Pyramiding of multiple genes generates rapeseed introgression lines with clubroot and herbicide resistance, high oleic acid content, and early maturity

Clubroot and herbicide resistance,high oleic acid(OA)content,and early maturity are targets of rapeseed(Brassica napus L.)breeding.The objective of this study was to develop new male-fertility restorer lines by pyramiding favorable genes to improve these traits simultaneously.Seven elite alleles for the four traits were introduced into the restorer line 621R by speed breeding with marker-assisted and phenotypic selection.Six introgression lines(ILs)were developed with four-to seven-gene combinations and crossed with two elite parents to develop hybrids.All ILs and their corresponding hybrids displayed high resistance to both clubroot pathotype 4 and sulfonylurea herbicides.Three ILs and their hybrids showed large increases in OA contents and four showed earlier maturity.These new ILs may be useful in rapeseed hybrid breeding for the target traits.

Comparison of pH Value, TBA Value, Glycogen and Lactic Acid Content in Muscle of Landrace and Yorkshire under Different Storage Time and Conditions

In order to assess the meat quality under different storage time and conditions, the pH value, TBA value, glycogen and lactic acid content in muscle of Landrace and Yorkshire were measured. The results showed that within 10 h post slaughtering, post slaughtering time had extremely significant influence on pH value and lactic acid content of Landrace, but had no significant influence on glycogen content and TBA value. On the other hand, post slaughtering time had extremely significant influence on pH value of Yorkshire, but had no significant influence on glycogen, lactic acid content and TBA value. The pH values of Yorkshire at 3, 5 and 10 h post slaughtering were 5.21% （ P 〈 0.01 ）, 5.64% （ P 〈 0.05 ） and 5.59% （ P 〈 0.05 ） higher than those of Landrace, respectively ; the lactic acid content in muscle of Landrace at 10 h post slaughtering was 56.04% （P 〈0.05） higher than that of Yorkshire; the TBA value in muscle of Yorkshire at 10 h post slaughtering was 89. 19% （P 〈 0.01 ） higher than that of Landrace; there was no significant difference in glycogen content between Yorkshire and Landrace. When pork stored at 4℃, storage time had no significant influence on pH value and glycogen content in muscle of Landrace, but had significant influence on drip loss, lactic acid content and TBA value ; storage time had no significant influence on pH value, glycogen content and TBA value in muscle of Yorkshire, but extremely significant influence on drip loss and significant influence on lactic acid content. The pH value in muscle of Yorkshire at 24 h post storage at 4℃ was 6.56% （P 〈0.05） higher than that of Landrace; the glycogen contents in muscle of Landrace at 24, 48, 72, 96, 120 and 144 h post storage at 4℃ were 84.90% （P〈0.05）,78.40% （P〈0.01）, 101.87% （P〈0.05）, 83.80% （P〈0.05）, 83.59% （P〈 0.05）and67.25% （P〈0.01）higher than those of York-shire, respectively; the drip loss of Landrace at 72 h post storage and TBA value at 144 h post storage were 55.70% （ P 〈 0.05 ） and 141.33 % （ P 〈 0.01 ） higher than those of Yorkshire, respectively; there was no significant difference in glycogen content between Yorkshire and Landrace. When pork stored at -20℃, storage time had significant influence on TBA value in muscle of Landrace, but had no significant influence on pH value, thawing water loss rate, glycogen and lactic acid content; storage time had no significant impact on pH value, thawing water loss rate, glycogen content and TBA value in muscle of Yorkshire, and had significant influence on lactic acid content. The pH value in muscle of Yorkshire at 24 h post storage at - 20℃ was 5.43 % （ P 〈 0.05 ） higher than that of I.andrace ; the glycogen contents in muscle of Landrace at 24, 48, 72, 96, 120 h post storage at - 20℃ were 85.08% （ P 〈0. 05 ）, 108.66% （ P 〈 0.01 ）, 72.69% （ P 〈 0.05 ）, 90.31% （ P 〈 0.01 ）, 70.38% （ P 〉 0.05 ） higher than those of Yorkshire, respectively ; the thawing water loss rates of Landrace at 24 and 72 h were 160.14% （ P 〈 0.05 ） and 74.32% （ P 〈 0.05 ） higher than those of Yorkshire, respectively ; there was no significant difference in lactic acid content and TBA value at the same time point between Yorkshire and Landrace.

