HPLC波长转换法同时测定天麻头痛片中天麻素、欧前胡素、阿魏酸、胡薄荷酮和11-羰基-β-乙酰乳香酸的含量

目的 建立同时测定天麻头痛片中天麻素、欧前胡素、阿魏酸、胡薄荷酮和11-羰基-β-乙酰乳香酸含量的HPLC方法。方法 采用Agilent Zorbax TC-C₁₈色谱柱,流动相为乙腈-0.5%磷酸溶液,流速为1.2 mL·min^-1,梯度洗脱,进样量为10 μL,柱温为35 ℃。结果 天麻素、欧前胡素、阿魏酸、胡薄荷酮和11-羰基-β-乙酰乳香酸检测浓度分别在1.25~25.00,0.50~10.00,10.00~200.00,0.50~10.00,0.50~10.00 μg·mL^-1内与峰面积积分值呈良好的线性关系;精密度、稳定性、重复性试验的RSD均≤1.0%;平均加样回收率分别为100.0%,99.4%,99.1%,98.9%,98.6%,RSD分别为1.33%,1.38%,0.79%,3.12%,1.26%(n=9)。结论 该方法简便、准确、重复性好,可为天麻头痛片的质量控制提供实验依据。     

甲基尿石素A对油酸诱导的Huh-7细胞脂质累积的改善作用及其机制研究

目的:研究甲基尿石素A对油酸诱导的人肝癌Huh-7细胞脂质累积的改善作用及机制。方法:采用油酸诱导建立细胞脂质累积模型。取Huh-7细胞,分为对照组(培养基)和模型组(1 mmol/L油酸)、低剂量药物组(1 mmol/L油酸+10μmol/L甲基尿石素A)和高剂量药物组(1 mmol/L油酸+20μmol/L甲基尿石素A),采用油红O染色法观察细胞中脂质累积情况;采用三酰甘油(TG)酶法测定细胞内TG含量,采用聚合酶链式反应(PCR)法检测细胞中脂肪酸合成酶(FASN)、胆固醇调节元件结合蛋白1(SREBP-1)、过氧化物酶体增殖剂激活受体α(PPAR-α)、PPAR-γ的m RNA表达水平;采用Western blotting法检测细胞中FASN的蛋白表达水平。结果:采用油酸诱导后,细胞膜周围出现大量脂滴累积;细胞内脂质及TG含量均显著升高,FASN、SREBP-1、PPAR-γ的mRNA表达水平及FASN的蛋白表达水平均显著升高,PPAR-α的mRNA表达水平显著降低(P<0.01)。采用甲基尿石素A干预后,细胞膜周围的脂滴明显减少;细胞中脂质及TG含量均显著降低,FASN、SREBP-1、PPAR-γ的mRNA表达水平及FASN的蛋白表达水平均显著降低(P<0.05或P<0.01)。结论:甲基尿石素A对油酸诱导的Huh-7细胞脂肪累积有一定的改善作用,其机制可能与下调FASN、SREBP-1、PPAR-γ等相关因子的表达,抑制脂肪酸从头合成、促进脂质代谢有关。     

土震素B类似物中A环2-烯-1-酮结构的合成方法改进及衍生物抗脑缺血活性

目的改进土震素B类似物A环2-烯-1-酮结构的合成路线,研究所合成的衍生物对新生大鼠原代培养海马神经元细胞缺糖缺氧损伤的保护作用.方法以2-烯-伊比林酯为原料、溴代丁二酰亚胺代替三氧化铬为氧化剂合成目标化合物,溴代副产物经水解、氧化也转化为目标化合物.以大鼠原代培养海马神经元细胞缺糖缺氧损伤模型测试所合成化合物的活性.结果与结论共合成4个未见文献报道的新化合物,经1H-NMR和MS确证其结构.体外试验表明:化合物6b对大鼠原代培养海马神经元细胞缺糖缺氧损伤有较强的保护作用.     

Opioid-mediated regulation of A11 diencephalospinal dopamine neurons: pharmacological evidence of activation by morphine

Dopamine (DA) neurons of the A11 diencephalospinal system represent the sole source of DA innervation to the spinal cord in mice, serving neuromodulatory roles in the processing of nociceptive input and movement. These neurons originate in the dorso-caudal diencephalon and project axons unilaterally throughout the rostrocaudal extent of the spinal cord, terminating predominantly in the dorsal horn. The density of A11 DA axon terminals in the lumbar region is greater in males compared to females, while in both sexes the activity of neurons terminating in the thoracic spinal cord is greater than those terminating in the lumbar region. The present study was designed to test the hypothesis that A11 DA neurons are activated by opioids. To test this hypothesis, male and female mice were systemically treated with agonists or antagonists acting at the μ-opioid receptor, and spinal cord concentrations of DA and its metabolite DOPAC were determined in the thoracic and lumbar spinal cord using high performance liquid chromatography coupled with electrochemical detection. Systemic administration of the μ-opioid agonist morphine led to a dose- and time-dependent increase in spinal cord DOPAC/DA ratio (an estimate of DA neuronal activity) in both male and female mice, with greater changes occurring in the lumbar segment. Blockade of opioid receptors with the opioid antagonist naloxone reversed the stimulatory effects of morphine on A11 DA neurons in both male and female mice, but had little to no effect on the activity of these neurons when administered alone. Present findings are consistent with the conclusion that spinal cord-projecting axon terminals of A11 DA neurons are activated by opioids in both male and female mice, most likely through a dis-inhibitory mechanism.     

