血小板-白细胞聚集体在全麻手术患者中的变化观察

［摘要］　目的　观察全麻手术患者术后血小板-白细胞聚集体（PLA）的变化,探讨全麻手术患者深部静脉栓塞发生的机理。方法　对112例全麻手术患者分别于麻醉诱导前、手术结束后、术后3 d采集标本,利用流式术检测血小板-粒细胞聚集体（PNA）和血小板-单核细胞聚集体（PMA）,观察PNA和PMA的变化。结果　112例全麻手术患者麻醉诱导前PNA为(19.32±3.48)%,PMA为(26.86±5.05)%;手术结束后PNA为(19.48±3.20)%,PMA为(31.86±5.31)%;术后3d PNA为(21.85±2.84)%,PMA为(77.69±5.72)%。麻醉诱导前的PNA和PMA与手术结束后、术后3 d比较差异均有统计学意义（P<0.01）。结论　全麻手术对患者PNA和PMA均有影响,尤其对PMA影响明显,因此PNA和PMA可以作为判断血小板活化程度和预防术后栓塞的敏感性观察指标。     

特发性血小板减少性紫癜患者外周血淋巴细胞亚群和血小板相关抗体变化及临床意义

用流式细胞术直接免疫法检测l72例早期特发性血小板减少性紫癜(ITP)患者外周血淋巴细胞亚群(CD3、CD4、CD8、CD19、C16、CD56)，ELISA法检测血小板表面血小板相关抗体(PAIgG、PAIgA、PAIgM)。结果显示，ITP组与正常对照组比较CD3、CD4／CD8显著降低(P＜0.01)，CD8增高(P＜0．05)，CD19增高极为显著(P＜0．01)；ITP血小板表面PAIgG、PAIgA、PAIgM显著高于对照组；CD19升高和CD4/CD8下降与PAIgG、PAIgA、PAIgM增高有显著的相关性。认为淋巴细胞亚群功能和比例失常、T细胞免疫调节机制紊乱在ITP的发病机制中起非常重要的作用。     

血小板单核细胞聚集体在脓毒症及合并ARDS患者中的表达

目的 探讨血小板单核细胞聚集体在脓毒症及合并急性呼吸窘迫综合征(ARDS)患者中表达及意义.方法 选取急诊科收治的脓毒症患者246例,其中脓毒症组122例,脓毒症合并ARDS组124例.脓毒症合并ARDS患者根据柏林新定义标准分为3个亚组:轻度组44例、中度组42例、重度组38例.流式细胞术检测患者外周血血小板单核细胞...     

肺结核病患者外周血淋巴细胞亚群的绝对数分析及临床意义

目的检测分析肺结核病患者外周血CD3^＋细胞,CD3^＋CD4^＋细胞,CD3^＋CD8^＋细胞,NK细胞（CD3^-/（CDr（16＋56^＋）,B细胞（CD19）数量及CD_3^＋CD_4^＋／CD3^_CD_8^＋比值的变化,并探讨其临床意义。方法采用多参数流式细胞术（FCM）检测21例肺结核患者外周血淋巴细胞亚群水平与15例健康者比较研究。结果肺结核病患者外周血CD3^＋细胞,CD3^＋CD4^＋细胞,CD3^＋CDR^＋细胞,NK细胞数量均比正常人明显减少（P〈0．01）,而B细胞数量没明显改变,CD3^＋CD4^＋／CD_3^＋CD_8^＋比值明显下降（P〈0．01）。结论肺结核病患者细胞免疫功能紊乱,监测结核病患者淋巴细胞亚群水平,对评价患者的细胞免疫功能及对疾病的免疫治疗均有重要的指导意义。     