Physiological Processes Contributing to the Difference in Grain Amino Acid Content Between Two Hybrid Rice(Oryza sativa L.)Cultivars

Improving grain amino acid content of rice(Oryza sativa L.)is essential for the health of consumers.This study was conducted to identify the physiological processes that contribute to the higher grain amino acid content in hybrid rice cultivar Lingliangyou 268 compared to Luliangyou 996.The results showed that total amino acid content in grains was 9%higher in Lingliangyou 268 than in Luliangyou 996.There was no significant difference in grain nitrogen(N)content between Lingliangyou 268 and Luliangyou 996,while ratio of amino acid to N was 6%higher in Lingliangyou 268 compared to Luliangyou 996.A total of 16 differentially expressed proteins related to amino acid metabolism(e.g.,erythronate-4-phosphate dehydrogenase domain containing protein)were identified in grains between Lingliangyou 268 and Luliangyou 996.The identified proteins were involved in 10 molecular functions.Six of the 10 defined functions were related to binding(heterocyclic compound binding,nucleoside phosphate binding,nucleotide binding,organic cyclic compound binding,protein binding,and small molecule binding)and the other 4 defined functions were catalytic activity,enzyme regulator activity,hydrolase activity,and transferase activity.These results indicate that the higher grain amino acid content in Lingliangyou 268 compared to Luliangyou 996 is attributed to increased efficiency of converting N to amino acid that results from altered expression of proteins related to amino acid metabolism.

Influence of citric acid content on magnetic properties of BaFe12O19 powder prepared by sol-gel auto-combustion method

BaFe12O19 powders with nanocrystaUine sizes were produced by sol-gel auto-combustion method. The precursors were prepared under the molar ratios of citric acid to the metal nitrate of 0.5, 1.0 and 1.5. Appropriate ethylene diamine （C2H8N2） was added in order to adjust pH of 7. The ions distribution of citric acid at different pH explains the effect of citric acid in the starting solution. The XRD patterns of the as-burnt powders and annealing powders show different phases for different citric acid content. In addition, the lattice constants （a, c） derived from X-ray diffraction pattern were changed from 0.58881 nm to 0.58997 nm and 2.32057 nm to 2.32296 nm respectively. The data from VSM indicated that the powder with high citric acid content took on good magnetic properties. Pure single BaFe12O19 of the specific maximum magnetization M（1 T）≈ 49.73 Am^2/kg, the specific remanent magnetization Mr ≈ 30.77 Am^2/kg and the coercive force He≈ 467 kA/m was produced when the molar ratios of citric acid to the metal nitrate was 1.5.

Determination and Analysis of Amino Acid Content in Maca from Different Habitats and Varieties

[Objectives]To determine and analyze amino acid content in Maca from different habitats and varieties.[Methods]Maca from two habitats and three colors were selected and a kind of turnip commonly used was to fake Maca.After optimizing the hydrolysis conditions,100 mg Maca samples were hydrolyzed in the concentration of 6 mol/L hydrochloric acid at 110℃for 24 h.The content range of amino acids in each sample was detected by HPLC.[Results]The average total content of amino acids in Maca was 9.52 g/100 g.Yunnan yellow Maca contained 16 kinds of amino acids,including 7 kinds of human essential amino acids,accounting for 17.2%of the total amino acids while Yunnan purple Maca held 16 kinds of amino acids,counting 7 kinds of human essential amino acids,accounting for 16.5%of the total amino acids.The amino acid ratio coefficient method also showed that the two kinds of Maca had higher nutritional value than the others.15 kinds of amino acids were detected in Yunnan black Maca,including 6 kinds of essential amino acids,accounting for 9.60%of the total amino acids.Among amino acids in Yunnan Maca,the highest content was Arg.A total of 12 kinds of amino acids,together with 6 essential human amino acids,were determined in Peru black Maca,accounting for 11.6%of the total amino acids.In Peru black Maca the highest concentration of amino acid was Pro.It was noteworthy that the amino acid species were comprehensive in Maca,and high level of amino acids resulted in high nutritional value.Its specific amino acid composition ratio was obviously different from Turnip,the counterfeit goods to deceive customers.[Conclusions]The method of automatic amino acid analyzer is simple and accurate,and can be used for the identification and content determination of amino acids in Maca as amino acid is an essential nutrient in Maca.

