饲料添加剂用黄芩甙提取工艺条件的优化

为了优化饲料添加剂用黄芩甙提取工艺，采用L₉(3⁴)正交试验设计方法，对浸提和酸沉工艺进行了优化试验研究，采用紫外分光光度法对提取物黄芩甙进行含量测定。结果表明：浸提较优工艺为A₃ B₃ C₂，即浸提时间1.5 h，浸提温度100℃，浸提pH 7.0；酸沉较优工艺为A′₂ B′₃ C′₁ D′₃，即保温时间1.0 h，保温温度80℃，保温pH 1.0，静置时间12 h。经两次正交试验设计所得优化提取工艺可行，提取率为91.56％，含量为94.12％。     

Inhibitory effect of naturally occurring flavonoids on the formation of advanced glycation endproducts

The objective of this study was to investigate the inhibitory effect of naturally occurring flavonoids on individual stage of protein glycation in vitro using the model systems of delta-Gluconolactone assay (early stage), BSA-methylglyoxal assay (middle stage), BSA-glucose assay, and G.K. peptide-ribose assay (last stage). In the early stage of protein glycation, luteolin, qucertin, and rutin exhibited significant inhibitory activity on HbA1C formation (p < 0.01), which were more effective than that of aminoguanidine (AG, 10 mM), a well-known inhibitor for advanced glycation endproducts (AGEs). For the middle stage, luteolin and rutin developed more significant inhibitory effect on methylglyoxal-medicated protein modification, and the IC50's were 66.1 and 71.8 microM, respectively. In the last stage of glycation, luteolin was found to be potent inhibitors of both the AGEs formation and the subsequent cross-linking of proteins. In addition, phenyl-tert-butyl-nitron served as a spin-trapping agent, and electron spin resonance (ESR) was used to explore the possible mechanism of the inhibitory effect of flavonoids on glycation. The results indicated that protein glycation was accompanied by oxidative reactions, as the ESR spectra showed a clear-cut radical signal. Statistical analysis showed that inhibitory capability of flavonoids against protein glycation was remarkably related to the scavenging free radicals derived from glycoxidation process (r = 0.79, p < 0.01). Consequently, the inhibitory mechanism of flavonoids against glycation was, at least partly, due to their antioxidant properties.     

Genetic diversity and sampling strategy of Scutellaria baicalensis germplasm resources based on ISSR

Destruction in wild resource and decline in genetic diversity of Scutellaria baicalensis are becoming a big problem urgently needed be dealt in China. To help efficiently utilize and conserve this medicinal plant resources, ISSR analysis was employed to reveal genetic diversity of 107 S. baicalensis accessions consisting of 82 wild and 25 cultivated germplasm resources from 6 populations in two main producing provinces in China. Only eighteen ISSR primers that resulted in accuracy and credibility, high polymorphism, well stability PCR products were selected. The maximal and minimal bands resulted from an ISSR primer are 35 and 18 respectively with a total of 480 bands were resolved in the agarose gels with an average of two alleles per locus. As indicated by the index of genetic diversity (Na = 1.96, Ne = 1.39, He = 0.25, I = 0.39), there were richness of genetic diversity in the collected samples and that the S. baicalensis germplasm had a large representativeness as primary collection. Cluster analysis by UPGMA and STRUCTURE placed the 107-germplasm resources into three linkage clusters. The samples from all the collection locations were clustered together, but the populations from one province were not clustered together. Compared to the RS and SW strategies, the LDSS proved to be more representative for core collection construct of S. baicalensis and the best sampling ratio was 10.3 % of the total germplasms. Finally, we found that the mini core collection by LDSS was composed of 11 accessions with high genetic diversity (Na = 1.82, Ne = 1.40, He = 0.25, I = 0.39). To verify the reliability of sampling strategy, the t test was used to evaluate the representativeness between core collections (constructed by LDSS), primary and reserve collection. Our results showed that no significant difference was found in the two important genetic parameters He and I (P > 0.05), thus suggesting that the LDSS will provide a rational sampling strategy for constructing a core collection of S. baicalensis germplasm resources in the future.     

