GC同时测定藏药七味铁屑丸中4种物质的含量

目的 建立同时测定藏药七味铁屑丸中土木香内酯、异土木香内酯、去氢木香内酯和木香烃内酯的GC法。方法 HP-5毛细管色谱柱（30 m×0.32 mm,0.25 μm）;FID检测器;柱温箱温度140℃,进样口温度220℃,检测器温度250℃;载气为氮气,流速为1.0 mL·min^-1,空气流速450 mL·min^-1,氢气流速45 mL·min^-1;进样量1 μL;分流比20︰1。结果 土木香内酯、异土木香内酯、去氢木香内酯和木香烃内酯的线性范围分别为7.120 2~71.202 2 μg·mL^-1（r=0.998 4）;5.478 5~54.785 1 μg·mL^-1（r=0.997 1）;8.195 3~98.344 1 μg·mL^-1（r=0.996 7）;6.395 4~76.744 8 μg·mL^-1（r=0.997 2）;平均回收率分别为98.11%（RSD=1.04%）,98.12%（RSD=1.09%）,98.39%（RSD=1.26%）,97.52%（RSD=1.25%）。结论 本方法可有效可靠地对七味铁屑丸中藏木香和木香进行定量控制。     

异土木香内酯体外抗病毒作用与机制研究

目的 研究异土木香内酯的体外抗病毒作用及机制。方法 CCK-8法检测0.125、0.500、1.000、2.000、4.000、8.000、16.000、32.000、64.000 μmol·L^-1的异土木香内酯处理24 h对人非小细胞肺癌细胞系（A549）细胞活力的影响;表达绿色荧光蛋白的水疱性口炎病毒（VSV-eGFP）以0.02的感染复数（MOI）感染A549细胞制备模型,流式细胞术检测共同孵育12 h的异土木香内酯5、10、20 μmol·L^-1对GFP阳性细胞率的影响;VSV感染（MOI=0.1） A549细胞,Western blotting检测异土木香内酯处理16 h对VSV病毒G蛋白表达的影响;分别使用甲型流感病毒（H1N1,MOI=0.1）、脑心肌炎病毒（EMCV,MOI=0.1）感染A549细胞,同时给药共同孵育8 h后,实时荧光定量PCR （qRT-PCR）检测土木香内酯对病毒RNA表达的影响;异土木香内酯分别以预处理12 h、病毒吸附过程中给药2 h、病毒吸附后给药10 h 3种不同方式给药,通过流式细胞术检测其对VSV-eGFP在A549细胞中复制的影响;异土木香内酯20 μmol·L^-1处理胚胎成纤维（MEF）细胞12 h,进行转录组测序分析;异土木香内酯5、10、20 μmol·L^-1处理MEF细胞12 h,qRT-PCR检测干扰素α1（Ifna1）、干扰素β1（Ifnb1）、干扰素诱导蛋白44（Ifi44）、干扰素刺激基因15（Isg15）的mRNA表达。结果 异土木香内酯对A549细胞的半数抑制浓度（IC₅₀）为51.01 μmol·L^-1;与模型组比较,流式结果显示异土木香内酯显著减少VSV-eGFP阳性细胞率（P<0.001）,但对病毒的吸附过程没有影响,药物预处理及吸附后给药显著抑制VSV病毒复制（P<0.01、0.001）;qRT-PCR结果显示异土木香内酯显著降低H1N1、EMCV病毒的mRNA水平（P<0.001）;Western blotting结果显示异土木香内酯显著抑制VSV的G蛋白表达（P<0.001）;转录组测序分析和qRT-PCR结果表明,异土木香内酯促进I型干扰素通路的激活,并诱导Ifna1、Ifnb1、Ifi44、Isg15（P<0.001）表达。结论 异土木香内酯通过促进基于干扰素通路的抗病毒天然免疫激活,发挥抑制病毒复制的作用。     

HPLC-DAD同时测定清感穿心莲片中3种成分的含量

目的 建立清感穿心莲片中穿心莲内酯、脱水穿心莲内酯和异丹叶大黄素3种成分的HPLC含量测定方法,为清感穿心莲片的质量控制提供参考。方法 采用HPLC-DAD法,Agilent Extend C₁₈色谱柱（4.6 mm×250 mm,5 μm）,以乙腈（A）-水（B）为流动相进行梯度洗脱,检测波长225 nm （穿心莲内酯）,254 nm （脱水穿心莲内酯）,323 nm （异丹叶大黄素）,柱温35℃,流速0.8 mL·min^-1。结果 穿心莲内酯、脱水穿心莲内酯和异丹叶大黄素分别在8.395~335.790 μg·mL^-1（r=0.999 9）,5.948~237.924 μg·mL^-1（r=0.999 9）和0.854~25.607 μg·mL^-1（r=0.999 9）内线性关系良好;平均加样回收率分别为97.2%,102.1%和98.2%,RSD分别为1.7%,3.0%和1.0%。对来自2个企业的14批次样品进行测定,穿心莲内酯、脱水穿心莲内酯和异丹叶大黄素含量范围分别为每片0.80~2.43,0.59~2.71,和0.045~0.275 mg。结论 本方法准确可靠,可用于清感穿心莲片的质量控制。     