Alleviation of Adverse Effects of Salt Stress in Wheat Cultivars by Foliar Treatment with Antioxidant 2—Changes in Some Biochemical Aspects, Lipid Peroxidation, Antioxidant Enzymes and Amino Acid Contents

Ascobin (compound composed of ascorbic acid and citric acid) is considered one of exogenous protectants which may alleviate the harmful effects of salinity stress. Pot experiments were performed at the screen greenhouse of National Research Centre, Cairo, Egypt to study the effect of foliar treatment of two cultivars of wheat plant with different concentrations of ascobin (0, 200, 400 and 600 mg/l) on some biochemical parameters, antioxidant enzymes, element contents and amino acid constituents of two cultivars of wheat plant grown under different salinity levels (0.0, 3000 and 6000 mg/l) in 2011/2012 and 2012/2013. Salinity with different concentrations levels increased phenolic compounds contents of the two wheat cultivars. The activities of antioxidant enzymes (SOD, CAT, POD, PPO, AXP and GR) dramatically increased due to salinity stress. Amino acid content was increased in cultivar Sids 1, while the content was decreased in cultivar Giza 168 in all salinity treatments. Increments in the above mentioned parameters compared to the untreated plants at normal and stressed conditions. The magnitude of increments was much more pronounced in response to 600 mg/l of ascobin. It could be concluded that, foliar treatment of wheat cultivars with ascobin could partially alleviate the harmful effect of salinity especially at the lower levels of salinity of the two cultivars of wheat at most of the studied parameters.

Association Analysis of the Amino Acid Contents in Rice

The main objective of the present study was to identify simple sequence repeat （SSR） markers associated with the amino acid content of rice （Oryza sativa L.）. SSR markers were selected by prescreening for the relationship to amino acid content. Eighty-four rice landrace accessions from Korea were evaluated for 16 kinds of amino acids in brown rice and ganotyped with 25 SSR markers. Analysis of population structure revealed four subgroups in the population. Linkage disequilibrium （LD） patterns and distributions are of fundamental importance for genome-wide mapping associations. The mean r^2 value for all intrachromosomal loci pairs was 0.033. LD between linked markers decreased with distance. Marker-trait associations were investigated using the unified mixed-model approach, considering both population structure （Q） and kinship （K）. A total of 42 marker-trait associations with amino acids （P 〈 0.05） were identified using 15 different SSR markers covering three chromosomes and explaining more than 40% of the total variation. These results suggest that association analysis in rice is a viable alternative to quantitative trait loci mapping and should help rice breeders develop strategies for improving rice quality.

Polyphenols,Ascorbic Acid and Carotenoids Contents and Antioxidant Properties of Habanero Pepper(Capsicum chinense)Fruit

Their high bioactive compounds content and importance as dietary antioxidants has increased interest in Capsicum fruit. The fruit of seven Capsicum chinense Jacq. var. habanero genotypes grown in Yucatan, Mexico, were analyzed to quantify their phenolic compounds, carotenoids and ascorbic acid contents, and to measure their free radical scavenging (ABTS assay) and antioxidant activities (β-carotene/linoleic acid assay). Phenolics (20.54 to 20.75 mg/100 g sample), carotenoids (1.00 to 1.26 mg/100 g sample) and ascorbic acid contents (187.24 to 281.73 mg/100 g sample) varied between genotypes. Trolox equivalent antioxidant capacity (TEAC) ranged from 1.55 to 3.23 mM/mg sample. During the 120-min decolorization trial, antioxidant capacity decreased over time in the studied genotypes. Values ranged from 36%to 57% β-carotene bleaching during the first 30 minutes. Fruit from all seven studied genotypes are good antioxidant sources and hold promise as natural ingredients in functional foods.

