正交试验优化酸枣仁中5种活性成分的超声离心提取工艺

目的 通过单因素和正交试验设计,对酸枣仁中5种活性成分的超声离心提取工艺进行优化。方法 选择甲醇浓度、液固比、提取时间3个因素,采用单因素和正交表L₉(3⁴)进行试验,以斯皮诺素、6’-阿魏酰斯皮诺素、酸枣仁皂苷A、酸枣仁皂苷B和白桦脂酸的峰面积为指标,进行综合评价。结果 酸枣仁最佳超声离心提取工艺为甲醇浓度80%、液固比7∶1(mL·g^-1)、提取时间40 min。结论 该提取工艺稳定可行,能够有效提取酸枣仁中活性成分,便于后续快速含量测定研究。     

同时快速测定酸枣仁中5种活性成分含量的研究

目的：简化样品处理及检测条件,建立同时快速测定酸枣仁中5种活性成分含量的方法。方法：采用《中国药典》方法、直接加热回流法、超声浓缩法和超声离心法提取酸枣仁中的活性成分,通过同一梯度洗脱程序,利用HPLC-DAD液相检测204 nm和335 nm下5种成分（包括斯皮诺素、6?-阿魏酰斯皮诺素、酸枣仁皂苷A、酸枣仁皂苷B和白桦脂酸）的峰面积,并进行方法学考查以及对9批酸枣仁药材的含量测定。结果：通过比较,超声离心法提取效率最高,速度最快,其结合HPLC-DAD液相检测法的方法学验证结果也符合试验要求,对9批酸枣仁药材进行含量测定,其中有7批符合《中国药典》标准。结论：本文建立的超声离心提取结合HPLC-DAD液相检测法,可以同时快速测定酸枣仁中5种活性成分含量,准确高效,切实可行。     

酸枣仁及其混淆品理枣仁的鉴别

酸枣仁为鼠李科植物酸枣（Ziziphus Jujuba Mill，Var，spinosa（Bunge）HuexH．F．chou）的干燥成熟种子，具有补肝、宁心、敛汗、生津功效，用于虚烦不眠，惊悸多梦，体虚多汗，津伤口渴等症。药理研究表明：本品具有镇静催眠，镇痛抗惊厥、降血脂及抗血小板聚集、增强免疫功能、降压、抗缺氧及抗心肌缺血、抗心律失常、抗衰老、抗辐射，降温和兴奋子宫等作用。     

UPLC-UV-ELSD测定不同产地炒酸枣仁饮片中酸枣仁皂苷A、酸枣仁皂苷B、斯皮诺素和6'-阿魏酰斯皮诺素的含量

目的 建立超高效液相色谱(UPLC)-紫外(UV)-蒸发光散射检测法(ELSD)同时测定不同产地炒酸枣仁饮片中酸枣仁皂苷A、酸枣仁皂苷B、斯皮诺素和6'-阿魏酰斯皮诺素的含量,对不同产地炒酸枣仁饮片进行质量评价.方法 采用Waters Acquity UPLC色谱系统,UV检测器,ELSD检测器,色谱柱为ACQUITY...     

天王补心丸中炒酸枣仁掺伪情况的研究

目的:建立天王补心丸中炒酸枣仁的混淆品炒理枣仁的检查方法,测定127批天王补心丸(水蜜丸、大蜜丸)样品,以考察样品中炒酸枣仁的投料状况,予相关生产企业及监管部门警示。方法:应用高分离度快速液相色谱-三重串联四极杆质谱(UPLC-QQQ/MS)技术,采用Agilent Eclipse C₁₈(100 mm×2.1 mm, 1.8μm)色谱柱,以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,流速0.2 mL·min^-1,柱温30℃,进样量1μL;采用电喷雾离子源(ESI),以多反应监测(MRM)模式进行正离子扫描。结果:在所设立的色谱条件下,欧鼠李叶碱在相应线性范围内线性关系良好(r>0.999),水蜜丸与大蜜丸中欧鼠李叶碱的加样回收率(n=9)分别为99.13%、97.16%,RSD均<3.3%。用所建立的方法测定127批天王补心丸样品,检出炒理枣仁15批。结论:天王补心丸中存在以炒理枣仁代替炒酸枣仁投料的情况。     

