鄂西北地区不同生长条件和品种柴胡中多糖含量比较

目的 测定鄂西北地区不同生长条件和品种柴胡中多糖的含量.方法采用蒽酮 硫酸比色法,以620 nm波长为检测波长,分别测定柴胡中可溶性多糖和粗多糖,计算总多糖含量.结果鄂西北地区野生竹叶柴胡多糖含量高于同地区种植柴胡,但低于山西产北柴胡和湖北南漳种植的三岛柴胡.结论不同生长条件的柴胡药材中柴胡多糖含量不同,但差异不明显;不同品种柴胡药材中的多糖含量有明显差异.     

鄂西北地区不同生长条件和品种柴胡中总皂苷含量比较

目的:比较鄂西北地区不同生长条件柴胡中柴胡总皂苷的含量。方法:使用可见分光光度法。结果:不同产地、不同品种柴胡中总皂苷含量有一定差异,同一区域种植和野生柴胡总皂苷含量无明显差异。结论:实行中药现代化必须大力推进规范化种植(GAP),鄂西北地区具有进行柴胡GAP种植的地理条件。     

鄂西北地区不同采收期竹叶柴胡根及地上部分总黄酮含量动态变化研究

目的：以鄂西北地区房县竹叶柴胡为样本,研究柴胡地下和地上部分总黄酮的动态变化规律,探讨竹叶柴胡的最佳采收期.方法：以柴胡总黄酮的含量为指标,采用紫外-可见光光度法进行分析,考察不同采收期柴胡中总黄酮的变化规律.结果：竹叶柴胡根中总黄酮含量随生长时间逐渐升高,6～11月总黄酮含量分别是0.394 7%,0.298 4%,0.578 9%,0.531 7%,0.952 3%,0.946 2%;地上部分总黄酮含量没有一定的变化趋势,但在6月起苔时和10月种子成熟时总黄酮含量最高,6～10月总黄酮含量分别是2.908 7%,2.336 0%,2.722 3%,2.160 8%,2.969 5%.综合分析,鄂西北地区竹叶柴胡的最佳采收期应在种子成熟期的10月.结论：确定了鄂西北地区竹叶柴胡的最佳采收期,为鄂西北地区柴胡药材的规范化种植（GAP）提供了一定的参考依据.     

醋柴胡的质量研究

目的:探讨柴胡醋炙前后主要有效成分的变化。方法:采用HPLC方法,测定柴胡及其醋炙柴胡中柴胡皂苷a和d的含量。结果:4种柴胡与其醋炙柴胡中柴胡皂苷a含量的变化率为-59.57%～-25.08%,柴胡皂苷d含量的变化率为-63.59%～-18.43%。结论:柴胡醋炙后,其中柴胡皂苷a、d含量显著降低。     

鄂西北地区不同生长条件柴胡中柴胡皂苷含量比较研究

目的:比较鄂西北地区不同生长条件柴胡中柴胡皂苷含量.方法:以Waters Atlantis(R)T3色谱柱、乙腈-水为流动相进行梯度洗脱,检测波长为210 nm.结果:柴胡皂苷a、d的线性回归方程分别为YSSa=6.042 5X+0.420 2(r=1.000 0),YSSd=4.787 2X+0.392 6(r=1.000 0).结论:鄂西北地区柴胡皂苷含量较高,种植柴胡与野生柴胡皂苷含量无明显差异,有利于进行柴胡GAP规范化种植.     