Effects of dietary fiber content and different fiber-rich ingredients on endogenous loss of fat and fatty acids in growing pigs

Background: Determination of the endogenous loss of fat(ELF) is used to adjust for the estimation of true total tract digestibility(TTTD) of fat in diets and ingredients.Any factor which affected ELF may further affect the digestibility of fat,including sources and concentrations of fat and fiber in the diet.There are some reports of determining the ELF using regression methods based on different levels of fat intake,while reports on effects of dietary fiber content and different fiber-rich ingredients in pig diets on ELF are very limited.Therefore,the objective of this study was to determine the effects of dietary fiber content and different fiber-rich ingredients on endogenous losses of fat and fatty acids at the end of ileum and throughout the entire intestinal tract in growing pigs.Methods: In Exp.1,the effect of fiber content on endogenous loss of fat was determined using six growing pigs(Duroc × Landrace × Yorkshire;27.6 ± 2.4 kg),fitted with a T-cannula at the end of ileum.The experimental design was a 6 × 6 complete Latin square design with six periods of feeding and six diets.The six experimental fat-free diets were formulated to include graded levels of neutral detergent fiber(NDF)(0,40,80,120,160 and 200 g/kg) and soybean hull(SH) was the only fiber source,providing 0,75,150,225,300 and 375 g/kg,respectively.Chromic oxide was included at4 g/kg in all diets as an indigestible marker.In Exp.2,six crossbred growing barrows(27.6 ± 1.6 kg) were used and the experimental design was the same as for Exp.1.The six fat-free diets were formulated to include six common fiber-rich ingredients and the concentration of NDF was 100 g/kg.The six fiber-rich ingredients were defatted rice bran(DRB),sugar beet pulp(SBP),rice hull(RH),corn germ meal(CGM),SH and wheat bran(WB) and they were fed at represented250,270,145,250,170 and 280 g/kg in the diet,respectively.Results: In Exp.1,the endogenous loss of fatty acids profile did not change as dietary NDF increased in growing pigs.The endogenous losses of fat,C16:0,C18:0,C18:1,C18:2,total unsaturated fatty acids(UFA) and total saturated fatty acids(SFA) in growing pigs at the end of ileum and throughout the entire intestinal tract increased linearly as NDF content of diets increased.The endogenous losses of fat,as well as C16:0 and C18:0 throughout the entire intestinal tract also increased quadratically as NDF content of diets increased.The ELF increased from 0.71 to 3.14 g/kg of dry matter intake(DMI) and 0.56 to 8.21 g/kg DMI at the end of ileum and throughout the entire intestinal tract in growing pigs,respectively.The ELF occurred in the hindgut except for the growing pigs fed 0 and 4% NDF in their diets.The endogenous losses of C16:0 and UFA occurred primarily in the upper regions of the gut and the greatest endogenous losses of C18:0 occurred in the hindgut.The endogenous losses of fat,individual SFA and total SFA throughout the entire intestinal tract were much greater than that at the end of ileum.However,the endogenous losses of individual UFA and total UFA were less throughout the intestinal tract than at the end of ileum.In Exp.2,the endogenous losses of fat at the end of ileum were greater in growing pigs fed CGM or WB diets.The endogenous loss of fatty acids profile changed to a slight degree at the end of ileum that the endogenous loss of UFA(particularly C18:1 and C18:2) in growing pigs fed CGM or WB diets were greater(P < 0.01) than that for the other four diets.The greatest(P < 0.01) endogenous loss of SFA(particularly C18:0) was in growing pigs fed the RH diet.The endogenous losses of fat,C16:0,C18:0 and SFA over the entire intestinal tract were much greater in growing pigs fed CGM or WB diets,whereas the lowest values were in growing pigs fed DRB diet.The ELF at the end of ileum in growing pigs fed CGM or WB diets were 3.50 or 4.17 g/kg DMI,respectively,and the ELF over the entire intestinal tract was 7.23 or 7.38 g/kg DMI.The contribution in percentage of ELF in the upper gut was greater than that in the hindgut of growing pigs fed DRB and RH diets,while the ELF in the upper gut and hindgut were equal in growing pigs fed SBP,CGM and WB diets.On the whole,the endogenous losses of C18:1 and C18:2 throughout the entire intestinal tract in growing pigs fed the six fiber-rich ingredients diets were less than losses at the end of ileum,whereas the endogenous loss of fat,C16:0,C18:0 and SFA were greater throughout the intestinal tract than at the end of ileum.Conclusion: The profile of loss in endogenous fatty acids did not change as dietary NDF increased in growing pigs and the endogenous losses of fatty acids(C16:0,C18:0,C18:1 and C18:2) fat,UFA and SFA increased linearly as NDF content increased in the diets of pigs.The endogenous losses of fat or fatty acids at the end of ileum were greater in growing pigs fed RH,CGM or WB diets.The endogenous losses of fat,fatty acids(C16:0 and C18:0) and SFA were greater over the entire intestinal tract in pigs fed CGM or WB diet,while these values were the lowest in growing pigs fed the DRB diet.The contribution in percentage losses of fat in the upper gut were greater than in the hindgut of growing pigs fed DRB and RH diets,while the contribution of losses of fat in the upper gut and hindgut were equal in growing pigs fed SBP,CGM and WB diets.In addition,the endogenous loss of individual or total UFA was less over the entire intestinal tract of growing pigs fed fiber diets than that at the end of ileum,and the greatest endogenous losses of fat,individual or total SFA were over the entire intestinal tract.Therefore,differences in fiber content and the nature of fiber-rich ingredients in diets of pigs have different effects to the endogenous losses of fat or fatty acids.Considering the requirement of fat or fatty acids of pigs,careful attention must be paid that the endogenous losses of fat and fatty acids when fiber ingredients are used in diets of pigs.