高效液相色谱法测定复方枣仁胶囊中左旋延胡索乙素含量

目的建立测定复方枣仁胶囊中左旋延胡索乙素含量的高效液相色谱（HPLC）法。方法采用Discovery C18（150mm×4．6mm,5／xm）,以甲醇-0．1％磷酸（63：37,pH=6．0）为流动相,检测波长为281nm,流速为1．0mL／min,柱温为30℃。结果左旋延胡索乙素质量浓度在63．52～635．2μg／mL范围内与峰面积线性关系良好,r=0．99998（n=7）,平均回收率为99．93％,RSD为0．7％（n=6）。结论HPLC法灵敏、快捷、准确,可用于复方枣仁胶囊中左旋延胡索乙素含量的测定。     

高效液相色谱法同时测定芝枣宁心胶囊中两组分含量

目的建立同时测定芝枣宁心胶囊中酸枣仁皂苷A、斯皮诺素含量的高效液相色谱法。方法采用Agilent Eclipse XDB C18柱(150 mm×4.6 mm,5μm),以乙腈(A)-水(B)为流动相,梯度洗脱,0 min(12%A)→8 min(19%A)→12 min(20%A)→26 min(36%A)→28 min(35%A)→32 min(100%A)→40 min(100%A),蒸发光散射检测器,漂移管温度为45℃,气体流量为4.00 bar,紫外检测器检测波长为335 nm。结果酸枣仁皂苷A进样量在0.31～1.55μg范围内与峰面积呈良好线性关系(r=0.998 8),平均回收率为98.67%,RSD为2.68%(n=9);斯皮诺素进样量在0.59～3.00μg范围内与峰面积呈良好线性关系(r=0.999 6),平均回收率为99.14%,RSD为2.31%(n=9)。结论该方法准确可靠、简单快速,可用于芝枣宁心胶囊的质量控制。     

HPLC-DAD-ELSD法同时测定酸枣仁中斯皮诺素、酸枣仁皂苷A和B的含量

目的:建立同时测定酸枣仁中斯皮诺素、酸枣仁皂苷A和B含量的HPLC-DAD-ELSD方法。方法:采用Grace BravaC18-BDS(250 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-水(B)为流动相进行梯度洗脱,流速1.0 mL.min-1,二极管阵列检测器的检测波长为335 nm,蒸发光散射检测器的漂移管温度为100℃,载气流速为2.9 L.min-1,柱温为25℃,。结果:斯皮诺素、酸枣仁皂苷A和B的线性范围分别为59.40~594.0μg.mL-1(r=0.9996),49.50~900.0μg.mL-1(r=0.9998),20.36~407.2μg.mL-1(r=0.9999);平均加样回收率(n=6)分别为96.9%(RSD=1.8%),101.2%(RSD=2.5%),100.3%(RSD=2.9%)。结论:该方法准确、可靠,在同一色谱条件下实现多指标成分的同时测定,为酸枣仁的全面质量评价提供参考。     

高效液相色谱法测定安神胶囊中酸枣仁皂苷A含量

目的建立测定安神胶囊中酸枣仁皂苷A含量的高效液相色谱法。方法色谱柱为Capcell pak mg C18柱(150 mm×4.6 mm,5μm),流动相为乙腈-水(40∶60),检测器为蒸发光检测器,流速为0.6 mL/min,进样量为10μL,柱温为30℃。结果酸枣仁皂苷A进样量在0.415 6～4.156 0μg范围内与峰面积积分值呈良好线性关系(r=0.998 4),平均加样回收率为98.81%,RSD=2.02%。结论该法简便快捷、结果准确、重复性好,可用于安神胶囊的质量控制。     

高效液相色谱法对枣仁安神颗粒中丹参酮Ⅱ_A含量测定的研究

目的提高枣仁安神颗粒的质量标准。方法采用薄层色谱法对枣仁安神颗粒进行鉴别;采用高效液相法测定其有效成分含量。结果薄层色谱法可鉴别出与酸枣仁、丹参、五味子中的特征斑点;高效液相色谱法测定丹参中的丹参酮ⅡA,在测定范围内线性良好,回收率在97%~101%之间。结论所建立的质量标准简便可行、重复性好,可作为枣仁安神颗粒的质量监控。     

两种不同形态的皂角刺的性状鉴定比较

目的：：比较两种不同形态的皂角刺的性状特征。方法：使用微距摄像进行鉴别。结果：同样是药典正品皂角刺,但是在性状上有较大差异。结论：在对皂角刺进行鉴别时要从多个方面论证,不能仅以性状作为依据。     

