Phenotypic and genetic characterization of patients with features of "nonclassic" forms of cystic fibrosis

OBJECTIVE: To determine which features of incomplete or "nonclassic" forms of cystic fibrosis (CF) are associated with deleterious CF transmembrane conductance regulator gene ( CFTR ) mutations, and to explore other etiologies for features not associated with deleterious CFTR mutations. STUDY DESIGN: Clinical features were compared between 57 patients with deleterious mutations in each CFTR and 63 with no deleterious mutations. The Shwachman Bodian Diamond syndrome gene ( SBDS ) was sequenced to search for mutations in patients with no deleterious CFTR mutations and steatorrhea to determine if any had unrecognized Shwachman-Diamond syndrome (SDS). RESULTS: The presence of a common CF-causing mutation, absence of the vas deferens, and Pseudomona aeruginosa in the sputum correlated with the presence of two deleterious CFTR mutations, whereas sweat chloride concentration, diagnostic criteria for CF, and steatorrhea did not. However, sweat chloride concentration correlated with CFTR mutation status in patients infected with P aeruginosa. One patient had disease-causing mutations in each SBDS . CONCLUSIONS: Presence of a common CF-causing mutation, absence of the vas deferens and/or P aeruginosa infection in a patient with features of nonclassic CF are predictive of deleterious mutations in each CFTR , whereas steatorrhea in the same context is likely to have etiologies other than CF transmembrane conductance regulator (CFTR) dysfunction.     

血脂康对不同血脂水平的急性冠状动脉综合征患者的干预作用

目的观察血脂康对不同血脂水平的急性冠状动脉综合征(ACS)患者的干预作用、方法采用双盲、随机、对照方法,将105例ACS患者分为治疗组和对照组．治疗组53例,采用常规西药治疗并加服中药血脂康1．2g／d,共12周,其中正常血脂(NBL)26例;高血脂(HL)27例;对照组52例,单用常规西药治疗,其中NBL25例,HL27例;同时设健康对照组40名;观察治疗前后肱动脉内皮依赖性舒张功能(FMD)变化,同时测定血清一氧化氮(NO)、内皮素-1(ET-1)、C反应蛋白(CRP)及血脂水平,、结果治疗前ACS患者FMD值和血清NO水平较健康对照组明显降低,ET-1、CRP水平较健康对照组显著升高,CRP、NO及FMD与血脂LDL-C存在显著的相关性。治疗12周后,治疗组FMD值和血清NO水平均有明显上升,ET-1、CRP水平显著降低。而且与对照组比较,差异有显著性(P＜0．05,P＜0．01)．血清TC、、TG、LDL-C水平治疗组均显著下降,HDL—C治疗组HL患者明显上升,NBL患者虽有升高,但差异无显著性结论早期应用中药血脂康治疗正常血脂和高血脂的ACS患者,均可改善内皮功能、拮抗炎症反应,具有稳定斑块作用。     

Beta2-glycoprotein I, anti-beta2-glycoprotein I, and fibrinolysis

β₂-glycoprotein-I (β₂GPI) is a phospholipid-binding plasma protein that consists of five homologous domains. Domain V is distinguished from others by bearing a positively charged lysine cluster and hydrophobic extra C-terminal loop. β₂GPI has been known as a natural anticoagulant regulator. β₂GPI exerts anticoagulant activity by inhibition of phospholipid-dependent coagulation reactions such as prothrombinase, tenase, and factor XII activation. It also binds factor XI and inhibits its activation. On the other hand, β₂GPI inhibits anticoagulant activity of activated protein C. According to the data from knockout mice, β₂GPI may contribute to thrombin generation in vivo. Phospholipid-bound β₂GPI is one of the major target antigens for antiphospholipid antibodies present in patients with antiphospholipid syndrome (APS). Binding of pathogenic anti-β₂GPI antibodies increases the affinity of β₂GPI to the cell surface and disrupts the coagulation/fibrinolysis balance on the cell surface. These pathogenic antibodies activate endothelial cells via signal transduction events in the presence of β₂GPI. Impaired fibrinolysis has been reported in patients with APS. Using a newly developed chromogenic assay, we demonstrated lower activity of intrinsic fibrinolysis in euglobulin fractions from APS patients. Addition of monoclonal anti-β₂GPI antibodies with β₂GPI also decreased fibrinolytic activity in this assay system. β₂GPI is proteolytically cleaved by plasmin in domain V (nicked β₂GPI) and becomes unable to bind to phospholipids, reducing antigenicity against antiphospholipid antibodies. This cleavage occurs in patients with increased fibrinolysis turnover. Nicked β₂GPI binds to plasminogen and suppresses plasmin generation in the presence of fibrin, plasminogen, and tissue plasminogen activator (tPA). Thus, nicked β₂GPI plays a role in the extrinsic fibrinolysis via a negative feedback pathway loop.     

