditions for polymer synthesis were optimized.These optimized results were applied into further studies in two-stage fed-batch with a 10L fermentor.The whole culture process consisted of two stages,that is,the cell growth and the copolymer production.The first stage was for the cell growing to a desired biomass and the second was for the copolymer synthesis.For producing PHA polymers,the selected 8 mM sodium decanoate was added into the broth by adopting a two-step adding method for avoiding of foaming when the biomass had approached 28.5g/L dry cell.The maximum P(HB-HH)production could be 17.55 g/L with a monomer ratio of 79.4%(W/W)3-HB and 20.6%(W/W)3-HH.The molecule constitute of the copolymer is poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)[P(HB-HH)] and its molecular weight(MW)is 1.4?105D.The results demonstrated that the employed S.fredii strain could be a potential candidate for industrial production of the copolymer.The fermentation parameters acquired in the experimental system offered some valuable references for studying large-scale production of the copolymer.

Four screening methods, colorimetric assay, blood-plate hemolysis method, surface tension activ- ity and oil spreading technique were introduced to isolate strains producing bio-demulsifiers from 6 different bacteria source samples. The results of various screening methods were evaluated in this paper. Seventeen demulsifying strains were obtained, which are qualified in demulsification test of kerosene model emulsions. Among them, 5 strains showed high demulsifying ability, achieving 70% plus demulsifying ratio within 24 hours. Petroleum-contaminated soil, excess sludge from biological process treating oilfield produced water and sludge from municipal wastewater treatment plant were the best among all tested sources. Due to the determination limit, the colorimetric assay and blood-plate hemolysis method are not competent to screen bio-demulsifiers strains. The measurement of surface tension and oil spreading method were easy but accu- rate methods to isolate bio-demulsifiers strains. Although demulsification test of model emulsion is an effec- tive technique to target strains with the capability of breaking emulsions, it is sophisticated and time-con- suming. Thus it is recommended to use surface tension and oil spreading methods in pre-screening and vali- date the results in demulsification test with kerosene model emulsions.

OBJECTIVETo investigate immune state in lung of BALB/c mice with ovalbumin (OVA) allergy and the effects of fulvotomentoside (Ful) on lungs of the mice and provide some clues for the mechanism that patients with food allergies were prone to asthma and observe the effects of the treatment with traditional Chinese medicine.

METHODNinety-six female BALB/c mice were randomly divided into 6 groups. Mice in group 1 and group 2 were sensitized intraperitoneally and challenged intragastrically with OVA and were exposed to phosphate buffer solution and OVA respectively by nebulized inhalation. Mice in group 3 and group 4 were treated with Ful, other processes were the same as the mice in group 1 and group 2, respectively. Mice in group 5 were not challenged intragastrically with OVA and other processes were the same as the mice in group 2. Group 6 was the control group. The number of total leukocytes and cell classification in bronchoalveolar lavage (BALF) were counted, and inflammatory characteristic of lung was scored by staining with hematoxylin and eosin. The protein expressions of transforming growth factor (TGF-β1), interleukin-6 (IL-6), interleukin-17 (IL-17A) in lung of the mice were detected by immunohistochemical method. The activation of neutrophils in lung was assayed by the level of myeloroxidase (MPO).

RESULTThere was no inflammatory cells infiltration in lung of the mice in group 1. Compared with group 6, numbers of total leukocytes and erythrocytes as well as the percentage of neutrophils and lymphocytes were increased in group 2. Inflammatory score and protein expressions of TGF-β1 [(75 437 ± 3 638) vs. (6 118 ± 1 978)], IL-6 [(121 650 ± 25 389) vs. (15 726 ± 9 360)], IL-17A [(252 105 ± 31 651)vs. (72 644 ± 12 285)] in lung were increased, too. Inflammatory score and TGF-β1 (11 054 ± 1 468), IL-6 (50 877 ± 11 744), IL-17A (137 864 ± 28 986) expressions in group 5 were lower than those in group 2. Eosinophils infiltration was significant in group 5. After the treatment with Ful, TGF-β1 expression did not change and IL-6, IL-17A expressions were decreased in lung of the mice that inhaled OVA. It was not enough for Ful to relieve the neutrophil aggregation and improve inflammatory reaction in lung.

CONCLUSIONThe expressions of TGF-β1, IL-6, IL-17A in lung of the mice with OVA allergy were increased markedly after they inhaled specific antigen, which caused serious inflammation that was induced by neutrophil infiltration in lung. Ful could decrease the expressions of IL-6, IL-17A to some extent, but it was not enough to improve pathologic state in lung.

