Simultaneous quantification of apolipoproteins A-I,E, and J in human plasma by LC-MS/MS for clinical application to diabetes mellitus complicated with cardiovascular disease

Apolipoproteins (Apos) play an important role in regulating plasma lipid concentration. Complex disorders of Apos are highly related with diabetes mellitus, cardiovascular and other diseases. Direct measures of lipoprotein fractions for risk assessment suffer from inaccuracy in the dyslipidemia and pathological states. Therefore, a reliable precise assay will be of high clinical utility. LC-MS/MS methods with multiple reaction monitoring modes have proven suitable for multiplexed quantification. We aimed to develop a simple, cost-effective and amenable LC-MS/MS assay for quantification of ApoA-I, ApoE and ApoJ in human plasma. Standards were constructed from substitute matrix and proteotypic peptides for external calibration and corresponding stable isotope labeled peptides were added as internal standards to remove matrix effects. Analytical validation of the assay included the assessment of linearity, accuracy (RE: −3.02% to 5.32%), intra-assay precision (RSD: 2.50% to 6.56%), inter-assay precision (RSD: 0.78% to 6.68%), spiking recovery rate (accuracy: 87.17% to 112.71%), matrix effect (accuracy: 88.03% to 114.87%), and reproducibility and repeatability of sample preparation (RSD: 1.95% to 7.26%). The performance of proteotypic peptides ApoA-I, ApoE and ApoJ was sufficient for triplex quantitation within a linear range from 16.26 to 1626.41 pmol mL⁻¹, 1.03 to 103.35 pmol mL⁻¹ and 0.86 to 86.46 pmol mL⁻¹ respectively. For all quantified peptides, the determination coefficient (R²) was >0.997. Besides, the validated LC-MS/MS method has been successfully applied to the quantification of plasma samples in diabetes mellitus and cardiovascular diseases. We anticipate that this assay may provide an alternative method for future clinical applications.

Simultaneous quantification of apolipoproteins A-I, E, and J in human plasma by LC-MS/MS.     

江苏省2012-2014年慢性HBV感染者HBeAg血清学转换后再逆转的特征分析

目的 了解慢性HBV感染者HBeAg血清学转换后发生HBeAg逆转的流行病学特征,为推进慢性HBV感染者的规范管理提供依据。方法 2012-2014年对在江苏省传染病防治示范区建立的已发生HBeAg血清学转换的慢性HBV感染者队列开展定期随访。通过Cox回归分析慢性HBV感染者HBeAg血清学转换后HBeAg逆转的特征及影响因素,通过重复测量数据方差分析方法分析不同HBeAg转归者HBV DNA的变化趋势。结果 2012年5 068例已发生HBeAg血清学转换的慢性HBV感染者［平均年龄（51.9±12.8）岁］随访至2014年时121例发生HBeAg逆转,观察9 359人年,HBeAg逆转率为1.3/100人年。HBeAg逆转率随年龄增长逐渐降低,从<20岁年龄组的4.7/100人年下降至≥60岁年龄组的1.0/100人年。HBV DNA<2 000 IU/ml的感染者HBeAg逆转率为1.1/100人年,低于≥200 000 IU/ml者（3.8/100人年）。通过Cox多元回归分析,相对于HBV DNA<2 000 IU/ml者,HBV DNA≥200 000 IU/ml的感染者HBeAg逆转可能性增大（HR=3.44,95% CI：1.91～6.20,P=0.000）。HBeAg逆转组的HBV DNA和ALT较HBeAg持续阴性组均上升更快（P=0.000）。结论 社区人群中已发生HBeAg血清学转换的慢性HBV感染者有一定比例会发生HBeAg逆转,低年龄段和高病毒载量者的HBeAg逆转率高。对已发生HBeAg血清学转换的慢性HBV感染者应定期随访管理。     

黑米花色素苷对大鼠血液生化指标和巨噬细胞功能影响的研究

目的观察黑米花色素苷(black rice anthocyanin,BA)对大鼠血液生化指标的影响及其对腹腔巨噬细胞(peritoneal macrophage,PM)与肺泡巨噬细胞(alveolar macrophage,AM)的激活作用,探讨其对大鼠的免疫调节作用。方法雌性SD大鼠24只,随机分为对照组和BA低(BAL,25mg/kgbw),中(BAM,50mg/kgbw),高(BAH,100mg/kgbw)剂量组4组。除对照组给生理盐水外,另三组分别给予相应剂量的BA,1/d连续30d。实验结束时,从股动脉取血,用生化方法测定巨噬细胞内乳酸脱氢酶(LDH)、酸性磷酸酶(ACP)、超氧化物歧化酶(SOD)活力、及丙二醛(MDA)含量,并进一步观察巨噬细胞摄取中性红的能力。结果BA组大鼠血液中尿素氮(UN)、尿酸(UA)、甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL)水平都有不同程度的降低,PM内ACP、LDH活力显著升高,PM和AM内SOD活力升高,MDA水平降低,而且能显著增强PM、AM的吞噬能力。结论BA对大鼠血液和巨噬细胞生化指标有重要的免疫调节作用。     

