Extraction and purification of the polysaccharides in Hippohaere rhamnoides L.

The different extraction technology and purification technology of Hippohpae rhamoides polysaccharides were researched in the paper.The best method of papain extraction were obtained,the ratio of papain 2%,pH at 5.5,temperature at 45℃and extraction time of 20 min were suitable for papain extraction.The highest content of Hippohpae rhamoides polysaccharides was 44.28 mg·g~(-1). The optimum process of ultrasonic extraction were obtained,namely extracted for 55 min at 480 W with the material ratio of 1:20. The highest content of Hippohpae rhamoides polysaccharides was 48.63 mg·g~(-1).The results showed that the ultrasonic and papain extraction together was the best method,the content was 54.30 mg·g~(-1).After the removing protein,pigment and dialysis.Two fraction were separated from the purified Hippohpae rhamoides by DEAE-cellulose chromatography,the main fraction was collected finally. The fraction was identified by Sepharose CL-4B gel filtration.Ultraviolet spectrometry,freeze-thawing analysis showed that fraction was purified.Its molecular weight was probably 109.4 ku.     

Optimization for Extraction Methods and Conditions of Polysaccharides from Trollius chinensis

Objective To determine the optimum extraction method and condition of poly-saccharides fromTrollius chinensis (TCP).Methods With the yield of TCP as index and water as solvent,reflux extraction,ultrasonic extraction and supercritical CO_2 extraction were employed to determine the efficient extraction method of TCP.Moreover,the conditions of reflux extraction were optimized by single factor test and orthogonal test.The content of polysaccharide was measured with phenol-sulfuric acid method.Results The data displayed that the crude polysaccharide(CPS)yield of regular reflux extraction was(11.7±0.12)%,superior to ultrasonic extraction and supercritical CO_2 extraction.The optimum conditions of reflux extraction were as follows:extraction temperature of 95℃,extraction time of 3hand the liquid-solid ratio of 20∶1.Under these conditions,the polysaccharide yield was (1.45±0.03)%,which was well in close agreement with the predicted value.Conclusion Regular reflux extraction was regarded as the optimal extraction method of TCP.The optimum conditions of reflux extraction were as follows:extraction temperature of 95℃,extraction time of 3hand the liquid-solid ratio of 20∶1.     

Extraction of Proteins from Apple Leaves for Two-Dimensional Electrophoresis Analysis

In order to develop an efficient method about protein extraction which is suitable for apple proteomic analysis, four protocols of total protein extraction from apple leaves, which are trichloroacetic acid/acetone precipitation method （TCA）, phenol extraction methanol/ammonium acetate precipitation method, Tris-HCl extraction method and modified Tris-HCl extraction method were compared. The results showed that the modified Tris-HCl extraction method was the most suitable method in protein extraction for two-dimensional electrophoresis （2-DE） from apple leaves based on the highest resolution and more informative spots of 2-DE gels with no apparent vertical or horizontal streaking.     

Protein Extraction Methods for Two-Dimensional Electrophoresis from Baphicacanthus cusia （Nees）Bremek Leaves-A Medicinal Plant with High Contents of Interfering Compounds

Protein extraction is a critical step for two-dimensional electrophoresis （2-DE）. Different plant samples require different and adaptive protein extraction protocols. The leaves of medicinal plant, Baphicacanthus cusia （Nees） Bremek are notoriously recalcitrant to common protein extraction methods due to high levels of interfering compounds （especially the secondary metabolites and pigments）. This study was aimed to establish a routine procedure for the proteomic analysis ofB. cusia leaves, and a new protocol for the protein extraction was developed by optimizing trichloroacetic acid （TCA）/ acetone extraction method. The efficiency of this protocol was demonstrated by comparison with 3 published protein extraction methods （chloroform/acetone, Mg/NP-40, Tris-base/acetone）. The results showed that the optimized TCA/ acetone precipitation extraction method gave a relatively high protein yield （9.263 mg g^-1 fresh weight）, high-resolution separation, clear protein profiles, the highest proteins spots （1 31 t protein spots）, and displayed less contamination in 2- DE gels. Therefore, the results suggested that the optimized TCA/acetone method was the most effective among the 4 methods for B. cusia leaves.     

