缺氧/复氧损伤人脐静脉内皮细胞黏附分子表达与新型植物雌激素α-玉米赤霉醇的影响

目的:探讨新型植物雌激素α-玉米赤霉醇对缺氧/复氧损伤后人脐静脉内皮细胞黏附分子表达的干预,并与内源性动物雌激素雌二醇的作用进行比较。方法:实验于2005-10/2006-01在首都医科大学完成。①健康产妇的婴儿脐带由首都医科大学附属宣武医院妇产科提供,产妇及其家属签署捐赠同意书。α-玉米赤霉醇(中国农业大学提纯,纯度99%以上,以乙醇作为溶剂),雌二醇(Sigma公司)。②脐静脉内皮细胞原代培养后,用质量浓度为1.25g/L的胰蛋白酶 0.1g/L的乙二胺四乙酸混合消化液进行传代,传至2～3代后用于实验。细胞以3×105/孔接种于24孔培养板,待生长至80%～90%融合后,设立10组:正常对照组、模型对照组、α-玉米赤霉醇1,10,100,1000nmol/L组、雌二醇1,10,100,1000nmol/L组,6孔/组。③除正常对照组外,其余各组细胞均复制缺氧/复氧损伤模型,置于体积分数为0.93的N2 0.05的CO2 0.02的O2缺氧环境中3h,然后恢复正常氧供应1h。α-玉米赤霉醇各组于造模前20min分别加入α-玉米赤霉醇,使细胞培养液中α-玉米赤霉醇终浓度分别达到1,10,100,1000nmol/L;雌二醇各组于造模前20min分别加入雌二醇,使细胞培养液中雌二醇终浓度分别达到1,10,100,1000nmol/L;正常对照组、模型对照组不予任何药物处理。④吸取各组上清液,按酶联免疫吸附试剂盒说明操作分别测定各孔可溶性E-选择素、细胞间黏附分子1、血管细胞黏附分子1的含量。结果:①缺氧/复氧后各组细胞E-选择素含量的比较:与正常对照组比较,缺氧3h/复氧1h后模型对照组脐静脉内皮细胞上清液中的E-选择素含量明显升高[(1.77±0.36),(5.62±0.74)pg/L,P<0.01]。与模型对照组比较,α-玉米赤霉醇1,10,100,1000nmol/L组E-选择素含量均明显降低[(5.62±0.74),(3.53±1.21),(3.16±0.94),(2.79±1.78),(2.18±0.75)pg/L,P<0.05或0.01],雌二醇1,10,100,1000nmol/L组E-选择素含量亦均明显降低[(5.62±0.74),(3.72±0.42),(2.99±0.61),(2.45±0.99),(2.34±0.70)pg/L,P<0.01],且呈剂量依赖性。相同质量浓度的α-玉米赤霉醇与雌二醇作用差异无显著性意义(P>0.05)。②缺氧/复氧后各组细胞间黏附分子1、血管细胞黏附分子1含量的比较:与E-选择素含量情况相似。结论:①人脐静脉内皮细胞给予α-玉米赤霉醇预处理后,可显著抑制由缺氧/复氧损伤所引起的黏附分子高表达,且呈剂量依赖性,保护内皮细胞免受炎症递质损伤。②与相同浓度的雌二醇抑制作用相似,是一种有一定应用前景的雌激素替代药物。     

Probiotic effects in allergic disease

The increasing prevalence of allergic disease has been linked to reduced microbial exposure in early life. Probiotics have recently been advocated for the prevention and treatment of allergic disease. This article summarises recent publications on probiotics in allergic disease, focusing on clinical studies of prevention or treatment of allergic disease. Studies employing the combined administration of pre‐natal and post‐natal probiotics suggest a role for certain probiotics (alone or with prebiotics) in the prevention of eczema in early childhood, with the pre‐natal component of treatment appearing to be important for beneficial effects. On the other hand, current data are insufficient to support the use of probiotics for the treatment of established allergic disease, although recent studies have highlighted new hope in this area. Probiotic bacteria continue to represent the most promising intervention for primary prevention of allergic disease, and well‐designed definitive intervention studies should now be a research priority.     

Inter-individual and inter-cell type variation in residual DNA damage after in vivo irradiation of human skin

Purpose

The aim of this study was to compare inter-individual and inter-cell type variation in DNA double-strand break (DSB) repair following in vivo irradiation of human skin.

Materials and methods

Duplicate 4 mm core biopsies of irradiated and unirradiated skin were collected from 35 patients 24 h after 4 Gy exposure using 6 MeV electrons. Residual DSB were quantified by scoring 53BP1 foci in dermal fibroblasts, endothelial cells, superficial keratinocytes and basal epidermal cells.

Results

Coefficients of inter-individual variation for levels of residual foci 24 h after in vivo irradiation of skin were 39.9% in dermal fibroblasts, 44.3% in endothelial cells, 32.9% in superficial keratinocytes and 46.4% in basal epidermal cells (p < 0.001, ANOVA). In contrast, the coefficient of inter-cell type variation for residual foci levels was only 11.3% in human skin between the different epidermal and dermal cells (p = 0.034, ANOVA). Foci levels between the different skin cell types were correlated (Pearson’s R = 0.855-0.955, p < 0.001).