Predictive value of expression of p16INK4A, retinoblastoma and p53 proteins for the prognosis of non-small-cell lung cancers

The predictive value of expression of p16INK4A, retinoblastoma (Rb) and p53 proteins for prognosis was evaluated in 76 patients with non-small-cell lung cancers (NSCLCs) that were potentially curatively resected between 1990 and 1995, using the results of immunostaining analyses of these proteins as reported in our previous study (Kinoshita et al, 1996). Of these NSCLCs, 22 (29%) lacked p16 protein expression and eight (11%) Rb protein, while 30 (39%) showed positive (altered) p53 protein expression. Survival of patients with p16-negative tumours was not significantly different from that of patients with p16-positive tumours (5-year survival rates 67% and 72% respectively, P = 0.8), nor was survival of patients with Rb-negative tumours significantly different from that of patients with Rb-positive tumours (5-year survival rates 42% and 69% respectively, P = 0.9). Moreover, survival of patients with p16/Rb-negative (either p16- or Rb-negative) tumours was not significantly different from that of patients with p16/Rb-positive (both p16- and Rb-positive) tumours (5-year survival rates 67% and 68% respectively, P = 0.7). In contrast, survival of patients with p53-positive (altered) tumours tended to be shorter than that of patients with p53-negative (unaltered) tumours (5-year survival rates 56% and 78% respectively, P = 0.06). In univariate analysis of potential prognostic factors, p16, Rb and p16/Rb proteins were not significant prognostic factors in the present cohort of potentially curatively resected NSCLCs. Altered p53 protein status tended to be a negative prognostic factor (P = 0.06 by the univariate analysis). These results indicate that loss of p16 protein alone, or in combination with loss of Rb protein, does not predict the clinical outcome of patients with resected NSCLCs.     

Antisense oligonucleotide eluforsen improves CFTR function in F508del cystic fibrosis

BackgroundCystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. In this study we assessed the effect of antisense oligonucleotide eluforsen on CFTR biological activity measured by Nasal Potential Difference (NPD) in patients with the most common mutation, F508del-CFTR.MethodsThis multi-centre, exploratory, open-label study recruited adults with CF homozygous or compound heterozygous for the F508del-CFTR mutation. Subjects received intranasal eluforsen three times weekly for 4 weeks. The primary endpoint was the within-subject change from baseline in total chloride transport (Cl-free+iso), as assessed by NPD. Secondary endpoints included within-subject change from baseline in sodium transport.ResultsIn the homozygous cohort (n = 7; per-protocol population), mean change (90% confidence interval) in Cl-free+iso was ?3.0 mV (?6.6; 0.6) at day 15, ?4.1 mV (?7.8; ?0.4, p = .04) at day 26 (end of treatment) and ? 3.7 mV (?8.0; 0.6) at day 47. This was supported by improved sodium transport as assessed by an increase in average basal potential difference at day 26 of +9.4 mV (1.1; 17.7, p = .04). The compound heterozygous cohort (n = 7) did not show improved chloride or sodium transport NPD values. Eluforsen was well tolerated with a favourable safety profile.ConclusionsIn F508del-CFTR homozygous subjects, repeated intranasal administration of eluforsen improved CFTR activity as measured by NPD, an encouraging indicator of biological activity.     

miR-34a/SIRT1在H2O2诱导人晶状体上皮细胞氧化应激中的表达

目的 观察微小RNA-34a(microRNA-34a,miR-34a)和沉默信息调节因子1(silent information regulator 1,SIRT1)在不同浓度H2O2诱导的人晶状体上皮细胞(SRA01/04细胞)氧化应激中的表达变化.方法 用不同浓度的H2O2(0μmol·L-1、100 μmol·L-1、200 μmol·L-1、300 μmol·L-1、400 μmol·L-1)处理细胞24 h后,用CCK-8检测细胞存活率,流式细胞仪检测细胞凋亡率和RT-PCR检测miR-34a/SIRT1的表达.结果 CCK-8检测结果显示:100～400 μmol·L-1 H2O2对SRA01/04细胞有增殖抑制作用,且呈剂量依赖关系,与0μmol·L-1 H202组相比差异均有统计学意义(均为P＜0.01);流式细胞仪凋亡检测结果显示:0μmol·L-1 H2O2组细胞凋亡率为(6.1±1.2)％;100 μmol·L-1、200μmol·L-1、300 μmol·L-1 H2O2组细胞凋亡率分别为(26.3±1.8)％、(32.5±2.2)％、(64.7±5.3)％,各组与0μmol·L-1H2O2组比较,差异均有统计学意义(均为P＜0.01).RT-PCR检测结果显示:不同浓度H2 O2处理SRA01/04细胞后,细胞中的miR-34a表达水平呈剂量依赖性升高,而SIRT1表达水平呈相应下降,与0μmol·L-1 H2O2组相比差异均有统计学意义(均为P ＜0.001).结论 在一定浓度H2O2诱导的人晶状体上皮细胞氧化应激中,miR-34a表达水平显著增加,而SIRT1表达水平显著下降.下调miR-34a表达可增加氧化应激人晶状体上皮细胞存活率.     

