同时测定血浆血管紧张素Ⅰ、Ⅱ和醛固酮的液相色谱 串联质谱法的建立与评价

目的 建立同时测定血浆中血管紧张素Ⅰ(AngⅠ)、血管紧张素Ⅱ(AngⅡ)和醛固酮(Aldo)的液相色谱串联质谱(LC-MS/MS)法。方法 将200μL血浆和温育液在37℃温育3 h后，加入同位素内标，用固相萃取法提取各激素。通过LC-MS/MS分析AngⅠ、AngⅡ和Aldo,最终用标准曲线定量。结果 AngⅠ、AngⅡ和Aldo的线性范围分别为0.2～50.0、0.002～0.500、0.02～5.00 ng/mL,定量限(LOQ)为0.2、0.002、0.02 ng/mL。3种分析物的回收率在91%～103%之间。方法灵敏度高、重现性好，变异系数(CV)分别为1.4%～4.6%(AngⅠ)、2.1%～5.1%(AngⅡ)、2.4%～5.2%(Aldo)。健康人血浆中AngⅠ、AngⅡ、Aldo的浓度范围分别为1.4～11.8、0.003～0.016、0.02～0.20 ng/mL。方法无明显基质效应和携带污染。结论 建立了一种可同时准确、快速测定血浆AngⅠ、AngⅡ和Aldo的LC-MS/MS方法。     

小分子化合物Salen-Mn的合成及其结构测定

目的合成一种化合物 ,并对其结构及活性进行测定。方法采用回流方法 ,在实验室合成化合物 ,并通过质谱、红外光谱、核磁共振氢谱分析法对其结构进行检测 ,并用黄嘌呤氧化酶法测定此化合物的超氧化物岐化酶 (SOD)酶活性。结果质谱分析确定此化合物分子量为 3 5 6,红外光谱分析此化合物含有的官能团有 OH、C =N和苯环的存在 ,核磁共振氢谱分析证明有金属顺磁性物质锰 (Mn)的存在 ,并具有一定的SOD酶活性。结论可合成一种低分子量的具有SOD酶活性的物质。     

液相色谱-串联质谱检测血浆3-甲氧酪胺方法的建立

目的建立一种稳定的检测血浆3-甲氧酪胺(3-MT)的液相色谱-串联质谱(LC-MS/MS)方法。方法对LC-MS/MS的分离条件(色谱柱、柱温、p H值)进行优化,建立检测血浆3-MT的方法,并对该方法的线性范围、回收率、精密度、最低检测下限和稳定性进行评价。结果以同位素氘代作为内标,采用BEH HILIC色谱柱进行分离。流动相为100 mmol/L甲酸铵缓冲液(p H值为3)和纯乙腈,梯度洗脱;柱温为35℃。LC-MS/MS检测3-MT的线性范围为5~1 000 pg/mL,定量检出限为5 pg/mL,天间和批间变异系数(CV)分别为6%和7%,回收率为97.1%~109.3%。由于3-MT在室温中稳定性较差,因此需冰浴送检。结论建立了检测3-MT的LC-MS/MS方法,能够灵敏、准确地检测血浆3-MT水平。     

除去乙醇中杂质的简易方法

手术室泡手用过的乙醇和临床浸泡器械用过的乙醇杂质较多,使乙醇混浊、沉淀,影响乙醇的质量。多年来我们利用活性碳的吸附作用和滤棒的机械过滤方法,除去乙醇中的悬浮物、沉淀物等杂质,使原来混浊,沉淀的乙醇变得外观无色澄明,再用酒精比重计调配浓度后即可重新使用。方法:将用过的乙醇放入适宜的容器内,再加入0.5%的活性碳搅拌,另取砂滤棒(G_3)用滤纸包好,外层再用纱布包好防止滤纸破坏,滤棒接上硬质胶管后就可放入上述溶液中,硬质胶管的另一端接上抽滤瓶,用减压法抽滤即可得到无色澄明的乙醇,调配浓度后可重新使用。     

纳米羟基磷灰石的制备及改性

背景：人工制得的羟基磷灰石具有较强的比表面,极易发生团聚.目的：解决羟基磷灰石团聚问题,使其分散性更好. 方法：以硝酸钙和磷酸二氢氨为原料,采用溶液共沉淀法制备纳米羟基磷灰石,分别应用聚乙二醇（2%,4%,5%,6%,8%）、聚乙烯醇（2%,4%,5%,6%,8%）、硬脂酸（2%,4%,5%,6%,8%）对其进行改性. 结果与结论：X射线衍射分析显示获得了纯度较高的纳米羟基磷灰石.在表面包覆改性纳米羟基磷灰石时,表面改性剂种类、活性剂浓度等都对其粒径大小产生影响.综合考察羟基磷灰石合成及改性过程中的影响因素主要得出以下结论：通过对羟基磷灰石改性,羟基磷灰石对聚乙二醇、聚乙烯醇、硬脂酸具有一定的选择性,聚乙二醇对羟基磷灰石改性最好,且浓度为5%时羟基磷灰石分散性最好.     