未经治疗的潜伏梅毒患者外周血淋巴细胞亚群的变化及意义

目的了解未经治疗的潜伏梅毒患者外周血淋巴细胞亚群的变化,并探讨其临床意义。方法对41例未经治疗的潜伏梅毒患者及29例健康对照者的抗凝全血,采用流式细胞仪检测相关淋巴细胞亚群（包括T淋巴细胞亚群、NK细胞及B细胞）,比较两组间的差异,并对其血清学滴度的变化进行相关性分析。结果潜伏梅毒患者CD4＋T淋巴细胞数、CD4＋/CD8＋比值均低于正常对照组（P〈0.01）,CD8＋T淋巴细胞数明显高于对照组（P〈0.01）;CD3＋、CD1＋9、CD1＋6CD5＋6与正常对照组相比无显著性差异（P〉0.05）;梅毒血清学滴度与CD4＋T淋巴细胞数量呈负相关。结论未经治疗的潜伏梅毒患者淋巴细胞亚群失衡,存在细胞免疫抑制。     

强直性脊柱炎患者外周血T细胞亚群的变化及意义

目的探讨强直性脊柱炎（AS）患者外周血中T细胞亚群的变化及其意义。方法采用流式细胞仪（FCM）检测外周血淋巴细胞表面CD3、CD4、CD8及其胞质内细胞因子IFN-γ和IL-4的表达。结果与正常对照组相比,As患者外周血Th细胞（CD;CD4＋）百分率无显著差异,Tc细胞（CD;CD;）明显升高（P〈0．05）;Th1细胞（CD3^＋CD4^＋IFN-γ^＋）百分率明显升高,Th2细胞（CD3^＋CD4^＋IL4^＋）无显著性差异;Te1细胞（CD3^＋CD8^＋IFN-γ^＋）百分率明显降低,T02细胞（CD3^＋CD8^＋IL4^＋）无显著性差异。结论AS患者外周血T细胞Th1／Th2、Tcl／Tc2亚群比例失衡,呈Th1、Tc2优势型。     

肝炎肝硬化及原发性肝癌患者外周血Ｔ淋巴细胞亚群分析

为了探讨慢性肝病合并原发性肝细胞癌患者细胞免疫功能紊乱的发病机理,我们运用流式细胞技术对39例肝炎肝硬化及35例慢性病毒性肝炎合并原发性肝癌患者外周Ｔ血淋巴细胞亚群进行了检测。现将结果分析如下。对象与方法1.对象:74例患者均为1997年1月～1997年6月我院住院病人,分为肝炎肝硬化组(39例)和原发性肝癌组(35例)。肝炎肝硬化的诊断标准符合1995年全国传染病与寄生虫病学术会议修订的《病毒性肝炎防治方案》(北京)标准,男性30例,女性9例,年龄为31～60岁,平均年龄(42.4±11.6)岁。39例肝炎肝硬化患者乙肝病毒标志物均为阳性,其中5例合并…     

免疫性血小板减少性紫癜35例患者外周血免疫细胞亚群的检测及其临床意义

目的 探讨免疫细胞亚群的变化在免疫性血小板减少性紫癜(ITP)发病机制中的作用及其临床意义.方法 应用流式细胞术检测35例ITP患者治疗前、后及20例正常对照者免疫细胞亚群各指标的变化,包括CD3+、CD4+、CD8+、CD56+、CD19+淋巴细胞及CD4+/CD8+比值.结果 ITP患者CD3+ T淋巴细胞百分比(61.58±6.45)%、CD4+ T淋巴细胞百分比(28.38±4.89)%、CD4+/CD8+比值(0.99±0.22)较对照组[(67.85±4.68)%、(38.00±3.37)%、1.54±0.13]均减低(P值均＜0.05),治疗后3项指标[(69.41±5.03)%、(38.17±3.18)%、1.60±0.15]均升高至正常水平;CD8+ T淋巴细胞百分比(29.20±4.50)%及CD19+ B淋巴细胞百分比(17.74±4.14)%较对照组[(24.82±2.93)%、(12.09±3.51)%]升高(P值均＜0.05),治疗后2项指标[(24.06±3.02)%、(10.90±3.55)%]均降至正常水平;ITP患者CD56+细胞百分比治疗前(15.80±2.85)%、治疗后(15.16±2.77)%与对照组(16.36±2.75)%差异无统计学意义(P＞0.05).结论 免疫细胞亚群紊乱参与了ITP的发病,对其检测可作为ITP的辅助诊断,在指导治疗方面可能有一定的意义.