Effects of Organic Fertilizer on Fruit Quality and Acidified Soil Chemical Properties in Yantai Orchard

Objective] This study aimed to investigate the improving effect of organic fertilizer on acidified soil as wel as their ef-fects on fruit quality and quantity in Yantai orchard. [Method] Plot experiment was conducted to investigate the effects of organic fertilizer on fruit yield and quality of Red Fuji and chemical properties of acidified soil. [Result] The apple yield in acidified soil applied with organic fertilizer al increased. Under the application of biological organic fertilizer, the apple yield was higher, and it was 8.92% higher than that in the control group. Under the mixed application of chemical fertilizer and biological organic fertiliz-er, the growth and development of apple trees were improved, and the total soluble solid (TSS) content, vitamin C (Vc) content and TSS-acid ratio in mature apples al increased. The application of organic fertilizer significantly reduced soil acidity. Compared with those in the control group, the soil pH value, organic matter content and alkali-hydrolyzable nitrogen content under the ap-plication of biological organic fertilizer were increased by 8.33%, 15.10% and 30.80%, respectively. [Conclusion] The application of biological organic fertilizer could improve the yield of apple in acidified soil.     

一种高效提取棉花细胞核的技术(英文)

[Objective] The experiment aimed to study an efficient method of Nuclei extraction of cotton and provided technical support for constructing large-insert genomic library and sequencing complete genome. [Method] The cotton cotyledons germinated in dark moisture chamber for one week were chopped with a sharp sterile scalpel in a Petri dish which contained ice-cold nucleus isolation buffer (10 mmol/L MgSO4, 5 mmol/L KCl, 0.5 mmol/L HEPES, 1 mg/ml DTT, 0.25% Triton X-100 and 2% PVP40), then the nuclei were collected after selected through 100, 50 and 30 μm nylon meshes and centrifugation. Meanwhile, the tender leaves and cotyledons with different germination time in dark were treated by grinding method and sharp scalpel method. [Result] The chopping with a sharp scalpel method was very simple and rapid, which did not need grind and mercaptoethanol treatment and the successful extraction rate was 100%.[Conclusion] An efficient method of nuclei extraction of cotton with simple, high efficiency, rapid reaction and poison free were established.     

流域不透水面及其变化信息提取(英文)

Impervious surface is one of the important parameters of valley water circular simulation,scientific estimation for which has significant and practical value for the urban water quantity and process simulation,diffuse pollution estimating and the forecast of climate changes.The objective of this research is to get the information of impervious surface and its dynamic change.Through the computer-assisted field method,the technologies of decision tree and data mining were applied to withdraw the impervious surface information in research region by the Landsat TM data in 1988,1994 and 2002.The results suggested that the accuracy of impervious surface information extraction in the study area arrived above 94.4% in 2002 image.On this basis,the mixed method was used to extract the location and the types of the impervious surface change.The overall accuracy of monitoring reached 89%,which meets the demand of the hydrological models.     

