Detection of apoptosis-associated microRNA in human apheresis platelets during storage by quantitative real-time polymerase chain reaction analysis

Background

Platelet transfusion is an essential part of the treatment of a variety of conditions such as thrombocytopenia and qualitative platelet disorders. As indicated in previous reports, during in vitro storage, platelets undergo morphological and physiological changes collectively known as the platelet storage lesion. Apoptosis is a programmed process of cell death, which has been considered as an important cause of platelet storage lesion under the common storage conditions in standard blood banks. Platelets are anucleate blood cells, but contain significant amounts of microRNA (miRNA, miR), which may play an important role in the regulation of gene expression. Drawing on previously published reports on cell apoptosis, we selected 49 miRNA for analysis to explore whether miRNA are of importance during the storage of platelets.

Materials and methods

We used quantitative real-time polymerase chain reaction analysis to determine the levels of expression of miRNA in apheresis platelets at different times of storage. Bioinformatics analysis was applied to explore target genes and the main functions of the selected miRNA.

Results

Our observations suggest that apheresis platelets contain large amounts of apoptosis-associated miRNA. The levels of expression of 25 miRNA remained high and ten of these miRNA showed different expression from that at day 0. Of these ten miRNA, hsa-miR-326, hsa-miR-96, hsa-miR-16, hsa-miR-155 and hsa-miR-150 were up-regulated, while hsa-miR-7, hsa-miR-145, hsa-miR-24, hsa-miR-25 and hsa-miR-15a were down-regulated. The markedly increased expression of hsa-miR-326 in all platelets is noteworthy (p<0.001).

Discussion

Since Bcl-xl and Bak1, members of the Bcl-2 family, are the targets of hsa-miR-326, our findings suggest that hsa-miR-326 may be involved in platelet apoptosis during storage.     

IL-35与炎症性疾病

IL-35是一种抑制性细胞因子,属于IL-12细胞因子家族的新成员,它是由调节性T细胞结构性分泌产生,由P35和EBI3两个亚基组成.研究发现,IL-35在自身免疫性疾病、炎症性疾病、肿瘤等治疗中有重要作用.在临床上,感染性疾病的控制一直是个难题,IL-35的应用也许为临床提供一种新的思路和方法.     

孟鲁司特钠对AECOPD患者MMP-9和TIMP-1水平的影响

目的通过采用酶联免疫吸附试验（ELISA）法测定中重度慢性阻塞性肺疾病急性加重期（acute exacerbations of chronic obstructive pulmonary disease,AECOPD）患者治疗前、后血清基质金属蛋白酶-9（matrixmetalloproteinase-9,MMP-9）和基质金属蛋白酶组织抑制剂-1（tissue inhibitor of metalloproteinases-1,TIMP-1）水平,观察孟鲁司特钠对中重度AECOPD患者血清MMP-9、TIMP-1水平的影响。方法选取中重度AECOPD患者63例为研究对象,随机分组：治疗组30例,给予常规治疗＋孟鲁司特钠10mg,1次／d,口服,疗程7～14d;对照组33例,给予常规治疗,疗程7～14d。采用双抗体夹心酶联免疫吸附法测定两组血清MMP-9和TIMP-1水平,并对其变化进行分析比较。结果①治疗组和对照组治疗前MMP-9水平比较差异无统计学意义,治疗组治疗后MMP-9水平[（22．02±6．34）μg／L]低于对照组治疗后[（26．31±8．23）μg／L]（P〈O．05）。治疗组和对照组治疗前MMP-9水平分别为[（27．59士7．71）μg／L]、[（27．74±7．96）μg／L]均高于治疗后E（22．02±6．34）,ug／L]、[（26．31±8．23）μg／L]（P〈0．05）。②治疗组和对照组治疗前TIMP-1水平比较差异无统计学意义,治疗组治疗后[（22．23±5．52）μg／L]TIMP-1水平高于对照组治疗后[（17．23±8．23）μg／L]（P〈O．05）。治疗组和对照组治疗前TIMP-1水平均分别为[（16．34±1．45）~g／L]、[（16．20±2．03）μg／L]低于治疗后[（22．23±5．52）μg／L]、[（17．23±2．45）μg／L]（P〈0．05）。结论孟鲁司特钠对中重度AECOPD患者血清MMP-9和TIMP-1水平有影响,推测孟鲁司特钠可减轻中重度AECOPD患者气道炎症并延缓气道重塑。     