Administration, Inhalation ; Animals ; Bronchoalveolar Lavage Fluid ; cytology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Food Hypersensitivity ; immunology ; metabolism ; pathology ; Immunohistochemistry ; Inflammation ; Interleukin-17 ; metabolism ; Interleukin-6 ; metabolism ; Lung Diseases ; immunology ; pathology ; prevention & control ; Mice ; Mice, Inbred BALB C ; Neutrophils ; drug effects ; immunology ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Ovalbumin ; adverse effects ; immunology ; Saponins ; pharmacology ; Transforming Growth Factor beta1 ; metabolism

Objective：This study was designed to investigate the apoptosis of HL 60 cells induced by the peptide from hemocyte of horseshoe crab (horseshoe crab hemocyte peptide). Methods： Cytotoxicity and cell viability were assayed by MTT method. Morphologic assessment of apoptosis was performed with fluorescence microscope; the apoptosis of the cells was confirmed by flow cytometry; cell cycle was analyzed by flow cytometry, changes of cell membrane were observed by scanning electron microscope. Results： HL 60 cells treated with horseshoe crab hemocyte peptide were showed apparently cytotoxicity with IC50 24 μ g/ml. The morphologic changes including reduction in volume, nuclear chromatin condensation, fluorescence strength were observed with fluorescence microscope. Treated with horseshoe crab hemocyte peptide 50- 100 μ g/ml for 6 h, apoptostic HL 60 cells were predominant. However the HL 60 cells were shown necrotic and apoptostic cells were reduced when treatment with horseshoe crab hemocyte peptide of 200 μ g/ml. SubG1 peak was detected by flow cytometry. Cell cycle analysis showed Phase G1 cells decreased and Phase G2 cells increased with horseshoe crab hemocyte peptide 50 μ g/ml treating for 0- 12 h. Ratio of apoptosis increase depended on concentrations of horseshoe crab hemocyte peptide in 25- 100 μ g/ml but it reduced in 200 μ g/ml, the HL 60 cells were shown necrotic mainly. Scanning electron microscope showed membrane of HL 60 cells was damaged by horseshoe crab hemocyte peptide treatment,cell membrane showed some holes. Conclusion： Peptide from hemocytes of horseshoe crab can induce apoptosis in HL 60 cell,it was early event,and may correlate with membrane damage. The HL 60 cell in Phase G1 seemed to be sensitive to the peptide.

Background:Glioblastoma is one of the most common malignant brain tumors.Conventional clinical treatment of glioblastoma is not sufficient,and the molecular mechanism underlying the initiation and development of this disease remains unclear.Our study aimed to explore the expression and function of miR-873a-5p in glioblastoma and related molecular mechanism.Methods:We analyzed the most dysregulated microRNAs from the Gene Expression Omnibus (GEO) database and examined the expression of miR-873-5p in 20 glioblastoma tissues compared with ten normal brain tissues collected in the Zhejiang Tongde Hospital.We then overexpressed or inhibited miR-873-5p expression in U87 glioblastoma cell lines and analyzed the phenotype using the cell counting kit-8 assay,wound healing assay,and apoptosis.In addition,we predicted upstream and downstream genes of miR-873-5p in glioblastoma using bioinformatics analysis and tested our hypothesis in U87 cells using the luciferase reporter gene assay and Western blotting assay.The differences between two groups were analyzed by Student's t test.The Kruskal-Wallis test was used for the comparison of multiple groups.A P ＜ 0.05 was considered to be significant.Results:The miR-873-5p was downregulated in glioblastoma tissues compared with that in normal brain tissues (normal vs.tumor,0.762 ± 0.231 vs.0.378 ± 0.114,t =4.540,P ＜ 0.01).Overexpression of miR-873-5p inhibited cell growth (t =6.095,P ＜ 0.01) and migration (t=3.142,P ＜ 0.01) and promoted cell apoptosis (t=4.861,P ＜ 0.01),while inhibition of miR-873-5p had the opposite effect.Mechanistically,the long non-coding RNA HOTAIRM1 was found to act as a sponge of miR-873-5p to activate ZEB2 expression in U87 cells.Conclusions:We uncovered a novel HOTAIRM1/miR-873-5p/ZEB2 axis in glioblastoma cells,providing new insight into glioblastoma progression and a theoretical basis for the treatment of glioblastoma.

Anesthesia, General ; Animals ; Central Venous Pressure ; physiology ; Heart Function Tests ; Hemodynamics ; physiology ; Respiration, Artificial ; Shock, Septic ; pathology ; Swine ; physiology ; Venae Cavae ; physiology

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.
Color ; Fabaceae ; chemistry ; enzymology ; growth & development ; metabolism ; Germination ; Peroxidases ; metabolism ; Plant Proteins ; metabolism ; Seeds ; chemistry ; enzymology ; growth & development ; metabolism ; Starch ; metabolism ; Superoxide Dismutase ; metabolism ; Time Factors

AIMTo establish RP-HPLC method for determination of plasma scutellarin concentration and study of the pharmacokinetic behavior of scutellarin in rat after ig administration of breviscapine and its beta-cyclodextrin complex (breviscapine-beta-CD).

