A new high-performance liquid chromatographic method with evaporative light scattering detector for the analysis of phospholipids. Application to Iberian pig subcutaneous fat

A new method for the analysis of phospholipids by normal-phase HPLC is described using a silica column. Addition of ammonia and triethylamine to a gradient based on chloroform/methanol/water promoted a good and rapid separation of phospholipid classes (20 min run). The use of an evaporative light scattering detector permitted an accurate analysis of a mixture of phospholipids. Calibration curves were linear within different range for each phospholipid class. The LOD and LOQ obtained were below 0.03 and 0.05 mg kg⁻¹ for all cases, respectively. Besides, a new method for the separation of phospholipids from total lipids before HPLC analysis by a solid-phase extraction (SPE) with Si cartridges has been developed. This methodology gave a good recovery ranging from 97 to 117%. The method was validated with a standard mixture of phospholipids. This method has been applied to characterize the phospholipid fraction of subcutaneous fat from Iberian pig. Cardiolipin, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylcholine, and sphingomyelin have been described for first time in these samples. The fatty acid composition of the different phospholipid classes and their HPLC electrospray ionization mass spectrometry have been used for characterizing the molecular species present in each one.     

基于PVDF薄膜传感器的猪肝动态压缩力学性能测试

瞬态冲击载荷作用下肝脏的力学响应是损伤生物力学的重要研究内容。本文提出了一种可用于软组织动态压缩力学特性测试的改进SHPB(分离式霍普金森压杆,Split Hopkinson Pressure Bar)方法。该方法采用PVDF(聚偏氟乙烯,Polyvinylidene Fluor)压电薄膜传感器测量实验过程中试件两端面的受力,以此来计算试件的应力,从而无需测量透射杆上的微弱透射信号。猪肝试样前后端面的PVDF压电信号对比表明,加载过程中试样达到了动态应力平衡状态。试样动态压缩中的惯性效应主要在加载的初始阶段对透射应力信号造成较大影响,在大变形阶段惯性效应引起的轴向应力较小。利用此方法对猪肝组织进行三种高应变率(1800s^-1,2500s^-1,3500s^-1)的动态压缩实验,并采用基于真实应变的惯性效应公式对实验数据进行修正计算。结果表明:猪肝组织在准静态与高应变率时的应力应变曲线都呈现出凹向上的非线性特征,即曲线初始阶段应力增长较缓慢,当应变达到15%后应力值则迅速增大;猪肝组织也具有明显的应变率效应,即随着应变率的增加,应力应变曲线的整体应力值也随之增大。最后,采用黏超弹性本构模型描述了猪肝组织的动态应力应变曲线。     

基于遗传算法-支持向量机的兔肝VX2肿瘤光谱鉴别

兔肝VX２肿瘤是一种快速生长的肿瘤模型,可以在多种器官如肝、肺、直肠等快速生长,常用于肿瘤研究。采用可见-近红外高光谱技术对四只兔子的兔肝VX２肿瘤和正常组织进行活体和离体的反射光谱检测,然后采用支持向量机分别实现了二分类(正常肝组织和肝VX2肿瘤组织)和四分类(未出血活体正常肝组织、未出血活体VX2肿瘤组织、出血离体正常肝组织和出血离体肝VX2肿瘤组织)。根据其光谱反射曲线的特征,选择了400～1 800 nm区间的数据为特征变量。为进一步提高分类准确率,分别采用5折交叉验证和遗传算法对支持向量机的核函数参数g和惩罚因子c进行了优化。其中5折交叉验证优化参数和分类结果为：二分类优化的惩罚参数c为4,核函数参数g为0.125 0,其校正集和预测集的准确率都达到了100%;四分类中优化出的参数c为8,g为0.121 1,其校正集和预测集的准确率分别达到了99.242 4%和93.333%。遗传算法优化参数和结果为：二分类中优化的参数c为0.845 6,g为0.062 5,其校正集和预测集的准确率同样都达到了100%;四分类中优化的参数c为5.5307,g为0.068 5,其校正集和预测集的准确率分别达到了99.242 4%和100%。结果显示两种优化方法都取得了很好的效果,遗传算法优化参数对四分类的分类更为精确。为进一步提升算法速度,采用间隔选取变量的方法来不断减少特征变量,最终每隔100 nm谱段选择一个变量,共选择14个谱段作为特征变量。采用遗传算法优化支持向量机参数并对其分类进行了研究,结果表明：二分类和四分类的校正集和预测集结果准确率均为99.242 4%,而且运行时间分别为11.4和20.0 s, 与选择全波段的运行时间：340.3和491.0 s相比, 说明多光谱技术可以进行肝VX2肿瘤组织和正常肝组织的鉴别,且分类准确率可达99%以上,而且运行时间缩短了很多。为未来多光谱技术在未来临床肿瘤诊断中实现肿瘤组织的快速实时在线检测和分类奠定了基础,显示出巨大的应用潜力。     

肝硬化合并低氧血症31例临床研究

为探讨肝硬化患者的血氧变化与临床关系，以肝炎后肝硬化患者31例为观察组（A组），采用血气分析仪测定其动脉血氧分压（PaO2)及动脉血氧饱和度（SaO2)，并以正常人15例作对照组（B组）进行比较结果表明，A组的低氧血症发生率为41％，B组为0。肝硬化患者低氧血症的发生率高，其严重程度与Child-Pugh分级有关。早期发现低氧血症并予以治疗有助于改善肝硬化患者的预后。     