LRP1-Dependent Endocytic Mechanism Governs the Signaling Output of the Bmp System in Endothelial Cells and in Angiogenesis

Rationale: Among the extracellular modulators of Bmp (bone morphogenetic protein) signaling, Bmper (Bmp endothelial cell precursor-derived regulator) both enhances and inhibits Bmp signaling. Recently we found that Bmper modulates Bmp4 activity via a concentration-dependent, endocytic trap-and-sink mechanism. Objective: To investigate the molecular mechanisms required for endocytosis of the Bmper/Bmp4 and signaling complex and determine the mechanism of Bmper's differential effects on Bmp4 signaling. Methods and Results: Using an array of biochemical and cell biology techniques, we report that LRP1 (LDL receptor-related protein 1), a member of the LDL receptor family, acts as an endocytic receptor for Bmper and a coreceptor of Bmp4 to mediate the endocytosis of the Bmper/Bmp4 signaling complex. Furthermore, we demonstrate that LRP1-dependent Bmper/Bmp4 endocytosis is essential for Bmp4 signaling, as evidenced by the phenotype of lrp1-deficient zebrafish, which have abnormal cardiovascular development and decreased Smad1/5/8 activity in key vasculogenic structures. Conclusions: Together, these data reveal a novel role for LRP1 in the regulation of Bmp4 signaling by regulating receptor complex endocytosis. In addition, these data introduce LRP1 as a critical regulator of vascular development. These observations demonstrate Bmper's ability to fine-tune Bmp4 signaling at the single-cell level, unlike the spatial regulatory mechanisms applied by other Bmp modulators.     

A giant cardiac lipoma associated with ventricular inversion and ventricular aneurysm: ultrasonography and CT imaging findings

Cardiac lipomas are rare, mostly asymptomatic, and usually found incidentally during noncardiac examinations; however, they also can be symptomatic, depending on their size and location. Here, we report a case of surgically proved pericardial lipoma that was big and for which cardiac structures were substantially altered. The combination of CT imaging and ultrasonography enabled a precise diagnosis in terms of localization, tissue characterization, and complications of the tumor. The origin of the tumor, however, remains undetermined despite a series of postoperative CT scan and ultrasound examinations.     

Accumulation of tissue advanced glycation end products correlated with glucose exposure dose and associated with cardiovascular morbidity in patients on peritoneal dialysis

ObjectivesAccumulation of tissue advanced glycation end products (AGEs) is a marker of cumulative glycemic and/or oxidative stress. Cutaneous AGEs levels measured by skin autofluorescence correlate well with cardiovascular outcomes in diabetes and hemodialysis (HD) patients. The present study aimed to compare tissue AGEs levels with peritoneal dialysis (PD) and HD patients and to evaluate the relationship between skin autofluorescence and cardiovascular morbidity in patients on PD.MethodsA total of 2388 maintenance dialysis patients (613 PD and 1775 HD) were enrolled in this cross-sectional study. Skin autofluorescence was measured non-invasively with an autofluorescence reader. Cardiovascular morbidity was defined as clinically diagnosed ischemic heart disease, heart failure, stroke or peripheral vascular disease from initiation of dialysis.ResultsMore than 90% of patients on both PD and HD had met current dialysis adequacy targets. Compared to HD group, PD patients receiving conventional glucose-containing dialyzate had significantly higher skin autofluorescence values in each category of age and dialysis duration, irrespective of the presence or absence of diabetes. In PD patients, skin autofluorescence values were strongly correlated with the duration of PD and glucose exposure dose and independently associated with cardiovascular morbidity. Multivariate analysis revealed that glucose exposure dose and skin autofluorescence were the strongest risk factors for cardiovascular morbidity in PD patients after adjustment by age, gender, and other classic- or uremic-related risk factors.ConclusionsAccumulation of tissue AGEs provides a potential link between PD exposure of metabolic stress and progression of cardiovascular disease in patients on PD.