鹰嘴豆芽素A抗HIV-1活性及抑制CD4~+淋巴细胞早期活化作用

目的研究鹰嘴豆芽素A(biochanin A,BioA)对HIV-1活性及CD4+淋巴细胞早期活化作用的影响。方法①以MT-2和H9/HIV-1ⅢB细胞或HIV-1病毒共培养建立HIV-1感染模型,以不同浓度的BioA干预该过程,观察细胞融合情况和测定培养上清中的p24抗原含量变化,以评价BioA的抗HIV-1活性。②以PHA刺激剂诱导人外周血CD4+淋巴细胞活化,利用流式细胞术检测不同浓度BioA对早期活化标志CD69分子的表达影响。结果①BioA在本实验所设浓度下均能减少由HIV-1介导的细胞融合数(EC50=5.1μmol.L-1,SI=39)及p24抗原的表达量(EC50=38μmol.L-1,SI=5.2)。②终浓度5、10、25、50μmol.L-1的BioA对CD4+淋巴细胞早期活化抗原CD69表达具有明显抑制作用(P<0.01),其中50μmol.L-1达到最大抑制率,能使活化率从(60.42±0.52)%降低到(19.70±0.38)%。结论BioA具有抗HIV-1介导的细胞融合和HIV-1复制的活性及抑制CD4+淋巴细胞早期活化作用。     

Improved synthesis and application of [11C]benzyl iodide in positron emission tomography radiotracer production

Positron emission tomography has increased the demand for new carbon‐11 radiolabeled tracers and building blocks. A promising radiolabeling synthon is [¹¹C]benzyl iodide ([¹¹C]BnI), because the benzyl group is a widely present functionality in biologically active compounds. Unfortunately, synthesis of [¹¹C]BnI has received little attention, resulting in limited application. Therefore, we investigated the synthesis in order to significantly improve, automate, and apply it for labeling of the dopamine D₂ antagonist [¹¹C]clebopride as a proof of concept. [¹¹C]BnI was synthesized from [¹¹C]CO₂ via a Grignard reaction and purified prior the reaction with desbenzyl clebopride. According to a one‐pot procedure, [¹¹C]BnI was synthesized in 11 min from [¹¹C]CO₂ with high yield, purity, and specific activity, 52 ± 3% (end of the cyclotron bombardment), 95 ± 3%, and 123 ± 17 GBq/µmol (end of the synthesis), respectively. Changes in the [¹¹C]BnI synthesis are reduced amounts of reagents, a lower temperature in the Grignard reaction, and the introduction of a solid‐phase intermediate purification. [¹¹C]Clebopride was synthesized within 28 min from [¹¹C]CO₂ in an isolated decay‐corrected yield of 11 ± 3% (end of the cyclotron bombardment) with a purity of >98% and specific activity (SA) of 54 ± 4 GBq/µmol (n = 3) at the end of the synthesis. Conversion of [¹¹C]BnI to product was 82 ± 11%. The reliable synthesis of [¹¹C]BnI allows the broad application of this synthon in positron emission tomography radiopharmaceutical development. Copyright © 2015 John Wiley & Sons, Ltd.     

Acute toxicity of monochlorophenols, dichlorophenols and pentachlorophenol in the mouse

Acute oral LD50 values were determined for 2-, 3-, and 4-chlorophenol, 2,3-, 2,4-, 2,5-, 2,6-, 3,4-, and 3,5 dichlorophenol and pentachlorophenol in male and female mice. LD50 values (mg/kg) ranged from 117 (females) and 177 (males) for pentachlorophenol to 2389 (females) and 2643 (males) for 3,5-dichlorophenol. It was found that 2-chlorophenol and 3-chlorophenol were considerably more toxic than the dichlorophenol series. Values for males and females were generally similar, the major differences being with pentachlorophenol and 2,5-dichlorophenol, where in both cases the female LD50 was lower.     