商城肥鲵的含肉率及其肌肉生化成分分析（英文）

[Objective] The ratio of flesh content and biochemical compositions in muscles of Pachyhynobius shangchengensis were researched in this experiment to provide basic referrences for studying nutritive physiological ecology of Pachyhynobius shangchengensis.[Method]The biochemical analytic method was used to detect the ratio of flesh content and biochemical compositions in muscle of Pachyhynobius shangchengensis.[Result]The ratio of flesh contents of female and male Pachyhynobius shangchengensis were 54.93% and 55.84% respectively.The contents of protein,total saccharides and fatty acid of female Pachyhynobius shangchengensis were 15.44%,1.70% and 0.68% respectively,while the same contents of male Pachyhynobius shangchengensis were 15.67%,1.89% and 0.53% respectively.The total content of 18 kinds of amino acids was 12.72%(the proportion in fresh weight),among which the content of 8 kinds of essential amino acids was 4.68%.The essential amino acids index(EAAI)was 72.68 and limiting amino acids was valine.There were 11 kinds of fatty acids in muscles of Pachyhynobius shangchengensis,among which the content of unsaturated fatty acids(80.02%)was higher than that of saturated fatty acids(19.98%).[Conclusion] Pachyhynobius shangchengensis was a kind of valuable natural resource belonging to the nutritious type of low fat content and high protein content.     

Optimization of Ultrasonic-Assisted Extraction Process of Polysaccharides from American Ginseng and Evaluation of Its Immunostimulating Activity

Ultrasonic-assisted extraction （UAE） of American ginseng polysaccharides （AGP） was investigated using response surface methodology. Three-factor-three-level Box-Behnken design was employed to optimize the ultrasonic power, extraction time and ratio of water to raw material to obtain a high AGP yield. The analysis of variance and response surface plots indicated that ultrasonic power was the most important factor affecting the extraction yield. The optimal conditions were ultrasonic power 400 W, extraction time 71 min, and ratio of water to raw material 33 mL g-1. Under these conditions, the yield of AGP was 8.09%, which was agreed closely to the predicted value. Gas chromatography （GC） analysis showed that AGP was composed of arabinose, rhamnose, galactose, glucose, and galacturonic acid. Fourier transform infrared spectra revealed the general characteristic absorption peaks of AGP. In addition, AGP exhibited good immunostimulating activities by up-regulating the production of nitric oxide and cytokines. Compared with hot water extraction, UAE required shorter extraction time and gave a higher extraction yield, without changing the structure and immunostimulating activity of AGP. The results indicated that UAE could be an effective and advisable technique for the large scale production of plant polysaccharides.     

Synthesis and Identification of SG-BSA and SG-OVA

Hapten sulfaguanidine （SG） was coupled with carrier protein bovine serum albumin （BSA） to form a full antigen SG- BSA by diazotization and glutaraldehyde methods. Ovalbumin （OVA） was used as protein carrier to couple with Hapten SG by glutaraldehyde method. Then, the immunogen and the coating antigen were purified by dialysis and gel exclusion chromatography. The conjugated ratio of SG to BSA in artificial antigen was 5.3 （using diazotization method） and 6.5 （glutaraldehyde method）, and the conjugated ratio of SG to OVA in coating antigen was 2.3 （glutaraldehyde method） by UV-visible spectrophotometer. The coupling was successful according to the analysis of sodium dodecyl sulphate polyacrylamide gel electrophoresis （SDS-PAGE）. BALB/c mice were immunized with the antigen （SG-BSA）, and the titers of antiserum were tested to be 1 ： 6 400 and 1 ： 400 after three periods of immunities by indirect ELISA, which further identified the success of the synthesis of both immunogen SG-BSA and coating antigen SG-OVA.     

Studies on Single Cell Culture in vitro in Wheat--The variation of grain protein content and its fractions from regenerated plants

On the basis of previous studies dealing with the variation of major agronomic and yield characteristics of regenerated plants derived from single cell culture in vitro of common wheat (Triticum aestivum L.Cultivar NE 7742),the grain protein content and its fractions from regenerated plants with stable agronomic characteristics were studied from 1992 to 1995.The results showed that the variation of grain protein content and its fractions in somaclones from single cell culture in vitro were very significant and the range was very wide (11.53-17.70%).Several types of variation were found in the studies,espercially the type with higher protein content than that of cultivar NE 7742(non-culture parent).Among them.-20.69% of lines the grain protein content was significantly higher than that of NE 7742 and combined with high yielding potential.The tendency of variation of the four protein fractions showed that the variation of albumin was not obvious and maintained the same level as NE7742,the content of gliadin increased in some somaclones and decreased in others.However,the percentages both globulin and glutenin tended to increase.The variation of total amount of structural protein and the ratio between globulin and glutenin tended to increase.the variation of total amount of structural protein and the ratio between globulin and albumm was mainly influenced by globulin under the condition of culture in vitro.The variation of total amount of storage protein and the ratio between gliadin and glutenin was mainly by glutenin.The results mentioned above bemonstrated that the induction and screening of somaclonal variation could be an effective way in wheat improvement in combining high protein content with high yield.     

