Animal Safety Test of Bacillus thuringiensis BT Protein

[Objectives]To determine the biological safety of BT protein from Bacillus thuringiensis(Bt)fermentation broth to mammals at high doses.[Methods]Healthy mice were randomly divided into 4 groups with 10 mice in each group.The experimental groups were fed with Bt fermentation supernatant at 0.2,0.6 and 1.0 mL/kg,respectively,once a day for 7 consecutive days.The blank control group was fed normally without intragastric administration.[Results]There was no significant difference in blood routine and blood biochemical analysis between the experimental group and the control group.After intragastric administration,the mice were dissected,and no obvious pathological changes were found;the heart,liver,spleen,lung and kidney were taken to make tissue sections,and no pathological changes were found by microscopic observation.[Conclusions]Mice can tolerate high doses of BT protein from B.thuringiensis fermentation broth.

Cyanidin-3-O-galactoside and Blueberry Extracts Supplementation Improves Spatial Memory and Regulates Hippocampal ERK Expression in Senescence-accelerated Mice

Objective To investigate whether the antioxidation and the regulation on the Extracellular Regulated Protein Kinases （ERK） signaling pathway are involved in the protective effects of blueberry on central nervous system. Methods 30 Senescence-accelerated mice prone 8 （SAMP8） mice were divided into three groups and treated with normal diet, blueberry extracts （200 mg/kg.bw/day） and cyaniding-3-O-galactoside （Cy-3-GAL） （50 mg/kg.bw/day） from blueberry for 8 weeks. 10 SAMR1 mice were set as control group. The capacity of spatial memory was assessed by Passive avoidance task and Morris water maze. Histological analyses on hippocampus were completed. Malondialdehyde （MDA） levels, Superoxide Dismutase （SOD） activity and the expression of ERK were detected. Results Both Cy-3-GAL and blueberry extracts were shown effective functions to relieve cellular injury, improve hippocampal neurons survival and inhibit the pyramidal cell layer damage. Cy-3-GAL and blueberry extracts also increased SOD activity and reduced MDA content in brain tissues and plasma, and increased hippocampal phosphorylated ERK （p-ERK） expression in SAMP8 mice. Further more, the passive avoidance task test showed that both the latency time and the number of errors were improved by Cy-3-GAL treatment, and the Morris Water Maze test showed significant decreases of latency were detected by Cy-3-GAL and blueberry extracts treatment on day 4. Conclusion Blueberry extracts may reverse the declines of cognitive and behavioral function in the ageing process through several pathways, including enhancing the capacity of antioxidation, altering stress signaling. Cy-3-GAL may be an important active ingredient for these biological effects.

Hydrolyzing Condition and Immunocompetence of Sheep Bone Protein Enzymatic Lysates

Utilizing collagen of sheep bone as material to get immunocompetent peptide, enzymatic hydrolysis conditions were optimized using quadratic regression general rotation design. The effect of temperature （T）, time （t）, enzyme/substrate （E/S） ratio, and substrate concentration （S） on the amount of tricarboxylix acid cycle （TCA） soluble peptides were investigated. The content of soluble peptide in the acquisition was measured by Folin-hydrozybebzebe method, and the correlation between soluble peptide content and immunocompetence was analyzed by SAS software. The best enzymatic hydrolysis condition was gotten from Design Expert 7.1.2 software. The optimal condition under which immunocompetent peptides could be prepared was 1 576 U g^-1 （E/S）, 64.05℃ （T）, 0.271 kg L^-1 （S）, and 7.22 h （t）. The correlation coefficient between TCA soluble peptides and the immuneocompetence was 0.045 〈r0.05 = 0.355, which indicated that they had no significant correlation. The result showed that the soluble peptide contained immunocompetent peptides which content was independent of immunocompetence in the hydrolasates.

