瑞香狼毒根部烷烃类提取物GC-MS分析及抑瘤活性初探

通过正己烷提取瑞香狼毒根部成分,采用气相色谱-质谱联用技术对瑞香狼毒根部烷烃类提取物(SRH)的成分进行分析,同时考察SRH对H22皮下荷瘤小鼠的抑瘤作用。结果表明:通过GC-MS分析,SRH共分离得到55个组分,结构确定的有44个,占总峰面积的97.73%,其中烃类化合物11个,芳香族化合物10个,酸类化合物有9个,酯类化合物有7个,醇类化合物有4个,其他类型化合物有3个。其中含量在1%以上的物质有(Z,Z)-9,12-十八烷二烯酸,邻苯二甲酸-2-乙基己酯,9,17-十八烷二烯,反式角鲨烯,γ-谷甾醇,邻苯二甲酸二丁酯,n-十六酸,硬脂酸,二十酸,菜油甾醇,占总峰面积90.74%,检测出的角鲨烯,亚油酸,菜油甾醇都具有一定的抗肿瘤活性,在瑞香狼毒抗肿瘤方面的研究未见报道;同时SRH在实验前期对肿瘤的生长起到了很好的抑制作用,尤其是SRH低剂量,其抑制率达到21.5%。该结果为瑞香狼毒根部成分的物质分析及抑瘤应用前景提供了一定的理论依据。     

瑞香狼毒根部烷烃类提取物GC-MS分析及抑瘤活性初探北大核心CSCD

通过正己烷提取瑞香狼毒根部成分,采用气相色谱-质谱联用技术对瑞香狼毒根部烷烃类提取物(SRH)的成分进行分析,同时考察SRH对H22皮下荷瘤小鼠的抑瘤作用。结果表明:通过GC-MS分析,SRH共分离得到55个组分,结构确定的有44个,占总峰面积的97.73%,其中烃类化合物11个,芳香族化合物10个,酸类化合物有9个,酯类化合物有7个,醇类化合物有4个,其他类型化合物有3个。其中含量在1%以上的物质有(Z,Z)-9,12-十八烷二烯酸,邻苯二甲酸-2-乙基己酯,9,17-十八烷二烯,反式角鲨烯,γ-谷甾醇,邻苯二甲酸二丁酯,n-十六酸,硬脂酸,二十酸,菜油甾醇,占总峰面积90.74%,检测出的角鲨烯,亚油酸,菜油甾醇都具有一定的抗肿瘤活性,在瑞香狼毒抗肿瘤方面的研究未见报道;同时SRH在实验前期对肿瘤的生长起到了很好的抑制作用,尤其是SRH低剂量,其抑制率达到21.5%。该结果为瑞香狼毒根部成分的物质分析及抑瘤应用前景提供了一定的理论依据。     

瑞香狼毒提取物对试验动物急性毒性及活性的初步研究

采用急性经口、眼刺激、皮肤试验方法,研究了瑞香狼毒乙酸乙酯萃取物的急性毒性作用,同时以番茄晚疫病菌为供试菌,研究了瑞香狼毒提取物的抑菌活性.初步研究表明,瑞香狼毒乙酸乙酯萃取物对小白鼠经口急性毒性为低毒,对白兔的急性皮肤刺激属无刺激性,对白兔的眼刺激属轻度刺激性,对鲫鱼的毒性属于中毒级;瑞香狼毒乙酸乙酯萃取物及其分段物流分B、c、D对番茄晚疫病菌有较好的抑制活性,这为瑞香狼毒乙酸乙酯萃取物进一步开发为产品提供了毒理学依据.     