不同施水量对黄芩干物质积累与分配的影响

为研究施水量对黄芩的影响，确定适宜黄芩生长的水分条件，在黄芩生长季节模拟降水量设4种不同施水量处理，测定黄芩生长形态指标和各器官干物质重量，计算黄芩各器官干重占总生物量大小。结果表明，施水量多的处理和施水量少的处理根干重、地上千重、总生物量较低，施水量不足对黄芩地上部的影响大于对根部的影响；中度和轻度水分胁迫处理黄芩根重、地上重、总生物量较高；黄芩比较耐旱，年降水量在321～450mm，黄芩生长季节降水量在250～350mm，黄芩地上部与地下部生长协调，各器官物质分配合理，产量高。     

不同施肥措施对黄芩根际微生物群落结构的影响

探究不同施肥措施对黄芩根际细菌群落结构和种间互作关系的影响,为黄芩的合理施肥和提高土壤微生物多样性提供理论依据。采用高通量测序技术对不施肥(CK)、单施化肥(NPK)、化肥配施生物有机肥(NPKM)3个处理的野外黄芩根际土壤样本细菌多样性、群落结构和网络关系进行测定和分析。结果表明,与CK和NPK处理相比,NPKM处理显著增加了细菌多样性,增幅分别为11.51% 和7.37%。与CK相比,NPK和NPKM处理均显著改变了细菌群落结构。不同处理中,变形菌门是丰度最高的门,其相对丰度均高于28%,此外,CK的优势菌群为寡营养型的酸杆菌门,NPK处理的优势菌群为富营养型的变形菌门和拟杆菌门,NPKM处理的优势菌群为能降解复杂有机物的放线菌门和绿弯菌门。网络分析的结果表明,黄芩根际土壤细菌各物种间的正相关性达64.32%,表明物种间主要呈现出互相合作的关系,与NPK处理相比,CK和NPKM处理的网络节点数分别增加了4.76% 和5.95%,而边数则分别增加了12.13% 和10.54%,说明仅施化肥不利于增强土壤微生物群落之间的联系和系统稳定性。综上所述,施用化肥,特别是化肥配施生物有机肥可以通过增强土壤细菌种间互作联系,优化群落组成,为黄芩生长提供更适宜的环境。     

氮磷钾对黄芩生长与有效成分累积的影响

探讨氮磷钾营养对黄芩生长动态、产量和有效成分累积的影响,为黄芩的合理施肥提供依据。通过大田试验,采用氮磷钾三因素二次饱和-D最优设计方案(310),定期采样,测定植株的生长指标、根中黄酮和黄芩苷的含量,计算产量,得到合理施肥量。结果表明:单施氮肥最有利于黄芩株高的生长;施磷肥可以促进黄芩根直径、根干重和产量的增加;磷钾配施可以显著提高根冠比和经济系数;单施磷肥最有利于黄芩根中黄酮和黄芩苷的累积。综合考虑各因素,氮磷钾配施优于单施和两两配施,N1P3K3(N 53.13 kg/hm2、P2O5224.89kg/hm2、K2O 224.89 kg/hm2)和N3P3K1(N 149.93 kg/hm2、P2O5224.89 kg/hm2、K2O 79.70 kg/hm2)为较优的两个施肥处理。     

Study of the chemical composition and antimicrobial activities of ethanolic extracts from roots of Scutellaria baicalensis Georgi