Regulation of hematopoietic cell signaling by SHIP-1 inositol phosphatase: growth factors and beyond

SHIP-1 is a hematopoietic-specific inositol phosphatase activated downstream of a multitude of receptors including those for growth factors, cytokines, antigen, immunoglobulin and toll-like receptor agonists where it exerts inhibitory control. While it is constitutively expressed in all immune cells, SHIP-1 expression is negatively regulated by the inflammatory and oncogenic micro-RNA miR-155. Knockout mouse studies have shown the importance of SHIP-1 in various immune cell subsets and have revealed a range of immune-mediated pathologies that are engendered due to loss of SHIP-1’s regulatory activity, impelling investigations into the role of SHIP-1 in human disease. In this review, we provide an overview of the literature relating to the role of SHIP-1 in hematopoietic cell signaling and function, we summarize recent reports that highlight the dysregulation of the SHIP-1 pathway in cancers, autoimmune disorders and inflammatory diseases, and lastly we discuss the importance of SHIP-1 in restraining myeloid growth factor signaling.     

X-连锁隐性视网膜色素变性的分子遗传学分析

目的探讨1个X-连锁隐性视网膜色素变性(X-linkedrecessiveretinitispigmentosa,XLRRP)家系先证者分子遗传学基础。方法应用聚合酶链反应-直接测序,检测与XLRP相关基因视网膜GTP酶调节因子(retinitispigmentosaGTPaseregulator,RPGR)基因的所有外显子和突变热区15号外显子开放阅读框(exonopenreadingframe15,ORF15)及其与内含子交界处序列。结果检测到2种新的同义突变,c.2166A>G(Glu722)和c.3396C>T(Asp1132),都位于ORF15;以及4种已知多态c.29-15G>A,c.469 63C>T,c.1227 67A>G和c.1675-101A>T。结论尚不能确定此XLRP家系的疾病相关基因。     

白藜芦醇对动脉粥样硬化兔心肌组织凋亡的影响

目的探讨白藜芦醇（Res）对动脉粥样硬化兔心肌组织线粒体途径凋亡的影响。方法健康雄性新西兰白兔70只,随机分为5组：正常组、病理对照组、病理＋低剂量Res组（4 mg·kg^-1·d^-1）,病理＋中剂量Res组（8 mg·kg^-1·d^-1）、病理＋高剂量Res组（16 mg·kg^-1·d^-1）。分笼饲养12周后,取左心室尖部心肌组织。H-E染色观察心肌组织形态学改变,原位末端标记法检测心尖部细胞凋亡率,半定量RT-PCR检测bax、bcl-2的mRNA变化。结果H-E染色下各组细胞形态学改变不明显;病理对照组细胞凋亡指数高于正常组（P〈0.01）和Res干预组（P〈0.01）,Res作用下凋亡指数下降呈剂量依赖性（P〈0.05）;病理对照组bax、bcl-2mRNA表达量较正常组升高（P〈0.01）,而加入Res干预后,bax的表达量明显下降,bcl-2表达升高（P〈0.01）。结论Res可以减少促凋亡基因bax的表达,增加抗凋亡基因bcl-2的表达,降低Bax/bcl-2比值,抑制心肌细胞的凋亡。     

Adenosine A2A receptors in ventral striatum, hypothalamus and nociceptive circuitry implications for drug addiction, sleep and pain

Adenosine A2A receptors localized in the dorsal striatum are considered as a new target for the development of antiparkinsonian drugs. Co-administration of A2A receptor antagonists has shown a significant improvement of the effects of l-DOPA. The present review emphasizes the possible application of A2A receptor antagonists in pathological conditions other than parkinsonism, including drug addiction, sleep disorders and pain. In addition to the dorsal striatum, the ventral striatum (nucleus accumbens) contains a high density of A2A receptors, which presynaptically and postsynaptically regulate glutamatergic transmission in the cortical glutamatergic projections to the nucleus accumbens. It is currently believed that molecular adaptations of the cortico-accumbens glutamatergic synapses are involved in compulsive drug seeking and relapse. Here we review recent experimental evidence suggesting that A2A antagonists could become new therapeutic agents for drug addiction. Morphological and functional studies have identified lower levels of A2A receptors in brain areas other than the striatum, such as the ventrolateral preoptic area of the hypothalamus, where adenosine plays an important role in sleep regulation. Although initially believed to be mostly dependent on A1 receptors, here we review recent studies that demonstrate that the somnogenic effects of adenosine are largely mediated by hypothalamic A2A receptors. A2A)receptor antagonists could therefore be considered as a possible treatment for narcolepsy and other sleep-related disorders. Finally, nociception is another adenosine-regulated neural function previously thought to mostly involve A1 receptors. Although there is some conflicting literature on the effects of agonists and antagonists, which may partly be due to the lack of selectivity of available drugs, the studies in A2A receptor knockout mice suggest that A2A receptor antagonists might have some therapeutic potential in pain states, in particular where high intensity stimuli are prevalent.