不同生产厂家赤芍配方颗粒中单萜苷类化合物含量比较

目的比较不同厂家生产的赤芍配方颗粒中单萜苷类化合物(吡啶芍药苷、牡丹皮苷F、氧化白芍苷、氧化芍药苷、10-羟基芍药苷、白芍苷、芍药苷、oxypaeonidanin、4-甲氧基-氧化芍药苷、没食子酰基芍药苷、4-甲氧基芍药苷、白芍苷R_1、paeonidanin、苯甲酰氧化芍药苷、苯甲酰芍药苷)的含量差异,为制定统一的质量标准奠定基础。方法采用Zorbax SB-Aq C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈和磷酸二氢钾缓冲盐(pH 2.8)溶液,梯度洗脱,体积流量1.0 mL/min,柱温30℃,检测波长260 nm。结果芍药苷、白芍苷和氧化芍药苷是赤芍配方颗粒中含量最高的3种单萜苷类化合物,且不同厂家生产的赤芍配方颗粒中主要单萜苷类化合物的含量差异较大。样品CSPFKL-KRT中芍药苷和氧化芍药苷的含量最高,样品中质量分数分别为73.214mg/g和16.935mg/g,白芍苷的质量分数最低,为2.343mg/g。而样品CSPFKL-XLS中芍药苷和氧化芍药苷的质量分数最低,样品中分别为26.327 mg/g和4.165 mg/g,白芍苷的质量分数最高18.893 mg/g。结论不同厂家生产的赤芍配方颗粒中主要单萜苷类化合物的含量差异较大,可能会对临床应用产生影响。建立统一的质量标准对于赤芍配方颗粒的质量控制具有重要作用。     

心衰宁颗粒对慢性心力衰竭大鼠心肌组织转化生长因子β1的影响

目的:探讨心衰宁颗粒对慢性心力衰竭大鼠心肌转化生长因子β1(TGFβ1)表达的影响。方法:将SD大鼠随机分为正常组、模型组、卡托普利组、心衰宁大、中、小剂量组,除正常组外,其余组采用阿霉素腹腔注射制作慢性心力衰竭大鼠模型,正常组注射等量生理盐水,每周1次,造模6周结束后,分别给予药物干预,正常组及模型组给予生理盐水,干预3周。干预结束后,超声心动图检测心功能,大鼠血清学检测pro BNP、TGFβ1水平;qRT-PCR、Western blot检测检测TGFβ1基因及蛋白表达水平。结果:与正常组比较,模型组左心室舒张末期内径(LVEDD)和左心室收缩末期内径(LVESD)明显增加(P0.01),左室射血分数(LVEF)降低(P0.01),血清TGF-β1、pro BNP明显升高(P0.01),TGF-β1基因及蛋白表达较正常组明显升高(P0.01);与模型组比较,心衰宁颗粒中、大剂量组能降低LVEDD、LVESD(P0.05),提高慢性心衰大鼠LVEF(P0.01),降低血清TGF-β1、pro BNP水平(P0.01),降低TGF-β1基因及蛋白表达水平(P0.05)。结论:心衰宁颗粒具有改善心功能,干预心肌重塑、抗心肌纤维化作用,其机制可能与抑制TGF-β1水平有关。     

中药配方颗粒和传统中药饮片调剂方式的应用对比分析

目的：对传统中药饮片调剂方式与中药配方颗粒调剂方式进行应用分析比较,优化调剂方法。方法统计并分析本院传统中药饮片与中药配方颗粒的使用数据(调剂出错率、处方数等),并随机抽取180例于该院就诊的患者及12名在该院从事配药的工作人员进行问卷调查,在患者选择和工作人员选择等方面对传统中药饮片调剂方式与中药配方颗粒调剂方式进行对比分析。结果与传统中药饮片调剂方式相比,中药配方颗粒调剂方式在出错率、取药方便程度、药品剂量准确度、卫生、食用方便程度、携带存储及提高工作效率、降低工作强度、减少药物浪费等方面具有显著优势（P<0.01）。结论中药配方颗粒调剂方式具有明显优势,但还存在许多不足,需要中医药工作者的不断完善与改进,发挥中医药优势。     

E2F1 is not essential for apoptosis induced by potassium deprivation in cerebellar granule neurons

Cerebellar granule neurons (CGNs) undergo apoptosis when deprived of depolarizing concentration of potassium. A key regulator of cell cycle, E2F1, was believed to play a role in CGN apoptosis induced by potassium deprivation. However, here we demonstrated that although E2F1 was upregulated in wild type CGNs following potassium deprivation, CGNs that derived from E2F1 knockout mice underwent apoptosis at a similar rate as the wild type. Analysis of the apoptotic neurons revealed no difference in the activation of caspase-3 in E2F1 null and wild type CGNs. Furthermore, knockdown of E2F1 expression by RNA interference failed to attenuate the apoptosis of CGNs induced by potassium deprivation. Taken together, our results suggested that E2F1 is not essential for apoptosis induced by potassium deprivation in CGNs.     