瑞香狼毒根的抑菌活性研究（Ⅰ）

以苹果干腐病菌、小麦赤霉病菌、番茄早疫病菌、南瓜枯萎病菌、玉米大斑病菌、烟草赤星病菌和辣椒疫霉病菌为供试菌,采用生物活性跟踪法、生长速率测定法和系统溶剂提取法对瑞香狼毒根中的杀菌活性物质进行了筛选和分离。结果表明,杀菌活性物质主要集中在乙酸乙酯提取物和甲醇粗提物中。从乙酸乙酯提取物中分离出两种杀菌活性物质AF1-4和AF1-5,同时还发现瑞香狼毒根中还存在增菌物质。     

瑞香狼毒根中活性物质的分离鉴定及作用机理

以瑞香狼毒根为研究材料,模式植物拟南芥为受体,采用实验室活体生物实验方法研究了瑞香狼毒根提取物及不同极性溶剂萃取物对7 d龄拟南芥幼苗的生长抑制作用;采用活性跟踪的化合物分离方法,分析了其中的活性化合物成分,并通过拟南芥DR5-GUS转基因株系,研究了单体化合物对拟南芥生长发育及根系生长素分布的影响.结果显示,瑞香狼毒根乙醇提取物对拟南芥有很强的生长抑制作用,其乙酸乙酯和氯仿萃取物的抑制活性最为显著,从氯仿萃取物中分离得到两种香豆素类化合物伞形花内酯(1)和西瑞香素(2),其中化合物1能够显著抑制拟南芥幼苗生长及根系发育,且明显降低了根部内源生长素的分布水平;化合物2也有较为明显的抑制活性.研究表明,氯仿和乙酸乙酯萃取物是瑞香狼毒植物毒活性的主要有效部位;伞形花内酯(1)和西瑞香素(2)是瑞香狼毒植物毒活性的有效成分,化合物1对拟南芥生长发育的抑制作用与生长素途径密切相关.     

青藏高原有毒植物瑞香狼毒根抑菌活性初步研究

以人体病原细菌大肠杆菌、金黄色葡萄球菌、深部真菌白色念珠菌、浅部真菌红色毛癣菌、石膏样毛癣菌、石膏样小孢子菌、絮状表皮癣菌为供试菌,采用药敏纸片法对瑞香狼毒活性成分的抑菌活性进行了初步研究。结果表明瑞香狼毒对人体常见病原细菌和真菌有抑制作用,乙酸乙酯萃取物在33．33g／L浓度下对细菌组的抑菌圈直径最高可达12．02mm,对真菌组最高可达9．04mm,并初测瑞香狼毒中的抑菌活性物质主要为中偏强极性成分。     

狼毒活性部位复合乳油功效

为增强瑞香狼毒制剂的杀虫药效、充分开发利用以瑞香狼毒为主的植物源农药,进行了瑞香狼毒杀虫活性部位与其它药物的复配实验。采用瑞香狼毒杀虫活性部位(LD)分别与烟碱乳油(YJR)、CT-901A(Bz)的不同质量比,在室内对家蝇、酢酱草茹叶螨(Petrbia hartiEwing)进行毒力测定,利用软件进行毒力回归分析。实验结果显示,LD和YJR、LD和Bz的两两复配及三者(LD YJR Bz)的复配能够起显著增效作用,特别是20:1:1(LD:YJR:Bz)、30:1:1(LD:YJR:Bz)对家蝇的毒力效果更加明显,LC50达到了0.005 mg/L,20:1(LD:Bz)、30:1(LD:Bz)、30:1:1(LD:YJR:Bz)对酢酱草茹叶螨(Petrbia hartiEwing)的毒力效果也同样显著,LC50也达到了0.005 mg/L。并对瑞香狼毒活性部位进行初步分离检测,其有效成分为黄酮类物质。     

