放线菌素D不能抑制α-双炔失碳酯诱导的白血病K562细胞凋亡

目的：研究放线菌素Ｄ（ＡｃｔＤ）对α－双炔失碳酯（α－ａｎｏｒｄｒｉｎ，ＡＮＯ）诱导的人白血病Ｋ５６２细胞凋亡的影响。方法：用光学显微镜观察细胞形态学变化；用流式细胞仪、琼脂糖凝胶电泳分别检测ＤＮＡ含量和ＤＮＡ断裂。结果：ＡＮＯ５０μｍｏｌ·Ｌ－１处理人白血病Ｋ５６２细胞２４ｈ引起大约１０％Ｋ５６２细胞产生凋亡。同时加入ＲＮＡ合成抑制剂ＡｃｔＤ０．００５μｍｏｌ·Ｌ－１不能抑制ＡＮＯ诱导的凋亡，相反地，ＡｃｔＤ加强ＡＮＯ的这一作用，使凋亡细胞从１０％上升到２０％。ＡｃｔＤ０．５μｍｏｌ·Ｌ－１本身可诱导约３２％的Ｋ５６２细胞凋亡。Ｓ期细胞对ＡＮＯ诱导的凋亡较敏感，而ＡｃｔＤ则较易诱导Ｓ期和Ｇ２－Ｍ期细胞凋亡。结论：ＡＮＯ诱导的Ｋ５６２细胞凋亡不依赖于新的ＲＮＡ合成。     

人参皂甙Rg1对老年人淋巴细胞蛋白质酪氨酸激酶活性的调控作用

选择７名健康老年人为对象，研究了人参皂甙Ｒｇ１对其淋巴细胞表面抗原。受体及蛋白质酪氨酸激酶（ＰＴＫ）活性的作用，用间接免疫荧光法测定了ＣＤ２５、ＣＤ４５ＲＡ及ＣＤ４５Ｒ阳性细胞百分率。单独ＰＨＡ（５μｇ／ｍｌ）培养７２小时分别为３８．３％±１７３％、４６．０％±１５．１％、５２６％±１４．４％；而用ＰＨＡ（５μｇ／ｍｌ）和Ｒｇ＿１（μｇ／ｍｌ）联合培养时则分别为５８．０％±１２．５％。６４．１％±１２．４％、７４．１％＋８．０％。两者相比分别皆有显著性（Ｐ＜０．０５），用ＥＬＩＳＡ法测定了经ＰＨＡ及ＰＨＡ＋Ｒｇ＿１培养３０分钟和７２小时细胞浆ＰＴＫ的吸光度值，ＰＨＡ组为０．１２０±０．０２０，ＰＨＡ±Ｒｇ＿１组为０．１３８±０．０１５，（Ｐ＜０００１）。结果证明，Ｒｇ＿１显著升高ＰＴＫ活性。据此得出几点结论，并讨论广Ｒｇ＿１对ＰＴＫ的作用机理。     

自发性高血压大鼠主动脉平滑肌细胞异常增殖和局部肾素—血管？…

目的：探讨ＳＨＲ大鼠主动脉平滑肌细胞（ＡＳＭＣ）异常增殖和肾素－血管紧张素系统９ＲＡＳ）的关系。方法：测定血管紧张素Ⅱ（Ａｎｇ）、卡托普利（Ｃａｐ）、沙拉新（Ｓａｒ）对培养的ＳＨＲ、ＷＫＹ ＡＳＭＣ增殖的Ａｎｇ、血管紧张素转化酶（ＡＣＥ）的影响，结果：Ａｎｇ在２％血清培养基中可刺激ＳＨＲ ＡＳＭＣ增生，ＳＨＲ ＡＳＭＣ分裂增殖能力比ＷＫＹ强，ＳＨＲ ＡＳＭＣ ＲＡＳ处于高功能状态。Ｃａｐ长期（４周     

三唑仑药物的固相萃取和气相色谱法的研究

本文建立了新型安眠镇静药三唑仑的气相色谱分析方法。采用固相柱ＧＤＸ４０３提取净化，ＧＣ／ＮＰＤ法分析送检样品中的三唑仑，并用ＳＫＦ５２５Ａ作为内标进行质量控制。进行了茶水中添加三唑仑的实验研究，确立了提取净化的最佳ｐＨ值范围和提取溶剂（淋洗剂）。结果表明，该方法灵敏度高，最小检测量小于５ｎｇ，２０ｍｌ茶水中添加１０μｇ三唑仑的回收率大于７０％。并报道了几例案例的应用情况。     

