7株野生虫草菌的鉴定及其菌丝体醇提取物对HepG2细胞的抑制活性

结合形态学与ITS序列分析对7株野生虫草真菌进行分类鉴定。MTT法分析它们的菌丝体醇提取物对肝癌HepG2细胞增殖的抑制活性。鉴定结果表明菌株MF7、MF9、MF14为细脚棒束孢Isaria tenuipes,菌株MF11、MF12、MF13为蝉棒束孢Isaria cicadae,菌株MF10为球孢白僵菌Beauveria bassiana;MTT结果显示分离到的3株细脚棒束孢和3株蝉棒束孢的菌丝体醇提取物对HepG2的抑制活性较差,IC₅₀均大于500μg/mL;球孢白僵菌MF10对HepG2细胞有一定抑制作用,IC₅₀值为221.6μg/mL,略强于蝙蝠蛾拟青霉发酵菌丝粉产品金水宝胶囊（IC₅₀=364μg/mL）和中华被毛孢发酵菌丝粉产品百令胶囊（IC₅₀=268.7μg/mL）。另外,发现供对比试验的3株蛹虫草菌株（MF1、MF5、MF15）对HepG2细胞均有较好的抑制作用,其中MF15的发酵菌丝体醇提取物活性最强,IC₅₀为55.56μg/mL,暗示蛹虫草发酵菌丝体具有重要的研究价值。     

越南槐内生真菌JXRPH-24的抗菌活性及其鉴定

以2株病原细菌金黄色葡萄球菌ATCC25923和耐甲氧西林金黄色葡萄球菌临床分离株为靶标菌,采用琼脂块法和改良的微量肉汤稀释法测定越南槐内生真菌JXRPH-24的抗菌活性;根据形态和分子特征鉴定该菌株的分类学地位。结果表明,该菌株的活菌块对金黄色葡萄球菌ATCC25923的抑菌圈直径与阳性对照相当;其代谢产物对这2株病原细菌的最小抑菌浓度均为10μg/mL,仅为阳性对照的16倍;基于形态和ITS序列分析未能鉴定该菌株。菌株JXRPH-24可能是一个潜在的新内生真菌,对参试金黄色葡萄球菌均具有强的抗菌活性,具有较大的开发应用潜力。     

毛头鬼伞子实体化学成分及抗糖尿病活性初探

从毛头鬼伞子实体中萃取得到乙醇、乙酸乙酯、石油醚3种有机提取物,采用α-葡萄糖苷酶活性抑制实验对3种有机提取物的抗糖尿病活性进行评价,结果显示,乙酸乙酯提取物对α-葡萄糖苷酶有较强的抑制活性。采用柱层析技术从乙酸乙酯提取物中分离纯化出10种化合物,经核磁等方法鉴定为：（1）顺,顺-9,12-十八(碳)二烯酸;（2）顺式-9-十八烯酸;（3）(22E,24R)-麦角甾烷-5,7,22-三烯-3β醇;（4）3β-5α-6α-22E-麦角甾-7,22-双烯-3,5,6-三醇-6-亚油酸酯;（5）3β-5α-6α-22E-麦角甾-7,22-双烯-3,5,6-三醇-6-油酸酯;（6）邻苯二甲酸二异丁酯;（7）对羟基苯乙醇;（8）4-羟基苯乙基乙酸酯;（9）3-(4-羟基-3-甲氧苯基)败脂酸;（10）N-反式-3,4亚甲二氧基肉桂酰基-3-甲氧基酪胺。对分离化合物的α-葡萄糖苷酶活性抑制实验结果显示,N-反式-3,4亚甲二氧基肉桂酰基-3-甲氧基酪胺对α-葡萄糖苷酶具有较强的抑制活性,其IC₅₀值为4.17mg/mL。     