Study on Anti-infection Effect of Polysaccharide from Agaricus blazei Murrill on Mice

In order to study the anti-infection effect of phoshporylated Agaricus blazei polysaccharide on mice, mice were drenched phoshporylated A. blazei polysaccharide for 14 d, and an A. blazei polysaccharide control group and a blank control group were also set. The mice in all the three groups were infected with Escherichia coli on the 15 th day and the 29 th day of the experiment, the body weight of the mice in each group was recorded, as well as the spleen index on the 29 th day of infection and the 36 th day of the experiment, and the anti- E. coli titer in serum was also detected. The results showed that phoshporylated A. blazei polysaccharide could promote and maintain the development of spleen, and improve the immune response of organisms, thereby resisting bacterial infection. Furthermore, phoshporylated A. blazei polysaccharide has better immunoregulation and anti-infection effects than A. blazei polysaccharide, and could play an important role in veterinary clinical treatment and prevention and control of diseases as a immunologic adjuvant or immunopotentiator.

Genome Polymorphisms Between Indica and Japonica Revealed by RFLP

Revealing the genome polymorphisms between indica and japonica subspecies; RFLP markers, which are located across 12 chromosomes of rice, were used to analyze indica-japonica differentiation in different rice varieties. At the same time, genome sequence variations of screened loci were analyzed by bioinformatics method. Twenty-eight RFLP probes, which can classify indica-japonica rice, were confirmed. Subspecies genome polymorphisms of screened loci were found by analyzing the publication of the genome sequences data of rice. The study indicated that these screened markers can be used for classifying indica-japonica subspecies. With the publication of the genome sequences of rice, marker polymorphisms between indica and japonica subspecies can be revealed by genome differentiation.

Effect of Calcination Temperature on Structure and O_2 Desorption Properties of Ce_xPr_(1-x)O_(2-δ) Mixed Oxides

A series of CexPr1-xO2-δ (x=0, 0.5, 0.9, 1.0) mixed oxide calcined at different temperatures were synthesized by sol-gel method and characterized by Raman, XRD and O2-TPD techniques. When x=0.9, only a cubic phase CeO2 is observed. When x=0.5, the compound was combined by Pr6O11 and CeO2 mixed oxides. For CexPr1-xO2-δ (x=0.5, 0.9)samples 465 cm-1 Raman peak is attributed to the Raman active F2g mode of CeO2. The broad peak at about 570 cm-1 can be linked to lattice defects resulting in oxygen vacancies. The crystallite size of the samples increased as increasing the calcined temperature. But the increased value of Ce0.9Pr0.1O2-δ and Ce0.5Pr0.5O2-δ is smaller than single CeO2 and Pr6O11 obviously. It reveals that the insertion of Pr atom into the ceria lattice could enhance the sintering resistance and thermal stability of the mixed oxides. Calcination temperatures had great effect on the peak intensity for CeO2 but less effect on Ce0.8Pr0.2O2-δ in Raman spectra, and it may be caused by the colors transformation of the mixed oxides. The result of O2-TPD experiment indicates that the formation of solid solution has elevation the stabilization and thermal stability of the mixed oxides.

Structure, Antibacterial Activity and Theoretical Study of the Forming Mechanism of Compound [(5-Methyl-3-oxo-2-phenyl-1H-pyrazol-4-yl)-thiophen-2-yl-methylene] Thiosemicarbazide

A new thiosemicarbazone compound derived from 1-phenyl-3-methyl-4-（2-thenoyl） pyrazolone-5 （HPMTP） and thiosemicarbazide has been synthesized and characterized by IR, H- NMR, MS, elemental analysis, UV and single-crystal X-ray diffraction. The compound crystallizes in monoclinic system, Cc space group with a = 7.5925（8）, b = 20.263（2）, c = 11.4669（13） A, r= 107.825（8）°, V = 1679.5（3） A3, Z = 4, R = 0.0316 and wR = 0.0687. The results of antibacterial activity test against Escherichia coli and Bacillus subtilis indicate that the compound possesses the same antibacterial activity as the contrast （Norfloxacin）. Theoretical study of the forming mechanism to the title compound at the RHF/6-31G（d） level shows that there are two steps. The distal amino group of thiosemicarbazide is added to the 4-carbonyl group of HPMTP which forms TM. Then a dehydration reaction occurs in TM and generates a stable product PC.