Oil Content and Nervonic Acid Content of Acer truncatum Seeds from 14 Regions in China

Acer truncatum is considered a promising species as a raw material to produce nervonic acid(NA). The goal of this study was to explore the variation of oil content and nervonic acid content of 138 accessions native to 14 regions in China as well as provide guidance for establishing plantations and improving the production of nervonic acid. A large range of variation was found in oil content, nervonic acid per gram of oil, and nervonic acid per gram of seed(17.81%–36.56%, 3.90%–7.85%, and 0.84%–2.31%, respectively). Accessions YS-6, ABZ-6 and DQTL-8 were found as the most promising potential oil sources; PQ-2 was considered the optimal germplasm, and JY-6, NJ-8, LF-6, and CC-8 were found as promising potential sources for producing NA. A very significant difference(P < 0.01) among 14 regions was observed. DQTL and YS regions are considered the optimal farming regions for crops with high oil content, while the CC and PQ regions are regarded as the most suitable regions for nervonic acid production. A geographical trend from South to North was observed in which amount of seed oil increased but no such trend in nervonic acid content was observed. The data collected in the study on oil content and nervonic acid in accessions of A. truncatum in various regions can be utilized for establishing plantations of promising genotypes through clonal means.

Protein Content and Amino Acid Composition in Grains of Wheat-Related Species

The protein content and amino acid composition for 17 wheat-related species （WRS） and three common wheats （control） were determined and analyzed, and the essential amino acids （EAAs） in WRS were evaluated according to FAO/WHO amino acid recommendations. The results showed that the mean protein content for WRS was 16.67%, which was 23.21% higher than that for the control. The mean contents （g 100 g^-1 protein） of most amino acids for WRS were lysine 2.74%, threonine 2.83%, phenylalanine 4.17%, isoleucine 3.42%, valine 3.90%, histidine 2.81%, glutamic acid 29.96%, proline 9.12%, glycine 3.59%, alanine 3.37%, and cysteine 1.57%, which were higher than those for the control. The contents of the other 6 amino acids for WRS were lower than those for the control. The materials （Triticum monococcum L., Triticum carthlicum Nevski, and Triticum turgidum L.） contained relatively high concentration of the most deficient EAAs （lysine, threonine, and methionine）. Comparing with FAO/WHO amino acid recommendations, the amino acid scores （AAS） of lysine （49.8%）, threonine （70.7%）, and sulfur-containing amino acids （74.8%） were the lowest, which were considered as the main limiting amino acids in WRS. It was observed that the materials with Triticum urartu Turn. （AA） and Aegilops speltoides Tausch. （SS） genomes had relatively high contents of protein and EAA. The contents of protein （16.91%）, phenylalanine （4.78%）, isoleucine （3.53%）, leucine （6.16%）, and valine （4.09%） for the diploid materials were higher than those for the other materials. These results will provide some information for selecting parents in breeding about nutrient quality and utilization of fine gene in wheat.

