一株蓝藻来源真菌Aspergillus sp.的次生代谢产物研究

目的 研究一株采自中国三亚蓝藻(Lyngbya majuscula)来源的真菌Aspergillus sp.的次生代谢产物的分离纯化、结构鉴定及其生物活性评价。方法 利用硅胶色谱、HPLC等色谱学分离方法对真菌Aspergillus sp.的次生代谢产物进行分离纯化,并结合NMR,MS等波谱学方法对分离得到的化合物进行结构鉴定;并采用CCK8法,用人肺癌细胞株A549和人乳腺癌细胞株MCF7评价了这6个化合物的细胞毒活性。结果 从真菌Aspergillus sp.的代谢产物中分离得到6个化合物：methyl asterric acid (1),methyl 3-chloroasterric acid (2),methyl 3,5-dichloroasterric acid (3),dihydrogeodin (4),benzomalvin B (5) 和benzomalvin E (6)。结论 化合物5和6为首次从Aspergillus属真菌中获得,化合物 1-6在40 μM浓度下没有显著的细胞毒活性。     

海南海桑内生真菌Bionectria ochroleuca HHS111023次生代谢产物的研究

目的 研究中国特有红树植物海南海桑内生真菌Bionectria ochroleuca HHS111023的次生代谢产物及其生物活性。方法 利用多种柱色谱技术对次生代谢产物进行分离纯化;通过核磁与质谱数据分析结合理化常数及文献比对,鉴定化合物的结构;分别采用纸片琼脂扩散法和MTT法对化合物的抗菌活性和细胞毒活性进行测试。结果 从海南海桑内生真菌Bionectria ochroleuca HHS111023的次生代谢产物中分离鉴定了7个化合物：Lasiodiplodin (1)、(R)-de-O-methyllasiodiplodin (2)、(5S)-5-hydroxylasiodiplodin (3)、 (5R)-5-hydroxylasiodiplodin (4)、methyl (E)-11,12,15-trihydroxyoctadec-13-enoate (5)、对羟基苯乙醇 (6)、对羟基苯乙酸甲酯 (7)。化合物1和2分别对白色念珠菌和青枯雷尔氏菌具有抗菌活性,且分别对SGC-7901和K-562具有体外细胞毒活性。结论 化合物1 ~ 4为首次从淡色生赤壳菌Bionectria ochroleuca次生代谢产物中分离得到,化合物5为首次分离自微生物次生代谢产物;海南海桑内生真菌Bionectria ochroleuca HHS111023能产生具有抗菌和细胞毒活性的化合物。     

深海来源的微生物抗肿瘤活性筛选及次级代谢产物的初步研究

目的 对来自深海的海水、海泥样品进行了微生物分离并通过抗肿瘤活性筛选获得活性菌株,并研究活性菌株c2b的次级代谢产物.方法 从样品中选择性分离得到真菌,并采用海虾生物致死法和人体慢性艇性白血病细胞(K562)为筛选模型对分离得到真菌的发酵产物进行抗肿瘤活性筛选;采用溶剂萃取、硅胶柱色谱及制备HPLC等分离手段对c2b菌株发酵产物的活性部位进行了活性追踪分离,通过理化性质及渡谱学手段进行化学结构鉴定,以SRB法评价了化合物的抗肿瘤活性.结果与结论 从深海来源的样品中共分离获得29株真菌,其中7株具有细胞毒活性;从c2b活性菌株的发酵产物中分离得到6个单体化合物(1～6),其化学结构分别鉴定为N-乙酰色氨(1),chrysogine(2),过氧化麦角甾醇(3),5,8-epidioxy-24-methylcholesta-6,22-dien-3β-ol(4),cerevisterol(5)和(4E,8E)-N-[(2'R,3'E)-2'-hydroxy-3'-hexadecenoyl]-1-O-β-D-glycopyranosyl-9-methyl-4,8-sphingadiene(6),其中化合物3,4对小鼠乳腺癌细胞(tsFT210)具有中等强度的细胞毒活性.     

黄河三角洲植物真菌的分离、活性菌株的筛选及活性产物的鉴定

目的 从黄河三角洲植物中分离真菌,筛选具有抗菌或抗肿瘤活性菌株,分离鉴定活性成分。方法 蔗糖密度梯度法分离菌株,对其发酵物进行抗菌活性和细胞毒活性筛选,采用硅胶柱色谱、凝胶柱色谱对发酵产物进行分离、纯化,运用核磁共振、质谱等手段鉴定化合物的结构。结果 从黄河三角洲的9种植物样品中分离纯化真菌136株,筛选得到具有抑菌活性菌株25株、具有细胞毒活性菌株17株,并从1株枝孢属真菌Cladosporium sp. OUCMDZ-2046的发酵产物中分离鉴定了1个对白色念珠菌和人乳腺癌细胞株MCF-7具有抑制作用的化合物：桔青霉素。结论 黄河三角洲的植物真菌具有开发为药用活性菌株的潜力。     

