探讨博来霉素(bleomycin,BLM)损伤所致小鼠肺纤维化发病过程中肺泡巨噬细胞的细胞周期抑制因子p21 (cell cycle inhibitor p21,p21)蛋白表达及其对巨噬细胞活化的调节作用.免疫荧光染色法检测肺纤维化组织中肺泡巨噬细胞中CD206的表达;RT-PCR (reverse transcription-polymerase chain reaction)法检测巨噬细胞活化标志蛋白的表达;巨噬细胞/成纤维细胞共培养法检测巨噬细胞对成纤维细胞活化及胶原收缩能力的影响;采用免疫荧光和免疫印迹法检测巨噬细胞中p21蛋白表达的变化;采用21敲低和过表达病毒感染肺泡巨噬细胞,RT-PCR和巨噬细胞/成纤维细胞共培养法检测改变p21表达对肺泡巨噬细胞促纤维化表型的调节作用.动物实验操作过程依照中国医学科学院、北京协和医学院药物研究所实验动物管理与动物福利委员会的要求执行.结果 显示:与对照组比较,模型组肺泡巨噬细胞CD206的表达增加、巨噬细胞活化标志蛋白表达增加、模型组肺泡巨噬细胞促进成纤维细胞的活化并增强其胶原收缩能力.模型组小鼠肺泡巨噬细胞中p21蛋白表达增加.敲低p21可显著抑制巨噬细胞促纤维化表型,而过表达对照小鼠肺泡巨噬细胞中21则促进其促纤维化表型的转化.以上研究结果表明,肺纤维化小鼠肺泡巨噬细胞中p21表达增加,p21可通过促进巨噬细胞活化参与肺纤维化发病.

经皮给药系统是目前制药工业的研究热点之一,因具有避免肝首过效应及肠胃灭活作用、维持稳定血药浓度避免峰谷波动性现象、延长有效作用时间减少用药次数、改善患者用药依从性等优点,越来越受到患者的青睐及研究人员的重视.渗透性改善技术是经皮给药技术发展中的关键技术.提高经皮给药渗透性的方法分为物理和化学方法,加热促渗方法是发展较为成熟的物理技术.本文从热促透方法及其促渗机制等方面综述加热促渗技术研究进展,为经皮给药系统相关研究提供借鉴和参考.

目的:首次研究何首乌中大黄素型单蒽酮的潜在肝细胞毒性及其作用机制.方法:本研究利用悬滴技术构建以人肝细胞HepaRG和人星型细胞HSC为基础的3D多细胞聚球体模型,用该模型评价何首乌中大黄素型单蒽酮成分的重复给药毒性,并与2D模型下单次给药毒性进行比较.进一步研究大黄素型单蒽酮对主要肝药酶和外排转运体表达的影响.结果:肝脏3D多细胞聚球体模型体外培养15 d仍然可以维持较好的肝脏功能.大黄素型单蒽酮2D培养给药48 h,IC50为1.091 μg· mL-1.肝脏3D悬滴模型下连续给药6d,同浓度大黄素型单蒽酮对肝细胞毒性显著性小于2D培养评价结果.RT-PCR结果显示大黄素型单蒽酮体外多次给药上调主要肝药酶CYP3A4,CYP2B6及外排转运体P-糖蛋白和多药耐药转运蛋白2(MRP2).结论:何首乌中的新化合物大黄素型单蒽酮具有明显的肝细胞毒性,体外3D模型多次给药引起主要肝药酶和外排转运体表达上调,为肝细胞降低大黄素型单蒽酮毒性提供了潜在机制,同时也提示服用含有大黄素型单蒽酮化合物的中药可能会产生药物相互作用.