维药唇香草的薄层色谱鉴别及其抗氧化活性成分筛选

目的建立维药唇香草的TLC定性鉴别方法,用TLC-DPPH法初步筛选唇香草药材中的抗氧化活性成分。方法以迷迭香酸、咖啡酸、地奥司明和蒙花苷为对照品,对展开剂和薄层板进行筛选,优化出最佳展开系统。结果在展开后的薄层板上喷以100mL·L~(-1)的二苯代苦味肼自由基(DPPH)乙醇溶液,初步筛选唇香草的抗氧化能力及抗氧化成分。唇香草TLC定性鉴别的结果显示,固定相为安徽良臣所产的硅胶G板,展开剂以氯仿-甲醇-水-甲酸-冰乙酸(15∶5∶0.5∶1∶1)的展开效果最好。TLC-DPPH抗氧化活性筛选结果显示,唇香草中至少有7个化学成分具有较好的抗氧化活性,包括咖啡酸和迷迭香酸。结论唇香草薄层色谱分析方法的建立为唇香草化学成分分析、资源优选和质量标准奠定基础。     

儿科医联体中基层医疗机构的儿童药品短缺问题分析与改进措施

目的:本调研旨在分析基层医疗卫生机构儿童药品短缺的现状和原因,并提出具有针对性的改进措施与建议.方法:通过收集2020年1～12月南京医科大学附属儿童医院所在的儿科医联体内各基层医疗卫生机构逐月上报的药品短缺信息,梳理短缺药品特点,分析用药短缺发生的影响因素.结果:基层医疗卫生机构儿童短缺药品分布在各个治疗领域,其中上...     

Mitochondrial Biology and Neurological Diseases

Mitochondria are extremely active organelles that perform a variety of roles in the cell including energy production, regulation of calcium homeostasis, apoptosis, and population maintenance through fission and fusion. Mitochondrial dysfunction in the form of oxidative stress and mutations can contribute to the pathogenesis of various neurodegenerative diseases such as Parkinson’s (PD), Alzheimer’s (AD), and Huntington’s diseases (HD). Abnormalities of Complex I function in the electron transport chain have been implicated in some neurodegenerative diseases, inhibiting ATP production and generating reactive oxygen species that can cause major damage to mitochondriaMutations in both nuclear and mitochondrial DNA can contribute to neurodegenerative disease, although the pathogenesis of these conditions tends to focus on nuclear mutations. In PD, nuclear genome mutations in the PINK1 and parkin genes have been implicated in neurodegeneration [1], while mutations in APP, PSEN1 and PSEN2 have been implicated in a variety of clinical symptoms of AD [5]. Mutant htt protein is known to cause HD [2]. Much progress has been made to determine some causes of these neurodegenerative diseases, though permanent treatments have yet to be developed. In this review, we discuss the roles of mitochondrial dysfunction in the pathogenesis of these diseases.     

(Z)-7,4'-Dimethoxy-6-hydroxy-aurone-4-O-β-glucopyranoside mitigates retinal degeneration in Rd10 mouse model through inhibiting oxidative stress and inflammatory responses

Abstract

Purpose: As an inherited retinal dystrophy characterised by progressive degeneration of photoreceptor cells, retinitis pigmentosa (RP) leads to partial or total blindness eventually. Possible causes of the photoreceptor cell death are oxidative stress and inflammatory responses. (Z)-7,4'-Dimethoxy-6-hydroxy-aurone-4-O-β-glucopyranoside (DHAG) is a novel compound with potent antioxidant properties. The aim of this study was to investigate whether DHAG could mitigate photoreceptor cell degeneration in an established mouse model of RP.

Materials and method: Rd10 mice were treated with DHAG daily by gavage from postnatal day 12 (P12) to P33. Retinal morphology was evaluated by haematoxylin and eosin staining. Apoptosis-positive cells were detected by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. Oxidative stress markers and inflammatory factors were measured by enzyme-linked immunosorbent assay (ELISA). Real-time polymerase chain reaction, immunostaining and western blot were applied to measure the gene and protein change to explore the underlying mechanisms.

Results: Results showed that DHAG significantly preserved the retinal morphology, reducing photoreceptor cell apoptosis, decreasing oxidative stress and inhibiting inflammatory responses in Rd10 mice. The mechanism might be related to inhibit the activation of P38 pathway.

Conclusions: This study showed the beneficial effects of DHAG, a compound possessing antioxidant properties, and provided scientific rationale to develop DHAG as a potential agent to treat RP.     

