Effects of Used Battery on Key Enzyme Activity during the Germination of Wheat Seeds

[Objective] This study aimed to explore the effects of used battery lixivium on wheat germination. [Method] The wheat seeds were treated with used battery lix- ivium at different concentrations to detect the change of activities of amylase, pro- tease, pyruvate dehydrogenase （PDH） and polyphenol oxidase （PPO） during the ger- mination period. [Result] The results showed that the used battery affected enzyme activity. With the increase of concentration of used battery lixivium, trends of the changes of amylase and protease activities were not different. The activities were en- hanced at low concentrations of lixivium, while were inhibited at high concentrations. The tends of changes of pyruvate dehydrogenase （PDH） and polyphenol oxidase （PPO） activities were not consistent with that of either amylase or protease, which showed continuous downward trends with the increasing concentration of used battery lixivium. [Conclusion] This study is of great practical significance for understanding the effects of used battery lixivium on the germination of wheat seeds.

Effects of Different Vegetable Planting Modes on Soil Microbial Flora and Enzyme Activity

To investigate the effects of different vegetable growing regions and planting modes on soil quality,soils in high,medium and low altitude areas of Guizhou were respectively sampled under different vegetable efficient planting modes,and the variations of soil microbial flora and enzyme activities were analyzed. The soil microbial count and total bacteria of the vegetable efficient cultivation mode were significantly higher than that of the control （traditional planting mode） in each planting area,and the microbial diversity index was also improved to varying de- grees.The soil phosphatase,catalase and urease activities of the vegetable efficient planting mode were higher than that of the control.The soil catalase and urease activities were higher than that of the control by 1.37-1.44 and 1.51-2.80 times. Application of vegetable efficient planting mode in different regions will help to im- prove the soil quality in a given period.

Detection and Quantitative Analysis of Dominant Bacteria On Aging Flue-cured Tobacco Leaves

[Objective] The aim was to construct the internal motivation of flued-cured tobacco natural aging. [Method] The variation of DNA copies of Bacillus megaterium, amylase activity, PPO, peroxydase and content of starch, neutro-protein, polyphenol in different aging period were investigated using Henan flue-cured tobacco leaves. [Result] Results showed that the DNA copies of Bacillus megaterium increased initially but decreased subsequently with the aging time. The DNA copies of Bacillus per gram of tobacco leaf surface reached the maximum number at 6 months aging. The activities of four enzymes showed the same variation trend as in DNA copies, and also reached the highest level at 6 months aging. Chemical ingredients, however, displayed a downward trend. The variation of enzyme activity and content of chemical composition were reflected in the changes of DNA copies in Bacillus megaterium. [Conclusion] The result indicate that Bacillus megaterium was the internal motivation of flue-cured tobacco natural aging.

Inhibitory effect of Cr(Ⅵ) on activities of soil enzymes

To evaluate the influence of various Cr(Ⅵ) concentrations (0.05, 0.25, 0.50, 1.00 and 2.00 g/kg) on the activity of soil enzymes, the activities of catalase, polyphenol oxidase, dehydrogenase, alkaline phosphatase in soils were investigated in the incubation experiment with a period of 35 d. The results indicate that all the tested Cr(Ⅵ) concentrations significantly inhibit dehydrogenase activity by over 70% after 35 d. The activity of alkaline phosphatase is slightly inhibited during the whole experiment except for on the day 7. Cr(Ⅵ) has no obvious effect on the activity of catalase in soil. On the contrary, Cr(Ⅵ) stimulates the activity of polyphenol oxidase. The results suggest that dehydrogenase activity can be used as an indicator for assessing the severity of chromium pollution.

Polyphenol Oxidase Activity and Inhibition in White Yam （Dioscorea Rotundata. Var. Laasirin） Chips as African Fries for Human Consumption

Polyphenol oxidase, a bi-functional enzyme, has been implicated in enzymatic browning of yam and other tubers in a negative way. The objective of this present study was to examine the activity of polyphenol oxidase in Dioscorea rotundata. Var. laasirin and the efficiency of heat and chemical treatments in inhibiting this enzyme. Crude Polyphenol Oxidase （PPO） of Dioscorea rotundata.Var. Laasirin was isolated and the kinetics studied using the lineweaver-burk plot. The activity of the enzyme was evaluated using spectrophotomeric method. Yam PPO catalyzes oxidation of various substrates with catechol being the most readily oxidized substrate. The Michaelis-Menten constant （Km） and maximum reaction velocity （Vmax） for yam PPO were 0.00037 and 0.3125 respectively. Inhibition data showed that the enzyme had least activity （71.70） when blanched at 95℃ for 7 mins with chemical treatment involving a combination of 0.5% Sodium metabisulphite （Food grade） and 0.5% Ascorbic acid （Food grade）. The activity was highest （83.02） when it was blanched at 95 ℃ for 7 rains. This study has shown that it is possible to inhibit polyphenol oxidase activity in white yam using the chemical pretreatments and processing conditions described in this study for possible adoption in the production of packaged frozen yam chips by food industries.