基于液相色谱-串联质谱技术快速检测6种蘑菇毒素及临床应用探讨

目的 基于液相色谱-串联质谱(LC-MS/MS)技术建立快速检测血液、尿液中6种蘑菇毒素的分析方法，并将其应用于临床实际中毒案例。方法 将0.1 mL血液、尿液经蛋白沉淀、氮吹复溶、超声过滤后用ACQUITY UPLC HSS T3液相色谱柱分离，质谱检测采用多反应监测模式(MRM)。结果 该分析方法经过验证，符合方法学验证要求，其定量结果准确可靠，并于9例患者血液和(或)尿液样品中检出目标毒素，血液样品中浓度为1.12～5.63 ng/mL,尿液样品中浓度为1.01～9.27 ng/mL,检出物多为γ-鹅膏毒肽(γ-AMA)。新鲜蘑菇样品中检出目标物浓度为1.48～41.54 ng/mg,两份新苦粉孢牛肝菌样品中均检出羧基二羟基鬼笔毒肽(PCD)16.51～41.54 ng/mg。结论 本研究中建立的LC-MS/MS方法前处理过程简单快速，具有较高的灵敏度，能同时检测多种蘑菇毒素类化合物，可为临床毒蕈中毒快速诊断提供策略和参考。     

粪便乳糖测定在婴儿腹泻中的临床应用

导致婴儿腹泻的原因之一是乳糖不耐受。为此,我们采用改良Rubnen法对粪便常规和培养阴性的腹泻患儿进行了粪便乳糖测定。一、对象20例腹泻婴儿,年龄在2～12个月之间,粪便呈蛋花汤样,培养阴性。对照组15例同龄健康婴儿。二、乳糖测定法和结果取新鲜粪便2g左右,立即加入37℃蒸馏水5ml,充分搅拌,静置后去除沉渣取上面液体置入试管中,再加入乙酸铅0.3g,溶解后酒精灯加热到沸腾1min左右,再加入7.14mol/L氢氧化铵2ml,溶液立即变成乳白液,继续加热3min左右,静置出现粉红色沉淀物即为阳性,无粉红色沉淀物为阴性。有微量粉红色沉淀,含量1～2g/L为(+);有少量粉红色沉淀·含量2～5g/L     

液相色谱-串联质谱法监测血清茶碱浓度及室间质量评价

目的对液相色谱-串联质谱(LC-MS/MS)检测茶碱进行方法学评价,探讨其在茶碱治疗药物监测(TDM)中的应用。方法在血清添加放射性核素内标茶碱-D6,经蛋白沉淀稀释后采用LC-MS/MS测定。以Capcell C18 MGⅢ(100 mm×2.0 mm,5μm)为分析柱进行反相色谱分离;以0.1%甲酸乙腈-0.1%甲酸水[20∶80(v/v)]为流动相,流速为0.3 m L/min;以电喷雾离子化串联四级杆质谱、正离子多反应监测进行定量检测。用建立的方法从2008年起连续参加卫生部临检中心茶碱TDM室间质量评价。结果 LC-MS/MS检测茶碱的线性范围为1~50μg/m L,批内和批间精密度分别为2.26%~6.65%和4.70%~6.84%,准确度分别为94.14%~104.00%。单个样品的监测分析时间为3.5 min。冻融(-30℃室温反复解冻3次)、室温放置24 h、自动进样器放置24 h、长期保存(-30℃放置28 d)的稳定性均良好。LC-MS/MS测定结果与室间质量评价靶值偏差为2.75%,斜率为1.04,相关系数(r2)为0.983。结论该LC-MS/MS采用放射性内标稀释,具有简单、快速、特异性和灵敏度较好的特点,连续6年测定结果符合全国室间质量评价要求,可用于茶碱的临床TDM。     

注射用灯盏细辛与乳酸左氧氟沙星存在配伍禁忌

在临床工作中发现,当病人输入5%葡萄糖250mL、灯盏细辛注射液30mL结束后,更换乳酸左氧氟沙星(来立信)100mL注射液时,输液管中立即出现混浊及沉淀物,即刻更换输液管后,液体澄清透明,病人无不良反应。对此现象进行了实验观察,将灯盏细辛与乳酸左氧氟沙星注射液各取5mL放于注射器中,即刻出现混浊及沉淀物,再向上述注射器中的液体加入5%碳酸氢钠5mL,液体立即澄清透明。临床实验表明,乳酸左氧氟沙星注射液pH值较低,灯盏细辛为纯中药制剂,在使用时避免与pH值过低的液体配伍。注射用灯盏细辛与乳酸左氧氟沙星存在配伍禁忌@鲍英华!150001$哈尔滨…     