环孢素抑制CD8+T细胞释放SCD8的实验研究

目的：探讨环孢素对ＣＤ８＾＋Ｔ细胞释放可溶性ＣＤ８（ＳＣＤ８）的影响。方法：在不同浓度的环孢素存在下，用ＰＨＡ刺激正常人淋巴细胞，然后用酶免疫测定法测定培养上清液的ＳＣＤ８浓度。结果：单加ＰＨＡ组的淋巴细胞在培养第３天后ＳＣＤ８显著增高，环孢素＋ＰＨＡ组在培养５ｄ后上清液中的ＳＣＤ８虽增加趋势，但不及加ＰＨＡ组显著（Ｐ〈０．０５），而且当环孢素浓度增至２５μｇ／ｍｌ时，培养上清液中的ＳＣＤ８减少不     

ABO血型不合的外周血干细胞移植及输血

探讨ＡＢＯ不合ＰＢＳＣＴ及其输血。方法采用血细胞分离机单采外周血干细胞去除红细胞。强化ＧＶＨＤ预防方案中ＭＴＸ用量及次数;选输合适血制品。结果例１主次要ＡＯＢ均不合,例２次要ＡＢＯ不合,输注ＣＳ３０００血细胞分离机单采ＣＤ３４＋细胞分别为１９０＊１６＾６／ＫＧ,２．０＊１０＾６／ＫＧ,其中红细胞残存量分别为１．４ｍｌ、２．６ｍｌ,残存率分别为３％,５．３％。移植后造血重建,完全植活,没输血溶血,也     

HPLC法测定甲磺酸加贝酯含量

目的：建立测定甲磺酸加贝酯含量的新方法。方法：以ＮＯＶＡＰＡＲＫ（１５０ｍｍ ×４ ．５ｍ ｍ ,６μｍ） 为色谱柱,甲醇水磷酸（５０∶５０∶０ ．２） 为流动相,检测波长为２３６ｎｍ ,外标法定量。结果：线形范围０ ．４ ～２ ．０μｇ ,ｒ ＝ ０ ．９９９７ ,回收率９９ ．６７ ％ ,日间ＲＳＤ 为０ ．６１ ％ 。结论：该法简便,可用以测定甲磺酸加贝酯含量。     

双酚A对大鼠睾丸Leyding细胞的毒性作用

双酚Ａ ,又名 4 二羟基二苯基丙烷 ,属低毒性 ,是制造聚碳酸脂、环氧树脂、聚酚酚树脂及聚氧树脂的重要原料。近年来有报道表明其具有雌激素样作用 ,影响生殖功能[1,2 ] 。睾丸Ｌｅｙｄｉｎｇ细胞是合成和分泌睾酮的细胞 ,并具有高度的调节性 ,本实验在建立睾丸Ｌｅｙｄｉｎｇ细胞离体原代培养的基础上 ,对双酚Ａ的Ｌｅｙｄｉｎｇ细胞毒性进行了初步的探讨。1　材料与方法1 1　实验动物　成年雄性ＳＤ大鼠 ,体重 2 0 0 ｇ左右 ,由中英合资西普尔 /必凯 (ＳＩＰＰＲ ＢＫ)实验动物有限公司提供。1 2　含双酚Ａ培养液的制备　将双酚Ａ与吐温 8…     

固定化小棘青霉菌生产磷霉素的研究

小棘青霉３１４９菌株的孢子经海藻酸钙包埋后用如下培养基培养：葡萄糖５．０％；蛋白胨２．０％；酵母膏０．３％；麦芽浸膏５．０％；（ＮＨ４）２ＳＯ４０．１％；ＭｇＳＯ．７Ｈ２Ｏ０．１％；ＦｅＳＯ４．７Ｈ２Ｏ０．０５％；ＮａＣｌ０．５％（ｐＨ６．５）控制培养温度为２８℃，当顺式丙烯磷酸加入量为０．３％时，振荡培养８天其转化率达４０％，在同样条件下高于游离菌丝细胞。固定化孢子的代谢曲线与游离菌丝细胞相比均     

Proliferation of aortic smooth muscle cells and renin-angiotensin system in SHR rats