ADENYLATE CYCLASE-MODULATING MEDIATORS OF INSULIN

Plasma membranes prepared from pig liver incubated with insulin (50—300 μU/ml) resulted in the release of at least two insulin chemical mediators. They appeared to modulate the activity of adenylate cyclase in liver plasma membrances of pig. One of them was fraction 1 of insulin mediator (M. W. about 3700—4000 dalton) which markedly stimulated the activity of the enzyme, the other was fraction 2 of insulin mediator (M. W. about 1000 dalton) which inhibited the enzyme activity. The results showed that the inhibitor of fraction 2 generated was significantly higher than that of fraction 1 when the membranes were incubated with insulin of low concentration (50—100 μU/ml). On the other hand, the generation of stimulator of fraction 1 from plasma membranes incubated with insulin of high concentration (200 μU/ml) was higher than that of fraction 2. So the ratio of yield between two mediators produced from the membranes was dependent on the concentration of insulin added. The results also showed that the     

Characterization and comparison of soluble and immobilized pig muscle aldolases

Pig muscle aldolase was insolubilized by covalent attachment to a polyacrylamide matrix containing carboxylic functional groups. The catalytic activity of the Akrilex C-aldolase was 2014 units/g solid, i.e., an activity loss of only about 5% relative to the initial activity. The pH optimum for catalytic activity shifted form 7.25 to 7.5 and the apparent temperature optimum from 313 to 318 K. The Michaelis constant of the insolubilized enzyme was significantly higher than that of the soluble aldolase. Heat- and urea-inactivation experiments revealed that the immobilization increased the stability of the enzyme.     

13C-Leucin-Atemtest zur Beurteilung der anabolen Verwertung von diätetischen Aminosäurengemischen bei Phenylketonurie

It is essential to establish whether and how environmental factors affect the reliability of [¹³C]methacetin breath test (¹³C-MBT). In 12 healthy volunteers (smokers), a standard ¹³C-MBT with 75 mg [¹³C]methacetin was performed twice in random order: on a control day without smoking and on another day with smoking two cigarettes antecedently. A considerable flattening of the curve of the momentary ¹³C recovery within the expiratory air was observed when the ¹³C-MBT was performed after smoking. The maximum of the momentary ¹³C recovery, D _max, decreased from 37.20±2.58 to 25.39±2.29% dose/h (p=0.00052). Moreover, the time to reach D _max was prolonged after cigarette smoking (26.5±3.1 vs. 16.5±1.9 min, p=0.0199). The curve of the cumulative ¹³C recovery on the cigarette smoking day appeared to be shifted downwards, and statistically significant differences relative to the control situation were found between the 24th and 75th minute following [¹³C]methacetin administration. Smoking cigarettes immediately prior to the ¹³C-MBT diminishes the ability of the liver to handle methacetin, and hence a possibility of such an interaction should be excluded in order to interpret the results of the test correctly.     

15Nmethacetin test to control liver function of infants with formerly very low birth weight

Abstract

Hepatic detoxification capacities of three groups of infants aged about two were estimated using the [¹⁵N]methacetin elimination test, as well as standard serum parameters:

1. Formerly hypotrophically born infants still too small for their age (n = 23)

2. Patients suffering from severe liver diseases (n = 15)

3. Patients without liver diseases (n = 14).

17 of the infants of group 1 showed ¹⁵N elimination rates as low as the rates of the liver-diseased patients of group 2. Compared to the infants of group 3, who had normal values, the findings reflect diminished hepatic monooxygenases activities in groups 1 and 2. On the other hand, the serum parameters of the infants of group 1 did not deviate from normal values estimated in group 3. Here only group 2 showed pathological values. Consequently, the [¹⁵N]methacetin test seems to be more sensitive in controlling hepatic parameters of growth retardations than the usual serum parameters used here. Further investigations have to answer the question to which proportion intrauterine malnutrition and postnatal effects, such as environmental living conditions, contribute to the retardation of liver function development in combination with growth retardation.     

磁共振弹性成像技术在肝纤维化检测中的研究

对肝脏组织的弹性应力分析在肝纤维化程度的精确定量评价方面是非常关键的,因此从弹性力学角度结合磁共振成像技术的磁共振弹性成像研究在肝脏疾病的早期诊断和治疗方面将具有重要意义和广泛应用. 该文针对弹性磁共振成像技术的理论与技术进行了全面的研究,开发了磁共振弹性图研究平台,主要包括：组织生物力学模型分析与建模,剪切波激励装置的开发,弹性成像序列的设计,弹性拟合算法研究与实现4个方面的工作. 基于上述工作,进行了相关体模和离体猪肝以及志愿者在体肝脏实验,实验结果表明,研究平台基本可以满足肝脏弹性成像研究的需要.     

HPLC profiling of phenolics and flavonoids of Adonidia merrillii fruits and their antioxidant and cytotoxic properties

In this study the antioxidant and cytotoxicity activity of the Adonidia merrillii fruits were investigated using different solvent polarities (methanol, ethyl acetate and water). The results showed that the total phenolic and flavonoid contents of the methanolic extract was higher compare with other extract with respective values of 17.80 ± 0.45 mg gallic acid equivalents/g dry weight (DW) and 5.43 ± 0.33 mg rutin equivalents/g DW. Beside that The RP-HPLC analyses indicated the presence of gallic acid, pyrogallol, caffeic acid, vanillic acid, syringic acid, naringin and rutin. In the DPPH, NO2 and ABTS scavenging assays, the methanolic extract exhibited higher antioxidant activity as compared to the ethyl acetate and water extracts. The extracts exhibited moderate to weak cytotoxic activity in the assays using human hepatocytes (Chang liver cells) and NIH/3T3 (fibroblasts cell) cell lines. The findings showed the Adonidia merrillii fruit extracts to possess considerable antioxidant and cytotoxicity properties. The fruit, therefore, is a potential candidate for further work to discover antioxidant and cytotoxic drugs from natural sources.