测定荆芥药材不同药用部位中铅(Pb)、镉(Cd)、镍(Ni)3种重金属元素的含量,比较分析荆芥不同部位的污染状况.对荆芥样品进行湿法消解,采用石墨炉原子吸收光谱法(GFAAS)测定样品中Pb、Cd、Ni含量,以质量分数2％磷酸二氢铵-1.67％硝酸镁作为基体改进剂测定Pb、Cd;以质量分数0.2％硝酸钯-2％磷酸二氢铵作为基体改进剂测定Ni.Pb、Cd、Ni的测定波长分别为283.3、228.8和232.0 nm,灯电流分别为10、6和25 mA;狭缝分别为0.7、0.7和0.2 L.Pb、Cd、Ni元素线性关系分别为Y=0.002 8X+0.002 7(r=0.998 9)、Y=0.034 8X+0.005 4(r=0.998 3)和Y=0.002 9X+0.004 3(r=0.998 4),回收率为98.2％～102.9％;荆芥梗中Pb、Cd和Ni的质量分数测定结果分别为0.55～0.95、0.05～0.11和0.1～1.22 mg/kg;荆芥穗质量分数测定结果分别为0.58～ 1.73、0.03～0.14和0.18～1.42 mg/kg.重金属元素Pb、Cd、Ni检测方法可靠,随着采收时间延长,重金属元素易由荆芥梗向荆芥穗中富集,为荆芥重金属限度质量标准制定以及临床用药部位的选择提供参考.

本研究旨在探究不同配比的18α、18β-甘草酸(18α、18β-GA)对P-糖蛋白(P-gp)转运功能和大鼠CYP3A酶活性的影响.建立Caco-2细胞单层模型,通过检测接收池罗丹明123溶液的荧光强度,考察不同配比的18α、18β-GA对P-gp转运功能的影响;利用大鼠的肝脏制作微粒体,通过温孵实验检测不同浓度及不同比的18α、18β-GL对CYP3A酶活性的影响.当18α-GA和18β-GA配比为6:4时,对P-gp的抑制作用最强;当两种异构体配比为0:10时,对P-gp的诱导作用最强.18α-GA浓度在50～100 μg/mL范围内,对CYP3A酶活性有明显抑制作用.18β-GA浓度在10～50μg/mL范围内,对CYP3A酶活性有明显诱导作用.在甘草酸总浓度为10μg/mL,18α、18β-GA比例为2:8时,对CYP3A酶活性的诱导作用最强,高于相同浓度的18β-GA单体.随着药物浓度及18α-GA比例增加,对CYP3A酶活性的抑制作用增强.当浓度为100μg/mL、18α与18β-GA配比为10:0时,对CYP3A酶抑制作用最强.18α、18β-GA的配比不同,对P-gp转运功能及对大鼠CYP3A酶活性的影响不同,这将为指导临床用药提供科学依据.

The objective of this paper was to investigate the effect of different proportions of 18α-and 18β-glycyrrhizic acid (Gly) on the morphology of organs in rats with alcoholic liver injury. Healthy male rats were randomly divided into ten groups, including normal control group, model group, positive control group (silymarin), 7 drug groups (18α-Gly and 18β-Gly group 10.83 mg/kg at ratios of 10:0, 8:2, 6:4, 5:5, 4:6, 2:8, 0:10). At the fourth week, important organs were collected and weighted, including liver, kidney and spleen, which were then stained by routine oil red O and hematoxylin-eosin. The histology of organs was observed by optical microscope. The results showed that 18α-Gly and 18β-Gly at the ratios of 2:8 significantly decreased organs indexes of respiratory and digestive system, obviously reduced muscle index and increased brain index, and meanwhile decreased cardiac index and spleen index. Additionally, 18α-Gly and 18β-Gly at the ratios of 4:6 and 2:8 reduced hepatocytes steatosis and lipid accumulation. 18α-Gly and 18β-Gly at the radio of 4:6 or less could decrease inflammation of the kidney, and meanwhile, alleviated injury to spleen demonstrated by the clear boundary of red and white pulp of spleen. Collectively, the optimal compatibility proportion of 18α-Gly and 18β-Gly on protecting organ injury induced by alcohol was 4:6 and 2:8.

中国"药食同源"的思想源远流长,随着国家对中医药产业的日益重视,伴随中医药产业走向世界,药食同源观点必将被世界所接受与认可.综述了药食同源文化来源、 应用现状和发展前景,总结了药食同源品种的质量标准、 检测方法、 企业状况和中药监管法律法规等环节的现状及面临问题,提出"采用多种检测方法、 完善质量标准、 重视科研创新和加强监督管理"等多方面监管措施,以推进药食同源产业的健康发展.