目的：探讨ＳＨＲ大鼠主动脉平滑肌细胞（ＡＳＭＣ）异常增殖和肾素血管紧张素系统（ＲＡＳ）的关系．方法：测定血管紧张素Ｉ（Ａｎｇ）、卡托普利（Ｃａｐ）、沙拉新（Ｓａｒ）对培养的ＳＨＲ、ＷＫＹＡＳＭＣ增殖和Ａｎｇ、血管紧张素转化酶（ＡＣＥ）的影响．结果：Ａｎｇ在２％血清培养基中可刺激ＳＨＲＡＳＭＣ增生．ＳＨＲＡＳＭＣ分裂增殖能力比ＷＫＹ强，ＳＨＲＡＳＭＣＲＡＳ处于高功能状态．Ｃａｐ长期（４周）干预显著抑制ＳＨＲＡＳＭＣ异常增殖和Ａｎｇ、ＡＣＥ活性，Ｓａｒ长期干预同样抑制ＳＨＲＡＳＭＣ的增殖和ＡＣＥ活性，但Ａｎｇ水平反而升高．Ｃａｐ短期（２４小时）干预不影响两种大鼠ＡＳＭＣＲＡＳ．结论：Ｃａｐ和Ｓａｒ长期干预通过减少ＳＨＲＡＳＭＣＡｎｇ生成或阻断Ａｎｇ和特异受体结合，抑制其异常增殖．     

Multiple dose study of interactions between artesunate and artemisinin in healthy volunteers

AIMS: To investigate whether coadministration of the antimalarials artesunate and artemisinin alters the clearance of either drug. METHODS: Ten healthy Vietnamese males (Group AS) were randomized to receive a single dose of 100 mg oral artesunate (pro-drug of dihydroartemisinin) on day -5 and then once daily for 5 consecutive days (days 1-5). Oral artemisinin (500 mg) was coadministered on days 1 and 5. Another 10 subjects (Group AM) were given 500 mg oral artemisinin on day -5 and then further doses on days 1-5. Artesunate 100 mg was given on days 1 and 5. Artemisinin and dihydroartemisinin plasma concentrations on days -5, 1 and 5 were quantified by h.p.l.c. with on-line postcolumn derivatization and u.v. detection. RESULTS: In Group AS, dihydroartemisinin oral clearance values (mean (95% CI)) were similar on day 1 (32 (22, 47)) l h(-1) and day 5 (38 (28, 51)) l h(-1) of daily artesunate administration but these mean values were approximately three fold higher compared with day -5 after a single dose (95 (56, 159)). In this group, artemisinin oral clearance increased from 196 (165, 232) l h(-1) on day 1-315 (241, 410) l h(-1) on day 5. In Group AM, dihydroartemisinin oral clearance on day 1 was 39 (34, 46) l h(-1) and increased 1.6 fold to 64 (48, 85) l h(-1) on day 5. In this group, artemisinin oral clearance increased sequentially (1.5 and 4.7 fold, respectively) from 207 (151, 285) l h(-1) on day -5-308 (257, 368) l h(-1) on day 1 and to 981 (678, 1420) l h(-1) on day 5. The increase in artemisinin oral clearance between days -5 and 1 (in the absence of artesunate) was similar to that between days 1 and 5 in Group AS subjects who took daily artesunate. Dihydroartemisinin was not a significant metabolite of artemisinin. CONCLUSIONS: Artesunate (dihydroartemisinin) did not alter the elimination of artemisinin. However, dihydroartemisinin elimination was inhibited by artemisinin. Artemisinin induced its own elimination even 5 days after a single oral dose. There was no evidence for the formation of dihydroartemisinin from artemisinin.     

Stimulation of furanochromone accumulation in callus cultures of Ammi visnaga L. by addition of elicitors

In order to check the possibility of producing secondary metabolites, in vitro cultures of A. visnaga callus were established. The best growth of A. visnaga callus was obtained on Murashige and Skoog medium (MS) containing 6-benzyladenine (BA) and alpha-naphtaleneacetic acid (NAA). The study was concentrated on the induction of production of secondary metabolites by exposing callus to abiotic elicitors: benzo(1,2,3)-thiadiazole-7-carbothionic acid S-methyl ester (BION) and a suspension of silica (SiO2) and biotic elicitors: autoclaved lysates of Enterobacter sakazaki and scleroglucan. GC analysis indicated that not-elicited callus of A. visnaga grown in darkness accumulated 2 times more visnagin than the one which was grown under a 16-h photoperiod. The highest accumulation of visnagin was observed in the callus culture elicited with scleroglucan or BION. Scleroglucan induced also the accumulation of khellin in A. visnaga callus. The presented work shows that biosynthesis of pharmacologically important secondary metabolites in A. visnaga cultures could be stimulated by application of elicitors.     