退化草地瑞香狼毒对小尺度群落组成及土壤理化性质的影响

过度放牧导致草原生态系统呈现明显的退化趋势,草原优良牧草减少,逐渐转变成以有毒植物为优势种的退化草原。作为草地退化的指示种,有毒植物对邻近植物及土壤环境的影响一直是生态学研究的重要科学问题之一。为了更好地探究有毒植物在退化草地生态系统恢复过程中的影响,本研究以海拉尔自然保护区内封育恢复中的羊草草原为研究对象,依据群落中瑞香狼毒(Stellera chamaejasme)枝条数将植物群落划分为3个水平:高密度瑞香狼毒植物群落、低密度瑞香狼毒植物群落、无瑞香狼毒植物群落,调查群落物种组成并测定了土壤理化指标。结果显示:(1)高密度瑞香狼毒小尺度范围内改变了群落物种组成,提高Shannon指数和Patrick指数,说明在退化草地恢复过程中瑞香狼毒的存在能够改变群落物种丰富度进而改变其物种多样性。(2)随着瑞香狼毒密度的增加,部分禾本科植物与瑞香狼毒的联结性质也发生了变化。早熟禾(Poa annua)、贝加尔针茅(Stipa baicalensis)与瑞香狼毒的联结性质由正联结变为负联结;洽草(Koeleria litvinowii)由无联结变为负联结;羊草(Leymus chinensis)、糙隐子草(Cleistogenes squarrosa)、羽茅(Achnatherum sibiricum)未发生改变。(3)瑞香狼毒能够改变群落中各科相对重要性。随着瑞香狼毒密度增加,豆科、蔷薇科重要值增大,菊科重要值降低,禾本科无明显变化。(4)瑞香狼毒对土壤全氮、pH有一定的影响。其中,低密度瑞香狼毒群落的土壤全氮含量显著高于其他两组,其土壤pH也显著高于无瑞香狼毒存在的对照组。土壤全碳、速效态的氮和磷在不同密度瑞香狼毒群落之间无明显差异。土壤理化性质小尺度的变化可能是推动退化草地植物恢复的基础。     

p53蛋白在周期调节蛋白A1变异引起的雄性小鼠生殖细胞凋亡中的作用

为研究p5 3蛋白在周期调节蛋白A1(cyclinA1)变异引起的雄性小鼠生殖细胞凋亡中的作用 ,以p5 3基因敲除的小鼠和周期调节蛋白A1基因敲除的小鼠杂交 ,获取同胎生单基因变异和双基因同时变异的雄性后代共 4组 12只 .比较它们的性腺和生殖细胞发育 ,并用TUNEL染色法观察和比较生殖细胞的凋亡情况 .在睾丸最大横切面上观察到 :周期调节蛋白A1变异组凋亡细胞最多 (348± 10 4个 ) ,明显高于p5 3 周期调节蛋白A1双基因变异组 (12 1± 38个 ) ,t=3 2 5 79,P =0 0 4 72 .p5 3变异组凋亡细胞最少 (45± 2 4个 ) ,配对t检验显示有非常显著性差异 ,t=8 4 0 13,P =0 0 0 35 .这一研究结果提示 ,p5 3基因可能在雄性生殖细胞的发育中起监视作用 ,并在周期调节蛋白A1变异引起发育异常时启动p5 3途径造成异常细胞的凋亡 .     

噬菌蛭弧菌代谢产物活性成分对小鼠免疫功能的影响

目的研究噬菌蛭弧菌代谢产物活性成分对小鼠免疫功能的影响。方法将噬菌蛭弧菌代谢产物的3种有机溶剂（石油醚、三氯甲烷、乙酸乙酯）提取物和提取后的剩余液体分别腹腔注射实验小鼠,分两次注射,共注射0.5 mL,注射后连续饲养28 d,每隔7 d小鼠采血检测离体白细胞吞噬活性（phagocytic activity）、吞噬细胞杀菌活性（bactericidal activity）、超氧化物歧化酶（superoxide dismutase,SOD）活性、血清凝集抗体效价、血红蛋白值和红细胞数的变化,以研究不同提取物对小鼠免疫功能的影响。结果石油醚提取物组和三氯甲烷提取物组小鼠血清SOD活性、血清凝集抗体效价、离体白细胞吞噬活性和吞噬细胞杀菌活性增强与对照组相比差异极显著（P〈0.01）;乙酸乙酯提取物组SOD活性和血清凝集抗体效价增强与对照组相比差异极显著（P〈0.01）,离体白细胞吞噬活性和吞噬细胞杀菌活性增强与对照组相比差异显著（P〈0.05）;石油醚提取物组的血红蛋白值（12.64 g/100 mL）和红细胞数（11.32×106/mL）最高。各项指标的峰值均出现在第7天~第21天。结论噬菌蛭弧菌代谢产物3种有机溶剂提取的活性物质具有增强小鼠免疫功能的作用。     