不同种植模式对延胡索产量及品质形成的影响

目的 研究不同种植模式下延胡索氮、磷、钾元素累积与分配对其产量、品质形成的影响,寻求不同种植模式下延胡索产量差异形成的因素。方法 试验设置延胡索单作、延胡索与水稻复种、延胡索与玉米复种3个处理组,通过测定不同种植模式下延胡索的农艺学性状、光合性能、生物量、产量及有效成分延胡索乙素含量,同时测定植株中各部位氮、磷、钾元素含量,分析植株氮、磷、钾元素含量对延胡索生长发育及产量品质的影响。结果 延胡索在复种条件下,其农艺性状、光合性能、收获时生物量、产量及有效成分含量更具优势,其中延胡索与水稻复种组效果最佳。延胡索植株的氮元素含量与块茎体积、产量、总生物量及净光合速率均呈显著正相关性;植株磷元素与总生物量成显著正相关;植株钾元素与延胡索乙素含量及净光合速率成显著正相关。结论 复种通过影响氮、钾在植株中的累积与分配,能显著提高延胡索农艺学性状、光合性能及药材的产量与品质。推荐在水田多采用水稻-延胡索复种模式、在旱地多采用玉米-延胡索复种;可以在全国的山区范围内推广玉米-延胡索复种的种植模式。     

产地对北柴胡皂苷类物质含量及急性毒性影响的实验研究

目的对不同产地北柴胡药材全组分、醇-水提取物中皂苷类物质的含量和急性毒性大小进行比较研究,探讨产地对柴胡皂苷类物质含量与毒性的影响。方法运用UV法、HPLC法、经典急性毒性实验法,进行北柴胡药材全组分、醇-水提取物柴胡总皂苷和皂苷a的含量测定及急性毒性比较研究。结果不同产地北柴胡药材全组分柴胡总皂苷含量大小依次为：河北柴胡〉内蒙柴胡〉山西柴胡〉河南柴胡,皂苷a的含量为：河北柴胡〉内蒙柴胡、山西柴胡〉河南柴胡,其最大给药量（MLD）大小为：河南柴胡〉山西柴胡〉内蒙柴胡〉河北柴胡;不同产地北柴胡药材醇-水提取物总皂苷含量大小依次为：河北柴胡〉内蒙柴胡〉山西柴胡〉河南柴胡,皂苷a的含量为：河北柴胡〉山西柴胡〉内蒙柴胡〉河南柴胡,其最小致死量（MTD）为：内蒙柴胡〉山西柴胡〉河南柴胡〉河北柴胡。河北产地的北柴胡药材全组分、醇-水提取物中柴胡总皂苷及皂苷a含量均较其他产地北柴胡药材含量高,其对应的全组分MLD、醇-水提取物MTD均较其他产地北柴胡药材的小,毒性最大。结论产地对柴胡皂苷类物质含量与毒性有一定的影响,且柴胡毒性大小与皂苷类物质含量有一定的相关性。     

HPLC法同时测定柴胡与春柴胡中皂苷类及黄酮类成分的含量

目的:建立HPLC法同时测定柴胡和春柴胡中2个皂苷类活性成分(柴胡皂苷a和柴胡皂苷d)、4个黄酮类活性成分(芦丁、槲皮素、山柰素和异鼠李素)的含量。方法:采用Agilent Zorbax C18(4.6 mm×250 mm,5μm)色谱柱,以0.1%磷酸水-乙腈为流动相,梯度洗脱,流速1 mL.min-1,检测波长203 nm(测定皂苷类成分)和360 nm(测定黄酮类成分),柱温35℃。结果:6个活性成分的峰面积与浓度的线性关系良好(r≥0.9994),线性范围宽;精密度试验的RSD为0.67%~1.9%;加样回收率为95.1%~104.3%。10批市售柴胡和春柴胡的黄酮类和皂苷类成分含量具显著差异:春柴胡中4个黄酮类成分平均含量约为柴胡中的7.5~72倍;春柴胡中柴胡皂苷a的平均含量为柴胡的1/10,而柴胡皂苷d的平均含量约为柴胡的3倍;柴胡与春柴胡中6个成分含量的相关系数为0.10~0.62。结论:本方法具良好的精密度、重复性和可靠性,可同时测定柴胡和春柴胡中皂苷类和黄酮类成分的含量并有效表征其差别。     

