CLINICAL ASSESSMENT OF GESTATIONAL AGE IN THE NEWBORN INFANT

ABSTRACT. Vogt, H., Haneberg, B., Finne, P. H. and Stensberg, A. (Department of Paediatrics, University of Bergen, and Department of Clinical Chemistry, Akershus Central Hospital, University of Oslo, Norway). Clinical assessment of gestational age in the newborn infant. An evaluation of two methods. Acta Paediatr Scand, 70:669,.–The scoring systems of Dubowitz et al. and Parkin et al. were evaluated in two selected materials of newborn infants referred to a neonatal unit. Our estimation of gestational age by Dubowitz scores tended to be too high for the extreme prematures. "Small-for-dates" were also overestimated, whilst "appropriate-for-dates" and infants with respiratory difficulties were underestimated. "Large-for-dates" fell close to the standard curve. Ninety-five per cent confidence limits were up to ±5 weeks for Dubowitz scores and nearly ±6 weeks for Parkin scores. Further statistical analysis displayed some limitations in the use of linear regression formulas for scoring systems based on external and/or neurological characteristics. Thus, the results obtained with these methods must be used with caution in some selected newborn materials.     

帕金森病患者Parkin基因的研究

目的 研究Parkin基因缺失在早发性和晚发性帕金森病（PD）患者中的分布情况，探讨其在不同亚型PD发病机制中的可能作用。方法 对63例PD患者分为早发性PD组和晚发性PD组。以提取的基因组DNA为模板，扩增Parkin基因2～5号外显子，然后行琼脂糖电泳，观察外显子的缺失分布。结果 63例PD患者中发现外显子2、4缺失各1例，外显子3缺失2例，这些缺失均出现于早发性PD组。结论 Parkin基因外显子缺失可能是我国早发性PD患者的致病基因之一。     

温肾养肝汤对MPTP所致帕金森病多巴胺能神经元损伤保护的分子机制研究

目的 探讨温肾养肝汤保护多巴胺(Dopamine,DA)能神经元,延缓帕金森病(Parkinson's disease,PD)进程的作用机制.方法 将50只小鼠随机分为正常组,模型组,温肾养肝汤高、低剂量组,金刚烷胺组,每组10只,除正常组外,其余组小鼠腹腔注射30 mg·kg-11-甲基-4-苯基-1,2,3,6-四...     

Midbrain neuronal cultures from parkin mutant mice are resistant to nitric oxide-induced toxicity

Nitric oxide (NO) is a modulator of differentiation and survival of dopamine (DA) neurons. NO may play a role in the pathogenesis of Parkinson's disease (PD) since its levels are increased in parkinsonian brains and it can nitrosylate and alter the function of key proteins involved in the pathogenesis of PD. NO producing neurons are spared in parkinsonian brains suggesting that toxicity by NO can be compensated. Furthermore, the neurotoxic or neurotrophic effects of NO on DA neurons depend on the balance between NO levels and the intracellular levels of glutathione (GSH). We have investigated the effects of NO-donating agents on midbrain neuronal cultures from parkin-deficient mice. Parkin mutations are the most common genetic deficit observed in hereditary parkinsonism. These mice have abnormal DA release and metabolism, increased production of free radicals and a compensatory elevation of GSH. Cultures from parkin knockout (PK-KO) mice were more resistant than those of wild type (WT) to the neurotoxicity by NO, and the difference of susceptibility applied equally to DA, GABA and total number of neurons, and to astrocytes. NO-induced cell death was mainly apoptotic and could be reduced by caspase inhibitors. Cultures from PK-KO had greater levels of GSH than WT and, after treatment with NO, greater levels of S-nitrosoglutathione. The differences in susceptibility disappear when the synthesis of GSH is inhibited or the GSH chelated with diethyl maleate. Our data show that, contrary to the expectations, and related to the enhanced production of GSH in parkin knockout mice, parkin-deficient dopamine neurons are less susceptible to toxicity by NO.     

Homozygous partial genomic triplication of the parkin gene in early-onset parkinsonism

Autosomal recessive mutations in the parkin gene are the predominant cause of familial, early-onset parkinsonism; missense mutations involving one or a few nucleotides, exonic deletions and duplications have been described. Here we report a family with two affected brothers. Direct sequencing of parkin did not detect mutations, but semi-quantitative analysis identified a novel exonic rearrangement of exons 2–4. Both patients were homozygous for unique genomic triplications of the parkin gene.     

