反复种植失败妇女“种植窗”时期子宫内膜组织的细胞骨架蛋白质的差异表达

目的探讨体外受精-胚胎移植治疗周期中反复种植失败(RIF)和已生育正常妇女"种植窗"时期(LH+6~LH+9d)子宫内膜组织蛋白质表达的差异。方法选择RIF妇女和正常对照妇女各4名分为实验组和对照组。采用胶内差异双向电泳、(2D-DIGE)和基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)技术鉴定子宫内膜组织的差异表达蛋白质。结果两组之间获得16个差异表达蛋白质(其中6个下调表达,10个上调表达),其中10个参与细胞骨架构成,分别是:波形蛋白、埃兹蛋白、Septin 2蛋白、radixin蛋白、纽蛋白、角蛋白8、角蛋白10、角蛋白18、角蛋白19及肌动蛋白。结论本研究结果证实"种植窗"时期RIF妇女子宫内膜组织存在多个细胞骨架蛋白质表达的异常,可能是子宫内膜细胞"质膜转换"缺陷和胚胎种植失败的重要原因。     

Amelioration of hyperglycemic and hyperosmotic induced vascular dysfunction by in vivo inhibition of protein kinase C and p38 MAP kinase pathway in the rat mesenteric microcirculation

BACKGROUND: Recently, we demonstrated in vivo effects of acutely induced hyperglycemia, diabetes and mannitol infusions on rat mesenteric microcirculation concerning leukocyte-endothelial-cell interaction (Sch?ffler et al. EJCI 28: 886-893, 1998). DESIGN: In this study we have investigated the possible involvement of the protein kinase C (PKC) and p38 MAP kinase cascade as signal transducing elements during hyperglycemic and osmotic stress in an in vivo rat model. RESULTS: Acutely induced hyperglycemia resulted in a significant increase in leukocyte adhesion. This effect could be mimicked by mannitol. Both PKC and p38 MAP kinase were involved in the effects mediated by glucose and mannitol. Acutely induced hyperglycemia resulted in a significant increase in leukocyte emigration. This effect could be imitated by mannitol. However, PKC and p38 MAP kinase were only involved under osmotic stimulation. The hyperglycemia-induced reduction in leukocyte rolling velocity seemed to be a glucose-specific effect, since mannitol did not influence leukocyte rolling velocity. This glucose effect on leukocyte rolling velocity was mediated by an activation of the PKC/p38 MAP kinase cascade. Both hyperglycemia and osmotic stimuli alone were able to reduce venular shear rate without recruitment of the p38 MAP kinase cascade. The observed reduction of shear rate seems to be mediated only by the osmotic effects of glucose via activation of the PKC system. CONCLUSION: The observed effects of glucose on adhesion, emigration and shear rate are due to osmotic effects. The PKC/MAP kinase cascade is involved as a signal transducing component. The reduction of leukocyte rolling velocity is a glucose-specific effect, mediated by the activation of both the PKC and the p38 MAP kinase cascade. Venular shear rate and leukocyte emigration can be influenced by glucose and mannitol due to different regulation mechanisms. It is concluded, that isoform-specific inhibitors of PKC and specific MAP kinase inhibitors represent a potential drug target for preventing microvascular dysfunction in diabetes.     

Platelet count and erythrocyte sedimentation rate are good predictors of Kawasaki disease: ROC analysis

Background and Aims: Kawasaki disease (KD) is the leading cause of acquired pediatric cardiac disease and requires a timely diagnosis. Available effective therapy is ideally administered within 10 days of illness diagnosis. Recent reports of several laboratory tests in KD have been published. In this study, we aimed to evaluate the sensitivity and specificity of several laboratory tests. Methods: We performed a retrospective study of consecutive patients diagnosed with KD from January to December 2008. We studied the sensitivity and specificity of several different tests [T‐cell subgroups, platelet count, erythrocyte sedimentation rate (ESR), and C‐reactive protein (CRP)] to predict KD using receiveroperator characteristic curve analysis. Results: No significant difference was demonstrated in T‐cell subgroups between patients with KD and referent patients (P>0.05). However, platelet count, ESR, and CRP were significantly higher in patients with KD than in referent patients (P<0.05). ESR showed a sensitivity of 93.9% and specificity of 83.3% with a cut‐off of 15 mm/hr (area under the curve [AUC], 89.1%; P=0.03). Platelet count showed a sensitivity of 70.6% and specificity of 75% with a cut‐off of 336.5×10⁹/l (AUC, 71.2%; P=0.03). Conclusions: These results indicate that platelet count and ESR are good predictors of KD. J. Clin. Lab. Anal. 24:385–388, 2010. © 2010 Wiley‐Liss, Inc.     