Path Choice on Rural Industrial Upgrading in the Process of Rural-urban Integration in Binhai New Area

With methods of reference research and field research, the current situations of rural industrialization in the process of rural-urban integration in Binhai new area are analyzed and some problems in the process of integration in the new area are pointed out, such as difficulties in breaking dual economic structure in urban and rural areas, imbalanced economic development in the area and inadequate development of the third industry. And relevant countermeasures are proposed: optimizing agriculture industrial structure and promoting agricultural industrialization to form urban agricultural model of Binhai new area focusing on facility agriculture, park agriculture and high-end agriculture and to realize the intensive growth of agriculture; rationally distributing rural industrial structure to form regional leading industry and playing the guiding role of industry to achieve industrial adjustment improvement; quickening the construction of the third industry centering on service to form the service system of modern countryside and the leading role of modern service industry so as to achieve rapid development of the third industry.

Study on Anti-infection Effect of Polysaccharide from Agaricus blazei Murrill on Chickens

In order to study the anti-bacterial infection effect of polysaccharide from Agaricus blazei Murrill on chickens, the experimental groups were orally administrated A. blazei polysaccharide at low dose and high dose, respectively, for 14 d continuously, and then, the chickens in various groups were infected with Escherichia coli or Pasteurella pneumotropica , so as to observe the clinical symptoms of chickens and record the change in body weight. Anatomy was performed 14 d later, and the organ indices were determined, so as to study the anti-bacterial infection effect of A. blazei polysaccharide on chickens. The results showed that after bacterial infection, the high-dose A. blazei polysaccharide group was significantly differed from other groups in changes of body weight and organ indices. It indicates that oral administration of high concentration of A. blazei polysaccharide could promote the development of poultry organs, thereby improving the immunity of organisms.

Construction and prokaryotic expression of the fusion protein Stx2B-IntiminC300 of EHEC O157: H7 and its immunoprophylactic potential

To construct and express the fusion protein Stx2B-IntiminC300 of EHEC O157 ： H7, and to further investigate its immunoprophylactic potential, the gene of Stx2B （stx2b） from EHEC O157：H7 chromosome was cloned into pMD18-T vector. Thereafter, the amplified gene was cloned into prokary- otic expression plasmid pET-28a （ ＋ ）-eaeC300, which was constructed previously. The recombinant pasmid pET-28a（ ＋ ）-stx2b-eaeC300 was transformed into E. coli BL21 （DE3）. After inducement, the protein Stx2B-IntiminC300 was successfully expressed and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE）, Western blotting and N-terminal amino acid residual sequencing. To evaluate its immunoprophylactic potential, it was primarily purified by ion-exchange chromatography and injected into 30 BALB/c mice with AI（OH）3 in the subscapular region. Ten days after the last booster vaccination, 20 mice were attacked with EHEC O157：H7 lysate and the protective efficacy was observed. In the present study, the gene of Stx2B-intiminC300 was successfully cloned into pET-28a （ ＋ ） vector. The results of SDS-PAGE and Western blotting assay showed that the fusion protein was successfully expressed in the inclusion body form, accounting for 25 % of total expression products, and its molecular weight was about 43 kDa. The result of the N-terminal amino acid residual sequencing showed that it was identical to that of the molecular designed. The purity was about 75 % after primary purification. Animal tests revealed that the fusion protein Stx2B-intiminC300 has elicited high titer of protective antibody relatively. These results demonstrate that the fusion protein Stx2B-IntiminC300 is successfully expressed in prokaryotic expression system and shows certain immunoprophylactic potential.

Research progress of micro(nano)plastics in marine survey in China

The extensive existence of microplastics in the marine environment and the various definite and indefinite harm to the living creatures have been paid much attention.In this paper,the definition,source and pollution of micro(Nano)plastics in marine environment are reviewed.The related literatures are retrieved by using big data platform,the distribution characteristics of micro(Nano)plastics in the marine environment,control measures and so on,and the characteristics of key words making,resource distribution and contribution rate of research institutions are comprehensively analyzed.It provides the technical support for the scientific management of micro(Nano)plastics in the future.