Scutellaria baicalensis Georgi (SBG), commonly named Huangqin, showed strong in vitro antimicrobial effects. However, limited research is available to systematically evaluate the effects of extraction methods on the phytochemical composition of SBG and its associated antimicrobial effects. In addition, limited studies have tested SBG as a natural antimicrobial agent on fresh produce such as tomatoes. In the current study, powered roots of SBG were extracted with 60, 80, and 100% ethanol, and their antiviral and antibacterial activities were evaluated. SBG ethanol extracts (SBGEEs) at 6.25 mg/mL showed limited antiviral activities against coliphage MS2 and hepatitis A virus. The SBG 80% ethanol extract (SBG80%EE) exhibited the lowest minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) against six foodborne pathogens. SBG80%EE had the highest contents of flavonoids and phenolic acids determined by high-performance liquid chromatography (HPLC). Among these bioactive compounds, ferulic acid had the lowest MIC and MBC values, 0.4 and 1.0 mg/mL, respectively, followed by baicalein and baicalin. Washing with SBG80%EE (12.5 mg/mL) resulted in >1 log reduction of Salmonella enterica serovars Typhimurium, Kentucky, Senftenberg, and Enteritidis on surface-inoculated grape tomatoes. None of SBGEE solutions changed the total phenolic content, color, or pH values of grape tomatoes. The quantification of these antimicrobial flavonoids and phenolic acids is important to maintain the quality and antimicrobial efficacy of SBG extracts. In addition, the application of SBG on tomatoes has provided valuable insights on the potential usage of this antimicrobial extract.     

黄芩分布区主要气候要素变化趋势时空特征分析

基于国内黄芩主要分布区确定的研究区内94个气象站点的气象观测数据,运用非参数统计检验Mann-Kendall、R/S等分析方法,分析了研究区1957-2007年气温、降水量和日照时数变化趋势的时空特征。结果表明：近50a来,研究区气温表现出显著的上升趋势,降水量、日照时数呈现出减少趋势,日照时数的变化趋势通过了0.01水平的显著性检验;Hurst指数分析表明,研究区未来气温、日照时数都表现出很强的持续性,即与过去近50a的变化趋势基本保持一致,降水量在东北段的减少持续性很强,华北段和西北段变化的持续性相对较弱;研究区气温、降水量、日照时数的变化趋势在空间分布上存在着一定的区域差异。     

Inhibitory effects of plant-derived flavonoids and phenolic acids on malonaldehyde formation from ethyl arachidonate

The antioxidant activities of naturally occurring plant compounds were measured in a lipid peroxidation system consisting of ethyl arachidonate and Fenton's reagent. Inhibitory effects of 24 plant-derived flavonoids and 5 phenolic acids on malonaldehyde (MA) formation from ethyl arachidonate were examined using gas chromatography (GC) with a nitrogen-phosphorus detector (NPD). Luteolin, which showed the strongest antioxidant activity, inhibited MA formation by 94% and 97% at the levels of 0.5 and 1.0 mM, respectively. The antioxidant activities of the flavones and flavonols decreased in the following order: luteolin > rhamnetin > fisetin > kaempferol > morin > quercetin. Among the flavanones tested, hesperitin, taxifolin, and naringenin exhibited appreciable antioxidant activities (61-84%) at the 1.0 mM level. The inhibitory effect of epigallocatechin gallate (82.5% at the 1.0 mM level) was the strongest among the flavan-3-ols tested. Ferulic acid had the most potent antioxidant activity (74.6% at the 1.0 mM level) of the phenolic acids tested.     

Epicatechin and catechin in cocoa inhibit amyloid beta protein induced apoptosis

To elucidate additional health benefits of cocoa phytochemicals on the neurotoxicity induced by amyloid beta protein (Abeta), PC12 cells were treated with toxic peptide (Abeta(25)(-)(35)) and the effects of epicatechin, catechin, and cocoa were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction, lactate dehydrogenase (LDH) release, and trypan blue exclusion methods. Significant increase in neuronal cell death was observed on PC12 cells treated with Abeta(25)(-)(35) (25 microM), while epicatechin and catechin and their mixture prevented the Abeta-induced neuronal cell death. Abeta treatment also led to the increased membrane instability of PC12 cells. The membrane protective effects of the phenolics determined by LDH release and trypan blue exclusion assays demonstrated that epicatechin, catechin, and their mixture protect cellular membrane from Abeta-induced cytotoxicity. In these three different cell viability assays, the mixture of epicatechin and catechin showed the highest protective effect and synergistic activity. The present results showed that the major flavonoids of cocoa, epicatechin and catechin, protect PC12 cells from Abeta-induced neurotoxicity, and suggest that cocoa may have anti-neurodegenerative effect in addition to other known chemopreventive effects.     