Cage size preference in rats in the laboratory

The size of an enclosure is an integral part of how well it accommodates a nonhuman animal's welfare; however, most enrichment studies concentrate on modifying the area inside the enclosure rather than enlarging it. It has been suggested that rats have little need for more cage space, but there is no empirical evidence about rats' need for space. This experiment provides preliminary evidence for the preferences of 5 male and 5 female albino rats using T-maze choices followed by 5 min dwelling times. The rats showed a moderate but significant preference for the larger of 2 cages [540 cm2 vs. 1,620 cm2; binomial z, p <.05]. When the rats shared the chosen cage with 4 familiar cage mates, their preference for the larger cage did not become any stronger [paired t[9] = -.820, p >.05]. The results suggest that rats should be given a somewhat larger space allowance but could share it with up to 4 other rats.     

Intrathecal substance P augments morphine-induced antinociception: Possible relevance in the production of substance P N-terminal fragments

The present study sought to examine the mechanism of substance P to modulate the antinociceptive action of intrathecal (i.t.) morphine in paw-licking/biting response evoked by subcutaneous injection of capsaicin into the plantar surface of the hindpaw in mice. The i.t. injection of morphine inhibited capsaicin-induced licking/biting response in a dose-dependent manner. Substance P (25 and 50 pmol) injected i.t. alone did not alter capsaicin-induced nociception, whereas substance P at a higher dose of 100 pmol significantly reduced the capsaicin response. Western blots showed the constitutive expression of endopeptidase-24.11 in the dorsal and ventral parts of lumbar spinal cord of mice. The N-terminal fragment of substance P (1–7), which is known as a major product of substance P by endopeptidase-24.11, was more effective than substance P on capsaicin-induced nociception. Combination treatment with substance P (50 pmol) and morphine at a subthreshold dose enhanced the antinociceptive effect of morphine. The enhanced effect of the combination of substance P with morphine was reduced significantly by co-administration of phosphoramidon, an inhibitor of endopeptidase-24.11. Administration of d-isomer of substance P (1–7), [d-Pro², d-Phe⁷]substance P (1–7), an inhibitor of [³H] substance P (1–7) binding, or antisera against substance P (1–7) reversed the enhanced antinociceptive effect by co-administration of substance P and morphine. Taken together these data suggest that morphine-induced antinociception may be enhanced through substance P (1–7) formed by the enzymatic degradation of i.t. injected substance P in the spinal cord.     

Poly ADP-ribosylation: A DNA break signal mechanism

Recent evidence obtained with transgenic knockout mice suggests that the enzyme poly(ADP-ribose)polymerase (PARP) does not play a direct role in DNA break processing [1, 2]. Nevertheless, inactivation of the catalytic or the DNA nick-binding functions of PARP affects cellular responses to genotoxins at the level of cell survival, sister chromatid exchanges and apoptosis [2, 3]. In the present report, we conceptualize the idea that PARP is part of a DNA break signal mechanism [4, 5]. In vitro screening studies revealed the existence of a protein family containing a polymer-binding motif of about 22 amino acids. This motif is present in p53 protein as well as in MARCKS, a protein involved in the regulation of the actin cytoskeleton. Biochemical analyses showed that these sequences are directly targeted by PARP-associated polymers in vitro, and this alters several molecular functions of p53- and MARCKS protein. PARP-deficient knockout mice from transgenic mice were found to exhibit several phenotypic features compatible with altered DNA damage signaling, such as downregulation and lack of responsiveness of p53 protein to genotoxins, and morphological changes compatible with MARCKS-related cytoskeletal dysfunction. The knockout phenotype could be rescued by stable expression of the PARP gene. - We propose that PARP-associated polymers may recruit signal proteins to sites of DNA breakage and reprogram their functions.     