Abstract：

Objective To explore the clinical significance of immunocyte subsets before and after immunosuppressive therapy in the peripheral blood of patients with immune thrombocytopenic purpura (ITP).MethodsThe percentages of immunocyte subsets in the peripheral blood of 35 patients with ITP and 20 healthy controls were detected by flow cytometry,including CD3+,CD4+,CD8+,CD56+,CD19+ lymphocytes and CD4+/CD8+.Results The percentages of CD3+ T lymphocyte (61.58 ± 6.45 ) %,CD4+ T lymphocyte (28.38 ±4.89)% and the ratio of CD4+/CD8+ 0.99 0.22 in patients with ITP were lower than those in healthy controls[( 67.85 ± 4.68 ) %,( 38.00 ± 3.37 ) %,1.54 ± 0.13,all P ＜ 0.05].After immunosuppressive therapy,the percentages of CD3+ T lymphocyte ( 69.41 ± 5.03 ) %,CD4+ T lymphocyte (38.17 ±3.18)% and the ratio of CD4+/CD8+ 1.60 ±0.15 recovered to control levels.The percentages of CD8+ T lymphocyte (29.20 ±4.50)% and CD19+B lymphocyte ( 17.74 ±4.14)% were higher than those in healthy controls[( 24.82 ± 2.93 ) % and ( 12.09 ± 3.51 ) %,all P ＜ 0.05].After the immunosuppressive therapy,the percentages of CD8+ T lymphocyte ( 24.06 ± 3.02 ) % and CD19+ B lymphocyte ( 10.90 ± 3.55 ) %recovered to control levels.There were no significant difference of the percentage of CD56+ lymphocyte among ITP patients ( 15.80 ± 2.85 )%,ITP patients after immunosuppressive therapy ( 15.16 ± 2.77 )% and healthy controls ( 16.36 ± 2.75 ) %.ConclusionThe aberrant immunocyte subsets are involved in the pathogenesis of ITP,and detection of immunocyte subsets might be helpful for the diagnosis and determination of therapeutic outcome of ITP.     

2型糖尿病患者血小板-白细胞聚集体与微血管病变部位及区域数量的关系

目的探讨T2DM患者不同部位及区域数量微血管病变外周血血小板-白细胞聚集体(PLA)水平的改变。方法检测T2DM患者外周血中血小板-中性粒细胞聚集体(PNA)及血小板-单核细胞聚集体(PMA),按照微血管病变部位,比较DR、糖尿病慢性肾脏疾病(CKD)和糖尿病神经病变(DN)患者以及不同损伤区域数量微血管病变患者PNA、PMA水平。结果糖尿病微血管病变患者PNA、PMA水平高于无血管病变患者和健康对照者(P0.01)。DR、CKD及DN患者PNA、PMA水平比较差异无统计学意义。合并3种微血管病变患者PNA、PMA水平高于合并1种微血管病变患者(P0.01),合并2种微血管病变患者PMA水平高于合并1种微血管病变患者(P0.05)。结论糖尿病微血管病变患者PLA水平升高,PLA水平与微血管病变部位无关,与微血管病变损伤区域数量有关,PLA水平与糖尿病微血管病变的广泛性一致。     

淋巴细胞亚群在成人特发性血小板减少性紫癜中的变化及临床意义

目的:探讨淋巴细胞亚群与成人特发性血小板减少性紫癜(ITP)的关系及临床意义。方法:采集52例成年ITP患者和21例正常对照者外周血标本,分离单个核细胞后,应用流式细胞术检测淋巴细胞亚群,并将ITP患者与对照组比较。结果:ITP组较对照组CD3、CD4降低,CD8升高,CD4/CD8降低,CD19升高,CD16+56降低,2组间差异有统计学意义(P<0.05或P<0.01)。结论:ITP的发病机制与细胞免疫和体液免疫均密切相关,纠正患者免疫紊乱有益于ITP的治疗,外周血淋巴细胞亚群变化对ITP的疗效评价及预后判断也有一定价值。     