维持性血液透析患者医院感染危险因素分析

目的对维持性血液透析(MHD)患者医院感染的相关危险因素进行分析,为控制和降低医院感染提供参考依据。方法回顾性分析2011年6月-2013年9月在医院接受MHD的137例患者临床资料,对发生医院感染患者的性别、年龄、低蛋白血症、贫血、糖尿病肾病(DN)、透析龄、心功能不全(HF)及PICC等感染相关危险因素进行单因素分析和logistic回归分析。结果 137例MHD患者中共有55例发生医院感染,感染率为40.1%;医院感染部位以呼吸道为主,占38.2%,其次为泌尿道、胃肠道和PICC相关感染,分别占21.8%、16.4%和14.5%;单因素分析显示,MHD患者年龄≥60岁、有贫血、低蛋白血症、DN、HF及应用PICC、透析龄≥1年与发生医院感染有相关性(P<0.05);多因素logistic回归分析显示,年龄≥60岁、有低蛋白血症、贫血、DN和应用PICC是发生医院感染的独立危险因素。结论 MHD患者医院感染危险因素较多,血透室应加强医院感染的培训和教育,规范管理,对感染危险因素进行针对性干预,可有效降低医院感染的发生。     

艾司西酞普兰与氟西汀治疗老年抑郁的对照研究

目的 评价草酸艾司西酞普兰与氟西汀治疗老年抑郁症的临床疗效及安全性.方法 将62例老年抑郁症患者随机分为艾司西酞普兰组31例,氟西汀组31例,观察6周.于治疗前和治疗1周、2周、4周、6周末采取汉密尔顿抑郁量表评定疗效,不良反应量表评定不良反映.结果 艾司西酞普兰组显效率77.4%,氟西汀组77.2%,两组比较差异无统计学意义（P＞0.05） 艾司西酞普兰组不良反应发生率35.48%,氟西汀组51.62%,两组比较差异没有统计学意义（P＞0.05）.结论 艾司西酞普兰与氟西汀治疗老年抑郁症疗效相当,但艾司西酞普兰起效比氟西汀起效更快更安全,适合作为老年抑郁患者的一线用药.     

中华猕猴桃的防癌作用——(七)阻断慢性萎缩性胃炎病人体内N-亚硝基脯氨酸合成

本文报导了中华猕猴桃汁阻断慢性萎缩性胃炎(CAG)病人体内N-亚硝基脯氨酸(NPRO)合成的作用。在11名CAG病人的自身对照研究中,病人食入300mg NaNO_3和500mg脯氨酸后,尿中NPRO从本底值26.3nmol升至63.9nmol;同时服4.1ml浓缩猕猴桃汁后,多数人尿中NPRO明显下降,阻断率达94.4％。此外,有2人体内NPRO合成水平很高,提示可能有较高的癌变危险性。     

Comparative effect of genistein and daidzein on the expression of MCP-1, eNOS, and cell adhesion molecules in TNF-��-stimulated HUVECs

We compared the effects of genistein and daidzein on the expression of chemokines, cell adhesion molecules (CAMs), and endothelial nitric oxide synthase (eNOS) in tumor necrosis factor (TNF)-α-stimulated human umbilical vascular endothelial cells (HUVECs). TNF-α exposure significantly increased expression of monocyte chemoattractant protein (MCP)-1, vascular adhesion molecule (VCAM)-1, and intercellular adhesion molecule-1. Genistein significantly decreased MCP-1 and VCAM-1 production in a dose-dependent manner, whereas CAM expression was not significantly lowered by genistein treatment. However, daidzein slightly decreased MCP-1 production. The effects of genistein and daidzein on MCP-1 secretion coincided with mRNA expression. Pre-treatment with either genistein or daidzein elevated eNOS expression and nitric oxide production disturbed by TNF-α exposure. A low concentration of isoflavones significantly inhibited nuclear factor (NF)κB activation, whereas a high dose slightly ameliorated these inhibitive effects. These results suggest that genistein had a stronger effect on MCP-1 and eNOS expression than that of daidzein. Additionally, NFκB transactivation might be partially related to the down-regulation of these mRNAs in TNF-α-stimulated HUVECs.     

Expansion of CD3+CD8+PD1+ T lymphocytes and TCR repertoire diversity predict clinical responses to adoptive cell therapy in advanced gastric cancer

The adoptive cell therapy (ACT) and delivery of ex vivo activated cellular products, such as dendritic cells (DCs), NK cells, and T cells, have shown promise for the treatment of gastric cancer (GC). However, it is unknown which cells can improve patient survival. This study was focused on the antitumour activity of a subset of these cellular products and their relationships with clinical outcomes. Nineteen patients were enrolled at the Capital Medical University Cancer Center, Beijing Shijitan Hospital, from June 1, 2013, to May 30, 2016. CD8⁺PD1⁺ T-cell sorting was carried out using flow cytometry, and the T-cell receptor (TCR) repertoire during ex vivo expansion for 15 days was analyzed by next-generation sequencing. After 15 days of culture, the number of CD8⁺ T cells had increased significantly, and the number of CD4⁺ T cells had increased correspondingly. After ex vivo expansion, CD8⁺ T cells exhibited significantly enhanced expression of PD-1, LAG-3, and TIM-3 but not 4-1BB. Survival analysis showed that patients with a pro/pre value of CD8⁺PD-1⁺ T cells >2.4 had significantly favorable overall survival (OS) (median OS time, 248 days versus 96 days, P=0.02) and progression-free survival (PFS) (median PFS time, 183 days vs. 77 days, P=0.002). The sorted CD8⁺PD-1⁺ T cells displayed enhanced antitumor activity and increased IFN-γ secretion after coculture with autologous tumor cell lines. TCR repertoire diversity was decreased after ex vivo expansion, which decreased the Shannon index and increased the clonality value. The prognosis of patients was significantly improved and was associated with the extent of CD8⁺PD-1⁺ T-cell expansion. In summary, this study showed that after ex vivo expansion for 15 days, CD8⁺PD-1⁺ T cells could be identified as tumor-reactive cells in patients treated for GC. Changing TCR species can predict the extent of CD3⁺CD8⁺PD1⁺ T-cell growth and the effect of ACT treatment.