白腐真菌Hypocrea lixii SCSIO 41520次级代谢产物及抗菌活性研究

对来源于深圳大亚湾近海水域软珊瑚中分离得到的白腐真菌Hypocrea lixii SCSIO 41520的次级代谢产物及抗菌活性进行研究。采用硅胶柱层析、凝胶Sephadex LH-20柱层析、半制备HPLC等色谱技术对其进行分离纯化,通过~1H NMR、~(13)C NMR、MS等波谱技术鉴定化合物结构。共分离鉴定了8个化合物,分别为4-(5,7-dimethoxy-4-oxo-4H-chromen-2-yl)heptanoic acid methyl ester(1)、5-methoxy eugenin(2)、rugulosin(3)、大黄素(4)、3-(5,7-dimethoxy-4-oxo-4H-chromen-2-yl) propanoic acid(5)、comazaphilone C(6)、kasanosin C(7)、comazaphilone E(8),其中化合物1是新化合物。纸片扩散法抗菌实验结果显示,该真菌发酵产物的乙酸乙酯粗提物及各分离组分均有一定的抗菌活性,单体化合物中,化合物3在25μg/disc的浓度下对金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌有显著抑制作用,抑菌圈直径分别为16、15、9 mm。     

致幻毒蘑菇卵囊裸盖菇化学成分研究初探

从一种采集于贵州省的致幻毒蘑菇——卵囊裸盖菇Psilocybe ovoideocystidiata中首次分离得到3种化合物,分别是3β-羟基-5α,8α-桥二氧麦角甾-6,22E-二烯(化合物1)、β-D-葡萄糖(化合物2)和腺苷(化合物3)。基于高分辨质谱与核磁共振谱数据以及相关文献比对确定以上3种化合物的结构,并首次推导出化合物2和3质谱裂解规律,其中重排与中性丢失在质谱裂解过程中起主导作用。利用UPLC-MS/MS法对卵囊裸盖菇的干燥子实体和新鲜子实体中的裸盖菇素和脱磷裸盖菇素进行检测,在干燥子实体中检测到裸盖菇素和脱磷裸盖菇素,但在-80 ℃保存6个月的新鲜子实体中未检测到裸盖菇素和脱磷裸盖菇素,推测可能是由于保存方法和提取方法的原因导致化合物发生变化。     

稳定表达人源GABAAR-CHO细胞株的建立

目的: 构建α₁亚基诱导表达、β₂和γ_2L亚基稳定表达的人源α₁β₂γ_2L-GABA_AR-CHO(Chinese hamster ovary)细胞株。方法: 从人cDNA文库中扩增α₁、β₂、γ_2L亚基编码基因,分别构建亚基表达载体;将三个亚基表达载体共转染CHO-K1细胞,通过抗性筛选、膜电位检测法进行稳定表达克隆筛选;通过qPCR、Western blot对亚基表达进行鉴定;以激动剂GABA、阳性变构调节剂地西泮(diazepam,Dia)、拮抗剂荷包牡丹碱(bicuculine)为工具药,采用全细胞膜片钳方法及膜电位检测法对稳定表达细胞的药理学功能进行鉴定。结果: 经克隆筛选获得表达量较高的α₁β₂γ_2L-GABA_AR-CHO并对其亚基表达鉴定,结果显示该细胞稳定表达α₁、β₂、γ_2L亚基,构建的α₁β₂γ_2L-GABA_AR-CHO细胞仅在加入四环素(tetracyclin)诱导的情况下表达α₁亚基并与β₂、γ_2L组装成具有功能活性的α₁β₂γ_2L-GABA_AR;对其进行全细胞膜片钳检测研究发现,GABA可对其产生激动效应,引起α₁β₂γ_2L-GABA_AR-CHO细胞产生氯离子通道特征性电流变化,Dia可剂量依赖性地增强GABA对α₁β₂γ_2L-GABA_AR的激动效应;在膜电位检测研究中,获得GABA激动效应EC₅₀为(177.72 ± 15.92)nmol/L,Dia变构效应EC₅₀为(3.63±0.52)μmol/L,拮抗剂Bicuculine拮抗效应IC₅₀为(538.83±29.55)nmol/L。结论: 通过采用诱导表达策略,成功构建了α₁β₂γ_2L-GABA_AR-CHO稳定表达细胞株,该细胞株具有对激动剂、阳性变构剂、拮抗剂特异性检测的药理学功能。     