The cyclin-dependent kinase Cdk5 controls multiple aspects of axon patterning in vivo

Cyclin-dependent kinase 5 (Cdk5) is one of a subfamily of Cdks involved in the control of cell differentiation and morphology rather than cell division. Specifically, Cdk5 and its activating subunit, p35, have been implicated in growth cone motility during axon extension. Both Cdk5 and p35 are expressed in post-mitotic neurons and are localized to growth cones [1] [2] [3] [4]. The Cdk5-p35 complex interacts with the Rac GTPase, a protein required for growth cone motility [5]. Studies using cultured neurons have suggested that Cdk5 activity controls the efficiency of neurite extension [3] [4]. Mutant mice lacking p35 exhibit subtle axon-guidance defects [6], but these mice have severe defects in neuronal migration [6] [7] [8], making it difficult to define precisely the role of the Cdk5-p35 complex in vivo. Here, we examined Cdk5 function in axon patterning in the Drosophila embryo. Although our data support the idea that Cdk5-p35 is involved in axonogenesis, they do not support the view that Cdk5 simply promotes growth cone motility. Instead, we found that disrupting Cdk5 function caused widespread errors in axon patterning.     

Amniotic fluid and plasma glycine/valine ratios in substrate deprived growth retarded fetal rats.

Characteristic profiles of the free amino acid concentration in umbilical cord blood of growth retarded newborns have been observed. We hypothesized that the amniotic fluid of growth retarded fetal rats would show an increase in the ratio between glycine and valine which would parallel the pattern observed in the cord blood of growth retarded neonates, thus providing an index for the antepartum identification of the substrate deprived growth retarded fetus. Six test and 6 control dams were tested. Four fetuses per dam, matched for uterine location were examined. Test animals were fasted for 72 hours. Sampling was performed on day 21 under anaesthesia. Fetal size was significantly reduced (P < 0.0001) in the test group. [T = 2.68 gs. +/- 0.28 vs. C = 3.67 gs. +/- 0.25]. Fetal plasma concentrations of glycine showed an increase in test animals (P < 0.01) while valine showed a significant reduction (P < 0.0001). Glycine (pm/microliters) T = 308 +/- 64 vs. C = 269 +/- 47, valine (pm/microliters) T = 424 +/- 79 vs. C = 671 +/- 218]. Amniotic fluid concentrations for both glycine and valine were significantly decreased (P < 0.0001) in test animals. [Glycine (pm/microliters) T = 710 +/- 124 vs. C = 931 +/- 178; valine (pm/microliters) T = 845 +/- 169 vs. C = 1,339 +/- 234]. The glycine/valine ratio was significantly increased (P < 0.01) in both fetal plasma and amniotic fluid in test animals [Plasma T = 0.74 +/- 0.18 vs. C = 0.43 +/- 0.13. Amniotic fluid T = 0.85 +/- 0.08 vs. C = 0.69 +/- 0.09]. Consistent with our hypothesis, the amniotic fluid concentrations generally parallel the observations made in the plasma. This finding could enhance the antepartum identification of the substrate deprived growth retarded fetus.     

Improving glycogenesis in Streptozocin (STZ) diabetic mice after administration of green algae Chlorella

