一个中国人Liddle综合征家系的SCNN1G基因新突变及其临床特征

目的 分析2个Liddle综合征家系上皮细胞钠通道编码基因SCNN1B及SCNN1G的基因突变.方法 收集2个临床诊断为Liddle综合征的家系,抽取先证者及其家系成员外周血基因组DNA,PCR扩增上皮细胞钠通道β亚单位编码基因SCNN1B和γ亚单位编码基因SCNN1G第13外显子,产物直接DNA测序进行基因突变检测.结果 例1 SCNN1B基因第13外显子的扩增片段经双向测序显示第564密码子存在CGA-TGA(R-X)杂合无义突变,其家系成员均未发现这一基因突变;例2 SCNN1G基因第567密码子存在CAG-TAG(Q-X)杂合无义突变,2个家系成员携带此突变基因,这一突变位点尚未在国内外报道过,50名无关正常人中未发现此突变基因.结论 对临床诊断的Liddle综合征患者及其亲属,进行基因突变检测有助于确定诊断及早期筛查出家系中的其他患者.编码人类肾小管上皮细胞钠通道γ亚单位基因SCNN1G第13外显子第567密码子CAG-TAG(Q-X)杂合无义突变可能会导致Liddle综合征.     

Histidine-rich protein genes and their transcripts in Plasmodium falciparum and P. lophurae

The presence of histidine-rich protein (HRP) related genes and gene products in Plasmodium falciparum was demonstrated using a synthetic pentahistidine-encoding oligonucleotide and a cloned HRP cDNA probe prepared from the avian parasite P. lophurae. In Northern blotting experiments, two knobby clones of P. falciparum were found to contain a 3500 nucleotide RNA species that hybridized with the oligonucleotide and HRP cDNA probes. As this component had the expected size for an mRNA encoding an 80-90 kDa protein and was absent from two knobless clones of P. falciparum, we concluded that it represented a 'knob protein' mRNA. Using the restriction enzyme EcoRI, three identical cross-hydribizing HRP gene fragments were found in the DNA of both knobby and knobless clones of P. falciparum. These fragments differed in size from those present in P. lophurae. These results suggest that the absence of knob protein mRNA in knobless clones is not due to loss of the corresponding gene(s).     

Isolation and partial characterization of the tegumental outer membrane of schistosomula of Schistosoma mansoni

Separation of the external membranes from freshly converted mechanical schistosomula of Schistosoma mansoni was achieved by osmotic shock under hypertonic conditions, followed by mechanical shearing and ultracentrifugation. Prior to treatment, the schistosomula were surface labeled by introduction of N-DNP-epsilon-aminocaproylphosphatidylethanolamine molecules into their lipid bilayer followed by anti-DNP antibodies and stained with either 125I-protein-A or ferritin labeled secondary anti-DNP antibodies. This label provided a membrane marker by which the purity of the preparation could be assessed at each stage. Fluorescence staining with FITC-conjugated secondary antibodies prior to treatment revealed that the homogeneously stained membrane of the intact schistosomula became swollen and ruptured after the osmotic shock. The isolated membrane pellet was intensely fluorescent. Electron microscopical examination revealed mostly vesicles, some of them with organized multilayer assembly. The vesicles were ferritin labeled, indicating that they originated from the outer surface membrane of the schistosomula. A 100 fold enrichment in the alkaline phosphatase activity and about 300 fold enrichment in acetylcholinesterase activity in the membrane preparations, as compared to the intact schistosomula, was found. The isolated tegument was analyzed by SDS-polyacrylamide gel electrophoresis. The pattern obtained showed three major bands, of molecular weights 69 000, 45 000 and 12 000 alongside with a large number of minor bands. Immunoprecipitation of the isolated 125I-labeled membrane antigens with antisera from chronically infected mice revealed these three major bands together with three other bands of molecular weight 38 000, 23 000 and 16 000.     