Improved production of galanthamine and related alkaloids by methyl jasmonate in Narcissus confusus shoot-clumps

Liquid-shake cultured shoot-clumps of Narcisus confussus were treated with the commonly used biotic elicitors methyl jasmonate, arachidonic acid, chitosan and salicylic acid. The effects of these compounds on the growth of the explants, as well as on the amount of the alkaloids released to the liquid culture medium and accumulated in the tissues at the end of the experiment were studied. The obtained results showed that, in general, high doses of these compounds had a negative effect on the growth of the explants, particularly the salicylic acid. On the contrary, the addition of methyl jasmonate, mainly at 25 microM, promoted the release of galanthamine and other related alkaloids to the liquid medium in proportions of up to 300% in relation to the control explants, and also their accumulation in tissues. The other elicitors studied did not have any interesting effects on the production of these Amaryllidaceae-type alkaloids.     

Extracellular heme enhances the antimalarial activity of artemisinin

Artemisinin exerts the antimalarial activity through activation by heme. The hemolysis in malaria results in the elevated levels of plasma heme which may affect the activity of artemisinin. We hypothesized that the extracellular heme would potentiate the antimalarial activity of artemisinin. Hemin (ferric heme) at the pathologic concentrations enhanced the activity of artemisinin against Plasmodium falciparum in vitro and increased the levels of the lipid peroxidation products in the presence of artemisinin. The antimalarial activity of artemisinin and potentiation by hemin was decreased by vitamin E. Hemin had no effect on the activity of quinoline drugs (chloroquine, quinine and mefloquine). Furthermore, the oxidative effect of hemin in the presence of artemisinin or quinoline drugs was studied using low-density lipoprotein (LDL) oxidation as a model. Artemisinin enhanced the effects of hemin on lipid peroxidation and a decrease of tryptophan fluorescence in LDL whereas the quinoline drugs inhibited the oxidation by hemin. In conclusion, the extracellular hemin enhances the antimalarial activity of artemisinin as a result of the increasing oxidative effect of hemin.     

Toxicokinetics and hydrolysis of artelinate and artesunate in malaria-infected rats

Comparative toxicokinetic (TK) and hydrolysis studies of intravenously administered two new antimalarial agents, artelinate (AL) and artesunate (AS), were performed in malaria-infected rats using three daily equimolar doses (96 micromoles/kg). The TK evaluation was related to select one drug for severe malaria treatment in U.S. Army. Drug concentration of AS with daily dose of 36.7 mg/kg was one-third less on day 3 than on day 1, which resembled its active metabolite, dihydroartemisinin (DHA), suggesting an autoinduction of hepatic drug-metabolizing enzymes for AS. The results were similar to other artemisinin drugs, but not for AL. TK parameters of AL were very comparable from day 1 to day 3 at same AS molecular dose at 40.6 mg/kg. AS is the prodrug of DHA with the DHA/AS ratio of 5.26 compared to the ratio of 0.01 for DHA/AL. Other TK parameters revealed that the total AUC1-3 days (84.4 microg.h ml-1) of AL was fivefold higher than that of AS (15.7 microg.h ml-1 of AS plus DHA). The elimination half-life of AL (7.1 h) was much longer than that of AS (0.36 h) or DHA (0.72 h). The remarkable alteration of the TK shape of AL may be caused by poor conversion rates to DHA and an enterohepatic circulation, which is confirmed by the present TK and tissue distribution studies. Compared to AS, higher drug exposure levels and longer exposure time of AL in the rat blood may be the cause of its increased toxicity.     

Extraction of pectinase from Candida isolated from textile mill effluent and its application in bio-scouring of cotton

Pectinase is an important enzyme in industrial biotechnology and is widely utilized in industries producing food and beverages. In textile chemical processing, pectinase is utilized for a pretreatment of cotton called bio-scouring where it helps to increase the absorbency of cotton fabric and makes it suitable for further coloration. Various products based on pectinase are available in the market; however, the extraction of pectinase from a novel source which can meet the requirements of bio-scouring remains the quest of research. The present work deals with the isolation of fungus (Candida) from textile mill effluent for the extraction of extracellular thermostable pectinase. Optimization of temperature and pH of pectinase activity was studied. The crude enzyme extract showed efficient activity under the broad range of temperature and pH. The bio-scoured cotton showed an acceptable level of absorbency (absorbency < 3 s). The colour values of the dyed fabrics, scoured using conventional and bio-scouring, were comparable. The effluent load was lower for a bio-scouring process as compared to that of traditional scouring.     