牛樟芝多糖对HepG2细胞酒精性氧化损伤的保护作用

研究了不同剂量（100、200和400μg/mL）的牛樟芝粗多糖（CP）和醇提物后的水提物（WEE）对酒精诱导的HepG2细胞氧化损伤的保护作用。研究结果表明：与模型组比较,各剂量组的CP和200、400μg/mL的WEE均能极显著提高HepG2细胞的细胞活力。100μg/mL的CP和WEE均能极显著降低细胞培养液的ALT水平;200和400μg/mL的CP和WEE均能显著降低细胞培养液的ALT、AST水平,同时提高胞内的CAT活力;200和400μg/mL的WEE及400μg/mL CP能明显提高胞内的SOD活力。此外,WEE各剂量组和400μg/mL CP中的胞内ROS水平显著下降。CP中含有甘露糖、鼠李糖、葡萄糖、半乳糖、岩藻糖5种单糖,摩尔比为1:0.1622:6.651:2.646:0.3929。WEE和CP能提高细胞的抗氧化应激能力,降低胞内ROS,对酒精诱导的HepG2细胞氧化损伤起到明显的保护作用,提示多糖是牛樟芝解酒保肝的重要活性成分之一。     

链霉菌Streptomyces sp. HS-NF-496中两个Anthrabenzoxocinone类化合物

对链霉菌Streptomyces sp.HS-NF-496的次级代谢产物进行了研究,分离得到两个化合物,经1D和2D NMR、CD、MS波谱分析及与文献数据比较,鉴定它们分别为(-)-(6R,16R)10-Cl-ABX(1)和(-)-(6R,16R)-ABX(2)。对两个化合物的细胞毒活性和抗菌活性评价实验结果表明:化合物1和2对人肺腺癌A549细胞,人结肠癌HCT-116细胞和人神经癌SF-268细胞表现出较弱的细胞毒活性;化合物1和2对绿脓杆菌,金黄色葡萄球菌,枯草芽胞杆菌和耐甲氧西林金黄色葡萄球菌表现出中等强度的抗菌活性。本文首次报道了化合物1的绝对构型。     

SARS-CoV-2重组S1和S蛋白疫苗诱导保护性免疫的研究*

目的:评价新型冠状病毒(SARS-CoV-2)重组S1蛋白和S蛋白疫苗对SARS-CoV-2的免疫保护效果。方法:将SARS-CoV-2重组S1蛋白和S蛋白分别联合氢氧化铝佐剂以0.1 μg/只、1 μg/只、5 μg/只、10 μg/只不同剂量接种6~8周BALB/c纯系健康雌性小鼠。第二次免疫后采血通过酶联免疫吸附试验(ELISA)检测血清中IgG抗体效价,通过假病毒中和试验比较免疫小鼠血清对SARS-CoV-2野生型株(WT)、英国株(B.1.1.7)、巴西株(P.1)、印度株(B.1.617.2)、Mu毒株(B.1.621)和南非株(501Y.V2-1)六种假病毒毒株中和活性效价,取脾细胞通过酶联免疫斑点技术(ELISpot)检测免疫小鼠的细胞免疫水平。结果:SARS-CoV-2重组S和S1蛋白都能诱导小鼠产生较强的IgG抗体水平。免疫S1蛋白的小鼠血清对SARS-CoV-2野生型株、英国株、巴西株有明显的中和活性,免疫S蛋白的小鼠血清除了对SARS-CoV-2野生型株、英国株、巴西株有明显中和活性之外,对印度株也有明显的中和活性,两种蛋白质免疫的小鼠血清均对野生型株中和效果最强。S蛋白免疫的小鼠脾细胞能够显著诱导出γ干扰素(IFN-γ)和白介素-4(IL-4)的产生。S蛋白诱导产生的IgG抗体、中和抗体、细胞免疫水平均高于S1。结论:SARS-CoV-2重组S蛋白疫苗能够诱导产生较强的保护性免疫应答。     