带官能团有机磷改性羟基磷灰石的可行性

背景:由于纳米羟基磷灰石粒子易团聚,由其制得的复合材料力学性能始终达不到皮质骨的水平。目的:通过新方法制备稳定的羟基磷灰石胶体,克服粒子团聚。方法:在0.05mol/L硝酸钙溶液38mL中分别加入等体积的2-氨乙基磷酸,3-膦酸基丙酸,L-O-磷酸丝氨酸溶液。将7.5mL的0.15mol/L磷酸三铵溶液分别加入到上述每种混合液中,在40,80℃各老化4h。所得的有机磷改性羟基磷灰石通过离心和水洗提纯,冷冻干燥得粉体。结果与结论:红外光谱和X射线衍射证实改性后的物质仍为羟基磷灰石,表面改性剂接枝量很少。Zeta电位分析表明3-膦酸基丙酸-羟基磷灰石、L-O-磷酸丝氨酸-羟基磷灰石可带上负电荷,2-氨乙基磷酸-羟基磷灰石可带上正电荷。3-膦酸基丙酸-羟基磷灰石、L-O-磷酸丝氨酸-羟基磷灰石的粉体在水中超声4h,得到碱性条件下稳定性较好(3d)的淡蓝色胶体;2-氨乙基磷酸-羟基磷灰石再分散胶体在酸性条件下的稳定性略好于中性条件,和碱性条件下相当,但都不理想(<3h)。表明提高羟基磷灰石胶体的稳定性还需探索新的方法。     

NO and HNO donors,nitrones, and nitroxides: Past,present, and future

The biological effects attributed to nitric oxide (^?NO) and nitroxyl (HNO) have been extensively studied, propelling their array of putative clinical applications beyond cardiovascular disorders toward other age‐related diseases, like cancer and neurodegenerative diseases. In this context, the unique properties and reactivity of the N‐O bond enabled the development of several classes of compounds with potential clinical interest, among which ^?NO and HNO donors, nitrones, and nitroxides are of particular importance. Although primarily studied for their application as cardioprotective agents and/or molecular probes for radical detection, continuous efforts have unveiled a wide range of pharmacological activities and, ultimately, therapeutic applications. These efforts are of particular significance for diseases in which oxidative stress plays a key pathogenic role, as shown by a growing volume of in vitro and in vivo preclinical data. Although in its early stages, these efforts may provide valuable guidelines for the development of new and effective N‐O‐based drugs for age‐related disorders. In this report, we review recent advances in the chemistry of NO and HNO donors, nitrones, and nitroxides and discuss its pharmacological significance and potential therapeutic application.     

Lipid messenger, diacylglycerol, and its regulator, diacylglycerol kinase, in cells, organs, and animals: history and perspective

Diacylglycerol kinase (DGK) metabolizes diacylglycerol (DG), a glycerolipid containing two acyl chains, to convert phosphatidic acid. DG is produced through phosphoinositide turnover within the membrane and is well known to act as a second messenger that modulates the activity of protein kinase C in the cellular signal transduction. Recent studies have revealed that DG also activates several proteins, including Ras guanine-nucleotide releasing protein and ion channels such as transient receptor potential proteins. Therefore, DGK is thought to participate in a number of signaling cascades by modulating levels of DG. Previous studies have disclosed that DGK is composed of a family of the isozymes, which differ in the structure, enzymological property, gene expression and localization, subcellular localization, and binding molecules. The present review focuses on the stories of phosphoinositide turnover and DG, including historical views, structural features, metabolism, and relevant cellular phenomena, together with the characteristics of DGK isozymes and the pathophysiological findings on animal studies using knockout mice and models for human diseases. Now it is being revealed that the structural and functional diversity and heterogeneity of and around DGK support the proper arrangement of the complex signal transduction machinery.     

Heat, cold, noise, and vibration

Exposure to a cold environment induces a number of physiological alterations, the most serious being hypothermia. This state can occur in all individuals, but the very young and the elderly are more susceptible. Environmental and industrially generated high ambient temperature can place further stress on aged individuals and workers, resulting in a complex symptom picture. Morbidity and death may result from such exposures. Causative factors have been identified. Noise exposure induces hearing losses above those secondary to the aging process. Psychophysiological effects during noise exposure are considered to result from the sympathetic activity secondary to a general stress reaction. Vibration from the use of power tools results in Raynaud's phenomenon. However, modification of power tools has reduced the symptoms associated with vibration exposure. Termination of exposure to vibration appears eventually to reduce symptoms related to white-finger spasms. Interaction between these stressors has not been clarified because of the complex effects of each. The need for additional information about the response to these stressors is evident.