Cerebellar cortical-layer-specific control of neuronal migration by pituitary adenylate cyclase-activating polypeptide

Migration of immature neurons is essential for forming the cortical layers and nuclei. Impairment of migration results in aberrant neuronal cytoarchitecture, which leads to various neurological disorders. Neurons alter the mode, tempo and rate of migration when they translocate through different cortical layers, but little is known about the mechanisms underlying this process. Here we show that endogenous pituitary adenylate cyclase-activating polypeptide (PACAP) has short-term and cortical-layer-specific effects on granule cell migration in the early postnatal mouse cerebellum. Application of exogenous PACAP significantly slowed the migration of isolated granule cells and shortened the leading process in the microexplant cultures of the postnatal day (P)0-3 cerebella. Interestingly, in the cerebellar slices of P10 mice, application of exogenous PACAP significantly inhibited granule cell migration in the external granular layer (EGL) and molecular layer (ML), but failed to alter the movement in the Purkinje cell layer (PCL) and internal granular layer (IGL). In contrast, application of PACAP antagonist accelerated granule cell migration in the PCL, but did not change the movement in the EGL, ML and IGL. Inhibition of the cAMP signaling and the activity of phospholipase C significantly reduced the effects of exogenous PACAP on granule cell migration. The PACAP action on granule cell migration was transient, and lasted for approximately 2 h. The duration of PACAP action on granule cell migration was determined by the desensitization of its receptors and prolonged by inhibiting the protein kinase C. Endogenous PACAP was present sporadically in the bottom of the ML, intensively in the PCL, and throughout the IGL. Collectively, these results indicated that PACAP acts on granule cell migration as "a brake (stop signal) for cell movement." Furthermore, these results suggest that endogenous PACAP slows granule cell migration when the cells enter the PACAP-rich PCL, and 2 h later the desensitization of PACAP receptors allows the cells to accelerate the rate of migration and to actively move within the PACAP-rich IGL. Therefore, endogenous PACAP may provide a cue that regulates granule cell migration in a cerebellar cortical-layer-specific manner.     

Mammalian Sir2-related protein (SIRT) 2-mediated modulation of resistance to axonal degeneration in slow Wallerian degeneration mice: a crucial role of tubulin deacetylation

It has been shown that Wallerian degeneration, an anterograde degeneration of transected axons, is markedly delayed in a mutant mouse called slow Wallerian degeneration (Wld(S)). These mice also show resistance to axonal degeneration caused by microtubule depolymerizing drugs, suggesting that axonal microtubules are stabilized. Here, we have focused on tubulin acetylation, a post-translational modification associated with microtubule stability. We found that the basal level of microtubule acetylation was increased in cultured cerebellar granule cells from Wld(S) mice. Nicotinamide but not 3-aminobenzamide, an inhibitor for poly(ADP)ribose polymerase, enhanced tubulin acetylation and resistance to axonal degeneration in cultured cerebellar granule cells from wild-type (WT) mice, suggesting that mammalian Sir2-related protein (SIRT) 2, a nicotinamide adenine dinucleotide (NAD)--dependent tubulin deacetylase, could modulate resistance to axonal degeneration. Indeed, the levels of NAD and SIRT2 were decreased in the cytoplasm from Wld(S) granule cells. Moreover, SIRT2 overexpression abrogated microtubule hyperacetylation and resistance to axonal degeneration in these cells. Conversely, SIRT2 knockdown by using a lentiviral vector expressing small interfering RNA, enhanced microtubule acetylation and resistance to axonal degeneration in WT granule cells. Taken together, these results suggest that SIRT2-mediated tubulin deacetylation is involved in both microtubule hyperacetylation and resistance to axonal degeneration in Wld(S) granule cells.     