The effects of soil sand contents on characteristics of humic acids along soil profiles

It is generally accepted that the compositions and properties of soil organic matter(SOM) are in?uenced by many factors. In order to reveal the effects of soil texture on characteristics and dynamics of SOM and its sub-fraction,humic acid(HA), along two soil pro?les, a yellow soil pro?le and a purplish soil pro?le, under the same climate and vegetation conditions were determined. Results indicate that the decomposition and humi?cation degrees of SOM and HA of the purplish soils are higher than those of the corresponding yellow soils indicated by A/O–A ratios of HAs, TOCs and HA yields of bulk soil samples, nevertheless, the development degree of the purplish soil is lower than that of the yellow soil. The variations of E_4/E_6 ratios of HAs along the soil pro?les indicate the overall molecular sizes of HAs decreased downward along the soil pro?les.A/O–A ratios of HAs decreased downward along both the soil pro?les indicate that humi?cation processes decrease downward along both the soil pro?les. Leaching of SOM shows signi?cant effects on the distribution and characteristics of HAs in the yellow soil pro?le but the purplish soil pro?le, which is consistent with the higher hydrophobicity of HAs in purplish soils, shows that the distribution characteristics of SOM along the soil pro?les are a complex result of the combination of soil texture and characteristics of SOM itself. The remarkably different sand contents are concluded tentatively as one of reasons to the different distributions and dynamics of HAs along the soil pro?les,however, to profoundly understand the evolution and transport of SOM along soil pro?les needs more researches.

Physicochemical properties, fatty acid and mineral content of some walnuts (Juglans regia L.) types

Some physical and chemical properties, mineral content and fatty acid compositions of kernel and oils of several walnut types (Büyük Oba, Kaman2, Kaman5) were determined. The oil yields from these kernels changed between 61.4% to 72.8%. The crude fibre contents of ker nels ranged between 3.77% and 3.80%. In add ition, crude protein contents of kernels ranged between 7.05% and 8.10%. While the peroxide values of kernel oils change between 3.18 meq/ Kg and 3.53 meq/Kg, acidity values ranged be tween 0.35% and 0.56%. The main fatty acids of walnut kernel oils were oleic, linoleic, linolenic and palmitic acids. Linoleic acid contents of ker nel oils varied between 49.7% and 55.5%. On the other hand, oleic acid contents ranged between 20.5% and 26.4%. As a result, the present study showed the walnut kernels of the researched species of walnut kernels from Turkey are a potential source of valuable oil which might be used for edible and other industrial applica tions.

Mapping loci controlling fatty acid profiles, oil and protein content by genome-wide association study in Brassica napus

Optimizing the profile and quantity of fatty acids in rapeseed(Brassica napus L.) is critical for maximizing the value of edible oil and biodiesel. However, selection of these complex seed quality traits is difficult before haplotypes controlling their contents are identified. To efficiently identify genetic loci influencing these traits and underlying candidate genes and networks, we performed a genome-wide association study(GWAS) of eight seed quality traits(oil and protein content, palmitic, stearic, oleic, linoleic, eicosenoic and erucic acids content). The GWAS population comprised 370 diverse accessions, which were phenotyped in five environments and genotyped using 60K SNP arrays. The results indicated that oil and protein contents generally showed negative correlations, while fatty acid contents showed positive or negative correlations,with palmitic and erucic acid contents directly affecting oil content. Seven SNPs on five chromosomes were associated with both seed oil and protein content, and five genes orthologous to genes in Arabidopsis thaliana were predicted as candidates. From resequencing data, besides known haplotypes in Bna A.FAE1.a and Bna C.FAE1.a, three accessions harboring a new haplotype conferring moderate erucic acid content were identified. Interestingly, in a haplotype block, one haplotype was associated with high palmitic acid content and low oil content, while the others showed the reverse effects. This finding was consistent with a negative correlation between palmitic acid and oil contents, suggesting historical selection for high oil content. The identification by this study of genetic variation and complex correlations of eight seed quality traits may be beneficial for crop selection strategies.