迷走神经刺激对难治性癫痫大鼠海马神经炎性反应及α7nAChR表达的影响*

目的: 探讨迷走神经刺激(VNS)对难治性癫痫(IE)模型大鼠海马神经炎性反应及α7nAChR表达的影响。方法: 80只成年雄性SD大鼠,SPF级,随机分为对照组、模型组、VNS组、甲基牛扁亭(MLA)+VNS组,其中对照组与MLA+VNS组分别20只,模型组与VNS组因模型制作失败与动物死亡,分别剩下15只和14只。除对照组之外,其余各组皆通过腹腔注射皮罗卡品建立氯化锂-皮罗卡品IE大鼠模型。对照组仅分离迷走神经,不采取电刺激;模型组不采取任何干预措施;VNS组在模型制作成功后7 d采取VNS,连续4周;MLA+VNS组先侧脑室给药MLA(3.4 μg/μl,5 μl),然后给予VNS,连续4周。观察并记录各组大鼠癫痫发作的次数与持续时间的变化;然后断头处死大鼠,快速分离海马并制备10%组织匀浆,离心并提取上清液,通过分光光度法测定上清液中AChE、ChAT活性;ELISA法检测TNF-ɑ、IL-6和IL-1β表达;Western blot检测海马组织α7nAChR蛋白表达;免疫荧光染色法检测海马组织α7nAChR与小胶质细胞共表达。结果: ①通过VNS治疗4周后,大鼠癫痫发作的频率以及持续的时间都明显低于模型组(P<0.01);MLA阻断后在给予VNS,大鼠癫痫发作的频率以及持续的时间也明显低于模型组,但高于VNS组(P<0.01)。②与对照组比较,模型组大鼠海马组织ChAT表达明显下降,AChE表达明显升高(P<0.01);与模型组比较,VNS组与MLA+VNS组大鼠海马组织ChAT表达明显升高,AChE表达明显降低(P< 0.01);与VNS组比较,MLA+VNS组大鼠海马组织ChAT、AChE表达无明显变化(P>0.05)。③与对照组比较,模型组大鼠海马组织TNF-ɑ、IL-6和IL-1β表达明显升高(P<0.01);与模型组比较,VNS组大鼠海马组织TNF-ɑ、IL-6和IL-1β表达明显降低(P<0.01);与VNS组比较,MLA+VNS组大鼠海马组织TNF-ɑ、IL-6和IL-1β表达明显升高(P<0.01)。④与对照组比较,模型组大鼠海马组织以及小胶质细胞上α7nAChR表达明显降低(P<0.01);与模型组比较,VNS组大鼠海马组织以及小胶质细胞上α7nAChR表达明显上调(P<0.01);与VNS组比较,MLA+VNS组海马小胶质细胞上共表达α7nAChR数量明显减少(P<0.01)。结论: VNS对IE大鼠有明显的治疗作用,其机制可能是通过直接激活海马小胶质细胞CAP,抑制海马神经炎性反应来实现的。     

北亚热带地带性森林土壤温室气体通量对土地利用方式改变和降水减少的响应

弄清土地利用和降水变化对林地土壤主要温室气体(CO₂、CH₄和N₂O)排放通量变化的影响, 是准确评估森林土壤温室气体排放能力的重要基础。该研究以常绿落叶阔叶混交林原始林、桦木(Betula luminifera)次生林和马尾松(Pinus massoniana)人工林为对象, 采用静态箱-气相色谱法研究了3种土地利用方式(常绿落叶阔叶混交林原始林、桦木次生林和马尾松人工林)和降水减少处理状况下森林土壤CO₂、CH₄和N₂O通量排放特征, 并探讨了其环境驱动机制。研究结果表明: 原始林土壤CH₄吸收通量显著高于次生林和人工林, 次生林CH₄吸收通量显著高于人工林土壤。人工林土壤CO₂排放通量显著高于原始林和次生林土壤。次生林土壤N₂O排放通量高于原始林和人工林, 但三者间差异不显著。降水减半显著抑制了3种不同土地利用方式下林地土壤CH₄吸收通量; 降水减半处理对原始林和次生林土壤CO₂排放通量均具有显著的促进作用, 而对人工林土壤CO₂排放通量具有显著的抑制作用; 降水减半处理促进了原始林和人工林林地土壤N₂O排放而抑制了次生林林地土壤N₂O排放。原始林和次生林林地土壤CH₄吸收通量随土壤温度升高显著增加, CH₄吸收通量与土壤温度均呈显著相关关系; 原始林、次生林和人工林土壤CO₂和N₂O排放通量与土壤温度均呈显著正相关关系; 土壤湿度抑制了次生林和人工林土壤CH₄吸收通量, 其CH₄吸收通量随土壤湿度增加显著减少; 原始林土壤CO₂排放通量与土壤湿度呈显著正相关关系。自然状态下, 原始林土壤N₂O排放通量与土壤湿度呈显著正相关关系, 原始林和次生林土壤N₂O排放通量与硝态氮含量呈显著相关关系。研究结果表明全球气候变化(如降水变化)和土地利用方式的转变将对北亚热带森林林地土壤温室气体排放通量产生显著的影响。     