Delayed wasting syndrome and alterations of liver gluconeogenic enzymes in rats exposed to the TCDD congener 3,3', 4,4'-tetrachloroazoxybenzene

A delayed wasting syndrome similar to that induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was observed in male Sprague-Dawley rats exposed to 3,3', 4,4'-tetrachloroazoxybenzene (TCAOB) and 3,3',4,4'-tetrachloroazobenzene (TCAB). After a slow growth period, all treatment animals (25 mg/kg, i.p., 2 doses per week) exhibited a starvation-like syndrome characterized by reduced food intake, dramatic loss of body weight and subsequent death. Although the growth of all major organs in the treatment animals was affected, the thymus appeared severely atrophied. The growth kinetics during the earlier phase were further analyzed using serially-killed rats receiving TCAOB. In addition, TCAOB was found to markedly depress the specific activity (mumol/min/g wet liver) of glucose-6-phosphatase, fructose-1,6-bisphosphatase, phosphoenolpyruvate carboxykinase, and pyruvate kinase in the liver. Significant changes in the levels of cytochrome P-450, glutamic-pyruvic transaminase and malic enzyme in the liver were also observed.     

Pharmacokinetic comparisons of Paeoniflorin and Paeoniflorin-6'O-benzene sulfonate in rats via different routes of administration

1.?The pharmacokinetics study of Paeoniflorin (Pae) and its acylated derivative (CP-25) was performed.

2.?The structure of CP-25 was identified by mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR). The rats were injected with CP-25(6, 12, 24?mg/kg) and orally treated with CP-25 (32, 64, 128?mg/kg), respectively. An high-performance liquid chromatography (HPLC) assay was developed to determine the plasma concentrations of Pae and CP-25.

3.?The results of MS and NMR showed that the acylated product was Pae-6'O-benzene sulfonate (CP-25). The plasma levels in oral CP-25 groups were detectable, whereas those of Pae in the oral groups (25 and 50?mg/kg) were undetectable. More specifically, the C_max values of oral CP-25 were 0.12, 0.19 and 0.44?μg/ml, and the corresponding t_1/2β of CP-25 were 1.44, 2.12 and 2.11?h, respectively. In addition, the t_1/2β values of intravenous CP-25 were 161.99, 152.81 and 153.76?min, respectively.

4.?Compared with the venous pharmacokinetics parameters of Pae, those of the t_1/2β, MRT, Vd and CL/F in the CP-25 groups increased noticeably. As expected, compared with oral parameters of Pae, those of t_1/2a, t_1/2β, AUC, MRT and Vd in the CP-25 group increased obviously. Finally, the absolute bioavailability of Pae and CP-25 were 3.6 and 10.6%, respectively.

5.?Our results indicate that CP-25 is characterized by improved absorption, well distribution, lower clearance, long mean residence time, and moderate bioavailability in rats.     

Cetiedil inhibition of calmodulin-stimulated enzyme activity

Cetiedil, an in vitro anti-sickling agent, inhibited calmodulin-stimulated cyclic 3':5'-nucleotide phosphodiesterase (EC 3.1.4.17) and Ca2+-ATPase (ATP phosphohydrolase, EC 3.6.1.3) activities. The drug had no effect on basal enzyme activities in the absence of calmodulin. The inhibition of phosphodiesterase was competitive with respect to the concentrations of both cAMP and calmodulin. Cetiedil did not inhibit calmodulin-stimulated enzyme activities by acting as a calcium chelator, since increasing the concentration of calcium did not reverse the inhibitory effect.     

呼吸科病房老年病人肺部感染常见病原菌及其耐药性分析

目的 对老年病人(年龄≥65岁)肺部感染的常见病原菌及其耐药性进行分析,为临床预防和治疗老年肺部感染提供依据。方法 回顾性收集中国医科大学附属盛京医院2013年1月至2015年12月老年病人感染的常见病原菌并对其耐药性进行分析。结果 共分离出318株病原菌,其中革兰阴性杆菌295例,占92.77%。其中鲍曼不动杆菌最多,占43.71%;其次是铜绿假单胞菌和肺炎克雷伯杆菌,分别占16.98%和12.89%。共分离出革兰阳性球菌20例,占6.29%。真菌3例,占0.94%。不同病原菌耐药程度不同,其中鲍曼不动杆菌对青霉素类、头孢类抗生素耐药率达90%~100%,对氨基糖苷类抗生素耐药率亦达38%~90%,仅替加环素耐药率为10.31%。铜绿假单胞菌耐药亦较严重。肺炎克雷伯杆菌耐药率相对乐观,临床可选药物较多。金黄色葡萄球菌对青霉素、头孢类抗生素几乎均耐药,对万古霉素、利奈唑胺、喹努普汀/达福普汀及替加环素最为敏感。结论 老年肺部感染以革兰阴性杆菌为主,临床应根据药敏实验结果合理应用抗生素来更好地控制疾病和防止产生耐药菌。