柴胡中柴胡总皂苷及柴胡皂苷a的含量测定

目的 建立柴胡中柴胡总皂苷及柴胡皂苷a的含量测定方法,并对样品进行含量测定.方法 采用紫外分光光度法,以柴胡皂苷a为对照品,在波长545nm处对样品中的总皂苷进行含量测定;采用高效液相色谱法,以C18色谱柱(4.6mmX250mm.5μm)、甲醇-水为流动相,流速为1mL·min<'-1>,检测波长为210nm,测定样品中柴胡皂苷a的含量.结果 柴胡总皂苷在30～70μg/mL、柴胡皂苷a在50.4～252μg/mL范围内呈良好的线性关系,r分别为09956、0.9991;平均回收率:柴胡总皂苷为99.31%、柴胡皂苷a为99.22%;RSD:柴胡总皂苷为1.25%,柴胡皂苷a为1.07%.结论 本研究所建立的紫外分光光度法测定柴胡总皂苷及高效液相法测定皂苷a含量的方法,简便、易行、快速,结果准确可靠,适用于柴胡中柴胡总皂苷及柴胡皂苷a的含量测定,并为在安全剂量范围内正确使用柴胡提供依据.     

大叶柴胡的毒性成分

从伞形科植物大叶柴胡Bupleurum logiradiatum Turcz.的根及根茎中分得四种新的多烯炔类化合物,分别命名为柴胡毒素(bupleurotoxin,Ⅰ)柴胡酮醇(bupleuonol,Ⅱ),乙酰柴胡毒素(acetyl-bupleufotoxin,Ⅲ)和柴胡炔醇(bupleurynol,Ⅳ)。根据理化性质和光谱数据确定柴胡炔醇(Ⅳ)为十七碳三烯-(2z,8 E,10 E)-二炔-(4,6)-醇-1;柴胡毒素(Ⅰ)为14-羟基柴胡炔醇;乙酰柴胡毒素(Ⅲ)为14-乙酰氧基柴胡炔醇;柴胡酮醇(Ⅱ)为14-羰基柴胡炔醇。柴胡毒素(Ⅰ)和乙酰柴胡毒素(Ⅲ)有毒性。     

Characterization of a basic phospholipase A2-homologeu myotoxin isolated from the venom of the snake Bothrops neuwiedii (yarará chica) from Argentina.

A basic protein was isolated by CM-Sephadex C-25 chromatography from the venom of Bothrops neuwiedii from Argentina, and named B. neuwiedii myotoxin I. This protein exerted local myotoxic and edema-forming effects in mice, with potencies comparable to other myotoxins isolated from Bothrops spp. venoms. When injected by i.v. route at doses up to 4.7 mg/kg of body weight, the toxin was not lethal. In vitro, the toxin had no detectable phospholipase A₂ activity on egg yolk phospholipids. B. neuwiedii myotoxin I appeared as a homodimer in sodium dodecylsulphate–polyacrylamide gel electrophoresis, with a subunit molecular weight of 15 kD. Gel immunodiffusion revealed a pattern of partial antigenic identity between the newly isolated myotoxin and myotoxin II from Bothrops asper venom. The sequence of B. neuwiedii myotoxin I was determined for the first 40 amino acid residues, showing high homology to several class II phospholipase A₂ myotoxins of the Lys-49 family from crotalids. Altogether, results suggest that this toxin is a new member of the Lys-49 phospholipase A₂-homologues with myotoxic, cytolytic, and edema-inducing activities.     

A study on the venom yield of venomous snake species from Argentina

A study on the venom yield of snakes from Argentina over a three year period was carried out on adult specimens of Bothrops alternatus (n=74); Bothrops neuwiedii (n=127); Bothrops ammodytoides (n=30); Bothrops moojeni (n=14); Bothrops jararaca (n=14); B. jararacussu (n=6); Crotalus durissus terrificus (n=120) and Micrurus spp. (n=6) as well as with 12 specimens of newborn C. d. terrificus kept in captivity. While for each species there was a positive correlation between venom yield and number of snakes milked, the correlation with the snake's body weights after individual milkings was even better, suggesting that the size of the snakes is more important in determining the venom yield than the number of snakes milked or the specimen's sex. Individual milkings indicated that, in addition to the snake size, when the amount of venom is normalized per 100 g body weight there is a species specific difference in venom yield. It follows the order B. jararacussu>B. moojeniB. jararacaB. alternatus>B. neuwiedii>Micrurus sppB. ammodytoides>C. d. terrificus. Although the venom yield per 100 g body weight of newborn C. d. terrificus specimens is 2-fold higher than that of adults, no correlation was observed between venom yield and body weight.     