网络课程质量保证是我国网络教育领域最受关注却一直难以解决的问题.网络课程的多元性与技术的快速发展使得对其进行系统评估和质量保证更难.本研究旨在通过对美国Quality Matters的深度探讨,分析其成为美国网络课程评价与质量保证领军者的核心竞争力:独特的课程质量评价标准体系、专家同行评审机制、多元的认证项目和扎根于院校的理念.QM具有宽广的国际视野并以教育技术研究为坚实基础,可为我国网络课程评价的顺利开展提供新思路.

当今国际社会人才的竞争愈演愈烈,STEM作为复合型创新人才培养的有效途径引起各国的重视.我国STEM课程实施过程中面临着教学模式、师资配备、政策支持等多方面挑战,BOPPPS教学模式强调以学习者为中心,注重学生实践与反馈,较传统的教学模式更适合STEM课程.通过以小学Scratch课程为例进行基于BOPPPS模式在STEM课程的设计探究,为解决当前STEM课程教学模式存在的问题提供新思路.

研究甘草酸18位差向异构体18α-甘草酸(18α-Gly)与18β-甘草酸(18β-Gly)不同配比和浓度对结肠癌(Caco-2)细胞P-糖蛋白(P-gp)功能的影响,选择最佳浓度比例.建立细胞模型,选择甘草酸总浓度为1∶ 10∶ 30∶ 60∶ 120∶240 μmol/L,18α-Gly与18β-Gly物质的量比分别为10∶0、8 ∶ 2、6 ∶ 4、5 ∶ 5、4 ∶ 6、2∶8、0∶10,根据细胞存活率,选择合适浓度比例,利用流式细胞仪测定细胞内荧光强度,寻找荧光强度最大的浓度比,即对P-gp抑制作用最强,P-gp功能最弱.研究结果表明甘草酸总浓度为1μmol/L时,随着18α-Gly比例的减少,荧光强度逐渐减弱,P-gp诱导作用逐渐增加.甘草酸总浓度为10 μmol/L和60 μmol/L时,随着两者物质的量比的变化,荧光强度变化明显;当总浓度为10 μmol/L,n(18α-Gly)∶n(18β-Gly)=4∶6时,荧光强度较强,抑制作用明显;当总浓度为60 μmol/L,两者物质的量比为5∶5时荧光强度最强,抑制作用最强.

In this paper,the authors focus on the problem of pharmaceutics experiment in pharmaceutical profession and put forward the significance of establishing comprehensive experiment.Moreover, the authors discuss the content of the comprehensive experiment,the specific requirements,grading methods and implementation experience,which emphasizes on the necessity of open comprehensive experimental in traditional pharmaceutics experimental teaching.

According to the information of HPLC, MS and NMR, the chromatographic and spectral characteristics of principal component isomers and the related substances in bulk drug of ammonium glycyrrhizate were studied. Firstly, according to the information of retention time, the validity of the substance's structure assignments of relative retention time of 1.2 in the European pharmacopoeia 7.0 version and British pharmacopoeia 2012 version was questioned. A high resolution electrospray ionization mass spectrometry(HR-ESI-MS) was used to confi rm the molecular weight and formula, and the molecular unsaturation was calculated. Moreover, the 1H NMR and 13C NMR signals were completely assigned by 2D-NMR techniques including correlation spectroscopy(COSY), heteronuclear singlequantum coherence(HSQC), nuclear overhauser enhanced spectroscopy(NOESY), and heteronuclear multiple-bonds correlation(HMBC). As a result, we concluded that the principal components of ammonium glycyrrhizate were 18α,20β-glycyrrhizic acid and 18β,20β-glycyrrhizic acid, the related substance A and the compound of relative retention time of 1.2 were(18β,20β)-24-hydroxyglycyrrhizinic acid and 18β,20α-glycyrrhizic acid, respectively.

Objective: To analyze the content differences and variation trends of the principal component isomers and related substances in the four generations of different glycyrrhizin preparations on the domestic and foreign markets. Methods: The condition of sample pretreatment was investigated,and the HPLC analytical method was established for the simultaneous separation and detection of 18α- glycyrrhizic acid,18β- glycyrrhizinic acid,related substance A and related substance B of ten brands of glycyrrhizin preparations. The labeled amount and composition ratio of 18α- Gly and 18β- Gly of glycyrrhizin preparations were calculated. The content differences and variation trends of each component in glycyrrhizin preparations were analyzed. Results: The labeled amount of ten brands of glycyrrhizin preparations accorded with the specification in the Chinese Pharmacopoeia. The content of principal components and related substances and the composition ratio of 18α- Gly and 18β- Gly showed significance difference among different brands of glycyrrhizin preparations. The total amount of each component in imported glycyrrhizin preparations was higher than that in domestic products. Conclusion: The method is simple,sensitive,and reproducible with accurate and reliable results,which can be used for content detection of various dosage forms of glycyrrhizinate and quality control. The analysis results can provide reference for the selection of clinical medicine and new- medicine development.