Extraction Process and Content Determination of Caffeic Acid in Laggera alata from Different Production Areas of Guangxi

[Objectives] To establish a method for determining the content of Laggera alata( D. Don) Sch. Bip. Ex Oliv. using caffeic acid the target component,and to compare the content of caffeic acid in the medicinal materials of L. alata in different production areas of Guangxi.[Methods]The content was determined by Inertsil~ODS-3 chromatographic column C_(18)( 4. 60 mm × 250 mm,5 μm,mobile phase: acetonitrile-0. 1% phosphoric acid( 22∶ 78),detection wavelength: 320 nm,flow rate: 1. 0 m L/min,column temperature: 30℃,and injection volume: 10 μL. [Results] The caffeic acid showed a good linear relationship in the range of injection volume of 0. 025 92-0. 259 2 μg( R =0. 999 5). The average recovery rate was 98. 33%( RSD = 1. 85%). L. alata in different production areas of Guangxi contained the caffeic acid,and there was a great difference in the caffeic acid. L. alata in Baise had the highest content of caffeic acid,while that in Guilin had the lowest content of caffeic acid. [Conclusions]This method can accurately determine the content of caffeic acid and is expected provide a scientific basis for the development and utilization of herbal medicine L. alata.

Effect of nitric oxide synthase inhibitor N^G-nitro-L-arginine on the content of amino acid in ischemic brain tissues of rats

BACKGROUND： Nitric oxide synthase （NOS） inhibrtors have been widely used to investigate the role of NO on cerebral ischemic injury, but the results are controversial. Moreover, it has been considered to aggravate the ischemic neuronal damage with the release of excessively excitatory amino acids （EAA） during cerebral ischemia. On the other hand, some inhibitory amino acid is suggested to be important for the neuronal protection against ischemic brain damage. Our study has recently showed that treatment with the NOS inhibitor NG-nitro-L-arginine （L-NA） reduced focal cerebral ischemic damage. The effect of L-NA on the contents of excitatory and inhibitory amino acid in the rat brain following cerebral ischemia is still unclear. OBJECTIVE： By evaluating the effect of NOS inhibitor, L-NA on the contents of aspartate, glutamate, glycine and γ-aminobutyric acid （GABA） in striatum, hippocampus and cortex in the rat brain following cerebral ischemia respectively, to investigate the beneficial effect of L-NA on cerebral ischemic injury and the possible mechanism. DESIGN： A randomized and controlled experiment SETTING ： Department of Pharmacology, Hebei Academy of Medical Sciences MATERIALS： A total of 42 male healthy SD rats （grade Ⅱ, weighting 250-300 g） were provided by the Experimental Animal Center of Hebei Province （Certification： 04036）. Aspartate, glutamate, glycine, GABA, L-NA and 2,3,5-triphenyltetrazolium chloride （TTC） were obtained from Sigma Chemicals Co, St Louis, MO, USA. HPLC-ultraviolet detector system consisted of Agilent 1100 HPLC. METHODS： The experiment was carried out in Department of Pharmrcology, Hebei Academy of Medical Sciences from June 2005 to June 2006. Rats were randomly divided into three groups： sham-operated group （n = 6）, ischemic group （n = 18）, L-NA group （n = 18）. The model of focal cerebral ischemia in rat was prepared with intraluminal line occlusion methods. In sham-operated rats, the external carotid artery was surgically prepared, but the filament was not inserted. Each group was further divided into 3 subgroups （n = 6 for each）： drugs were administrated at 2, 6 and 12 hours after the middle cerebral artery occlusion （MCAO） respectively. L-NA （20 mg/kg, ip） was administrated, twice a day, for 3 consecutive days. Same volume of normal saline was administrated in ischemic and sham operation groups. The changes of infarcted volume and the contents of amino acids were respectively assayed. Image analysis software was used for the measurement of the infarcted area. The results were expressed as a percentage of the infarcted volume of cerebral/volume of whole brain （IV%） in order to control for edema formation. The contents of aspartate, glutamate, glycine and GABA in striatum, hippocampus and cortex in the rat brain following cerebral ischemia were respectively measured by HPLC method. All data were analyzed with one-way ANOVA and Dunnett＇s test. MAIN OUTCOME MEASURES： （1） The volume of cerebral infarction; （2） The contents of aspartate, glutamate glycine and GABA in brain tissue after cerebral ischemia. RESULTS ： All 42 rats were involved in the final analysis. （1） Infarcted volume： Volume was 0 in sham-operated group. When L-NA was administrated at 2 and 6 hours after MCAO, the infarcted volume was （20.13±3.59）% and （23.12±5.84）% in L-NA group, which was not similar to that in ischemic group [（22.10±3.98）%, （25.38± 5.37）%, P〉 0.05]. However, the infarcted volume was markedly decreased compared with that of ischemic group when L-NA was administrated at 12 hours after MCAO [（26.11±3.55）% and （37.15±3.58）%, P 〈 0.01]. Changes of amino acid content： At 2 and 6 hours after ischemia, the contents of aspartate, glutamate, glycine and GABA in striatum, hippocampus and cortex in ischemic group were significantly increased compared with those in sham-operated group （ P〈 0.05-0.01）. However, contents in L-NA group were similar to those in ischemic group （P 〉 0.05）. At 12 hours after ischemia, the contents of aspartate [（0.21 ±0.06）, （0.36±0.05）, （0.29±0.12） mg/g] and glutamate [（0.55±0.06）, （0.78±0.10）, （0.52±0.10） mg/g] in striatum, hippocampus and cortex in L-NA group were significantly decreased compared with those in ischemic group [（0.49±0.17）, （0.63± 0.03）, （0.51±0.15） mg/g; （0.98±0.30）, （1.15±0.15）, （0.93±0.15） mg/g, P〈 0.05-0.01]. Glycine in hippocampus was （0.40±0.07） mg/g, which was higher than that in ischemic group [（0.21±0.07） mg/g, P 〈 0.05]. GABA in striatum, hippocampus and cortex was （0.93±0.10）, （0.62±0.12） and （0.81 ±0.10） mg/g, respectively, which was higher than that in ischemic group [（0.60±0.08）, （0.37±0.17）, （0.59±0.10） mg/g, P 〈 0.05-0.01]. CONCLUSION ： It may be concluded that L-NA have beneficial effect on ischemic cerebral injury in ischemic later stage in rats. The possible mechanism is that L-NA can decrease the contents of aspartate and glutamate, increase the contents of glycine and GABA.