Neutralizing potency of horse antibothropic Brazilian antivenom against Bothrops snake venoms from the Amazonian rain forest

Neutralization of lethal toxicity (50% effective dose; ED₅₀), hemorrhagic (minimum hemorrhagic dose; MHD) and hemolytic activity (PLA₂) and levels of antibodies, measured by enzyme-linked immunosorbent assay (ELISA), were investigated to test the potency of horse antibothropic serum (ABS) against Bothrops venoms from the Amazonian rain forest. ABS neutralized the lethal activity with a potency (mg of venom neutralized per 1 ml of antivenom) of 5.5, 3.7, 1.6, 1.3 and 6.5, respectively, for B. jararaca (reference venom for assessing the ABS potency in Brazil), B. atrox, B. brazili, B. bilineatus smaragdinus and B. taeniatus venoms. The volume of antivenom (μl) that neutralized one MHD of B. jararaca, B. atrox, B. brazili, B. bilineatus smaragdinus and B. taeniatus venoms was 5, 7.71, 7.76, 8.3 and 5, respectively. ABS neutralized the PLA₂ activity with a potency of 6.2, 3.2, 1.4, 2.6 and 5 respectively, for B. jararaca, B. atrox, B. brazili, B. bilineatus smaragdinus and B. taeniatus venoms. ELISA reactivity of ABS against the separate venoms was found to be quite variable. The reactivity against B. jararaca venom was higher than against other Bothrops venoms. In conclusion, the assays described here suggest that Brazilian Bothrops polyspecific antivenom is not very efficient in neutralizing the effects of venom from some Amazonian Bothrops species.     

Neutralization of myotoxic activity of Bothrops venoms by antisera to purified myotoxins and to crude venoms

A phospholipase myotoxin (MOO-1) and a non-phospholipase myotoxin (JSU-5) were studied for their antigenic cross-reactivity and neutralization by different antisera. Antisera against JSU-5 and MOO-1 reacted equally with both myotoxins in ELISA assays. The specificity of these antisera was also similar, recognizing the same 14,000–18,000 mol. wt components in the venoms of Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi and Bothrops pradoi. Using creatine kinase assays, JSU-5 myotoxicity was completely neutralized by B. jararacussu antivenom or anti-JSU-5 antibodies and partially neutralized by B. moojeni antivenom or anti-MOO-1 antibodies. MOO-1 myotoxicity was completely neutralized by antisera against JSU-5 and MOO-1 and B. jararacussu antivenom, and only partially neutralized by B. moojeni antivenom. B. jararacussu venom induced high titres of antibodies against purified myotoxins. This antiserum completely inhibited the myotoxicity of the homologous venom and significantly reduced the myotoxicity of the remaining myotoxin-containing venoms. It is suggested that B. jararacussu venom is a good immunogen to induce antibodies against myotoxins present in the venoms of the different species of Bothrops.     

Molecular mechanisms of the metabolite 4-hydroxytamoxifen of the anticancer drug tamoxifen: use of a model microorganism

A strain of the thermophilic eubacterium Bacillus stearothermophilus was used as a model system to identify membrane mediated cytotoxic effects of 4-hydroxytamoxifen, following previous studies with tamoxifen. With this experimental approach we attempted to further clarify tamoxifen and 4-hydroxytamoxifen membrane interactions often evoked as responsible for their multiple cellular effects. Bacterial growth and the oxygen consumption rate provided quantitative data of the cytotoxic action of hydroxytamoxifen. The effects of hydroxytamoxifen on the physical properties of bacterial lipid membrane preparations were also evaluated by fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene. Cultures of B. stearothermophilus grown in a complex medium containing hydroxytamoxifen in the concentration range of 1 to 7 μ exhibited progressively longer lag adapting periods, decreased specific growth rates and lower growth yields, as compared to control cultures. Hydroxytamoxifen also affected the electron redox flow of B. stearothermophilus protoplasts and induced significant perturbation of the structural order of bacterial lipid dispersions. We concluded that the bacterial model provides useful information about the nature and repercussion of membrane physical interactions of this lipophilic drug, on the basis of an easy and economic methodology.     