Inhibitory effect of red koji extracts on mushroom tyrosinase

Red koji has been recognized as a cholesterol-lowering diet supplement because of it contains fungi metabolites, monacolins, which reduce cholesterol synthesis by inhibiting HMG-CoA reductase. In this study, water extracts of red koji were loaded onto a C(18) cartridge, and the acetonitrile eluate was collected as test fraction. Red koji water extracts and its C(18) cartridge acetonitrile eluent had total phenols concentrations of 5.57 and 1.89 mg/g of red koji and condensed tannins concentrations of 2.71 and 1.20 mg/g of red koji, respectively. Both exhibited an antioxidant activity and an inhibitory activity to mushroom tyrosinase. The higher antioxidant activity of the red koji acetonitrile eluent was due to the existence of a high percentage of condensed tannins. The results from the kinetic study for inhibition of mushroom tyrosinase by red koji extracts showed that the compounds in the extracts competitively inhibited the oxidation of tyrosine catalyzed by mushroom tyrosinase with an ID(50) of 5.57 mg/mL.     

Inhibitory effect of protein hydrolysates on calcium carbonate crystallization

Protein hydrolysates, prepared by enzymatic digestion of soybean protein and egg white albumin using several proteases, inhibited the crystal growth of calcium carbonate. Each hydrolysate showed different inhibitory activities, suggesting the key role of peptide structures in the inhibition. The deamidation of protein hydrolysates by glutaminase increased not only the inhibitory activity toward the crystal growth of calcium carbonate but also the resistance of the hydrolysates against peptic digestion. Furthermore, the addition of sodium chloride, citric acid, or lactose into the reaction mixture enhanced the inhibitory activity. The protein hydrolysates inhibited both nucleation and crystal growth of calcium carbonate and also affected the crystal morphology.     

Inhibitory effect of low partial pressures of acetylene on nitrification

The inhibiting effect of C₂H₂ on nitrification was investigated in two agricultural soils. Nitrification was totally inhibited at 10 Pa partial pressure of C₂H₂ which is lower than previously reported for soils. There were no differences in rates of nitrate production between flasks without C₂H₂ and flasks with C₂H₂ at 0.01 Pa, while there was an effect on nitrification at 0.1 PaC₂H₂. At 0.1 Pa no inhibition was noted during the first 3 days; after this period nitrification was partially inhibited. The inhibiting effect did not cease until 7 days after removal of C₂H₂. The sensitivity of nitrification to low concentrations of C₂H₂ should be noted when denitrification rates are determined by the use of the acetylene inhibition method (usually C₂H₂ at 10kPa).     

Inhibitory effect of yuzu essential oil on the formation of N-nitrosodimethylamine in vegetables

The inhibitory effect of yuzu (Citrus junos Tanaka) essential oil on the formation of N-nitrosodimethylamine (NDMA) in the presence of vegetables (31 species) or saliva was investigated by HPLC. Most vegetable extracts enhanced the formation of NDMA. However, the formation ratio of NDMA in vegetable extracts was decreased by yuzu oil in the range of 59 to 22%. In the presence of yuzu oil and saliva, its ratio ranged between 62 and 24%. These results indicated that yuzu oil inhibited the formation of NDMA even in vegetables and saliva. The contents of ascorbic acid, nitrate, and nitrite in the 31 vegetable species were 0.3-65 mg/100 g, 3-581 mg/100 g, and 10-750 microg/100 g, respectively. Ascorbic acid and nitrite had little effect on the inhibition or formation of NDMA at their intact levels. Nitrate accelerated the formation of NDMA, and the addition of saliva further enhanced it. The mechanism of inhibition of NDMA formation by alpha-terpinene was studied. It was assumed from the results of LC-MS that a new compound formed by the reaction of alpha-terpinene with nitrite would be a derivative of alpha-terpinene with dinitroso groups. The molecular weight of this compound was 194. It is suggested that terpene hydrocarbons in citrus essential oils would contribute to the decrease of NDMA formation.     

