Conjugated linoleic acid increases intracellular ROS synthesis and oxygenation of arachidonic acid in macrophages

OBJECTIVE: Conjugated linoleic acids (CLAs) have potential antiatherosclerotic properties: they may inhibit atherosclerotic processes by reducing the intensity of inflammatory processes. However, in vivo studies have shown that the application of trans-10, cis-12 CLA in obese men increased their oxidative stress. The objective of this study was to determine whether CLA can lead to an increase in oxidative stress and to isoprostane synthesis in macrophages. METHODS: Monocytes from peripheral blood and human monocytic leukemia cells were used in this study. Monocytes were differentiated to macrophages, and were incubated with 30 microM cis-9, trans-11 CLA and trans-10, cis-12 CLA or linoleic acid for 2 days. In some experiments the inhibitors of peroxisome proliferator-activated receptor-alpha (PPAR-alpha) or respiratory chain were added. After incubation, synthesis of reactive oxygen species (ROS), total cellular concentration of adenosine triphosphate, concentration of 8-epi-prostaglandin F2 alpha, activity of cytoplasolic phospholipase A2 (cPLA2), activity of mitochondria, and expression of mRNA of PPAR-alpha were measured. RESULTS: In cells cultured with CLAs intercellular ROS synthesis increased. In this condition the mitochondrial energy potential was high, and the inhibitors of the respiratory chain and PPAR-alpha reduced ROS concentration. At the same time, the cPLA2 activity was abolished. In contrast, 8-iPF2 alpha III synthesis increased in CLA cells. CONCLUSION: Cultivation of cells with CLA leads to an increased ROS synthesis, partly by PPAR-alpha mechanism. An increase in ROS concentration and inhibition of cPLA2 activity can stimulate oxygenation of arachidonic acid and contribute to an increase in 8-epi-PF2 alpha III level and in the apoptosis process in macrophages.     

Angelman syndrome: clinical findings and follow-up data of 14 patients

The diagnosis of Angelman syndrome (AS) is based on the clinical features, behavior, EEG findings, and genetic abnormalities. The physical, clinical and behavioral aspects appear to attributable to localized central nervous system (CNS) dysfunction of the ubiquitin ligase gene, UBE3A, located at 15q11.2. The features of AS frequently become apparent at 1-4 years of age, and the average age at diagnosis is 6 years. Angelman syndrome was considered in the differential diagnosis of 30 patients who were referred to the Medical Genetics Department of Istanbul Medical Faculty between 1995 and 2005. The diagnosis was confirmed in 14 patients (8 female, 6 male) by detecting the presence of deletion through the use of fluorescence in situ hybridization (FISH) technique in all, while high-resolution banding technique (HRBT) detected only seven of the deletions. The patients' ages at the time of diagnosis ranged from 2 to 12 (mean 4.10+/-2.59) years. We report here on 14 patients with definite diagnosis of AS who displayed the characteristic clinical features of the syndrome and additional findings not previously reported, along with the follow-up data concerning neuromotor development and seizures.     

Catch-up growth in appropriate- or small-for-gestational age preterm infants

The aim was to evaluate postnatal growth of preterm infants in childhood and to determine factors that have an effect on catch-up growth (CUG). Ninety-six (42F, 54M) preterm born children with a gestational age of 32.6+/-2.9 weeks and birth weight of 1815+/-668 g were evaluated at age 4.7+/-1.1 years. Preterm children with birth weight and/or length below 10th percentile were accepted as small-for-gestational age (SGA) and those above as appropriate-for-gestational age (AGA). Height SDS was similar (-0.5+/-1.0) in preterm AGA and SGA children. Both groups had low body mass index (BMI) SDS (-0.6+/-1.4 and -1.0+/-1.5, respectively). Of the preterm SGA children, 65.8% showed a CUG in height and 3.8% catch- down growth. These rates were 24.6% and 33.5% in preterm AGA children. CUG in height was best explained by birth length and mother's height and CUG in weight by birth weight and mother's weight. In conclusion, although most of the preterm SGA children show CUG, they reach a compromised height in childhood. A number of preterm AGA children show a catch-down growth.     

骨肉瘤细胞中Polo样激酶3的表达和定位

目的:构建新型人源真核表达载体3* Flag-hPlk3并同时检测其在骨肉瘤细胞U2OS中的表达及定位.方法:以GST-hPlk3作为模板,利用聚合酶链反应扩增人源Plk3目的基因eDNA全长,并将其克隆到含有3* Flag标签的真核表达载体中.进一步将构建的重组质粒进行酶切和测序分析鉴定,并转染至U2OS骨肉瘤细胞系中,提取细胞总蛋白进行Western blot检测.同时利用荧光共聚焦激光扫描显微镜观察3*Flag-hPlk3在U2OS细胞系中的定位,进一步利用免疫沉淀方法纯化人源Plk3蛋白.结果:hPlk3基因cDNA全长成功构建于含有3* Flag标签的真核表达载体中,Western blot检测3*Flag-hPlk3融合蛋白表达,分子量约为74kDa.3 * Flag-hPlk3在骨肉瘤细胞系U2OS中主要定位于细胞质及核周,并成功纯化了hPlk3蛋白.结论:成功的构建了含有3* Flag标签的人源Plk3真核表达质粒,同时鉴定3*Flag-hPlk3融合蛋白的表达,最终成功纯化hPlk3蛋白.3*Flag-hPlk3蛋白主要定位在细胞质和核周.     

hSesn3基因真核表达载体的构建及在骨肉瘤细胞中的蛋白表达和定位

目的:通过构建人Sesn3真核表达载体同时证实融合蛋白在细胞中的表达及定位.方法:提取Hela工具细胞的mRNA,反转录至cDNA.进一步通过PCR来扩增人Sesn3基因的cDNA全长片段,并将其亚克隆于pEGFP-C1真核表达载体中,进而将构建的重组质粒进行酶切及测序鉴定,转染到MG-63骨肉瘤细胞中,获取细胞总蛋白,通过Western blot方法检测表达.进一步利用激光共聚焦扫描显微镜方法观察MG-63细胞内pEGFP-hSesn3的定位,最后利用免疫沉淀方法纯化人源hSesn3蛋白.结果:hSesn3基因cDNA全长片段克隆于真核表达载体pEGFP-C1中,酶切鉴定片段为1 410 bp,并成功测序.Western blot检测到了GFP-hSesn3融合蛋白表达,分子量约为79 kDa.在MG-63细胞中pEGFP-hSesn3主要定位于细胞质,进一步成功纯化了hSesn3蛋白.结论:成功构建了hSesn3基因cDNA全长的真核表达载体,在MG-63细胞中pEGFP-hSesn3蛋白主要定位于细胞质,最终成功纯化了hSesn3蛋白.     

成批重度烧伤患者相关感染的个体化治疗

成批重度烧伤患者的成功救治是考验科室对突发事件应急能力的重要标准之一。成批烧创复合伤患者救治的过程中,烧伤脓毒症、肺部感染、局部创面感染以及肠源性感染等一系列感染问题严重地威胁着患者的生命。除了对重度烧伤患者的常规治疗以外,本文着重介绍了针对患者不同感染情况进行的个体化治疗方案,并且取得良好效果。成批重度烧伤患者的治疗,需要针对具体情况进行调整,才能最大程度地提高救治成功率。     

An rs13293512 polymorphism in the promoter of let-7 is associated with a reduced risk of ischemic stroke

Journal of Thrombosis and Thrombolysis - The expression of let-7 family members was differentiated in ischemic stroke (IS), functioning as an important regulating molecular in the pathophysiology...