针对中国国家药品标准(WS1-XG-2002)中,甘草酸单铵盐含量测定方法分析时间长、效率低、不能实现对甘草酸主成分异构体18α-甘草酸(18α-Gly)和18β-甘草酸(18β-Gly)的有效分离,以及未对有关物质进行准确定量分析和质量控制等问题,改进并建立甘草酸单铵盐原料药及不同剂型制剂中有关物质及含量的检测方法.对色谱条件进行优化,采用Durashell-C18柱(4.6 mm×250 mm,5μm),以0.01 mol/L高氯酸铵(氨水调节pH至8.20)-甲醇(48∶52,V/V)为流动相,流速0.80 mL/min,检测波长254 nm,柱温50℃,进样量10 μL.18α-Gly,18β-Gly,甘草酸单铵盐杂质A和杂质B在0.5～100.0 μg/mL内线性关系良好(r=0.999 9),检出限分别为0.10,0.10,0.15,0.15μg/mL,平均回收率(n=3)为98.06％～101.61％.与中国国家药品标准(WS1-XG-2002)收载的甘草酸类制剂质量标准比较,本实验建立的方法能够实现对主成分异构体的有效分离,并能真实、准确地检测各有关物质的含量,方法精密度,重现性良好,灵敏度高,结果准确可靠,可用于甘草酸单铵盐原料药及其不同剂型制剂的检测分析及质量控制.

A chemicals derivative-synchronous fluorescence method for determination of the content of γ-aminobutyric acid in injection has been described.The ramification of γ-aminobutyric acid has been synchronous fluorescence scanned to examine the factors which affect the intensity of systematic fluorescence.The optimum experiment conditions: 8.0×10-3 mol/L borax buffer solution at pH=9.6,the combination reagents of 1.0×10-5 mol/L OPA-2.86×10-5 mol/L β-MCE were derivatization reagent,the derivative time was 60 min.The band-pass width of Ex,Em were 5.0 nm,λem=455.0 nm,Δλ=120.0 nm,The temperature of the solution under test was Less than 25 ℃.The results showed that the peak shape was the best and the fluorescence intensity was maximum gained under these conditions by the synchronous fluorescence spectra.The linear ranges were 2.50 ~50.00 μg/L(0.999 0) for γ-aminobutyric acid.The detection limit of γ-aminobutyric acid was 0.79 μg/L.The method which had high sensitivity,easy operations and low costs could be used to determine the content of γ-aminobutyric acid in injection.

A micellar electrokinetic chromatography (MEKC)sweeping technique was developed for the determi-nation of folidol in vegetables.The effects of the concentration of SDS, the concentration of NaH2PO4, the pH of running buffer,the concentration of organic additive,the concentration of solution to dilute sample, voltage and injection pressure on enrichment effect were discussed.The effect of additiving the activated carbon to sample on recovery was discussed. 20 mmol/L NaH2PO4+120 mmol/L SDS(pH 6.2),Methanol 30%(volume ratio) was used as running buffer,the voltage was 7 kV,the injection pressure was 2 406.25 kPa ×s.The recovery was not effected by 1.8 g activated carbon adding to the vegetable.The calibration curves had good linearity in the range of 0.06 mg/L-2 mg/L for folidol,the correlation coefficients was 0.998 9,the detection limit was 0.038 mg/L. At spikedlevels of 0.32 mg/kg-0.48 mg/kg,the recoveries were 90.30 %-90.52 %,the relative standard deviations was 1.01%-1.44%(n=4).The MEKC method based on on-line Sweeping technique can be used to detect the residuals of trace pesticides in vegetables.