Monocyte subsets and monocyte-platelet aggregates in patients with unstable angina

Monocyte subsets and monocyte-platelet aggregates (MPAs) play important role in atherosclerosis and thrombosis. We aimed to determine their changes in patients with unstable angina (UA). In this cross-sectional case–control study, Global Registry of Acute Coronary Events (GRACE) score was determined in 95 UA patients without elevated troponin level. Thirty age-and-sex matched stable coronary heart disease (CHD) subjects served as control group. The classical (CD14++CD16?, Mon1), the intermediate (CD14++CD16+, Mon2) and the non-classical (CD14+CD16++, Mon3) monocytes, as well as subset-specific MPAs, were measured by flow cytometry. Compared with stable CHD patients, UA patients had increased Mon2 and Mon3 counts (all P < 0.001). For UA subjects, compared with GRACE score-determined low risk patients (GRACE score ≤108, n = 70), intermediate-to-high risk patients (GRACE score >108, n = 25) had higher counts of Mon2 and total MPAs, as well as Mon1- and Mon2-associated MPAs (all P < 0.001). Adjusted binary logistic regression analysis revealed that increased counts of Mon2 subset (for per 5 cells/μL increase, OR 1.186, 95 % CI 1.044–1.347, P = 0.009), Mon2 MPAs (for per 5 cells/μL increase, OR 1.228, 95 % CI 1.062–1.421, P = 0.006) and total MPAs (for per 5 cells/μL increase, OR 1.072, 95 % CI 1.010–1.137, P = 0.022) independently associated with GRACE score-determined intermediate-to-high risk UA patients. In UA patients with intermediate-to-high risk (determined by GRACE score), counts of Mon2 subset, Mon2-associated MPAs and total MPAs are increased, which are independent of traditional risk factors.     

A sensitive flow cytometric assay for circulating platelet–leucocyte aggregates

A whole blood flow cytometric method for studies of platelet–leucocyte aggregates (PLAs) in vivo , involving neither fixation nor centrifugation, is described. With this method, PLAs in the total leucocyte population (PLA/L) were 15.3 ± 8.5% in 36 healthy volunteers. Blocking antibodies had little effect on PLAs in the absence of in vitro stimulation, suggesting that the aggregates were preformed in vivo . Fixation with formaldehyde or paraformaldehyde increased PLA/L significantly. Similarly, prefixation of the blood or red blood cell lysis, with repeated washing and centrifugation, caused artefactual 3–5-fold increases in PLAs ( P  < 0.05). ADP, thrombin, platelet activating factor (PAF) or N-formyl-methionyl-leucyl-phenylalanine (fMLP) each increased PLA formation in unfixed whole blood dose-dependently; additive effects were found when they were combined. Experiments with blocking MAbs suggested that different ligand–receptor systems mediate PLA formation by different agonists. PLA formation by the platelet agonist ADP was inhibited by P-selectin blockade, but enhanced by GPIIb/IIIa blockade (which inhibits platelet–platelet interactions). PLA formation by the leucocyte agonist fMLP was inhibited by GPIIb/IIIa blockade, suggesting linking via fibrinogen. Platelet–leucocyte aggregate analysis by this whole blood method appears to reflect in vivo conditions, and enables investigations of the mechanisms involved in their formation.     

Two-dimensional flow cytometric analysis of intrahepatic lymphocyte subsets from patients with chronic hepatitis

Peripheral and intrahepatic lymphocyte subsets were analyzed in 22 patients with chronic hepatitis by two-dimensional flow cytometry. Activated T cells in the liver significantly increased compared with those in the peripheral blood. Helper T cells increased, but the CD4+ cells decreased due to a marked decrease of suppressor inducer T cells. CD8+ cells increased due to a increase of both cytotoxic T and suppressor T cells. Fc-receptor-positive cells, which increased significantly, were not NK cells but Fc-receptor-bearing T cells. In comparison with immunohistochemical methods, flow cytometric analysis enables more objective quantitation and simpler two-color staining of intrahepatic lymphocytes. Our findings using this method suggest that activated T cells and helper T cells have important roles in hepatitis and that hepatocellular injury may be generated not only by cytotoxic T cells but also by Fc-receptor-bearing T cells.     