甲型乙型肝炎联合疫苗的制剂安全性试验研究

目的评价甲型乙型肝炎联合疫苗是否具有肌肉刺激性、急性毒性和过敏性反应。方法家兔后肢股四头肌im 0.5mL甲型乙型肝炎联合疫苗,每天给药1次,连续给药2 d,停药2和21 d后对注射部位进行肉眼和病理组织学检查;小鼠后肢im 0.2 mL甲型乙型肝炎联合疫苗,给药1次后观察小鼠的不良反应情况;豚鼠后肢隔日im 0.5 mL甲型乙型肝炎联合疫苗,连续注射3次,分别于末次致敏后第14和21 d iv给予1.0 mL甲型乙型肝炎联合疫苗进行激发,观察豚鼠30 min内是否出现过敏反应。结果甲型乙型肝炎联合疫苗可导致注射部位肌肉局灶性炎细胞浸润,停药21 d后肌肉组织无异常;在本实验条件中,未见对小鼠有明显的毒性反应;豚鼠全身主动过敏反应阴性。结论甲型乙型肝炎联合疫苗无全身主动过敏性反应且毒性较低,对肌肉有轻度的刺激反应,但其刺激作用可恢复。     

昂丹斯琼杂质的小鼠急性毒性和细胞毒性研究

目的研究昂丹斯琼杂质的急性毒性和细胞毒性。方法预实验确定用药剂量,正式实验120只动物随机分组（雌雄各半,每组20只）,给药后连续观察14d,记录动物反应及死亡情况。应用成纤维细胞（L929）、狗肾细胞（MDCK）及人脐静脉内皮细胞（HUVEC）观察化合物的细胞毒性。结果昂丹斯琼杂质1灌胃给药对小鼠的LD50及95％置信限为1798．8±178．2mg·kg^-1,解剖未见明显内脏损伤。昂丹斯琼杂质2静脉注射给药对小鼠的LD50及95％置信限为49．63±7．63mg·kg^-1,静注后出现呼吸急促、抽搐等现象,解剖未见明显内脏损伤。昂丹斯琼杂质32000mg·kg^-1。灌胃给予小鼠,每天1次,连续给药3天,动物全部存活且无明显中毒表现,解剖未发现明显内脏损伤。昂丹斯琼杂质1,2,3均对成纤维细胞（L929）有一定的细胞毒作用（P〈0．05）。昂丹斯琼杂质3显著促进人脐静脉内皮细胞的增殖（P〈0．05）。结论昂丹斯琼杂质1和杂质2对小鼠和细胞具有一定毒性,昂丹斯琼杂质3毒性低。本研究为制定昂丹斯琼质量标准提供实验依据,提高临床使用安全。     

Acute and subchronic toxicity of xylo-oligosaccharide in mice and rats

Xylo-oligosaccharide (XOS) is sugar oligomers composed of a β-1,4-linked xylopyranosyl backbone that are obtained by either chemical or, more commonly, enzymatic hydrolysis of xylan polysaccharides extracted from plant cell wall. In this study, acute and subchronic toxicity of XOS in mice and rats have been evaluated, respectively. In the acute study, no obvious clinical signs of toxicity or mortality were observed in mice at the dosage of 32?g/kg BW XOS, excepting transient unformed stools were observed. In the subchronic study, XOS was evaluated in rats with dietary administration at concentrations of 0 (control), 0.9, 2.9, 8.8 and 10% for 13 weeks. Measurements included clinical observations, body weight, food consumption, food conversion efficiency, hematology, blood chemistry, gross necropsy, organ weight and histopathology. Under the conditions, no treatment-related changes were noted in behavior or appearance of the rats and no mortalities occurred. No toxicological findings were found in food consumption, food conversion efficiency, hematology, clinical biochemistry or organ weights in either sex. It is concluded, therefore, that the high dose level, at which the female and male rats consumed about 11.51 and 14.95?g XOS/kg bw/d, respectively, is the no observed adverse effect level (NOAEL) of this 13-week toxicity study.     

Biological effects of cocaine derivatives I: Improved synthesis and pharmacological evaluation of norcocaine.

An improved synthesis of norcocaine, a metabolite of cocaine, is reported. Following intravenous administration to a rhesus monkey, respiratory effects were similar to those observed following cocaine treatment. In addition, clonic convulsions, hypothermia, and mydriasis resulted. Norcocaine could be responsible for part of the pharmacological activity of cocaine.     

Design,synthesis and evaluation of novel non-ATP competitive CHK1 inhibitors as chemotherapy sensitizing agents

A series of urea-based compounds containing purine moieties were designed and synthesized as novel non-ATP competitive CHK1 inhibitors. The biologicalevaluation showed that several target compounds exhibited more potent inhibitory effects against CHK1 than the lead compound. In addition, one particular compound displayed synergistic effects with gemcitabine against HT29 cells.     

一种克拉霉素关键中间体的合成工艺优化

目的研究克拉霉素关键中间体——（2＇4,″-O-双三甲基硅基）-红霉素A-9-[O-（1-乙氧基-1-甲乙基）]肟的＂一锅法＂合成工艺。方法以9-（E）-红霉素肟为原料,在内酰胺盐酸盐的催化下,与2-乙氧基丙烯进行醚化反应,再进行硅烷化,即经＂一锅法＂得到克拉霉素关键中间体。结果与结论目标物的结构经质谱、核磁共振谱确证。该合成路线收率良好（两步总收率为88%）、环境友好,为工业化生产克拉霉素提供了一种新的方法。