Greenhouse gas fluxes of typical northern subtropical forest soils: Impacts of land use change and reduced precipitation

Aims It is important to study the effects of land use change and reduced precipitation on greenhouse gas fluxes (CO₂, CH₄ and N₂O) of forest soils. Methods The fluxes of CO₂, CH₄ and N₂O and their responses to environmental factors of primary forest soil, secondary forest soil and artificial forest soil under a reduced precipitation regime were explored using the static chamber and gas chromatography methods during the period from January to December in 2014. Important findings Results indicate that CH₄ uptake of primary forest soil ((-44.43 ± 8.73) μg C·m^-2·h^-1) was significantly higher than that of the secondary forest soil ((-21.64 ± 4.86) μg C·m^-2·h^-1) and the artificial forest soil ((-10.52 ± 2.11) μg C·m^-2·h^-1). CH₄ uptake of the secondary forest soil ((-21.64 ± 4.86) μg C·m^-2·h^-1) was significantly higher than that of the artificial forest ((-10.52 ± 2.11) μg C·m^-2·h^-1). CO₂ emissions of the artificial forest soil ((106.53 ± 19.33) μg C·m^-2·h^-1) were significantly higher than that of the primary forest soil ((49.50 ± 8.16) μg C·m^-2·h^-1) and the secondary forest soil ((63.50 ± 5.35) μg C·m^-2·h^-1) (p < 0.01). N₂O emissions of the secondary forest soil ((1.91 ± 1.22) μg N·m^-2·h^-1) were higher than that of the primary forest soil ((1.40 ± 0.28) μg N·m^-2·h^-1) and the artificial forest soil ((1.01 ± 0.86) μg N·m^-2·h^-1). Reduced precipitation (-50%) had a significant inhibitory effect on CH₄ uptake of the artificial forest soil, while it enhanced CO₂ emissions of the primary forest soil and the secondary forest soil. Reduced precipitation had a significant inhibitory effect on CO₂ emissions of the artificial forest soil and N₂O emissions of the secondary forest (p < 0.01). Reduced precipitation promotes N₂O emissions of the primary forest soil and the artificial forest soil. CH₄ uptake of the primary forest and the secondary forest soil increased significantly with the increase of soil temperature under natural and reduced precipitation. CO₂ and N₂O emission fluxes of the primary forest soil, secondary forest soil and artificial forest soil were positively correlated with soil temperature (p < 0.05). Soil moisture inhibited CH₄ uptake of the secondary forest soil and the artificial forest soil (p < 0.05). CO₂ emissions of the primary forest soil were significantly positively correlated with soil moisture (p < 0.05). N₂O emissions of primary forest soil and secondary forest soil were significantly correlated with the nitrate nitrogen content (p < 0.05). It was implied that reduced precipitation and land use change would have significant effects on greenhouse gas emissions of subtropical forest soils.     

Crystallographic and NMR studies of antiinfective tricyclic guanidine alkaloids from the sponge Monanchora unguifera

Three tricyclic guanidine alkaloids, including 1,8a;8b,3a-didehydro-8β-hydroxyptilocaulin (1), 1,8a;8b,3a-didehydro-8-hydroxyptilocaulin (2) and mirabilin B (3), were identified from the marine sponge Monanchora unguifera. 1,8a;8b,3a-Didehydro-8-hydroxyptilocaulin (2) is a new stereoisomer of 1, the structure of which was elucidated by spectroscopic analysis, comparison of its spectral data with those of 1, and confirmed by X-ray analysis. Compounds 1 and 2 co-crystallized in an unusual perfect order and packed around an approximate inversion center. A mixture of 1 and 2 is active against the malaria parasite Plasmodium falciparum with an IC₅₀ value of 3.8 μg/mL while mirabilin B (3) exhibited antifungal activity against Cryptococcus neoformans with an IC₅₀ value of 7.0 μg/mL and antiprotozoal activity against Leishmania donovani with an IC₅₀ value of 17 μg/mL.     

