血浆同型半胱氨酸及其相关硫醇物在全血中的稳定性研究

目的 观察含EDTA的全血样本放置时间、保存温度以及不同稳定剂对血浆同型半胱氨酸(homocysteine,Hcy)及其相关硫醇物水平的影响.方法 用含EDTA、EDTA-氟化钠(EDTA-NaF)、EDTA-3-Deazaadenosine(EDTA-3DA)的试管收集17名健康成人静脉血,置于碎冰上(0～4 ℃)或室温中(25 ℃)保存,放置0、3、6、24、48 h后分离血浆.用HPLC法测定血浆总同型半胱氨酸(tHcy)、总半胱氨酸(total cysteine,tCys)、总半胱氨酰甘氨酸(total cysteinylglycine,tCysGly)、总谷胱甘肽(total glutathione,tGSH)浓度.设定全血样本0 h分离血浆所测硫醇物浓度为基础值.结果 EDTA管在室温中放置3、6、24、48 h,tHcy分别增加38.5%、64.2%、141.9%、225.4%;tCysGly、tGSH在3 h分别增加20.0%、37.9%,tCys则降低3.5%.EDTA管在碎冰上保存,各硫醇物浓度6 h内增加不超过5%.EDTA-3DA和EDTA-NaF管在室温放置3 h,与各自基础值相比,血浆tHcy、tCys、tCysGly、tGSH浓度差异无统计学意义(EDTA-3DA管:F值分别为0.01、0.94、0.09、0.01,P值均>0.05;EDTA-NaF管:F值分别为0.85、0.04、0.03、0.02,P值均>0.05).结论 EDTA抗凝血浆,所测tHcy及其相关硫醇物浓度呈时间、温度依赖性增加.血浆tHcy等硫醇物测定的分析前处理条件必须标准化.EDTA-3DA和EDTA-NaF管可使血浆tHcy、tCys、tCysGly、tGSH在室温中至少稳定3 h.

Abstract：

Objective To investigate the effects of different stabilizers, time and temperature before centrifugation on the stability of homocysteine (Hcy) and other related thiols levels involving EDTA-containing whole blood.Methods Blood was drawn from 17 healthy adults and collected into tubes containing EDTA, EDTA plus NaF and EDTA plus 3-deazaadenosine(3DA),then stored on crush ice(0-4 ℃) immediately or at room temperature(25 ℃).Plasma was separated at 0, 3, 6, 24 and 48 hours, respectively.The levels of plasma total Hcy (tHcy), total cysteine (tCys), tatal cysteinylglycine (tCysGly) and tatal glutathione (tGSH) were measured by HPLC.The plasma immediately separated was used as baseline sample. Results In EDTA tubes stored at room temperature, tHcy levels increased by 38.5%, 64.2%, 141.9%, 225.4% for 3, 6, 24, and 48 h, respectively.The levels of tCysGly and tGSH increased by 20.0% and 37.9% within 3 h, however, tCys decreased by 3.5%.The levels of the thiols increase by less than 5% up to 6 h in EDTA tubes stored on crush ice.In EDTA-3DA and EDTA-NaF tubes, no statistical differences were observed in the plasma levels of tHcy, tCys,tCysGly and tGSH compared with their respective baseline values at room temperature for 3 h(EDTA-3DA tubes:F=0.01,0.94,0.09,0.01,all P>0.05;EDTA-NaF tubes:F=0.85,0.04,0.03,0.02,all P>0.05).Conclusions The EDTA-plasma levels of tHcy and other related thiols are time and temperature-dependent. There is a strong need for standardization of blood sample collection and processing in tHcy and other thiols assays. The plasma concentrations of tHcy, tCys, tCysGly and tGSH were stable for 3 h at least in the EDTA-3DA and EDTA-NaF tubes kept at room temperature.     