线粒体自噬在百草枯中毒大鼠模型肺纤维化中的作用研究

目的:建立百草枯(paraquat,PQ)中毒大鼠模型,观察不同阶段肺线粒体膜电位(JC-1染色)、调控线粒体自噬关键性蛋白PINK1、Parkin及活性氧(reactive oxygen species,ROS)的变化,探讨线粒体自噬是否参与PQ中毒肺纤维化的发生。方法:成年雄性SD大鼠42只,随机分为正常对照组(n=6)、模型组(n=36),模型组下设2 h、12 h、1 d、3 d、7 d和14 d共6个时间点,每个时间点各6只大鼠。使用20%PQ溶液50 mg/kg一次性灌胃大鼠建立PQ中毒模型。通过流式细胞仪检测血红细胞ROS浓度;HE染色和Masson染色观察PQ中毒后肺组织病理损害;JC-1染色检测肺组织线粒体膜电位变化;Western blot检测PINK1、Parkin蛋白变化。结果:与对照组相比,随着PQ中毒时间的延长,肺纤维化病理评分较对照组升高,差异有统计学意义(P<0.05);PQ组ROS荧光阳性强度率比值在中毒后2 h显著升高,中毒后12 h达高峰后逐渐下降(P<0.05),中毒后14 d与对照组相比差异无统计学意义(P>0.05);PQ组肺组织JC-1红绿荧光比均较对照组降低,Western blot提示随中毒时间延长,PINK1及Parkin蛋白表达均升高,差异有统计学意义(P<0.01)。PQ组肺组织JC-1红绿荧光比与PINK1及Parkin、肺纤维化病理评分均呈负相关(r=-0.890,P<0.01;r=-0.845,P<0.01;r=-0.794,P<0.01)。在PQ中毒12 h内,血红细胞ROS荧光强度阳性率与JC-1红绿荧光比呈负相关(r=-0.712,P<0.01),与PINK1及Parkin、肺纤维化病理评分呈正相关(r=0.571,P<0.01;r=0.484,P<0.01;r=0.602,P<0.05)。结论:PQ中毒导致大鼠产生了明显氧化应激,诱发了线粒体自噬。ROS的产生与线粒体自噬的发生密切相关,共同参与了PQ大鼠肺纤维化的发生发展。     

环孢素A对百草枯中毒大鼠线粒体自噬导致的肺纤维化作用研究

【摘要】 目的 建立百草枯（PQ）中毒大鼠模型,探讨环孢素A(CsA)对PQ中毒大鼠肺线粒体自噬的作用及机制。方法 成年雄性SD大鼠54只,随机分为正常对照组（n=12）、PQ组（n=24）,CsA大、中、小剂量组（n=18）。PQ组下设1天、3天、7天和14天4个时间点,每个时间点每个剂量组各6只大鼠。使用20%PQ溶液50 mg/kg一次性灌胃建立大鼠PQ中毒模型,并以自噬发生明显的时间点作为CsA干预时间点。各CsA干预组在给予20%PQ溶液灌胃的前3天开始分别予环孢素5 mg/(kg·d)、10 mg/(kg·d)、15 mg/(kg·d)灌胃。通过Masson染色观察PQ中毒后肺组织病理损害,JC 1染色检测肺组织线粒体膜电位变化,Western blot检测调控线粒体自噬关键性蛋白PINK1、Parkin的水平变化。结果 与正常对照组相比,随着PQ中毒时间的延长,PQ中毒大鼠肺纤维化病理评分升高,肺组织JC 1红绿荧光比降低,PINK1及Parkin蛋白表达均升高,差异有统计学意义(P＜001)。与PQ组比较,环孢素A组大鼠肺组织肺纤维化病理评分、PINK1及Parkin蛋白均下降,差异有统计学意义(P＜0.05),JC 1红绿荧光比升高,差异有统计学意义(P＜005)。结论 环孢素A可通过影响PINK1/Parkin蛋白,减轻百草枯中毒引起的线粒体自噬,改善肺纤维化。     