红树林来源耐酸真菌Aspergillus fumigatus OUCMDZ5210次生代谢产物的研究

目的 对源自泰国红树林底泥中的耐酸真菌Aspergillus fumigatus OUCMDZ 5210的次生代谢产物进行化学成分和生物活性的研究。方法 综合利用硅胶柱色谱、凝胶柱色谱和半制备高效液相色谱等多种方法对耐酸真菌Aspergillus fumigatus OUCMDZ5210的次生代谢产物进行分离;采用核磁共振谱（NMR）、质谱（MS）、紫外光谱（UV）等现代波谱学技术对分离得到的化合物进行结构鉴定;以人慢性髓性白血病细胞K562及人结肠癌细胞HCT116两株肿瘤细胞为研究模型,采用MTT法和CCK-8法对所分离的化合物进行细胞毒活性评价。结果 从耐酸真菌Aspergillus fumigatus OUCMDZ5210的次级代谢产物中分离得到6个吲哚二酮哌嗪类生物碱：tryprostatins B(1),fumitremorgin C(2),spirotryprostatin A(3), spirotryprostatin.B(4), 6-methoxyspirotryprostatin B (5), 8,9-dihydroxyspirotrypr-ostatin A(6)以及1个酰胺类化合物cephalimysin B(7)。结论 生物活性初步评价结果显示,化合物3和6对人结肠癌细胞HCT116以及人慢性髓性白血病细胞K562具有不同程度的抑制活性。     

盐碱地真菌棘孢曲霉Aspergillus aculeatus的化学成分研究△﹡

目的 对一株具有抗肿瘤活性的滨州盐碱地真菌棘孢曲霉Aspergillus aculeatus的活性次级代谢产物进行研究。方法 采用硅胶柱层析、LH-20凝胶色谱和半制备高效液相色谱分离纯化代谢产物,通过理化性质、波谱手段结合X-ray单晶衍射技术鉴定化合物,采用MTT法测试所得化合物对肿瘤细胞A549的增殖抑制活性。结果 从棘孢曲霉Aspergillus aculeatus中分离得到8个化合物(1～8),分别鉴定为5-Hydroxymethyl-2-furoic acid (1)、Isorhodoptilometrin (2)、Secalonic acid F (3)、Emodin (4)、Protocatechuic acid (5)、Altechromones B (6)、Cyclo (L-Tyr-L-Leu) (7)和Cyclo (L-Phe-L-Ala) (8)。化合物3对A549细胞具有一定的增殖抑制活性,其IC50值为14.9 μM。结论 首次研究了来源于盐碱地真菌棘孢曲霉Aspergillus aculeatus的活性次级代谢产物,并首次报道secalonic acid F对A549肿瘤细胞增殖抑制活性。     

海藻真菌的分离及其抗肿瘤活性筛

目的从海藻中分离海藻真菌并通过抗肿瘤活性筛选获得活性菌株。方法从样品中选择性分离得到真菌,并采用海虾生物致死法和人体慢性髓性白血病细胞K562为筛选模型对所得真菌的发酵产物进行抗肿瘤活性筛选。结果从黄海海岸潮间带采集到的28种海藻中分得221株真菌,经筛选得到具有海虾生物致死及细胞毒活性菌株各8株。结论实验结果表明,海藻真菌是寻找抗肿瘤活性物质的重要资源。     

南海深海沉积环境来源真菌Aspergillus sp. SCSIO F063的次生代谢产物研究

目的 采用单株菌多次级代谢产物(OSMAC)策略对一株南海深海沉积环境来源真菌的次生代谢产物进行分离、鉴定及活性研究。方法 通过改变培养基组成并筛选合适的发酵条件,采用硅胶柱层析、反相ODS柱层析、半制备高效液相等色谱学方法对真菌Aspergillus sp. SCSIO F063的发酵产物进行化学分离,利用NMR, MS等波谱学技术并结合文献进行化合物的结构鉴定,并对化合物进行初步的抗氧化活性测试。结果 从菌株SCSIO F063中新增分离鉴定5个单体化合物： 6-O-methyl-averythrin(1), (2,4-dichlorophenyl)-2,4-dichlorobenzoate(2),dibutyl phthalate(3),folipastatin(4),di-(2-ethylhexyl)-phthalate(5)。结论 改变培养基组成可以刺激该菌株产生不同类型的化合物,化合物1-5为首次从真菌SCSIO F063中分离得到。     

南海沉积环境来源真菌Eurotium sp.SCSIO F452的次生代谢产物研究

目的对1株南海沉积环境来源真菌的次生代谢产物进行分离、鉴定及活性研究。方法采用溶剂萃取、硅胶柱层析、凝胶柱层析等方法对真菌Eurotiumsp.SCSIO F452的发酵产物进行化学分离,通过NMR、MS等波谱学技术并参阅文献进行化合物结构鉴定,采用SRB法评价化合物的细胞毒活性。结果从菌株SCSIO F452中分离鉴定6个单体化合物,分别为:isodihydroauroglaucin(1)、flavoglaucin(2)、tetrahydroauro-glaucin(3)、2-(1,1-dimethyl-2-propen-1-yl)-1H-indole-3-carboxaldehyde(4)、neoechinulin A(5)和methyl lino-leate(6)。化合物1-5对4种肿瘤细胞系表现出不同强度的细胞毒活性。结论苯甲醛衍生物1-3是真菌SC-SIO F452的优势代谢产物,细胞毒活性较强,具有潜在的研究价值。     

表观遗传试剂诱导海洋真菌Aspergillus versicolor DJ013产生次级代谢产物的研究

目的 对一株来源于浙江东极岛海域沉积物的海洋真菌Aspergillus versicolor进行表观遗传试剂2-hexyl-4-pentynoic acid诱导并研究其次级代谢产物。方法 采用溶剂萃取、柱层析色谱及制备色谱等方法对化合物进行分离纯化,通过LC-MS和NMR等方法进行结构鉴定,采用滤纸片扩散法测定抗菌活性。结果 在表观遗传试剂2-hexyl-4-pentynoic acid诱导下,从Aspergillus versicolor分离得到三个化合物,经结构鉴定为curvularin(1)、cyclo-(L-Trp-L-Phe)(2)和diorcinol (3)。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

---

Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.