Study on DNA Barcoding of Four Common Cyprinidae Species

This study aimed to analyze the partial sequence of COI gene in Pengze crucian carp,gold crucian carp,Ukraine carp and common carp,with silver carp and tilapia fish as two outgroups.DNA fragments 814 bp in length were amplified and analyzed.Statistical analysis of sequence compositions and analysis of intraspecific and interspecific genetic distances were performed by using Mega4.1 software.The phylogenetic tree of COI gene was constructed with neighbor-joining method.According to the results,15 fish individuals could be divided into five groups,which was consistent with the results of traditional taxonomy.Furthermore,the partial sequence of COI gene was a good DNA barcode to identify different species and exhibited certain applicability in the analysis of interspecific phylogenetic relationships of Cyprinidae species.

ECMpy 2.0:A Python package for automated construction and analysis of enzyme-constrained models

Genome-scale metabolic models(GEMs)have been widely employed to predict microorganism behaviors.However,GEMs only consider stoichiometric constraints,leading to a linear increase in simulated growth and product yields as substrate uptake rates rise.This divergence from experimental measurements prompted the creation of enzyme-constrained models(ecModels)for various species,successfully enhancing chemical pro-duction.Building upon studies that allocate macromolecule resources,we developed a Python-based workflow(ECMpy)that constructs an enzyme-constrained model.This involves directly imposing an enzyme amount constraint in GEM and accounting for protein subunit composition in reactions.However,this procedure de-mands manual collection of enzyme kinetic parameter information and subunit composition details,making it rather user-unfriendly.In this work,we’ve enhanced the ECMpy toolbox to version 2.0,broadening its scope to automatically generate ecGEMs for a wider array of organisms.ECMpy 2.0 automates the retrieval of enzyme kinetic parameters and employs machine learning for predicting these parameters,which significantly enhances parameter coverage.Additionally,ECMpy 2.0 introduces common analytical and visualization features for ecModels,rendering computational results more user accessible.Furthermore,ECMpy 2.0 seamlessly integrates three published algorithms that exploit ecModels to uncover potential targets for metabolic engineering.ECMpy 2.0 is available at https://github.com/tibbdc/ECMpy or as a pip package(https://pypi.org/project/ECMpy/).

DNA Polymorphism Among Yewei B, V20B, and Oryza minuta J. S. Presl. ex C. B. Presl.

The new cytoplasmic male sterile （CMS） line Yewei A and its maintainer line Yewei B, with better agronomic characteristics, have been developed from a mutant of V20B （a rice maintainer line） through transformation of genomic DNA of wild rice （Oryza minuta J. S. Presl. ex C. B. Presl.）. Analysis of molecular markers, DNA sequences, and Southern blot revealed that high DNA polymorphism exists between the mutant and its receptor, indicating that the special DNA fragment from O. minuta may be integrated into the genome of Yewei B. Therefore, transformation of genomic DNA from distant relatives to the plant of a target receptor may open an avenue for creating a new rice germplasm.

Purification efficiency of ecological spur dikes for river pollutants in different geometric arrangements:Experiments and numerical modeling

The traditional ecological spur dike is improved,by using the biological fillers such as the plant materials,combined with microorganisms,to obtain a larger water purification capacity.Most of the researches on spur dikes focused on the hydrodynamic force and the migration and the diffusion of the pollutants,but not so much on the effects of the spur dike materials and the arrangement parameters on the water purification capacity.In this paper the biological zeolite is used to make ecological spur dikes.The laboratory flume experiment shows that compared with the natural zeolite spur dike,the purification effect of the biological zeolite spur dike is significantly improved,and the average removal rate of the NH3-N is kept at 6.8%after 24 h compared with 3.5%of the natural zeolite spur dike.The NH3-N removal rates of a single spur dike,the counterpart spur dikes and the staggered spur dikes decrease with the increase of the inlet flow,and the staggered spur dikes have the best purification effect.Secondly,a numerical model based on the MIKE21 is verified by the experimental results and applied for the Tangjiali Section of the Xutangqiao River in Xinwu District,Wuxi City,Jiangsu Province.The orthogonal test shows that the length of the spur dike has the most important effect on the interception of pollutants and the purification,followed by the angle of the spur dike and the spacing of the spur dikes has the least important effect.The geometric arrangement parameters of the ecological spur dike group with the highest efficiency on the pollutant interception and the purification are:60°(angle),6.7 m(length)and 20 m(spacing).