Inhibition of Polyclonally Induced Immunoglobulin Secretion by Aurothiomalate

The influence of sodium aurothiomalate on the secretion of immunoglobulins by normal human lymphocytes in vitro was investigated by means of a reverse hemolytic plaque forming cell (PFC assay, Aurothiomalate inhibited the FFC response induced by pokeweed mitogen (PWM) and by Epstein-Barr virus (EBV) in a dose dependent manner. The inhibition was irreversible, as pre-incubation for 2 h with the drug followed by extensive washing and further culture in gold salt–free medium still caused an inhibition of the PFC response to PWM and to EBV. Cell proliferation was not significantly affected, suggesting that the inhibition of PFC formation was not due to cytotoxicity. Pre–incubation of monocytes/macrophages (Mø's), T lymphocytes and B lymphocytes with the gold compound prior to culture with PWM showed that M0's and B cells were highly sensitive, whereas T lymphocytes where resistant to the drug. The findings indicate that aurothio-malate inhibits the polyclonally induced PFC response by interfering with accessory Me function and by affecting the B lymphocyte itself.     

Oral Sodium Cromoglycate Treatment of Atopic Dermatitis Related to Food Allergy

Thirty-two children and two adults with chronic atopic dermatitis related to food allergy entered this double-blind crossover study comparing oral sodium cromoglycate (200–1600 mg/24 h) with placebo. Each treatment period comprised 6 weeks: 1 weeks on elimination diet, and 2 weeks on a normal. i.e. unrestricted diet. The diagnosis of food allergy was made after clinical improvement with elimination diet and relapse after challenge. Overall analysis of skin symptoms evaluated by means of clinical assessments and diary cards, opinions of treatment, and use of concomitant medication gave no evidence of any difference between sodium cromoglycate and placebo. Unusual symptoms were reported by 18 patients. In one case the patient was withdrawn during the sodium cromoglycate period because of side effects. The majority of symptoms with both treatments were stomach problems. Overall analysis of the laboratory data gave no significant differences between treatments.     

Proteases and their inhibitors as prognostic factors for high-grade serous ovarian cancer

Ovarian carcinoma is one of the most lethal malignancies, but only very few prognostic biomarkers are known. The degradome, comprising proteases, protease non-proteolytic homologues and inhibitors, have been involved in the prognosis of many cancer types, including ovarian carcinoma. The prognostic significance of the whole degradome family has not been specifically studied in high-grade serous ovarian cancer. A targeted DNA microarray known as the CLIP-CHIP microarray was used to identify potential prognostic factors in ten high-grade serous ovarian cancer women who had early recurrence (<1.6 years) or late/no recurrence after first line surgery and chemotherapy. In women with early recurrence, we identified seven upregulated genes (TMPRSS4, MASP1/3, SPC18, PSMB1, IGFBP2, CFI – encoding Complement Factor I – and MMP9) and one down-regulated gene (ADAM-10). Using immunohistochemistry, we evaluated the prognostic effect of these 8 candidate genes in an independent cohort of 112 high-grade serous ovarian cancer women. Outcomes were progression, defined according to CA-125 criteria, and death. Multivariate Cox proportional hazard regression models were done to estimate the associations between each protein and each outcome. High ADAM-10 expression (intensity of 2–3) was associated with a lower risk of progression (adjusted hazard ratio (HR): 0.51; 95% confidence interval (CI): 0.29-0.87). High complement factor I expression (intensity 2–3) was associated with a higher risk of progression (adjusted HR: 2.30, 95% CI: 1.17–4.53) and death (adjusted HR: 3.42; 95% CI: 1.72–6.79). Overall, we identified the prognostic value of two proteases, ADAM-10 and complement factor I, for high-grade serous ovarian cancer which could have clinical significance.     

Formation of direct-acting mutagens from mixtures of N-nitrosomorpholine and carboxylates by UVA irradiation