Effect of Ultrasonic Treatment on the Extracting Activity of Kid Rennet

Using 2-3 weeks kid abomasums as materials, the ultrasound intensity, extracting time, NaC1 concentration and pH value in extracting solution and ratio of abomasums to extracting solution were studied by ultrasonic method. The results showed that the main factor affecting kid rennet activity during extraction was ultrasound intensity, and then NaCl concentration, extracting time and ratio of abomasums to extracting solution in order. Kid rennet activity reached peak with ultrasound intensity 30 W cm^-2, extraction time 40 min, NaCl concentration 8%, pH value 3.0, ratio of abomasums to extracting solution 1:15.     

Design of Manure Gas Exhaustion to Reduce Ammonia Concentrations in Loose-Housing Systems for Laying Hens

Uniformity of air flow in extraction openings in exhaust air channels for manure gas exhaustion is determined by the distribution of pressure. The areas required in extraction vents and in extraction ducts are determined by the uniformity of air flow desired along the duct and by the loss of pressure that can be accepted. The area ratio between the vents and the cross section of the exhaust air duct will have a strong influence on both uniformity of flow and loss of pressure. The following ventilation properties were studied： Uniformity of air flow; Variations in static pressure along a duct; Air velocity at different distances from the vents. The area ratio should be about 1 for uniform exhaustion. The studies showed that the relative variation in air velocity is independent of the level of the ventilation rate. The uniformity of the exhaust distance is influenced in about the same way by the area ratio as the air velocity in the exhaust vents. Thus, it is important that the area ratio is not too high if a good exhaust function should be guaranteed. The studies also demonstrated that the uniformity of the exhaust distance is independent of the ventilation flow rate. The exhaust ventilation range is, maximally 0.3 m from the vents. The static friction coefficient was on average 0.80.     

紫鸭跖草细胞中铜的分配和化学形态特征研究(英文)

[Objective] The aim of this study was to reveal the mechanism of Cu2+ hyper-accumulator in Setcreasea purpurea Boom from the angle of distribution characteristics and binding form of Cu2+ in tissue cells.[Method]The distribution characteristics of Cu2+ in subcells of Setcreasea purpurea Boom was studied by the technique of differential centrifugation,and the binding form of Cu2+ in roots and leaves of Setcreasea purpurea Boom was also investigated by the sequential chemical extraction method and the enzymolysis method.[Result]Cu2+ in roots mainly distributed in the cell wall which is accounting for one third of the Total Cu2+ in roots,while Cu2+ in leaves mainly distributed in the chloroplast which is accounting for a quarter to the Total Cu2+ in leaves.Under the high concentration of Cu2+ or the extended treatment duration,the translocation of Cu2+ in root cells into the cell wall increased but the translocation of Cu2+ in root cells into the plastid decreased,while the translocation of Cu2+ in leaf cells into the chloroplast increased but the translocation of Cu2+ in leaf cells into the cell wall decreased.Cu2+ in leaves was mainly combined with amino acid,small molecular polymeric pigments,protein and polysaccharide,while Cu2+ in roots was mainly combined with cell wall substances such as cellulose and membrane-bound protein.[Conclusion]The distribution characteristics and binding form of Cu2+ in cells is possibly one of the dominant mechanisms for Cu2+ hyper-accumulator in Setcreasea purpurea Boom.     