Conclusions

Patient-specific factors appear to be more important than cell type-specific factors in determining residual foci levels following in vivo irradiation of human skin.     

Quality of life,work productivity impairment and healthcare resources in inflammatory bowel diseases in Brazil

BACKGROUND Inflammatory bowel diseases(IBD) have been associated with a low quality of life(QoL) and a negative impact on work productivity compared to the general population.Information about disease control,patient-reported outcomes(PROs),treatment patterns and use of healthcare resources is relevant to optimizing IBD management.AIM To describe QoL and work productivity and activity impairment(WPAI),treatment patterns and use of healthcare resources among IBD patients in Brazil.METHODS A multicenter cross-sectional study included adult outpatients who were previously diagnosed with moderate to severe Crohn's disease(CD) or ulcerative colitis(UC).At enrolment,active CD and UC were defined as having a Harvey Bradshaw Index≥8 or a CD Activity Index≥220 or calprotectin 200 μg/g or previous colonoscopy results suggestive of inadequate control(per investigator criteria) and a 9-point partial Mayo score≥5,respectively.The PRO assessment included the QoL questionnaires SF-36 and EQ-5 D-5 L,the Inflammatory Bowel Disease Questionnaire(IBDQ),and the WPAI questionnaire.Information about healthcare resources and treatment during the previous 3 years was collected from medical records.Chi-square,Fisher's exact and Student's t-/Mann-Whitney U tests were used to compare PROs,treatment patterns and the use of healthcare resources by disease activity(a=0.05).RESULTS Of the 407 patients in this study(CD/UC:64.9%/35.1%,mean age 42.9/45.9 years,54.2%/56.6% female,38.3%/37.1% employed),44.7%/25.2% presented moderate-to-severe CD/UC activity,respectively,at baseline.Expressed in median values for CD/UC,respectively,the SF-36 physical component was 46.6/44.7 and the mental component was 45.2/44.2,the EQ-visual analog scale score was 80.0/70.0,and the IBDQ overall score was 164.0/165.0.Moderate to severe activity,female gender,being unemployed,a lower educational level and lower income were associated with lower QoL(P 0.05).Median work productivity impairment was 20% and 5% for CD and UC patients,respectively,and activity impairment was 30 %,the latter being higher among patients with moderate to severe disease activity compared to patients with mild or no disease activity(75.0% vs 10.0%,P 0.001).For CD/UC patients,respectively,25.4%/2.8% had at least one surgery,38.3%/19.6% were hospitalized,and 70.7%/77.6% changed IBD treatment at least once during the last 3 years.The most common treatments at baseline were biologics(75.3%)and immunosuppressants(70.9%) for CD patients and 5-AS A compounds(77.5%) for UC patients.CONCLUSION Moderate to severe IBD activity,especially among CD patients,is associated with a substantial impact on QoL,work productivity impairment and an increased number of IBD surgeries and hospitalizations in Brazil.     

RAGE interacts with the necroptotic protein RIPK3 and mediates transfusion-induced danger signal release

RBC transfusion is associated with increased morbidity and mortality in critically ill patients. Endothelial cell necroptosis and subsequent damage-associated molecular pattern (DAMP) release has been identified as a mechanism of injury following RBC transfusion. Mounting evidence implicates the pro-inflammatory pattern recognition receptor, Receptor for Advanced Glycation End Products (RAGE), in initiating cell death programmes such as necroptosis. Here, we demonstrate the role of RAGE in endothelial necroptosis, as deletion of RAGE attenuates necroptotic cell death in response to TNFα, LPS or CpG-DNA. We show direct interaction of RAGE with the critical mediator of necroptosis, Receptor Interacting Protein Kinase 3 (RIPK3), during necroptosis. Furthermore, we observe decreased plasma High Mobility Group Box 1 (HMGB1) and RIPK3 levels in RAGE deficient mice compared to WT mice post-transfusion, substantiating the role for RAGE in transfusion-induced DAMP release in vivo. Collectively, these findings underscore RAGE as an essential mediator of regulated necrosis and post-transfusion DAMP release. Further studies to understand the role of RAGE and the necroptotic pathway in transfusion-induced organ injury may offer key targets to mitigate transfusion-related risks, including the risk of ARDS, in susceptible hosts.     

Cushing syndrome: pituitary CT scanning

Proper treatment of patients with Cushing syndrome depends on knowledge of whether the condition derives from a pituitary or adrenal adenoma or an ectopic tumor that secretes adrenocorticotropic hormone (ACTH). Fifty-seven patients with Cushing syndrome were prospectively evaluated to determine whether computed tomography (CT) of the pituitary gland is useful in diagnosing or excluding an ACTH-producing adenoma and in predicting the site of the adenoma within the pituitary gland. Findings on CT scans were normal in 35 of 50 patients with surgically confirmed Cushing disease and in seven patients with ectopic ACTH-producing tumors. Of the 15 patients with Cushing disease for whom CT results were abnormal, eight had macroadenomas and seven had microadenomas. This study demonstrates a low sensitivity (30%) and low diagnostic accuracy (39%) of CT in the detection of ACTH-producing pituitary tumors and indicates the need for alternative methods of confirming the pituitary origin of excessive ACTH secretion in most patients.