妊娠相关血浆蛋白A的原核表达及蛋白纯化

目的构建原核表达PAPP-A重组蛋白,并对蛋白质进行纯化。方法根据DNAstar软件分析获得的编码PAPP-A特异抗原表位,利用Primer Premier 5.0软件设计引物,以PAPP-A c DNA-质粒转化菌液为模板,进行PCR扩增。PAPP-A的DNA和p ET42a质粒分别双酶切,经琼脂糖凝胶电泳及胶回收后进行连接反应,转化至大肠埃希菌Top10,筛选阳性克隆经测序鉴定正确后,转化至大肠埃希菌BL21,诱导产生PAPP-A重组蛋白,利用镍柱初步纯化及离子交换层析柱进一步纯化,用SDS-PAGE及DEAE层析鉴定重组抗原蛋白。结果筛选出抗原表位集中区,在大肠埃希菌BL21中高效表达了重组的PAPP-A蛋白,经过镍柱及离子交换层析纯化后,PAPP-A浓度为0.22 g/L,DEAE层析分析证实其纯度较高。结论成功筛选出PAPP-A的抗原表位集中区,并制备了重组抗原蛋白,为PAPP-A的后续临床研究提供依据。     

Decreased small dense LDL levels in Gilbert's syndrome

Objective

To investigate the role of small dense low density lipoprotein cholesterol (sd-LDL-C) in the mechanism of decreased incidence of cardiovascular disease in Gilbert's syndrome (GS).

Design and methods

sd-LDL-C, ox-LDL, and high sensitive C reactive protein (hs-CRP) levels were investigated in subjects with GS (n = 42) and compared to healthy controls (n = 52).

Results

Age, gender and body mass index (BMI) distributions were similar between the two groups. sd-LDL-C, ox-LDL and hs-CRP levels were lower in GS than the healthy controls (p < 0.001, p < 0.001 and p = 0.001, respectively). Unconjugated bilirubin was negatively correlated with sd-LDL-C, ox-LDL and hs-CRP (r = −0.594, p < 0.001; r = −0.249, p = 0.016 and r = − 0.373, p < 0.001 respectively). In addition, sd-LDL-C was positively correlated with ox-LDL (r = 0.307, p = 0.003).

Conclusions

The findings of this preliminary study suggest that reduced sd-LDL-C, ox-LDL and hs-CRP levels may have a role in preventing atherosclerosis in subjects with GS.     

阿托伐他汀对脑梗死患者C反应蛋白的影响

目的研究阿托伐他汀对脑梗死患者C反应蛋白及血脂的影响。方法56例脑梗死患者给予阿托伐他汀10mg口服,1次/d,疗程4周,观察治疗前后血清C反应蛋白及血脂浓度的变化。结果使用阿托伐他汀治疗后,患者血清C反应蛋白、胆固醇、甘油三酯、低密度脂蛋白显著降低,高密度脂蛋白明显上升。结论阿托伐他汀对脑梗死患者具有良好的调脂、抗炎作用。     

Characterization and gene transfer in mesenchymal stem cells derived from human umbilical-cord blood

It has been shown that the stromal-cell population found in bone marrow can be expanded and differentiated into cells with the phenotypes of bone, cartilage, muscle, neural, and fat cells. However, whether mesenchymal stem cells (MSCs) are present in human umbilical-cord blood (UCB) has been the subject of ongoing debate. In this study, we report on a population of fibroblastlike cells derived from the mononuclear fraction of human UCB with osteogenic and adipogenic potential, as well as the presence of a subset of cells that have been maintained in continuous culture for more than 6 months. These cells were found to express CD29, CD44, CD90, CD95, CD105, CD166, and MHC class, but not CD14, CD34, CD40, CD45, CD80, CD86, CD117, CD152, or MHC class II. We also compared gene expression after gene transfer using lenti- and adenoviral vectors carrying the green fluorescence protein to the MSCs derived from UCB because a reliable gene-delivery system is required to transfer target genes into MSCs, which have attracted attention as potential platforms for the systemic delivery of therapeutic genes. The lentiviral vectors can transduce these cells more efficiently than can adenoviral vectors, and we maintained transgene expression for at least 5 weeks. This is the first report showing that UCB-derived MSCs can express exogenous genes by way of a lentivirus vector. These results demonstrate that human UCB is a source of mesenchymal progenitors and may be used in cell transplantation and a wide range of gene-therapy treatments.