The basic principles and characteristics of two separation modes of capillary electrophore-sis (CE) were introduced. The advantages of CE on-line concentration technique , the detection principles of on-line stacking and on-line sweeping for micellar electrokinetic chromatography (MEKC) were discussed. This review summarized the research progress of two separation modes and two on-line concentration techniques on analysis to trace exogenous medicine in food.Meanwhile ,the applied prospect of on-line concentration technique for CE was expected.

A simple, fast and sensitive analytical method for the simultaneous separation and detection of 18alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B by RP-HPLC and drug quality standard was established. The structures of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate have been confirmed. Reference European Pharmacopoeia EP7.0 version, British Pharmacopoeia 2012 version, National Drug Standards of China (WS 1-XG-2002), domestic and international interrelated literature were referred to select the composition of mobile phase. The experimental parameters including salt concentration, pH, addition quantities of organic solvent, column temperature and flow rate were optimized. Finally, the assay was conducted on a Durashell-C18 column (250 mm x 4.6 mm, 5 microm) with 0.01 mol x mL(-1) ammonium perchlorate (add ammonia to adjust the pH value to 8.2) -methanol (48 : 52) as mobile phase at the flow rate of 0.8 mL x min(-1), and the detection wavelength was set at 254 nm. The column temperature was 50 degrees C and the injection volume was 10 microL. The MS, NMR, UV and RP-HPLC were used to confirm the structures of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate. Under the optimized separation conditions, the calibration curves of 18 alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B showed good linearity within the concentration of 0.50-100 microg x mL(-1) (r = 0.999 9). The detection limits for 18alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B were 0.15, 0.10, 0.10, 0.15 microg x mL(-1) respectively. The method is sensitive, reproducible and the results are accurate and reliable. It can be used for chiral resolution of 18alpha-glycyrrhizinic acid, 18Pbeta-glycyrrhizinic acid, and detection content of principal component and related substances of raw material drug of ammonium glycyrrhizinate. It is concluded that the separation of principal component isomer of raw material drug of ammonium glycyrrhizinate and the validity of the substance's structure assignments of retention time being 1.2 in the European pharmacopoeia EP7.0 version, British pharmacopoeia 2012 version remains open to question. It may be of practical value for the quality control of raw material drug, preparation, and Chinese herbal medicine of ammonium glycyrrhizinate.

An analytical method for the simultaneous determination of 18alpha-glycyrrhizic acid, 18beta-glycyrrhizinic acid, related substances A and B and drug quality standard by reversed-phase high performance liquid chromatography (RP-HPLC) was established. The assay was carried out on a Durashell-C18 column (250 mm x 4.6 mm, 5 microm) with 10 mmol/L ammonium perchlorate (the pH value was adjusted to 8.20 with ammonia)-methanol (48:52, v/v) as mobile phase at a flow rate of 0.80 mL/min, and the detection wavelength was set at 254 nm. The column temperature was 50 degrees C and the injection volume was 10 microL. Under the separation conditions, the calibration curves of the analytes showed good linearities within the mass concentrations of 0.50 -100 mg/L (r > 0.999 9). The detection limits for 18alpha-glycyrrhizic acid, 18beta-glycyrrhizinic acid, related substances A and B were 0.15, 0.10, 0.10, 0.15 mg/L, respectively. The average recoveries were between 97.32% and 99.33% (n = 3) with the relative standard deviations (RSDs) between 0.05% and 1.06%. The method is sensitive, reproducible, and the results are accurate and reliable. The method can be used for the determination of principal components and related substances of ammonium glycyrrhizinate for the quality control of raw material drug of ammonium glycyrrhizinate.

衰老是复杂的生命过程,机体衰老时不仅身体外表发生了变化,体内亦发生了许多生理和组织学的变化,国内外对此进行了大量研究,并发现了许多关键指标,为抗衰老措施提供了依据. 1.免疫衰变与抗衰老 免疫功能的衰退是造成机体衰老的重要因素,通过改善免疫系统功能,可以延缓衰老的进程.实验研究显示,衰老动物胸腺和脾脏的重量减轻,胸腺指数降低,T淋巴细胞因子活性降低,巨噬细胞的吞噬作用降低,自然杀伤细胞(NK)活性亦降低.通过升高动物胸腺指数,降低抑制性T淋巴细胞数量,升高辅助性T淋巴细胞的比值,可以延缓衰老的进程.