Screen and effect analysis of immunostimulants for sea cucumber,Apostichopus japonicus

Immunostimulants may improve disease resistance of aquaculture animals by promoting the nonspecific immunity response of the organisms. Five types of saccharides, including chitosan, yeast polysaccharide, burdock oligosaccharide, seaweed polysaccharide and lentinus edodes polysaccharide, were screened for potential use as immunostimulants by using spectrophotometry. The saccharides were injected into Apostichopusjaponicus, a sea cucumber, and the lysozyme and superoxide dismutase （SOD） activities of the coelomic fluid and epidermal slime were monitored in six consecutive days. The results show that the lysozyme activity of the animal＇s coelomic fluid was significantly stimulated on day 2, day 4 and day 6 after the injection of the saccharides （P〈0.05）. The effects of chitosan and yeast polysaccharide were the most notable. The lysozyme activity of the epidermal slime was significantly increased by chitosana, yeast polysaccharide, seaweed polysacchafide, and burdock oligosaccharide on day 1 and day 2 after the injection （P〈0.05）. The SOD activity of the coelomic fluid was significantly promoted by the saccharides on day 2 and day 4 post-injection （P〈0.05）, while the SOD activity of the epidermal slime increased on day 2. These findings indicate that chitosan and yeast polysaccharide are the most effective immunostimulants and potential healthy anti-disease feedstuff for A. japonicus.

Effect of Dopamine Injection on the Hemocyte Count and Prophenoloxidase System of the White Shrimp Litopenaeus vannamei

Effects of dopamine injection on the hemocyte count, phenoloxidase activity, serine proteinase activity, proteinase in-hibitor activity and α2-macroglobulin-like activity in L. vannamei were studied. Results showed that dopamine injection resulted in a significant effect on the parameters measured (P < 0.05), while no significant difference was observed in the control group (0.85% NaCl). In the experimental groups,the hemocyte count reached the minimum in 3 h;granular and semi-granular cells became stable after 12 h and hyaline cells and the total hemocyte count became stable after 18 h. Phenoloxidase activity reached the minimum in 6 h, and then became stable after 9 h. Serine protease activity and proteinase inhibitor activity reached the minimum in 3 h, and α2-macro-globulin-like activity reached the maximum in 3 h,and all the three parameters became stable after 12 h. The results suggest that the activating mechanisms of the proPO system triggered by dopamine are different from those triggered by invasive agents or sponta-neously activated under a normal physical condition.

Physical, Chemical and Biochemical Changes of Sweetsop （Annona squamosa L.） and Golden Apple （Spondias citherea Sonner） Fruits during Ripening

This study aimed at the physical, chemical and biochemical changes during ripening of Sweetsop （Annona squamosa L.） and Golden Apple （Spondias citherea Sonner） fruits during ripening as important features to better understand their postharvest handling. It was carried out physical analysis such as firmness and chemical analysis such as total chlorophyll, total carotenoids, soluble solids, pectins and titrable acidity and biochemical analysis such as pectin methyl esterase, polygalacturonase, cellulase, and peroxidase and polyphenoloxidase activities in crude extract. Fruits were harvested at different stages of ripening. Experimental design was completely randomized and was carried out analysis of variance and Tukey tests, Total chlorophyll was decreasing in later stages of ripening, total soluble solid contents increased as the fruits ripen, while the acidity expressed percentage of citric acid decreased during fruits ripening. The loss of firmness and soluble solids content increased as the fruit get ripped stage, while the content of pectin decreased. Activity was observed for pectin methyl esterase and polygalacturonase enzymes during all stages of maturation, presenting the highest activity for both enzymes in the mature state. No cellulase activity detected at any stage during the ripening of these fruits. Activity of the enzyme polyphenoloxidase and peroxidase, associated with pulp browning was higher in the last stages of ripening of these fruits. Physical, chemical and biochemical patterns during ripening were different according to fruit species suggesting differential postharvest handling requirements.