基于正交试验优选益母草配方颗粒提取工艺

目的:运用正交试验优化益母草配方颗粒工艺提取,为益母草配方颗粒的生产工艺提供理论依据。方法:以料液比、提取次数和提取时间3个因素设计L9(34)正交试验,以水苏碱含量、益母草碱含量与浸膏得率三者的综合考察指标作为评价指标,优选益母草配方颗粒提取工艺。结果:优选工艺为料液比1∶12,提取2次,每次1.5 h,优化后的提取工艺经3个产地每个产地进3次平行验证试验,水苏碱的提取含量在34.29~35.93 mg·g^-1,益母草碱的提取含量在2.88~3.01 mg·g^-1,浸膏得率在25.11%~26.42%,3个评价指标的RSD均小于2.0%,以及三者的综合考察值在0.93~0.97。结论:优选的提取工艺方法稳定可靠,水苏碱、益母草碱及浸膏的提取率较高,为规范和标准化益母草配方颗粒生产研究工艺提供一定的参考依据。     

复元醒脑颗粒的提取工艺研究

目的 研究复元醒脑颗粒的提取工艺及其工艺优化。方法 采用常规提取方法结合药效学实验确定初步提取工艺,使用正交试验法对提取工艺进行优化。结果 乙醇回流提取后,水煎煮提取、乙醇沉淀,过滤,浓缩制得浸膏。最佳乙醇回流提取方案为提取2次;乙醇浓度为50%;料液比：第1次8倍,第2次6倍;提取时间：第1次120 min,第2次80 min。最佳水煎煮提取方案为提取2次;提取时间：第1次120 min,第2次80 min;提取温度为100℃;液料比：第1次10倍,第2次8倍。最佳乙醇沉淀工艺为乙醇浓度80%,乙醇用量为2倍,提取时间为16 h。结论 采用本优化工艺提取的浸膏制成的复元醒脑颗粒,质量符合《中华人民共和国药典》2015年版（四部）（0104颗粒剂）中的要求。     

加味大黄?虫颗粒对精索静脉曲张模型大鼠精子线粒体结构的影响

目的:探索加味大黄?虫颗粒对精索静脉曲张模型大鼠精子线粒体结构的影响。方法:40只雄性大鼠随机选出10只作为假手术组,剩余30按照Turner方法制造左侧精索静脉曲张模型大鼠,造模完成后,按照随机数字表法分为加味大黄?虫颗粒组、模型组、迈之灵组,每组各10只。加味大黄?虫颗粒组给予加味大黄?虫颗粒灌胃; 迈之灵组予迈之灵片,假手术组和模型组分别予等剂量0.9%生理盐水灌胃。8周后处死所有大鼠,取出左侧睾丸组织,完成精液质量分析、睾丸组织病理检测。结果:灌胃8周后,加味大黄?虫颗粒组精子浓度、总活率、前向运动精子、精子直线速度(VSL)、精子曲线速度(VCL)、平均路径速度(VAP)、精子头侧摆幅度(ALH),明显高于模型组(P<0.05)。模型组睾丸组织病理报告生精小管管腔空化,各级生精细胞破坏严重,精子膜结构破坏,精子线粒体肿胀、溶解。而加味大黄?虫颗粒组中睾丸组织的病理损伤得到不同程度的改善,精子线粒体结构完整,形态大小规则。结论:加味大黄?虫颗粒在一定程度上修复精索静脉曲张大鼠睾丸组织的病理损伤,进而起到保护精子线粒体结构与功能的作用,为精子的发育、活动提供稳定的能量代谢环境,有助提高精子活力及增加受精能力。     

反相-高效液相色谱法测定黄龙止咳颗粒中盐酸麻黄碱和盐酸伪麻黄碱的含量

目的：建立反相-高效液相色谱法（RP-HPLC）测定黄龙止咳颗粒中盐酸麻黄碱和盐酸伪麻黄碱含量的方法.方法：色谱柱：AgilentZORBAXSB-C18色谱柱（4.6mm×250mm,5μm）;流动相：乙腈-磷酸二氢钾溶液（4∶96）（取磷酸二氢钾6.8g,加水1000ml溶解后,加三乙胺1ml,用磷酸调节pH值至3.0）;检测波长：210nm;柱温：35℃.结果：盐酸麻黄碱、盐酸伪麻黄碱进样量分别在0.0040～0.0480μg（r=0.9996）、0.0237～0.2844μg（r=0.9999）范围内线性关系良好;平均回收率分别为96.2％、97.5％,RSD分别为1.97％、1.81％.结论：本方法简便、准确,适用于黄龙止咳颗粒中盐酸麻黄碱和盐酸伪麻黄碱的含量测定.