UPLC波长切换法同时测定小陷胸汤中4种指标性成分的含量

目的 基于UPLC波长切换法构建小陷胸汤中腺苷,巴马汀,小檗碱,葫芦素B等4种指标性成分的含量测定方法。方法 固定相为Poroshell 120 EC-C₁₈ (3.0 mm×150 mm,2.7 μm)色谱柱;流动相为A相为0.1%甲酸-水,B相为0.1%甲酸-乙腈,梯度洗脱（0~6 min,B：5%→8%;6~9 min,B：8%→35%;9~12 min,B：35%→50%）,检测波长0~10 min,262 nm;10~12 min,234 nm,柱温46℃,流速0.6 mL·min^-1。结果 小陷胸汤中4个成分腺苷、巴马汀、小檗碱、葫芦素B分别在0.017～0.17,0.073～0.73,0.15～1.50 ,0.01～0.10 mg?mL^-1范围内呈良好线性关系;平均加样回收率分别为100.0%、100.6%、101.2%、99.1%,RSD均小于3.0%（n=6）;4种成分的含量分别为0.052~0.061、0.24~0.30、0.49~0.59、0.033~0.045 mg?g^-1。结论 该方法精密度高、重复性好、稳定性好、简便高效,可用于小陷胸汤的质量控制。     

HPLC-CAD法检测小建中合剂中6个活性成分及稳定性研究

目的 利用高效液相色谱联合电喷雾检测器（HPLC-CAD），建立同时测定小建中合剂中芍药内酯苷、芍药苷、甘草苷、甘草酸、桂皮醛及6-姜辣素含量的检测方法，并对小建中合剂稳定性进行研究。方法 采用Thermo Hypersil GOLD C₁₈色谱柱（250 mm×4.6 mm，5 μm），以80%乙腈（A）-10 mmol·L^-1甲酸铵（甲酸调pH值至4.0）（B）为流动相，梯度洗脱，体积流量1.0 mL·min^-1，柱温35℃，CAD雾化器温度为35℃，过滤常数为5.0，采集频率为10 Hz，进样量20 μL。同时测定小建中合剂在加速及长期试验下各成分含量的变化。结果 各成分分离度均良好，阴性无干扰；芍药内酯苷、芍药苷、甘草苷、甘草酸、桂皮醛及6-姜辣素的线性范围分别为2.01～40.16、4.21～84.20、1.18～23.64、0.60～12.04、0.70～14.06及1.30～25.90 μg·mL^-1（r≥0.999 1）；平均加样回收率（n＝6）为97.55%～98.46%，RSD≤3.0%。稳定性研究结果表明，芍药内酯苷、芍药苷、甘草苷、桂皮醛及6-姜辣素在加速条件下各成分质量浓度有明显下降，提示本品种不宜在高温下长时间放置。结论 经方法学验证，建立的HPLC-CAD法具有良好的灵敏度及准确度，为小建中合剂的全面质量控制提供科学依据，本品应置阴凉处贮藏。     

60Co-γ射线辐照灭菌对香砂枳术丸中5种有效成分的影响

目的 建立同时测定香砂枳术丸中木香烃内酯、去氢木香烃内酯、柚皮苷、橙皮苷及新橙皮苷5种活性成分含量的测定方法,考察⁶⁰Co-γ射线辐照前后有效成分含量的变化。方法 采用Agilent-C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相为乙腈-0.5%磷酸溶液,梯度洗脱,检测波长为225 nm,流速为1.0 mL·min^-1,柱温为35℃。分别选择辐照剂量2,5,8 kGy对香砂枳术丸进行辐照,测定辐照前后香砂枳术丸中5种有效成分的含量,并进行成组t-检验考察其显著情况。结果 木香烃内酯、去氢木香烃内酯、柚皮苷、橙皮苷及新橙皮苷分别在0.060 6~3.032 5,0.025 8~1.290 0,0.037 3~1.865 0,0.044 2~2.210 0及0.033 6~1.680 0 μg内有良好线性,平均加样回收率分别为100.6%,101.0%,99.7%,100.2%和99.5%,RSD分别为0.5%,1.3%,0.8%,1.0%和0.7%。经过2,5,8 kGy辐照后,香砂枳术丸所含的有效成分木香烃内酯、去氢木香烃内酯、柚皮苷、橙皮苷及新橙皮苷均有变化,经成组t-检验,在>5 kGy后,木香烃内酯及去氢木香烃内酯辐照前后含量有统计学意义（P<0.05）。结论 所建立的香砂枳术丸活性成分测定方法回收率高、重复性好、简单实用,可作为香砂枳术丸质控方法。在辐照剂量≤ 5 kGy时,各成分变化不显著,为香砂枳术丸辐照灭菌手段提供了参考。     