农杆菌介导Txpam基因转化烟曲霉TMS-26及其产紫杉醇效果评价

为研究红豆杉紫杉醇合成途径限速酶基因功能及其对内生真菌烟曲霉TMS-26发酵产紫杉醇的影响,以曼地亚红豆杉愈伤组织制备cDNA作为模板扩增苯丙氨酸氨基变位酶基因（Txpam）,构建重组质粒pGEX-4T-1-Txpam,转入大肠杆菌中进行异源诱导表达,经亲和层析纯化,获取重组酶TxPAM并验证其酶活性。构建pCAMBIA1302-Txpam质粒,转化农杆菌感受态细胞,利用农杆菌介导的转化体系获得转化子并优化转化条件,结合插入片段携带的分子标记和目的基因进行转化子验证,同时培养转化菌株并检测紫杉醇产量。结果表明：纯化获取的重组酶TxPAM,经HPLC检测具有将α-苯丙氨酸催化为β-苯丙氨酸的功能;在最优转化条件下,转化子数目达到471个/10⁶个孢子;根据基因hyg和Txpam的克隆以及测序结果,说明成功构建了基因工程菌株,通过对其发酵条件进行优化,紫杉醇产量达到721.87μg/L。     

Bioactive metabolites from the endophytic fungus Ampelomyces sp. isolated from the medicinal plant Urospermum picroides

Extracts of cultures grown in liquid or on solid rice media of the fungal endophyte Ampelomyces sp. isolated from the medicinal plant Urospermum picroides exhibited considerable cytotoxic activity when tested in vitro against L5178Y cells. Chromatographic separation yielded 14 natural products that were unequivocally identified based on their ¹H and ¹³C NMR as well as mass spectra and comparison with previously published data. Six compounds (2, 4, 5, 7, 9 and 11) were natural products. Both fungal extracts differed considerably in their secondary metabolites. The extract obtained from liquid cultures afforded a pyrone (2) and sulfated anthraquinones (7 and 9) along with the known compounds 1, 3, 6 and 8. When grown on solid rice medium the fungus yielded three compounds 4, 5 and 11 in addition to several known metabolites including 6, 8, 10, 12, 13 and 14. Compounds 4, 8 and 10 showed the strongest cytotoxic activity against L5178Y cells with EC₅₀ values ranging from 0.2–7.3 μg/ml. Furthermore, 8 and 10 displayed antimicrobial activity against the Gram-positive pathogens, Staphylococcus aureus, S. epidermidis and Enterococcus faecalis at minimal inhibitory concentrations (MIC) of 12.5 μg/ml and 12.5–25 μg/ml, respectively. Interestingly, 6 and 8 were also identified as constituents of an extract derived from a healthy plant sample of the host plant U. picroides thereby indicating that the production of bioactive natural products by the endophyte proceeds also under in situ conditions within the host plant.     

长白山阔叶红松林生态系统光能利用率的动态变化及其主控因子

生态系统光能利用率(LUE)反映了植被通过光合作用利用光能吸收和固定大气中CO₂的能力, 是表征生态系统生产力的重要指标。选取长白山温带阔叶红松(Pinus koraiensis)林生态系统为研究对象, 利用涡度相关通量观测数据, 采用直角双曲线方程获取了生态系统光合作用的表观量子效率(ε); 基于总生态系统初级生产力(GEP)与下垫面入射光合有效辐射(Q)的比值得到生态光能利用率(LUE_eco)。研究表明: 在季节尺度上, ε与LUE_eco均表现出显著的单峰变化特征, 并主要受到土壤温度和归一化植被指数(NDVI)的调控, 同时, ε和LUE_eco都受到GEP的显著影响, 而与Q的相关性较弱或无显著相关关系, 但散射辐射的增加在一定程度上有助于提高生态系统的LUE。ε与LUE_eco存在显著的线性正相关关系, 但ε明显高于LUE_eco。2003-2005年, ε与LUE_eco每年最大值的平均值分别为(0.087 ± 0.003)和(0.040 ± 0.002) μmol CO₂·μmol photon^-1, 年际间变异度分别为4.17%和4.25%, 而不同年份之间最大差异均达到8%或8%以上, 从而对模型模拟结果产生明显影响。因此, 在基于光能利用率模型的模拟研究中, 最大LUE的年际变异需要在参数反演和优化中给予重要考虑。     