Inhibitory effect of fruit extracts on the formation of heterocyclic amines

Natural extracts have attracted considerable attention for development into effective inhibitors against the formation of genotoxic heterocyclic amines (HAs) in processed foods. In this study, four fruit extracts (apple, elderberry, grape seed, and pineapple) were evaluated for their effects on HA formation in fried beef patties. Apple and grape seed extracts were found to be the most effective in both the degree of inhibition in the formation of individual HAs (2-amino-3,8-dimethylimidazo [4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx), and 2-amino-1-methyl-6-henylimidazo [4,5-b]pyridine (PhIP)) and in the reduction of total HA content (approximately 70% relative to the control). Activity-guided analysis of apple extract using model systems (PhIP- and MeIQx-producing models) showed that the proanthocyanidins, phloridzin and chlorogenic acid were responsible for reducing the amount of HAs formed. Proanthocyanidins were identified as the dominant inhibitors because they were strongly active against HA formation in both the PhIP and MeIQx model systems. For phloridzin, the inhibitory effect was observed only on the formation of PhIP. In contrast, chlorogenic acid, although effective against the formation of MeIQx, significantly enhanced the formation of PhIP. This is the first report showing the inhibitory activities of apple phenolics on the formation of heterocyclic amines. The findings provide valuable information for the development of effective strategies to minimize HA content of cooked meats and to identify several new natural products that may have new applications in the food industry.     

Inhibitory effect of betel quid on the volatility of methyl mercaptan

Betel quid, a popular natural masticatory in Taiwan, is mainly composed of fresh areca fruit, Piper betle (leaf or inflorescence), and slaked lime paste. People say that halitosis disappears during betel quid chewing. In this study, the removal of mouth odor during betel quid chewing was discussed by using a model system which measured its inhibition on the volatility of methyl mercaptan. Results showed that crude extracts of betel quid (the mixture of areca fruit, Piper betle, and slaked lime paste) and extracts of the mixture of areca fruit and slaked lime paste exhibited marked effects on the volatility of methyl mercaptan, and the inhibition function increased when increasing amounts of slaked lime paste were added. The same condition (increased inhibition) was also found by replacing the slaked lime paste with alkaline salts (calcium hydroxide, potassium hydroxide, or sodium hydroxide). Areca fruit, the major ingredient of betel quid, contained abundant phenolics. However, the crude phenolic extract of areca fruit did not show any inhibitory activity on the volatility of methyl mercaptan. Great inhibitory activity occurred only when the crude phenolic extract of areca fruit was treated with alkali. Further studies by using gel filtration determined that the effect probably came from the oxidative polymerization of phenolics of areca fruit after alkaline treatment.     

Interactions between flavonoids and proteins: effect on the total antioxidant capacity

Flavonoids are potent antioxidants. It is also known that flavonoids bind to proteins. The effect of the interaction between tea flavonoids and proteins on the antioxidant capacity was examined. Their separate and combined antioxidant capacities were measured with the Trolox equivalent antioxidant capacity (TEAC) assay. It was observed that the antioxidant capacity of several components of green and black tea with alpha-, beta-, and kappa-casein or albumin is not additive; that is, a part of the total antioxidant capacity is masked by the interaction. This masking depends on both the protein and the flavonoid used. Components in green and black tea, which show the highest masking in combination with beta-casein, are epigallocatechin gallate and gallic acid. The results demonstrate that the matrix influences the efficacy of an antioxidant.     