METHODSMobile phase composed of methanol and acetate buffer. The column was Shim-pack C18. Twelve rats randomized into 2 groups were separately given breviscapine and breviscapine-beta-CD at single dose of 10.8 mg.kg(-1). Drug in plasma was extracted and determined by HPLC. The pharmacokinetic parameters were calculated by 3P97 software.

RESULTSLinearity was obtained over the range of 10-400 ng.mL(-1). The recovery was 95.32%-98.81%. C(max) and AUC(0-12 h) of breviscapine were (154 +/- 18) ng.mL(-1) and (710 +/- 126) ng.h.mL(-1). For breviscapine-beta-CD, C(max) and AUC(0-12 h) were (328 +/- 31) ng.mL(-1) and (1,093 +/- 200) ng.h.mL(-1), respectively. There were significant differences of AUC(0-12 h) between breviscapine and breviscapine-beta-CD (P < 0.01).

CONCLUSIONThe assay method was suitable for the determination of scutellarin plasma concentration in rat. Brevescapine-beta-CD showed greater absorption compared with that of brevescapine.

Animals ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Drug Compounding ; Erigeron ; chemistry ; Flavonoids ; blood ; isolation & purification ; pharmacokinetics ; Male ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; beta-Cyclodextrins ; blood ; pharmacokinetics

OBJECTIVETo investigate the effects of fulvotomentoside (Ful) on inflammatory factors and antiinflammatory factors in intestine of ovalbumin (OVA)-sensitized BALB/c mice, and to explore the mechanisms of its anti-food allergy effect.

METHODTwenty-four female BALB/c mice aged 6 weeks fed with ovalbumin-free feed were randomly divided into 3 groups, food allergy (FA) group, Ful group and normal saline (NS) group. Mice in FA and Ful groups were sensitized intraperitoneally two times with OVA and challenged intragastrically with OVA. Mice in Ful group were treated with 200 mg/kg of Ful by subcutaneous injection once daily for 22 days. The mice in FA and NS groups were used as positive control and negative control, respectively, and were treated with normal saline solution by subcutaneous injection for 22 days. Just 48 hours after the last challenge, the mice in each group were sacrificed and specimens of jejunum were taken. The mRNA expressions of transforming growth factor β1 (TGF-β1), interleukin-6 (IL-6), interleukin-17A (IL-17A) and forkhead box P3 (Foxp3) in jejunum were detected by reverse transcription-PCR (RT-PCR). The protein expressions of TGF-β1, IL-6, and IL-17A in jejunum were detected by immunohistochemical method. The activation of neutrophils in jejunum was assayed by the levels of MPO.

RESULTThe expressions of TGF-β1, IL-6, IL-17A mRNA [(0.370 ± 0.013), (0.475 ± 0.015), (0.541 ± 0.013)] and related protein [(53,075.70 ± 20,727.06), (256,881.66 ± 36,561.79), (435,064.25 ± 69,911.48)] in jejunum were increased and the Foxp3 mRNA [(0.231 ± 0.014) vs. (0.365 ± 0.015)] expression was decreased in group FA. After the treatment with Ful, IL-6 and IL-17A mRNA [(0.196 ± 0.005), (0.204 ± 0.008)] and protein [(114,040.30 ± 20,295.25), (218,200.74 ± 30,077.69)] expressions were decreased and Foxp3 mRNA (0.578 ± 0.021) expression was increased, and no change of TGF-β1 expression was unchanged. There were no significant differences of the levels of MPO among the three groups.

CONCLUSIONInflammatory reaction which was characterized by the increase of IL-6 and IL-17A expressions was found in intestine of ovalbumin-sensitized BALB/c mice. Ful could decrease overexpression of IL-6 and IL-17A, and increase the expression of specific transcription factor Foxp3 of regulatory T cells significantly in intestine. It may be one of the mechanisms that Ful improved intestinal inflammation.

Animals ; Female ; Food Hypersensitivity ; metabolism ; Forkhead Transcription Factors ; metabolism ; Inflammation ; Interleukin-17 ; metabolism ; Interleukin-6 ; metabolism ; Intestines ; drug effects ; metabolism ; Mice ; Mice, Inbred BALB C ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Ovalbumin ; adverse effects ; Saponins ; pharmacology ; Transforming Growth Factor beta1 ; metabolism