固相萃取-超高效液相色谱-串联质谱法测定新鲜块菌子实体中α-雄烷醇

本研究建立了一种固相萃取-超高效液相色谱-串联质谱的检测方法,用于检测新鲜块菌子实体中α-雄烷醇(5α-雄甾-16-烯-3α-醇)的含量。新鲜块菌样品经无水乙醇提取,Qasis HLB柱萃取富集后,采用超高效液相色谱-串联质谱进行分析定量。方法学验证结果表明该方法的回收率为88.49%-92.22%;检出限为0.120 9 ng/mL,定量限为0.398 9 ng/mL。该方法简便、精确,适用于新鲜块菌中α-雄烷醇含量的测定。     

Dynamics and regulations of ecosystem light use efficiency in a broad-leaved Korean pine mixed forest,Changbai Mountain

Aims Ecosystem light use efficiency (LUE) reflects the ability of CO₂ uptake and light utilization via photosynthesis, which is a key parameter in ecosystem models to evaluate ecosystem productivity. The objectives of this study were to: (1) compare the differences of LUE derived from different methods; (2) elucidate the seasonal dynamics of LUE and its regulatory factors; and (3) evaluate the maximum LUE (LUE_max) and its variability based on eddy-covariance (EC) flux.Methods Using the flux data from an EC tower during 2003-2005 at a broad-leaved Korean pine (Pinus koraiensis) mixed forest, Changbai Mountain, two types of LUE indicators were generated from: 1) the apparent quantum yield (ε) estimated with rectangular hyperbolic curve, and 2) the ecological light use efficiency (LUE_eco) calculated as the ratio between gross ecosystem productivity (GEP) and photosynthetically-active radiation (Q).Important findings The seasonal variation of ε and LUE_eco appeared a unimodal pattern within a year, with the variations significantly dominated by soil surface temperature and Normalized Difference Vegetation Index (NDVI). A positive correlation between GEP and LUE was found for both ε and LUE_eco, with the effect of Q on LUE relatively weak. The increase in diffusion radiation appeared favorable for enhanced LUE. Generally, there was a significant positive relationship between ε and LUE_eco, while ε was higher than LUE_eco, especially during the mid-season. The annual maximum value of ε and LUE_eco was (0.087 ± 0.003) and (0.040 ± 0.002) μmol CO₂·μmol photon^-1 over the three years, respectively. The interannual variability of LUE_max for ε and LUE_eco was 4.17% and 4.25%, respectively, with a maximum difference of >8%, likely resulted from considerable uncertainty in model simulations. Our results indicated that the inversion and optimization of maximum LUE should be taken seriously in the application of LUE models.     

In vitro antiplasmodial activity of extract and constituents from Esenbeckia febrifuga, a plant traditionally used to treat malaria in the Brazilian Amazon

Esenbeckia febrifuga (Rutaceae) is a plant traditionally used to treat malaria in the Brazilian Amazon region. Ethanol extract of stems displayed a good antiplasmodial activity against Plasmodium falciparum strains W-2 (IC₅₀ 15.5±0.71 μg/ml) and 3 D7 (IC₅₀ 21.0±1.4 μg/ml). Two coumarins (bergaptene 1 and isopimpinellin 2), five alkaloids (flindersiamine 3, kokusaginine 4, skimmiamine 5, γ-fagarine 6 and 1-hydroxy-3-methoxy-N-methylacridone, 7), besides a limonoid (rutaevine 8), have been isolated for the first time from this species. Antiplasmodial activity of compounds 3, 5–8 has been evaluated in vitro against P. falciparum strains (W-2 and 3D7) and the furoquinolines 5 and 6 were the most potent displaying IC₅₀ values <50 μg/ml; flindersiamine (3) showed a weak activity while alkaloid 7 and rutaevine (8) were inactive (IC₅₀>100 μg/ml).