Reactive protein, plasminogen activator inhibitor type-1 （PAI-1） levels, PAI-1 promoter 4G/5G polymorphism and acute myocardial infarction

Objective To investigate the relationship between CRP, plasminogen activator inhibitor type 1 （PAI-1） levels, PAI-1 gene promoter 4G/5G polymorphism and the type of acute myocardial infarction （ST elevation myocardial infarction, STEMI vs the non-ST elevation Myocardial infarction, NSTEMI）. Methods One hundred seventy-six consecutive patients with AMI were included for the study, of whom 60 had STEMI and 56 had NSTEMI, and 60 adults without cardiovascular and cerebrovascular disease were selected as controls. Blood samples were obtained from patients within 6 h of AMI and the plasma PAI-1, CRP, and the gene polymorphism were measured. Results Plasma levels of PAI- 1 and CRP were higher in AMI groups, compared those in the control group, and plasma levels of PAI-1 were significantly higher in patients with STEMI compared to those with NSTEMI （80.12ng/ml VS.73.01ng/ml, P 〈0.01）, while CRP levels were not significantly different between patient with STEMI and NSTEMI （3.87 ± 0.79 mg/ml VS.4.01 ±0.69mg/ml, P〉0.05）. PAI-1 levels presented a significant correlation with CRP levels in the NSTEMI subjects. However, PAI-1 and CRP levels could explain the lack of a significant relationship between them in control and STEMI subjects.The frequencies of 4G/4G genotype in the AMI group were higher than those in the control group and higher in patient with STEMI than in patient with NSTEMI. Plasma levels of PAI-1 in subjects with 4G/4G genotype were significantly increased as compared to those in subjects with 4G/5G and 5G/5G genotype. Conclusions Plasma PAI-1 levels were associated with different myocardial infarction type, and PAI-1 promoter 4G/5G polymorphisms and CRP may be related to plasma PAI-1 levels

Bone Marrow Urokinase Plasminogen Activator Receptor Levels are Associated with the Progress of Multiple Myeloma

Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors(u PAR) in bone marrow fluid and bone marrow tissue from multiple myeloma(MM) patients and assess association of u PAR level with prognosis of MM.Methods u PAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow(control) were examined by ELISA.Furthermore,u PAR expression in bone marrow tissue was investigated by RT-PCR and Western blot,respectively.The distribution of u PAR in MM cells was examined using immunofluorescence staining.The pathological changes in different stages of MM patients were studied by HE staining.Results u PAR level in bone marrow fluid of MM patients(1.52±0.32 μg/ml) was found to be higher than that in the control group(0.98±0.15 μg/ml).Interestingly,u PAR protein(0.686±0.075 vs.0.372±0.043,P<0.05) and m RNA(2.51±0.46 vs.4.46±1.15,P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage.The expression of u PAR in MM bone marrow was confirmed by immunofluorescence staining.Moreover,HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma.Conclusion Our study reveals that u PAR expression is positively correlated with the development and progress of MM.

Cause and control of Radix Ophiopogonis browning during storage

In the storage of Radix Ophiopogonis, browning often happens to cause potential risk with regard to safety. Previously few reports investigate the browning of Radix Ophiopogonis. In this research, the causes and mechanisms of the browning of Radix Ophiopogonis were preliminarily elucidated. Content determination by high-performance liquid chromatogra- phy （HPLC） and spectrophotometry, enzyme activity determination by colorimetry, and morphological observation by electron mi- croscopy were performed in the present study. Uniform design and three-dimensional response surfaces were applied to investigate the relationship between browning and storage factors. The cortex cell wall of browned Radix Ophiopogonis was ruptured. Compared with the normal Radix Ophiopogonis, cellulase and polyphenol oxidase enzymes were activated, the levels of 5-hydroxymethylfurfural （5-HMF）, total sugars, and reducing sugars were increased, while the levels of polysaccharides and methylophiopogonanone A were de- creased in browned Radix Ophiopogonis. The relationship between the storage factors and degree of browning （Y） could be described by following correlation equation： Y= - 0.625 4 ＋ 0.020 84 × X3 ＋ 0.001 514×X1×x X2 - 0.000 964 4 × X2 × X3. Accompanied with browning under storage conditions, the chemical composition of Radix Ophiopogonis was altered. Following the activation of cellulase, the rupture of the cortex cell wall and the outflow of cell substances flowed out, which caused the Radix Ophiopogonis tissue to become soft and sticky. The main causes of the browning were the production of 5-HMF, the activation of polyphenol oxidase, Maillard reactions and enzymatic browning. Browning could be effectively prevented when the air relative humidity （HR）, temperature, and moisture content were under 25% RH, 12 ℃ and 18%, respectively.