Characteristic of hyaluronic acid derivative films cross-linked by polyethylene glycol of low water content

Objective:To test the characteristics of hyaluronic acid(HA)derivative cross-linked by polyethylene glycol films of low water content.Methods:The cross-linked HA film with 200μm thickness was got at atmospheric pressure at 25℃for 5 d.After dried,cross-linked films of 10 mm×10 mm were weighed and immersed in phosphate buffered saline (PBS pH 7.45)at 37℃for 24 h.Then the solution fraction and water content were estimated.Meanwhile,cross-linked HA derivative films were immersed in phosphate buffered saline(PBS:pH 7.45)at 37℃for determined time and then implanted subcutaneously in the back of white rats to test in vitro or in vivo degradation characteristic.Results and Conclusion:HA hydrogel cross-linked by polyethylene glycol with water content is as low as 60% and this kind of HA derivative has a slow degradation rate.

Content Determination of Chlorogenic Acid and Luteoloside in Flos Lonicerae

The contents of chlorogenic acid and luteoloside in Flos Lonicerae from Binhai New Area and Jinnan District of Tianjin were determined to provide basis for the quality identification of this medicinal material. The content of chlorogenic acid was determined by HPLC. In Flos Lonicerae from Binhai New Area and in Flos Lonicerae harvested at different stages from Jinnan District,the contents of chlorogenic acid were 3. 804%,5. 507%( three green stage),4. 855%( silver flower stage) and 4. 220%( golden flower stage),respectively,and the contents of luteoloside were 5. 53%,12. 405%( three green stage),14. 370%( silver flower stage) and 0. 917%( golden flower stage),respectively. The contents of chlorogenic acid in Flos Lonicerae from Jinnan District were higher than that from Binhai New Area. Among different stages,the content of chlorogenic acid was highest in three green stage,followed by that in silver flower stage. As the flowers bloomed,the content of chlorogenic acid in the medicinal material showed a significant downward trend. In Flos Lonicerae from Jinnan District,the content of luteoloside was highest in silver flower stage and lowest in golden flower stage.