Previously, we found that a directly mutagenic compound is produced from N-nitrosopiperidine (NPIP) in phosphate buffer on exposure to near-ultraviolet light (UVA) and we identified its structure as α-hydroxy-N-nitrosopiperidine phosphate ester. In the present study, we show that a similar photoactivation of an N-nitrosamine can take place with carboxylates in place of phosphate. When a neutral solution of a mixture of N-nitrosomorpholine (NMOR) and sodium acetate was irradiated with UVA, the solution became directly mutagenic towards Salmonella typhimurium TA1535. O⁶-Alkyl-guanine-DNA alkyltransferase-deficient strains of S. typhimurium showed remarkably higher mutagenesis responses to this mutagen than the proficient strains. Citrate, succinate, and several other biological carboxylates were also effective in producing the mutagens. Since a treatment of the “NMOR plus acetate” photoproduct with carboxylic ester hydrolase resulted in a loss of the mutagenicity, the active principle is suggested to be an acetate-esterified derivative of NMOR. The role of the esters as intermediates in the photomutagenesis of nitrosamines is discussed. Environ. Mol. Mutagen. 31:163–168, 1998 © 1998 Wiley-Liss, Inc.     

孟鲁司特钠和细菌溶解产物对支气管哮喘豚鼠气道重塑及TGF-β1、Smad7表达的影响

目的 探讨白三烯受体拮抗剂孟鲁司特钠（MK）和（或）细菌溶解产物（OM-85BV）干预下,对支气管哮喘豚鼠气道重塑及转化生长因子-β1（TGF-β1）、Smad7水平变化的影响及其相关性。方法 将40只Hartley雄性豚鼠随机分成正常对照组、哮喘组、MK组、OM-85BV组和MK+OM-85BV组,每组8只。经腹腔内注射10%卵清蛋白（OVA）致敏并雾化吸入1% OVA激发以制备哮喘气道重塑模型,正常对照组以生理盐水替代;在雾化吸入激发阶段,MK组、OM-85BV组和MK+OM-85BV组给予相应的药物混悬液灌胃,正常对照组和哮喘组给予等量的生理盐水灌胃。激发阶段结束后24 h内,取豚鼠支气管肺泡灌洗液（BALF）,用ELISA法测定BALF中TGF-β1、Smad7含量;并处死豚鼠,取肺组织病理切片观察气道重塑程度,采用图像分析技术测定肺内支气管基底膜周径（Pbm）、总管壁面积（Wat）及平滑肌面积（Wam）。采用Pearson直线相关对两变量间进行相关分析。结果 哮喘组、MK组、OM-85BV组和MK+OM-85BV组肺组织病理切片显示支气管平滑肌、肺泡壁均较正常对照组明显增厚,标准化的支气管总管壁面积（Wat/Pbm）及平滑肌面积（Wam/Pbm）均较正常对照组增大,TGF-β1水平均高于正常对照组,Smad7水平均低于正常对照组（均P < 0.05）;MK组、OM-85BV组和MK+OM-85BV组肺组织病理切片显示病理损害程度较哮喘组有所改善,Wat/Pbm、Wam/Pbm均较哮喘组降低,TGF-β1水平均低于哮喘组,Smad7水平均高于哮喘组,且MK+OM-85BV组较MK组、OM-85BV组改善得更多（均P < 0.05）。TGF-β1与Smad7表达水平呈负相关;TGF-β1表达水平与Wat/Pbm及Wam/Pbm分别呈正相关;Smad7表达水平与Wat/Pbm及Wam/Pbm分别呈负相关（均P < 0.01）。结论 MK和（或）OM-85BV干预哮喘豚鼠后能减轻气道重塑,其中MK联合OM-85BV干预效果最好;其机制可能是降低TGF-β1和提高Smad7含量,从而改善TGF-β1和Smad7表达水平的失衡,最终减轻气道重塑。     

Effects of ADH on the activity and function of the renin-angiotensin-aldosterone system in infants and in children

In babies ranging in age from 1 to 25 weeks and in children between 1 and 14 years, plasma renin activity and urinary aldosterone activity were determined in relation to urinary sodium excretion. A reciprocal correlation was found demonstrating that the hyperactivity of the renin-angiotensin-aldosterone system is stimulated in infants by a low sodium intake. A second stimulus was observed in the influence of the hypothalamo-neurohypophyseal system, when the plasma renin activity was suppressed by administration of antidiuretic hormone and sodium excretion increased due to a decreased aldosterone activity.Our study suggests that there exists a feedback between the renin-angiotensin-aldosterone system and ADH release and that this feedback plays an important role in the regulation of water and electrolyte balance in the young infant.Supported by Deutsche Forschungsgemeinschaft.