短刺小克银汉霉菌对维拉帕米转化的能力

目的　研究微生物模型对药物维拉帕米的代谢转化能力 ,并与人体内药物代谢进行比较。方法通过菌株筛选 ,确定以短刺小克银汉霉菌AS 3.15 3为转化菌株 ,采用液相色谱 质谱联用技术对代谢产物进行检测 ,并考察了微生物转化体系中影响代谢物产率的主要因素。结果　该霉菌转化维拉帕米形成C -N键和C -O键断裂的 10种氧化代谢产物。发现以pH 6 .5 ,底物浓度 0 .0 75 % ,转化反应持续 96h等条件 ,代谢产物的总产率 >95 % ,其中 1种主要产物N 去短链烷基维拉帕米的产率 >6 5 %。结论短刺小克银汉霉菌AS 3.15 3对维拉帕米具有与人体类似而且广泛的代谢途径 ,是研究人体药物代谢适宜的体外模型     

黑曲霉AS 3.739对莪术二酮的羟基化修饰

目的筛选对莪术二酮具有羟基化修饰作用的菌株。方法通过二级发酵法培养菌株,应用TLC和HPLC法检测转化结果 ,通过硅胶柱层析法分离纯化羟基化产物,应用光谱学方法鉴定产物的化学结构。结果从数十株丝状真菌筛选得到黑曲霉AS 3.739对莪术二酮具有羟基化修饰作用,并分离、纯化和鉴定了2种主要产物1、2。结论用黑曲霉(Aspergillus niger)AS3.739对莪术二酮实现了羟基化修饰,2种产物分别鉴定为2β-羟基莪术二酮(1)和3α-羟基莪术二酮(2),其中主产物2为首次用曲霉(Aspergillus)菌属转化获得。     

醋毒毛花苷元对间离体浦肯野纤维动作电位时程的影响及与细胞外钾 …

AIM: To study the relation between the effect of acetylstrophanthidin on action potential duration (APD) and the extracellular potassium concentration. METHODS: Effect of acetylstrophanthidin (AS 0.15 mmol.L-1) on APD at different extracellular potassium concentrations was studied at the stimulation cycle lengths of 990 and 690 ms in sheep isolated cardiac Purkinje fibers using the standard microelectrode technique. RESULTS: At [K+]o 4.0 mmol.L-1, the biphasic effect of AS on APD appeared obviously. Both APD50 and APD90 were lengthened within the first 10 min of drug exposure. After 10 min, they were shortened at all pacing cycle lengths. On the other hand, at [K+]o 5.4 mmol.L-1, AS only shortened APD markedly without lengthening effect on it. The biphasic and monophasic effects of AS on APD were found at [K+]o 4.0 mmol.L-1 and 5.4 mmol.L-1, respectively. CONCLUSION: The effect of AS on APD was related to the concentration of [K+]o.     

短刺小克银汉霉AS 3.153菌株对安非他酮的代谢转化

目的寻找适用体外转化安非他酮的微生物菌株及采用此菌株制备吗啉环羟基化安非他酮纯品的可能性。方法首先采用液相色谱-质谱测定4种小克银汉霉对安非他酮的转化能力,确定菌株,再利用最佳菌株,采用高效液相色谱-多级质谱联用法检测转化液中安非他酮的代谢产物,并针对代谢产物2(M2)的转化进行了培养基初始pH值6水平(分别为6.0,6.5,7.0,7.5,8.0和8.5),底物浓度4水平(分别为0.025,0.05,0.1和0.2g·L-1),转化时间5水平(分别为72,96,120,144和168h)等转化条件的单因素考察,以及采用四因素三水平的正交试验设计对安非他酮进一步优化转化条件。结果根据液相色谱和质谱数据,经短刺小克银汉霉AS3.153转化,安非他酮主要形成单羟基化安非他酮和单羟基化安非他酮的硫酸结合物等3种转化产物,对M2进行分离纯化,根据质谱和核磁共振数据,确证代谢产物M2为吗啉环羟基化安非他酮。M2的最优转化条件为采用短刺小克银汉霉AS3.153培养基、初始pH8.0、底物浓度0.1g·L-1和转化时间168h时转化产率最高。结论短刺小克银汉霉AS3.153对安非他酮的转化与人体代谢结果相同,说明此菌株适宜作为研究人体药物代谢的体外模型,采用此模型及相应的优化转化条件可以制备M2纯品。