基于多指标成分测定昆仙胶囊的体外溶出度

目的 建立昆仙胶囊多指标成分的溶出度测定方法，以有效控制其内在质量。方法 采用Agilent Zorbax SB-C₁₈色谱柱(250 mm×4.6 mm，5 μm)，以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱，流速为0.8 mL·min^–1，柱温33℃，检测波长270 nm。采用篮法，以900 mL pH 6.8的磷酸盐缓冲液为溶出介质，转速100 r·min^–1，测定昆仙胶囊中朝藿定A、朝藿定B、朝藿定C及淫羊藿苷的溶出度，并通过相似因子(f₂)法对不同批次样品的溶出曲线进行相似性比较。结果 朝藿定A、朝藿定B、朝藿定C及淫羊藿苷在各自范围内线性关系良好(r=0.999 9)，平均回收率分别为100.9%(RSD=1.29%)，102.1%(RSD=1.18%)，100.8%(RSD=1.30%)及99.9%(RSD=0.92%)，不同批次昆仙胶囊中4种成分的溶出曲线相似度较高，相似因子f₂均>67。结论 该法简便、准确、可行，专属性良好，可为昆仙胶囊的质量评价提供实验依据。     

HPLC法测定仙灵骨葆胶囊中5种成分的含量

目的 采用多波长HPLC法建立仙灵骨葆胶囊中淫羊藿苷、朝藿定C、川续断皂苷Ⅵ、补骨脂素及异补骨脂素的含量测定方法。方法 采用月旭Ultimate^® XB-C₁₈柱,流动相采用乙腈-水系统,梯度洗脱;柱温：30 ℃;检测波长：淫羊藿苷、朝藿定C、川续断皂苷Ⅵ为212 nm,补骨脂素、异补骨脂素为246 nm。结果 5种成分均能达到基线分离,各成分的进样量分别在：淫羊藿苷0.008 2~0.328 μg（r=0.999 5）、朝藿定C 0.055 6~2.224 μg（r=0.999 6）、川续断皂苷Ⅵ 0.144 1~5.764 μg（r=0.999 6）、补骨脂素0.005 4~0.215 2 μg（r=0.998 0）、异补骨脂素0.006 6~0.265 6 μg（r=0.998 5）范围内线性关系良好,平均回收率分别为97.59%、98.58%、98.11%、97.86%、98.22%,RSD均小于2.0%。结论 该方法操作简便、分离良好、数据准确、灵敏度高,可用于仙灵骨葆胶囊的多指标成分定量测定。     

淫羊藿苷对脂多糖诱导成骨细胞骨架F-actin损伤的保护作用

目的 考察淫羊藿苷对成骨细胞骨架F-actin损伤的保护作用及其对相关信号通路的调控。方法 采用脂多糖诱导大鼠成骨细胞损伤模型。实验分为空白对照组、模型组和0.1,1,10 μg·mL^-1淫羊藿苷组,MTT法观察淫羊藿苷对成骨细胞活性的影响,TRITC-Phalloidin荧光染色观察淫羊藿苷对细胞骨架的影响,ELISA法测定各组成骨细胞中F-actin的含量,RT-PCR法测定细胞骨架Rho A和Cofilin的mRNA表达量。结果 与空白对照组比较,100 μg·mL^-1脂多糖能显著降低成骨细胞存活率（P<0.01）;与模型组比较,1~10μg·mL^-1淫羊藿苷预处理后能显著提高细胞存活率（P<0.05或P<0.01）;而0.1 μg·mL^-1淫羊藿苷则无显著影响。与模型组比较,1~10μg·mL^-1淫羊藿苷能显著抑制成骨细胞F-actin解聚（P<0.05）,抑制Rho A的mRNA表达（P<0.05或P<0.01）,0.1~10 μg·mL^-1淫羊藿苷能抑制Cofilin的mRNA表达（P<0.01）。结论 淫羊藿苷对脂多糖诱导的细胞骨架F-actin损伤具有保护作用,其机制与抑制细胞骨架相关因子Rho A和Cofilin的mRNA表达有关。     

补肾温肺微乳中淫羊藿苷的大鼠在体肠吸收特征研究

摘 要 目的： 研究补肾温肺微乳中淫羊藿苷的大鼠不同肠段的吸收特性,并考察P 糖蛋白( P gp)对其肠吸收的影响。方法: 采用大鼠在体单向肠灌流实验,采用HPLC法测定淫羊藿苷的含量,分别研究肠段不同吸收部位、不同淫羊藿苷浓度、 P gp抑制药维拉帕米对淫羊藿苷单体及其在补肾温肺微乳中吸收的影响。结果: 淫羊藿苷单体在各肠段的吸收速率常数(Ka)和表观吸收系数(P_app)差异显著(P<0.01);在0.10～0.40 ml·min^-1流速内和0.1～1.0 mg·ml^-1质量浓度内,十二指肠吸收速率常数和表观吸收系数无显著差异;补肾温肺微乳中淫羊藿苷的肠灌流试验结果与淫羊藿苷单体相比略小,但无显著差异; P gp抑制药对淫羊藿苷的肠吸收有影响,加入0.1 mmol·L^-1盐酸维拉帕米后,Ka和P_app分别为（2.13±0.66）×10^-2h·cm^-1和（0.23±0.051）×10^-6 cm·s^-1,与不加P gp 抑制药组相比较差异显著(P<0.01)。结论: 淫羊藿苷属于难吸收化合物,淫羊藿苷和补肾温肺微乳中淫羊藿苷在大鼠不同肠段的吸收具有相似的吸收特性;其在大鼠肠内的吸收受 P gp外排转运影响。因此,推测淫羊藿苷可能是P gp的底物。     