慢性乙型肝炎及乙型肝炎性肝硬化患者色氨酸代谢变化及其相关性分析

目的 探讨慢性乙型肝炎(CHB)和乙型肝炎性肝硬化(HBC)患者色氨酸(Trp)代谢变化及相关性.方法 选取2015年10月至2016年1月在重庆医科大学附属第一医院住院的CHB 81例和HBC 40例患者为研究对象,40例健康者为对照.采用高效液相色谱-荧光检测法(HPLC-FD)对各组受试者血浆中Trp及代谢产物犬尿氨酸(Kyn)、5-羟色胺(5-HT)水平进行测定,实时荧光定量PCR检测HBV-DNA病毒复制数,分析3组间血清学指标差异,采用Pearson相关分析和Logistic回归分析筛选危险因素.结果 与对照组比较,CHB组、HBC组受试者血浆中Trp和5-HT水平明显降低(P<0.01);HBC组受试者血浆中Kyn/Trp明显高于其他两组(P<0.01).HBC组Kyn/Trp与A/G呈负相关(r=-0.686,P<0.01),CHB组Kyn/Trp与Log DNA呈正相关(r=0.784,P<0.01).Logistic回归分析显示Kyn/Trp为CHB和HBC的危险因素.结论 检测血浆中Trp及其代谢产物可作为辅助指标评估乙型肝炎进展情况.     

GnRH-a联合GnRH-ant超短方案对卵巢低反应患者疗效的Meta分析

目的 系统评价促性腺激素释放激素激动剂(GnRH-agonist,GnRH-a)联合促性腺激素释放激素拮抗剂(GnRH-antagonist,GnRH-ant)超短方案对卵巢低反应患者治疗的有效性.方法 检索MEDLINE、EMBASE、CENTRAL、中国生物医学数据库和万方数据库,检索时间截至2017年7月.纳入关于GnRH-a联合GnRH-ant超短方案治疗卵巢低反应患者的随机对照试验或临床对照试验,由2位评价员根据纳入与排除标准独立进行文献筛选、资料提取和质量评价后,采用RevMan 5.3软件进行Meta分析.结果 共纳入6篇文献,336个周期;GnRH-a联合GnRH-ant超短方案与对照组比较,Gn用药总量(WMD=6.32,P=0.09,95％ CI:-0.92 ～ 13.55)、Gn用药时间(WMD=0.38,P=0.07,95％CI:-0.04 ～0.81)、胚胎移植数(WMD=0.24,P=0.49,95％ CI:-0.44～0.91)方面差异无统计学意义,其获卵数(WMD=0.29,P=0.40,95％ CI:-0.38～0.96)、受精率(WMD=4.39,P=0.21,95％CI:-2.41 ～ 11.18)、临床妊娠率(OR=1.72,P=0.38,95％ CI:0.52 ～5.71)较对照组稍升高,但差异均无统计学意义.结论 GnRH-a联合GnRH-ant超短方案对卵巢低反应患者的疗效与对照组促排卵方案比较无明显差异,且在获卵数、受精率、临床妊娠率上稍优于其他促排卵方案.     

血清胆汁酸代谢轮廓分析在妊娠肝内胆汁淤积症诊断中的应用

目的:探讨血清胆汁酸代谢轮廓分析在妊娠肝内胆汁淤积症(Intrahepatic cholestasisofpregnancy,ICP)诊断中的作用.方法:应用柱前衍生HPLC-荧光检测法测定ICP患者,乙肝孕妇和正常孕妇(各11例)血清胆汁酸谱(5种游离型和4种甘氨结合型),结合主成分分析法(Principal component analysis,PCA)和偏最小二乘判别分析法(Partial least-squares discriminant analysis,PLS-DA)对血清胆汁酸谱进行分析.结果:血清胆汁酸代谢轮廓分析可区分ICP患者,乙肝孕妇和正常孕妇,其模型的正确诊断率和预测能力分别为97.0%和75.3%.结论:血清胆汁酸代谢轮廓分析有助于ICP的诊断和鉴别诊断.     

川芎嗪对应激家兔血液流变学的影响

川芎嗪（Ligustrazin，LGT）是中药川芎的有效成分之一，有活血化淤作用。有资料表明，川芎嗪常用于心脑血管疾病、糖尿病等多种高粘滞综合征的治疗。情绪紧张、激动可引起高血压等心血管疾病，与交感一肾上腺系统的激活有关。本研究采用束缚应激方法观察应激家兔血液流变学指标和血小板聚集功能及去甲肾上腺素的变化，初步探讨使用川芎嗪后在血流变学方面的抗应激损伤作用。