Effects of Different Protein Levels on the Growth Performance and Metabolic Rate of Nutrition in Broilers

The objective of this study is to determine the effect of different protein levels on the growth performance and metabolic rate of nutrition in broilers. Total 360 healthy and weight closed local broilers of 3 weeks were chosen and then divided into three groups randomly by one factor. Each group contains three handlings, each handling consists of 40 broilers. The period of experiment was 7 weeks. The metabolic experiment was performed at the 7th week. Three different protein levels were used in broilers＇ diet. The levels of protein were 19%, 17.5% and 16%. The results showed that the different levels of protein in crude dietary had significant difference between 19% group and the other two groups. The average daily weight gain and daily efficiency were significantly higher than that of the other two groups （P〈0.05）. Compared to protein levels of 19% and 16%, there was no significant difference in the metabolism of calcium （P〉0.05）, and the metabolic rate of the impact of phosphorus was significantly different （P〈0.05）. The result showed that when protein level was 19%, the growth of the local broiler was the best.     

Determination of Rotenone Residues in Foodstuffs by Solid-Phase Extraction （SPE） and Liquid Chromatography/Tandem Mass Spectrometry （LC-MS/ MS）

We developed a novel approach to determine rotenone residues in foodstuffs, by integrating solid-phase extraction （SPE） and liquid chromatography/tandem mass spectrometry （LC-MS/MS） technologies, to achieve high sensitivity and selectivity. In our method, the solvent extraction with n-hexane-dichloromethane （50：50, v/v） and cleanup with florisil SPE cartridges using ethyl acetate-ethyl ether （25：75, v/v） as eluents provided adequate recovery of rotenone. The detection of rotenone was then carried out by LC-MS/MS using acetonitrile-water with the 0.1% formic acid （w/v） as the mobile phase. The multiple reaction monitoring （MRM） scheme employed in the approach involved the transitions of the precursor ion to three selected product ions, in which one pair for quantification was m/z 395.3 〉 213.2 and the other two pairs for identification were m/z 395.3 〉 192.2 and 395.3 〉 367.0. The limits of quantification （LOQs） of the method ranged from 0.001 to 0.005 mg kg-1 depending on the matrix. Intra- and inter-day precisions （relative standard deviations, RSDs） for rotenone were less than 7.1 and 14.8%, respectively. Results from repetitive analysis suggested good reproducibility of the method for rotenone residue detection. The recoveries at three concentrations （LOQ, 10LOQ and 100LOQ） ranged from 79.3-118.3% in cabbage, potato, onion, carrot, apple, orange, banana, lichee, tea, and Shiitake mushroom. The proposed procedure was then applied to the analysis of 129 real samples collected from Xiamen, Fujian Province, China. The existence of rotenone was found in two tea products with concentrations of 0.012 and 0.016 mg kg-1, respectively. The method has great potential for routine analysis of monitoring rotenone residue in foodstuffs.     

Peat1蛋白及其3个缺失突变体的表达与热稳定性分析（英文）

[Objective] The research aimed to reveal the functions of NAC and UBA domains in Peat1's thermal stability.[Method] Fusion expression vectors of Peat1 protein and the 3 deletion mutants were constructed.The recombinant plasmids were induced by IPTG and the target proteins(Peat1,Peat1-△CD99,Peat1-△ND49 and Peat1-△ND108)were expressed obtained by AKTA and its thermal stability was analyzed.[Result] The research found that 3 deletion mutants have good thermal stability like Peat1.[Conclusion] The research demonstrated that the coexistence of NAC or UBA domains is not necessary to thermal stability of Peat1 protein,and they may give the protein particular stability structure seperately.     