Inhibitory effect of blueberry polyphenolic compounds on oleic acid-induced hepatic steatosis in vitro

Nonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases worldwide and is closely associated with metabolic syndromes, such as obesity, diabetes, and insulin resistance. Nonalcoholic fatty liver (NAFL), also called simple steatosis, is the initial phase of NAFLD, which is accompanied the characteristic pathological overaccumulation of lipids without inflammation. To prevent NAFLD from reaching the NAFL stage through dietary therapy, in the present work, wild Chinese blueberries (Vacciniun spp.) were selected for their well-known benefits in inhibiting metabolic syndrome. After being purified from wild Chinese blueberries, polyphenol-rich extracts were subsequently separated into three fractions, namely, anthocyanin-rich fraction, phenolic acid-rich fraction, and ethyl acetate extract. The inhibition of oleic acid (OA)-induced triglyceride (TG) deposition in HepG 2 cells was referred to as the potential activity of preventing NAFL. Biochemical indicators, such as cytotoxicity, TG level, levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and intracellular reactive oxygen species, were used to evaluate the analogous pathological stage of NAFLD. The results show that OA ≤ 1.0 mM exhibits a dose-dependent induction of TG accumulation, and no inflammation was observed based on the changes in ALT and AST levels. Therefore, 1.0 mM OA was used to simulate an in vitro fatty liver. Blueberry polyphenol-rich extract efficiently inhibited OA-induced TG accumulation in HepG2 cells, and the phenolic acid-rich fraction performed efficiently. Seven phenolic acids were subsequently identified using a high-performance liquid chromatography assay, and the main types were caffeic, chlorogenic, ferulic, p-coumaric, and cinnamic acids. These phenolic acid standards also displayed good efficiency in inhibiting TG accumulation in HepG2 cells. These results imply that wild Chinese blueberries have a potential preventive effect on NAFLD in its early stage, and phenolic acids are the most efficient component.     

生物质炭介导生防微生物抑制辣椒疫霉的作用

生物质炭可有效防控土传病害,筛选并鉴定出生物质炭介导下的生防微生物,可为研究生物质炭防病机理和强化生物质炭防病效果提供理论依据。本研究首先进行秸秆生物质炭防控辣椒疫病盆栽试验,利用定量PCR和平板计数明确生物质炭在防控辣椒疫病时可富集的已知生防微生物,再通过选择性培养基初筛和定殖复筛筛选出生物质炭可富集的潜在生防微生物菌株,最后研究各菌株在土壤中对辣椒疫霉的抑制作用。结果表明,秸秆生物质炭使根际辣椒疫霉数量显著降低95.1%、辣椒疫病发生率显著降低91.1%,并使具有生防功能的木霉菌、青霉菌、曲霉菌、芽孢杆菌、假单胞菌和鞘氨醇单胞菌数量显著增加2.22倍、4.09倍、3.89倍、2.45倍、1.45倍和1.30倍。通过平板初筛得到可能被生物质炭富集的22株潜在生防菌株。定殖复筛剔除部分假性生物质炭介导菌株,获得可明确被生物质炭富集的2株木霉菌、3株青霉菌、2株曲霉菌、3株芽孢杆菌、3株假单胞菌、3株链霉菌和2株鞘氨醇单胞菌。木霉菌（TR1和TR3）、青霉菌（PE1）、曲霉菌（AS1和AS2）、芽孢杆菌（BA1、BA2和BA3）、假单胞菌（PS1和PS3）、链霉菌（ST1、ST4和ST5）13个菌株可显著削减土壤辣椒疫霉数量。其中,所有木霉菌和曲霉菌菌株（TR1、TR3、AS1和AS2）及芽孢杆菌（BA1和BA2）、假单胞菌（PS1和PS3）和链霉菌（ST1）9个菌株与生物质炭具有显著的协同抑制辣椒疫霉效果。因此,防控辣椒疫病时,木霉菌、曲霉菌、芽孢杆菌、假单胞菌和链霉菌是生物质炭介导下的主要防病微生物。