大环骨架结构的倍半萜化合物——Ⅵ.马兜铃新内酯的化学结构测定

自绵毛马兜铃(Aristolochia mollissima Hance)根茎中分得九个化合物,其中已报道的七个化合物是尿囊素、马兜铃内酯、绵毛马兜铃内酯、β-谷甾醇、马兜铃酸A、9-乙氧基马兜铃内酯和9-乙氧基马兜铃内酰胺,本文报道结晶K₃的化学结构,经光谱分析(IR,¹H-NMR,¹³C-NMR,2D-NMR和MS),化学反应及X-衍射晶体分析,确证K₃为一个新骨架结构的倍半萜化合物,命名为马兜铃新内酯。     

HPLC同时测定藤珠胃康颗粒中4个皂苷类成分

目的 建立HPLC同时测定藤珠胃康颗粒中4个皂苷类成分（人参皂苷Re、人参皂苷Rb₁、人参皂苷Ro和竹节参皂苷Ⅳa）的方法。方法 采用Hypersil GOLD C₁₈色谱柱（250 mm×4.6 mm,5 μm）,流动相为乙腈-0.2 %磷酸水溶液,梯度洗脱（-5~0 min,19%乙腈;0~14 min,19%→26%乙腈;14~22 min,26%→29%乙腈;22~30 min,29%乙腈;30~40 min,29%→35%乙腈;40~55 min,35%乙腈）,体积流量1.0 mL·min^-1,检测波长203 nm,柱温30 ℃。结果 人参皂苷Re在0.080 4~1.608 μg（r=1）,人参皂苷Rb₁在0.108 8~2.176 μg（r=0.999 9）,人参皂苷Ro在0.288 8~5.776 μg（r=0.999 9）,竹节参皂苷Ⅳa在0.176 0~3.520 μg（r=0.999 9）内线性关系良好;平均回收率（n=6）分别为99.41%,101.92%,99.76%,100.31%,RSD值分别为2.22%,2.07%,0.33%,0.64%。结论 本实验所建立的方法简单,专属性强,重复性好,可用于藤珠胃康颗粒中人参皂苷Re、人参皂苷Rb₁、人参皂苷Ro和竹节参皂苷Ⅳa的测定。     

HPLC同时测定瑞香狼毒中瑞香素、伞形花内酯和东莨菪内酯的含量

目的 建立HPLC同时测定瑞香狼毒中瑞香素、伞形花内酯和东莨菪内酯含量的方法。方法 样品经无水乙醇回流。采用Synergi 4u Polar-RP 80R柱（4.6 mm×250 mm,5 μm）,流动相为甲醇-1%冰醋酸溶液,等度洗脱,流速：1.0 mL·min^-1,检测波长：324 nm,柱温：30℃。结果 瑞香素在0.012~0.250 μg（r=0.999 9）、伞形花内酯在0.149~2.981 μg（r=0.999 9）、东莨菪内酯在0.012~0.239 μg（r=0.999 9）均呈良好的线性关系;瑞香素、伞形花内酯和东莨菪内酯的加样回收率分别为99.0%（RSD=0.94%）,98.7%（RSD=0.77%）和98.3%（RSD=1.16%）。结论 3种香豆素成分在35 min内达到基线分离,该方法分析时间短、稳定性和回收率良好,对瑞香狼毒药材的质量控制具有一定的参考价值。     

脑心通胶囊联合阿替普酶治疗急性脑梗死的临床研究

目的 研究银杏内酯滴丸中的主要成分银杏内酯A、B在健康受试者体内的药动学特征,为制定合理的临床给药方案提供依据。方法 采用随机、开放的试验设计,10例健康受试者单次口服银杏内酯滴丸后,按预定时间点采集血样,肝素锂抗凝,离心分离血浆。采用LC-MS/MS法测定血浆样品中银杏内酯A、B的开闭环总质量浓度,以及银杏内酯A、B闭环质量浓度,并应用WinNonlin 6.3软件非房室模型计算药动学参数。结果 健康受试者单次口服银杏内酯滴丸后,银杏内酯A闭环及开闭环总量的t_max分别为（3.05±1.40）、（3.40±1.22）h,C_max分别为（84.3±32.8）、（92.2±35.0）ng/mL,C_max比值为91.4%,AUC_0～t分别为（636±183）、（753±205）ng·h/mL,AUC_0～t比值为84.5%,t_1/2分别为（13.00±10.30）、（12.90±8.49）h;银杏内酯B闭环及开闭环总量的t_max分别为（3.15±1.42）、（3.35±1.25）h,C_max分别为（74.10±31.50）、（148.00±60.10）ng/mL,C_max比值为50.1%,AUC_0～t分别为（627±202）、（1 410±431）ng·h/mL,AUC_0～t比值为44.5%,t_1/2分别为（13.20±5.83）、（13.7±5.83）h。结论 健康受试者口服银杏内酯滴丸后,银杏内酯A、B吸收速率适中,消除速率适中,在人血浆中银杏内酯A主要以闭环形式存在,而银杏内酯B以开、闭环2种形式存在、暴露量相当。     

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.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

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.