HPLC法测定肌瘤化消颗粒中淫羊藿苷和丹参酮ⅡA

目的 建立测定肌瘤化消颗粒中淫羊藿苷和丹参酮II_A的HPLC法。方法 采用高效液相色谱法,Diamonsil^TM C₁₈色谱柱（250 mm×4.6 mm,5 μm）;流动相：乙腈-水,梯度洗脱;检测波长为270 nm;柱温为40℃;体积流量为1.0 mL/min;进样量10 μL。结果 淫羊藿苷在7.5～50 μg、丹参酮II_A在3～20 μg与峰面积的线性良好（r=0.999 9）。平均回收率分别为100.2%、100.1%,RSD值分别为0.9%、1.3%（n=9）。结论 本法操作简便,重现性好,可有效控制肌瘤化消颗粒的质量。     

基于Micro-CT技术分析淫羊藿苷和朝藿定C对糖皮质激素性骨质疏松症小鼠骨组织微结构的影响

目的 应用Micro-CT技术观察淫羊藿苷和朝藿定C对糖皮质激素性骨质疏松症（GIOP）小鼠模型骨组织骨量、骨微结构的影响。方法 8周龄C57/BL6雄性小鼠随机分为4组：对照组、模型组、淫羊藿苷（200 mg/kg）组和朝藿定C（200 mg/kg）组,除对照组外,其他3组小鼠im地塞米松5 mg/kg,0.125 mg/次,每周3次,制备GIOP模型,对照组im等体积的生理盐水,造模同时,每天ig给药1次,连续60 d后取材,采用micro-CT方法对胫骨近端骨微结构进行三维分析;HE染色病理切片观察胫骨近端骨组织病理形态。结果 骨量参数：与对照组比较,模型组骨矿物质含量（BMC）、骨矿物质密度（BMD）、组织矿物含量（TMC）、组织矿物质密度（TMD）均显著下降（P<0.05、0.01）;与模型组比较,淫羊藿苷组、朝藿定C组BMC、BMD、TMC、TMD指标均显著提高（P<0.05）;其中朝藿定C组各指标均较淫羊藿苷组显著升高（P<0.05）。与对照组比较,模型组相对骨体积（BV/TV）、骨小梁数量（Tb.N）、骨小梁厚度（Tb.Th）指标均显著下降（P<0.05）,骨小梁分离度（Tb.Sp）、结构模型指数（SMI）均显著提高（P<0.05）;与模型组比较,淫羊藿苷、朝藿定C组BV/TV、Tb.N、Tb.Th显著升高,Tb.Sp、SMI显著下降（P<0.05）,其中朝藿定C组各指标均较淫羊藿苷组改善显著（P<0.05）。HE染色病理切片示,模型组骨小梁数目明显减少,稀疏断裂,大部分不能连接成网状,骨髓腔明显增大,骨小梁结构出现较大的空白区域;淫羊藿苷及朝藿定C组小鼠骨小梁明显宽厚,数目也显著增加,骨小梁断裂较少,骨小梁光滑,接近对照组,其中朝藿定C组增加较淫羊藿苷组明显。结论 地塞米松诱导的骨质疏松小鼠模型成功建立,朝藿定C和淫羊藿苷抗骨质疏松活性明显,主要通过增加骨量和改善骨小梁微结构来最终提高骨强度,其中朝藿定C抗骨松作用更强;Micro-CT技术与传统的检测方法相比,在中药干预GIOP骨微结构参数分析上具有便捷、高效,经济,图像多维、全面,准确的优势。     

扶正胶囊质量标准的提高

摘 要 目的：建立扶正胶囊的质量标准。方法: 采用 TLC 法对扶正胶囊中黄芪、甘草进行定性鉴别;采用中国药典2015年版进行扶正胶囊微生物限度检查的方法学验证;采用 HPLC 法测定淫羊藿中淫羊藿苷的含量,色谱柱：Agilent TC C18(2) (250 mm×4.6 mm, 5 μm),以乙腈 水(30 ∶〖KG-*2〗70)为流动相,流速：1.0 ml·min^-1,检测波长：270 nm,进样量：5 μl。结果:薄层色谱斑点清晰,在与对照品色谱相应的位置上,显相同颜色的斑点,阴性样品无干扰;可采用平皿法和直接接种法进行扶正胶囊微生物限度检查;淫羊藿中淫羊藿苷在0.101～1.008 μg范围内线性关系良好(r=0.999 7),平均回收率为99.36%,RSD为0.81%(n=6)。结论:本方法操作简便,专属性高、重复性好,可作为扶正胶囊的质量控制方法。     