Neutralization of proteases from Bothrops snake venoms by the aqueous extract from Casearia sylvestris (Flacourtiaceae).

Aqueous extract from Casearia sylvestris leaves, a typical plant from Brazilian open pastures, was able to neutralize the hemorrhagic activity caused by Bothrops asper, Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi and Bothrops pirajai venoms. It also neutralized two hemorrhagic metalloproteinases from Bothrops asper venom. Proteolytic activity on casein induced by bothropic venoms and by isolated proteases, including Bn2 metalloproteinase from B. neuwiedi venom, was also inhibited by the C. sylvestris extract in different levels. The -fibrinogen chain was partially protected against degradation caused by B. jararacussu venom, when this venom was incubated with C. sylvestris extract. We also observed that this extract partially increased the time of plasma coagulation caused by B. jararacussu, B. moojeni and B. neuwiedi venoms. C. sylvestris extract did not induce proteolysis in any substrate assayed.     

赤红壤在不同农作制中养分平衡的研究

在广州的赤红壤上进行六种农作制的五年试验表明,所有衣作制的氮平衡均盈余,其中水稻连作的农作制氮盈余较少,而含豆科作物者尤其是牧草大绿豆混播及双季稻冬种绿肥盈余最多,后者年亩增加氮素约10kg;各农作制氮盈余的原因是具有较高的氮肥效率和土壤全氮量显著增加。磷平衡在所有农作制中皆为亏损;磷肥效率低,多数衣作制投入的磷约有1/3移至耕层以下。钾平衡在含水稻的衣作制中亏损艮大,旱作者接近平衡或盈余;钾亏损的原因是含水稻的农作制钾的淋失量大,平均达投入的44％:土壤经种植五年后钾含量一般只能维持原有水平。养分平衡状况表明,供试六种衣作制中,油一茨和牧草混豆科作物两个农作制最有利于养分平衡,亏损较少,其次是油—稻和稻—肥两个农作制亦较有利于养分平衡。试验还说明稻秆和豆科作物基秆回田是维持农作制中氮平衡、钾平衡的重要手段。     

In vitro toxicity of cereulide on porcine pancreatic Langerhans islets

Cereulide is a K⁺ ionophore cytotoxic and mitochondriotoxic to primary cells and cell lines of human and other mammalian origins. It is a heat-stable, highly lipophilic (log K_ow 5.96) peptide (1152 g mol⁻¹) produced by certain strains of Bacillus cereus, a bacterium connected to emetic food poisonings. In this study the pancreatic toxicity of purified cereulide, and cereulide-containing bacterial extracts, was studied using fetal porcine Langerhans islets in culture. Exposure to 1 ng ml⁻¹ of purified cereulide caused necrotic cell death of the islet cells impairing their insulin content within 2 days. Cell extracts of cereulide-positive B. cereus strains connected to food poisoning or isolated from foodstuffs were toxic, corresponding to their measured cereulide content. Extracts of B. cereus strains producing or not producing the B. cereus diarrheal toxin, but no cereulide, were tolerated by the porcine islet cultures up to concentrations 1000-fold higher compared to extracts from strains containing cereulide, and up to exposure times of 7 d. Cereulide thus was identified as the B. cereus-produced substance toxic towards porcine fetal Langerhans islets and beta cells.     

Collagenolytic activity of snake and spider venoms

The venoms of the snakes B. atrox, B. Jararaca, C. durissus, A. piscivorus, and the spiders P. fera, L. erythrognatha possess proteolytic activity which attacks gelatine, casein and azocoll. Denatured (heat, urea) collagen is digested too. However, native collagen is not attacked by the enzymes present in the venoms investigated. Following subcutaneous injection of A. piscivorus venom, a statistically significant increase in urinary hydroxyproline excretion occurred. This finding indicates a degradation of collagen by animal venoms in vivo. The mechanism of this action remains unknown.