Relationship Between Leaf C/N Ratio and Insecticidal Protein Expression in Bt Cotton as Affected by High Temperature and N Rate

Expression of insecticidal protein for transgenic Bacillus thuringiensis （Bt） cotton is unstable and related to nitrogen metabolism. The objective of this study was to investigate the relationship between leaf carbon nitrogen ratio （C/N） and insecticidal efficacy of two Bt cotton cultivars. C/N ratio and Bt protein content were both measured at peak square period and peak boll period respectively under 5-7 d high temperature and different nitrogen fertilizer rates on the Yangzhou University Farm and the Ludong Cotton Farm, China. All plants were grown in field. The results showed that the C/N ratio enhanced slightly and the Bt protein content remained stable at peak square period, but significant increases for the C/N ratio and decreases markedly for the leaf Bt protein concentration were detected at the peak boll period. The similar patterns at the two growth periods were found for the leaf C/N ratio and Bt protein content by different N fertilizer treatments. When nitrogen rate was from 0 to 600 kg ha-l, the C/N ratio was reduced by 0.017 and 0.006 for Sikang 1 and Sikang 3 at peak square period, compared to the 1.350 to 1.143 reduction for Sikang 1 and Sikang 3 at peak boll period, respectively. Correspondingly, the leaf Bt protein contents were bolstered by 2.6-11.8 and 26.9-36.9% at the two different growth periods, respectively. The results suggested that enhanced C/N ratio by high temperature and nitrogen application may result in the reduction of inseetiocidal efficacy in Bt cotton, especially in peak boll period.     

Treatment of Chinese Traditional Medicine Wastewater by Photosynthetic Bacteria

The influence factors treating wastewater of Chinese traditional medicine extraction by photosynthetic bacteria are tested and discussed. The results indicate that the method of photosynthetic bacteria can eliminate COD and BOD from wastewater in high efficiency. And it also has high load shock resistance. On the conditions of slight aerobic and semi-darkness, treating wastewater of Chinese traditional medicine extraction, the method has better efficiency to eliminate COD and BOD from the wastewater than those by anaerobic illumination and aerobic darkness treatments. After pretreatment of hydrolytic acidization, the removal rate of COD in the wastewater reached more than 85%, and that rate of BOD reached more than 90% in the treating system of photosynthetic bacteria, it may be more feasible and advantageous than traditional anaerobic biological process to treat organic wastewater using PSB system.     

Detection of Tocopherol in Oilseed Rape （Brassica napus L.） Using Gas Chromatography with Flame Ionization Detector

The variation among Chinese genotypes of Brassica napus L.for seed tocopherols content and their analysis using gas chromatography has not been comprehensively reported till to date.In the present study,the tocopherol contents of four Chinese genotypes of Brassica napus L.,namely,Gaoyou 605,Zhejiang 619,Zheshuang 758,and Zheshuang 72,were evaluated using three modified sample preparation protocols(P1,P2,and P3) for tocopherol extraction.These methods were distinguished as follows.Protocol one(P1) included the evaporation of solvent after extraction without silylation.Protocol two(P2) followed the direct supernatant collection after overnight extraction without drying and silylation.Protocol three(P3) included trimethylsilylation with N,O-bis(trimethylsilyl) trifluoroacetamide.Genotypic comparison of tocopherol and its isoforms revealed that Gaoyou 605 was dominant over the other genotypes with(140.5±10.5),(316.2± 9.2),and(559.1±24.3) μg g-1 of seed meal α-,γ-,and total(T-) tocopherol,respectively,and a 0.44±0.04 α-to γ-tocopherol ratio.The comparison of the sample preparation protocols,on the other hand,suggests that P3 is the most suitable method for the tocopherol extraction from Brassica oilseeds and for the analysis of tocopherols using gas chromatography flame ionization detector(GC-FID).Trimethylsilylation is the key step differentiating P3 from P1 and P2.Variations detected in tocopherol contents among the Chinese rapeseed(B.napus) genotypes signify the need to quantify a wide range of rapeseed germplasm for seed tocopherol dynamics in short and crop improvement in long.