加校正因子的自身对照法测定淫羊藿饮片中6种成分含量

摘 要 目的： 采用加校正因子的自身对照法测定淫羊藿饮片中6种成分含量。方法: Agilent ZORBAX Eclipse XDB-C₁₈(150 mm×4.6 mm,5 μm)色谱柱,流动相为乙腈 水梯度洗脱,检测波长268 nm,柱温30℃。通过加入自身内标物色谱峰峰面积增加值建立校正因子,用相对保留时间和光谱对照法确定其他成分位置,采用HPLC色谱法对其他成分进行定量分析。结果:淫羊藿饮片中朝藿定A、朝藿定A1、朝藿定B、朝藿定C、宝藿苷Ｉ相对于淫羊藿苷的保留时间分别为0.750,0.810,0.865,0.939,1.651;校正因子分别为0.998 6,0.998 7,0.998 8,0.989 4,0.985 6。结论: 加校正因子的自身对照法可以用于相同类型化学环境成分的定量测定,该方法简便、快捷,适合中药多成分的定量分析。     

UPLC-MS/MS测定大鼠血浆中胡蔓藤碱乙和胡蔓藤碱丁及其药动学与生物利用度研究

目的 建立一种检测大鼠血浆中蔓藤碱乙和胡蔓藤碱丁的UPLC-MS/MS法，并研究大鼠口服和舌下静脉给药方式下胡蔓藤碱乙和胡蔓藤碱丁的药动学差异。方法 大鼠分别灌胃和舌下静脉注射蔓藤碱乙和胡蔓藤碱丁，在一定时间内取血，离心获得血清；士的宁为内标，通过UPLC-MS/MS来测定血清中蔓藤碱乙和胡蔓藤碱丁的浓度，绘制药-时曲线，计算药动学参数。结果 经灌胃给药5 mg·kg^–1，胡蔓藤碱乙的t_1/2为(5.7±1.2) h，AUC_(0-t)为(59.6±20.1) ng·h·mL^–1，CL/F为(89.1±30.5) L·h^–1·kg^–1，C_max为(18.3±5.3) ng·mL^–1；经舌下静脉给药1 mg·kg^–1后，胡蔓藤碱乙的t_1/2为(1.5±0.3) h，AUC_(0-t)为(140.2±37.4) ng·h·mL^–1，CL为(7.6±2.3) L·h^–1·kg^–1，C_max为(114.9±36.0) ng·mL^–1。经灌胃给药5 mg·kg^–1，胡蔓藤碱丁的t_1/2为(4.7±4.1) h，AUC_(0-t)为(44.3±5.1) ng·h·mL^–1，CL/F为(100.3±11.7) L·h^–1·kg^–1，C_max为(13.0±4.0) ng·mL^–1；经舌下静脉给药0.1 mg·kg^–1后，胡蔓藤碱丁的t_1/2为(1.1±0.4) h，AUC_(0-t)为(72.9±19.1) ng·h·mL^–1，CL为(1.4±0.4) L·h^-1·kg^–1，C_max为(43.7±6.8) ng·mL^–1。胡蔓藤碱乙的生物利用度为8.5%；胡蔓藤碱丁的生物利用度为1.2%。结论 胡蔓藤碱乙、胡蔓藤碱丁吸收迅速，半衰期较短。     

HPLC同时测定坤泰胶囊中毛蕊花糖苷、芍药苷和黄芩苷含量

目的 建立HPLC同时测定坤泰胶囊中毛蕊花糖苷、芍药苷和黄芩苷含量的方法，为完善现有质量标准提供参考。方法 采用Phenomsil C₁₈（4.6 mm×250 mm，5 μm）色谱柱；以乙腈为流动相A，以0.1%乙酸为流动相B，采用梯度洗脱；流速为1.0 mL·min^-1；柱温为35℃；毛蕊花糖苷的检测波长为334 nm，芍药苷检测波长为230 nm，黄芩苷的检测波长为280 nm。结果 毛蕊花糖苷在3.27~32.68 μg·mL^-1内线性关系良好，平均回收率为98.4%，RSD为0.68%（n=6）；芍药苷在17.65~176.51μg·mL^-1内线性关系良好，平均回收率为99.2%，RSD为1.08%（n=6）；黄芩苷在48.78~487.77 μg·mL^-1内线性关系良好，平均回收率为97.7%，RSD为0.87%（n=6）。结论 该方法简便、快捷、结果准确、重复性好，可应用于坤泰胶囊的质量控制。     

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.

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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.

     

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.     

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.     

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.