Chlorella, a type of unicellular fresh water algae, has been a popular foodstuff in Japan and Taiwan. Studies have shown the hypoglycemic effects of Chlorella in alloxan-induced and Streptozocin (STZ)-induced diabetic animals. However, the mechanisms by which Chlorella treatment affects blood glucose homeostasis have not been studied. Diabetes in ICR mice was induced by injection of STZ. Lipogenesis in vivo was measured by incorporating 3H-H2O into lipids in brown and white adipose tissues. Glucose uptake in the liver and soleus muscles was measured by assaying 2-deoxy-D-[1,2-3H] glucose levels. The effects of Chlorella on serum non-esterified fatty acids (NEFA) were measured with commercial assay kits. Insulin-stimulated lipogenic rates in brown and white adipose tissues were unaffected by Chlorella. However, Chlorella increased 2-deoxyglucose uptake in the livers and soleus muscles in normal and STZ mice compared to that in their respective controls (p < 0.01). In addition, fasting NEFA levels were lower in Chlorella-treated STZ mice compared to H2O-treated STZ mice (p < 0.005). The current results suggest that the hypoglycemic effects of Chlorella are due to an enhancement of glucose uptake in the liver and in soleus muscles. The improved insulin sensitivity after Chlorella treatment could be also due to lower NEFA levels, since insulin sensitivity is usually blunted by elevated NEFA in diabetes.     

Studies on the Isolation,Identification and Bioactivities of Daucosterol in the Roots ot Paeonia lactiflora

A white crystalline powder substance (m.p. 290–292℃; [a]23D=-48.3˚) was iso- lated from the roots of Paeonia lactiflora. It was identified by chemical and spectral analysis as β-sitosterol-D-glucoside, i.e. daucosterol. It amounts to 4×10-5 of the dry material Bioassays showed that daucosterol increased the fresh weight of cucumber cotyledons, promoted the elongation of wheat coleoptiles, and delayed the destruction of chlorophyll. Daucosterol also induced the initiation of root and leaf of isolated cucumber cotyledons. These phenomena showed that daucosterol was an active phytosetroid and exhibited both auxin-like and kinetin-like activities.     

Sleep patterns of day-working, evening high-schooled adolescents of São Paulo, Brazil

Children who grow up in developing countries of the world must work to help financially support their families, and they must also attend school. We investigated the impact of work on the sleep of working vs. nonworking high school students. Twenty-seven S?o Paulo, Brazil, public high school students (eight male and eight female working students plus six nonworking female and five nonworking male students) 14-18 yrs of age who attended school Monday-Friday between 19:00 to 22:30h participated. A comprehensive questionnaire about work and living conditions, health status, and diseases and their symptoms was also answered. The activity level and rest pattern (sleep at night and napping during the day) were continuously assessed by wrist actigraphy (Ambulatory Monitoring, USA). The main variables were analyzed by a two-factor ANOVA with application of the Tukey HSD test for multiple comparisons, and the length of sleep during weekdays vs. weekends was compared by Student t-test. Working students went to sleep earlier weekends [F(1,23)=6.1; p=0.02] and woke up earlier work days than nonworking students [F(1,23) = 17.3; p = 0.001]. The length of nighttime sleep during weekdays was shorter among all the working [F(1,23)= 16.7; p <0.001] than all the nonworking students. The sleep duration of boys was shorter than of girls during weekends [F(1,23)= 10.8; p <0.001]. During weekdays, the duration of napping by working and nonworking male students was shorter than nonworking female students. During weekdays working girls took the shortest naps [F(1,23)= 5.6; p = 0.03]. The most commonly reported sleep complaint during weekdays was difficulty waking up in the morning [F(1,23) = 6.5; p = 0.02]. During weekdays, the self-perceived sleep quality of working students was worse than nonworking students [F(1,23) = 6.2; p = 0.02]. The findings of this study show that work has negative effects on the sleep of adolescents, with the possible build-up of a chronic sleep debt with potential consequent impact on quality of life and school learning.     

The comparison of immobility time in experimental rat swimming models

Rat swimming models have been used in studies about stress and depression. However, there is no consensus about interpreting immobility (helplessness or adaptation) in the literature. In the present study, immobility time, glucose and glycogen mobilization, corticosterone and the effect of desipramine and diazepam were investigated in two different models: swimming stress and the forced swimming test. Immobility time was lower in swimming stress than in the forced swimming test. Both swimming models increased corticosterone levels in comparison with control animal levels. Moreover, swimming stress induced higher corticosterone levels than the forced swimming test did [F(2,14)=59.52; p<0.001]. Liver glycogen content values differed from one another (swimming stress相似文献