Effects of Earthworms and Ryegrass on the Removal of Fluoranthene from Soil

Earthworms can promote the bioremediation of contaminated soils through enhancing plant growth and microorganism development. The individual and combined effects of earthworms and ryegrass （Lolium multifloram Lain.） on the removM of fiuoranthene from a sandy-loam alluvial soil were investigated in a 70-d microcosm experiment. The experiment was set up in a complete factorial design with treatments in four replicates： without earthworms or ryegrass （control, CK）, with earthworms only （E）, with ryegrass only （P）, and with both earthworms and ryegrass （EP）. The residual fluoranthene, microbial biomass C, and polyphenol oxidase activity in the soil changed significantly （P 〈 0.01） with time. In general, the residual concentration of fluoranthene in the soil decreased sharply from 71.8-88.7 to 31.7-37.4 mg kg-1 in 14 d, and then decreased gradually to 19.7-30.5 mg kg-1 on the 70th d. The flu- oranthene concentration left in the soil was the least with both earthworms and ryegrass, compared to the other treatments at the end of the experiment. Half-life times of fluoranthene in the E, P, and EP treatments were 17.8%-36.3% smaller than that of CK. More fluoranthene was absorbed by earthworms than ryegrass. However, the total amounts of fluoranthene accumulated in both the ryegrass and earthworms were small, only accounting for 0.01%-1.20% of the lost fluoranthene. Therefore, we assumed that microbial degradation would play a dominant functional role in fluoranthene removal from soil. We found that earthworms significantly increased microbial biomass C and polyphenol oxidase activity （P 〈 0.01） in the presence of ryegrass at the end of the experiment. Furthermore, microbial biomass C and polyphenol oxidase activity were significantly （P 〈 0.05） and negatively related to the residual fluoranthene concentration. This implied that earthworms might promote the removal of fluoranthene from soil via stimulating microbial biomass C and polyphenol oxidase activity.

Effects of Maize Residue Quality and Soil Water Content on Soil Labile Organic Carbon Fractions and Microbial Properties

Investigating the effects of residue chemical composition on soil labile organic carbon （LOC） will improve our understanding of soil carbon sequestration. The effects of maize residue chemical composition and soil water content on soil LOC fractions and microbial properties were investigated in a laboratory incubation experiment. Maize shoot and root residues were incorporated into soil at 40% and 70% field capacity. The soils were incubated at 20 ℃ for 150 d and destructive sampling was conducted after 15, 75, and 150 d. Respiration, dissolved organic carbon （DOC）, hot-water extractable organic carbon （HEOC）, and microbial biomass carbon （MBC） were recorded, along with cellulase and β-glucosidase activities and community-level physiological profiles. The results showed that the cumulative respiration was lower in root-amended soils than in shoot-amended soils, indicating that root amendment may be beneficial to C retention in soil. No significant differences in the contents of DOG, HEOC and MBC, enzyme activities, and microbial functional diversity were observed between shoot- and root-amended soils. The high soil water content treatment significantly increased the cumulative respiration, DOC and HEOC contents, and enzyme activities compared to the low soil water content treatment. However, the soil water content treatments had little influence on the MBC content and microbial functional diversity. There were significantly positive correlations between LOC fractions and soil microbial properties. These results indicated that the chemical composition of maize residues had little influence on the DOC, HEOC, and MBC contents, enzyme activities, and microbial functional diversity, while soil water content could significantly influence DOC and HEOC contents and enzyme activities.

Construction of self-assembled nanogel as mulitenzyme mimics for bioresponsive tandem-catalysis imaging

The self-assembled phospholipid-or cytosolassociated multienzyme complexes constitute necessary components of the foundation of life.As a proof of concept,metalcoordinated supramolecular nanogels (MCSGs) have been designed,with the self-assembly of di-lysine coordinated iron(Fe(Lys)_(2))-functionalized peptide gelators on the interface by an in situ amidation-induced protonation process.The monoatomic and highly dispersed active centers of Fe(Lys)_(2) offered the nanogel mimics with excellent reaction rates due to the high density and nano compartmental structure similar to the natural matrix-associated multienzyme complex.SiO_(2)@MCSGs show both superoxide dismutase (SOD) activity and peroxidase (POD) activity,and the higher activities compared with the activity of free Fe(Lys);molecules can be detected.After loading the substrate 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate)(ABTS),SiO_(2)@MCSGs;can responsively convert O^(-)_(2) in the tumor microenvironment into H_(2)O_(2) intermediates and then tandem catalyze the oxidization of ABTS for contrast photoacoustic (PA) imaging of tumor by the SOD-POD mimic activity,showing their great potential as the efficient enzymatic agents for pathological theranostics.