高迁移率族蛋白1通过PINK1/Parkin介导的线粒体自噬促进骨髓干细胞的趋化及成骨分化

目的 探讨高迁移率族蛋白1（HMGB1）通过PTEN诱导假定激酶1（PINK1）/帕金蛋白(Parkin)介导的线粒体自噬对骨髓干细胞的趋化及成骨分化的影响。方法 将人骨髓间充质干细胞（hBMSCs）分为7组：control组、siRNA-NC组、siRNA-HMGB1组、pcDNA-NC组、pcDNA-HMGB1组、pcDNA-HMGB1+siRNA-NC组、pcDNA-HMGB1+siRNA-PINK1组。采用Transwell检测细胞迁移能力;茜素红染色检测各组细胞中钙结节数目;ELISA检测骨桥蛋白（OPN）、碱性磷酸酶（ALP）的含量;RT-qPCR检测各组细胞中成骨细胞特异性转录因子（Osterix）、HMGB1及Runt相关转录因子2（RUNX2）、转录因子CCAAT/增强子结合蛋白（C/EBPα）、过氧化物酶体增殖物激活受体γ（PPARγ）水平;透射电镜检测线粒体自噬小体数目;Western Blot检测hBMSCs中Parkin及微管相关蛋白1A/1B-轻链3（LC3Ⅱ/Ⅰ）、选择性自噬接头蛋白62（P62）和自噬关键分子酵母Atg6同系物（Beclin1）等线粒体自噬相关蛋白的表达。结果 HMGB1通过PINK1/Parkin介导的线粒体自噬对hBMSCs的趋化作用研究表明：与pcDNA-NC组相比,pcDNA-HMGB1组HMGB1表达、细胞迁移率、线粒体自噬数目及自噬相关蛋白Beclin-1、LC3B-Ⅱ/Ⅰ的表达增加,Parkin、P62等蛋白的表达降低（P＜0.05）;与siRNA-NC组相比,siRNA-HMGB1组HMGB1表达、细胞迁移率、线粒体自噬数目及Beclin-1、LC3B-Ⅱ/Ⅰ等表达降低,Parkin、P62表达增高（P＜0.01）。HMGB1通过PINK1/Parkin介导的线粒体自噬对hBMSCs的成骨分化作用研究结果显示：与pcDNA-NC组相比,pcDNA-HMGB1组钙结节数量、Osterix及RUNX2含量、ALP及OPN的表达升高（P＜0.01）,PPARγ、C/EBPα的表达降低（P＜0.05）;与pcDNA-HMGB1+siRNA-NC组相比,pcDNA HMGB1+siRNA-PINK1组钙结节数量、Osterix及RUNX2含量、ALP及OPN的表达降低（P＜0.05）,PPARγ、C/EBPα的表达升高（P＜0.05）。结论 HMGB1高表达能通过PINK1/Parkin介导的线粒体自噬促进hBMSCs的趋化、成骨细胞分化及抑制成脂细胞分化,可能是骨质疏松症的潜在治疗靶点。     

Substantia nigra hyperechogenicity correlates with clinical status and number of <Emphasis Type="Italic">Parkin</Emphasis> mutated alleles

To further evaluate (1) transcranial sonography (TCS) for (pre)clinical diagnosis of Parkinson's disease (PD) and (2) to examine asymptomatic carriers of Parkin mutations we investigated substantia nigra (SN) hyperechogenicity in PD patients and unaffected subjects with and without Parkin mutations. The area (aSN) of the hyperechogenic SN were calculated bilaterally and study subjects were assigned to high versus low value groups. Eleven of the (affected and unaffected) mutation carriers had previously undergone 18-fluoro-dopa-(FDOPA)-PET scans. Fifty-eight individuals were investigated, including 24 with clinically definite and 34 without symptoms or signs of PD. Of the patients, three had one mutated and six had two mutated Parkin alleles. Of the unaffected subjects, 13 carried a single Parkin mutated allele. After dichotomization, 21 subjects had high and 37 subjects low values of mean aSN. Regarding the clinical status, 13 (62%) of the individuals with a high mean aSN had PD,while 26 (70%) of the study subjects with low values did not show signs of PD (p = 0.0393). Similarly, probands with high mean aSN values more frequently carried Parkin mutations (58%) than probands with low values (27%, p = 0.0234). A negative correlation between FDOPA uptake in the posterior putamen and maximum aSN was found in the group of mutation carriers (r = –0.809, p = 0.0234). In conclusion, hyperechogenicity of the SN is found in both idiopathic and Parkin-associated PD. Further strengthening the notion of a potential relationship between SN hyperechogenicity and Parkin mutational status, a larger aSN was associated with an increasing number of mutated alleles in our study. The authors have no financial or other interests to disclose.     

Responsiveness to distracting stimuli, though increased in Parkinson's disease, is decreased in asymptomatic PINK1 and Parkin mutation carriers

Patients with Parkinson's disease (PD) are more sensitive than healthy controls to response-triggering by irrelevant flanking stimuli in speeded choice-response tasks. This increased responsiveness may either indicate a lack of executive control or reflect compensatory efforts to cope with the reduced internal motor drive. Of interest in this context is whether responsiveness is already enhanced in the presymptomatic stage of PD. To address these questions, we studied a group of non-manifesting carriers of heterozygous Parkin and PINK1 mutations while they performed a choice-response task with response-compatible or incompatible flankers. These mutation carriers may be considered a model for pre-clinical PD because the mutant allele leads to a latent nigrostriatal dysfunction and may increase the risk for PD. For comparison, we studied groups of medicated patients with idiopathic PD and of healthy persons age-matched to the mutation carriers and to the patients.Measurements of reaction time, error rate, and the lateralized readiness potential of the EEG provided converging evidence that the mutation carriers were less responsive to distracting flankers than their healthy control group. In contrast, PD patients were more distractible by flankers than their control group, which replicated previous results. Mutation carriers also showed a smaller N2 component of the event-related EEG potential in trials with incompatible flankers relative to their control group, which might indicate reduced inhibitory control.We hypothesize that faulty executive control is the primary deficit, reflected by the reduced N2 component in the mutation carriers. To compensate for this deficit, mutation carriers change their strategy of speed-accuracy trade-off, in order to dampen the excitability of their lateral motor system. Disease progression might prevent symptomatic PD patients from using this compensatory mechanism, leading to increased disinhibition of their lateral motor system.