冠心病患者血浆中单核细胞趋化蛋白-1的意义

目的:观察冠心病患者血浆中单核细胞趋化蛋白-1(MCP-1)的浓度,探讨其诊断冠心病的意义及临床价值。方法:将122例患者分为非冠心病组(30例)、稳定型心绞痛组(32例)、不稳定型心绞痛组(27例)和急性心肌梗死组(33例),应用酶联免疫吸附法检测患者血浆中MCP-1、组织因子(TF)的浓度。结果:①血浆中MCP-1、TF的浓度均为:非冠心病组<稳定型心绞痛组<不稳定型心绞痛组<急性心肌梗死组(均P<0.05)。②血浆中MCP-1的浓度与患者的性别、吸烟、冠心病家族史、高血压和糖尿病、TG、HDL-C、尿素氮、肌酐无明显相关性(均P>0.05),与年龄、体质指数、TC、LDL-C、尿酸、心肌肌钙蛋白I、肌酸激酶同工酶、TF呈正相关性(均P<0.05)。③以血浆中MCP-1的浓度作为诊断急性冠状动脉综合征的指标,ROC曲线分析结果显示MCP-1的诊断价值较大,曲线下面积0.939(P<0.05),最佳临界值为320.5ng/L,其特异度和灵敏度分别是86.7%和88.0%。结论:血浆中MCP-1在判断冠状动脉粥样硬化斑块易损性方面有一定的临床意义,可能是冠心病的一个独立危险因素,可作为诊断急性冠状动脉综合征的理想指标。     

老年冠心病患者血小板-白细胞聚集体的表达及其意义

目的:探讨老年冠心病患者血小板-白细胞聚集体(PLA)的表达及其意义.方法:选择我院内科2010年1月至2012年12月期间收治的冠心病患者162例,分为稳定型心绞痛(SAP)组(92例),急性冠脉综合征(ACS)组(70例),选择同期非冠心病患者80例为正常对照组.比较3组间PLA、血小板-中性粒细胞聚集体(PNA)、血小板淋巴细胞聚集体(PLyA)和血小板-单核细胞聚集体(PMA)的水平,采用Spearman相关分析分析PLA与有关临床指标的相关性.结果:与正常对照组和SAP组比较,ACS组PLA[(2.23±1.20)％比(3.25±2.16)％比(4.36±2.65)％]、PMA[(15.3±7.6)％比(23.5±19.7)％比(32.4±19.6)％]、PNA[(1.85±0.89)％比(2.14±1.56)％比(3.64±4.65)％]和PLyA(1.38±0.78)％比(1.42±0.87)％比(2.06±1.28)％]水平显著升高(P均＜0.01),且SAP组PLA、PMA水平显著高于正常对照组(P＜0.01).Spearman相关性分析显示:冠心病患者PLA与血糖、甘油三酯呈正相关(r1=0.680,P1=0.030;r2=0.738,P2=0.015),与高密度脂蛋白胆固醇呈负相关(r=-0.722,P=0.018).结论:PLA、PMA水平升高提示可能有老年冠心病,甚至是急性冠脉综合征.     

Reduction of monocyte-platelet interaction and monocyte activation in patients receiving antiplatelet therapy after coronary stent implantation

BACKGROUND: Monocyte activation induces different procoagulant and proadhesiveinflammatory responses and thus may play a role in thromboticcomplications after coronary interventions. Monocyte-plateletinteraction may trigger these effects inducing monocyte activation. AIMS: To characterize the effect of antiplatelet vs anticoagulationtherapy on monocyte-platelet interaction and monocyte functionafter intracoronary stenting. METHODS AND RESULTS: Immediately before, and during the first 12 days after successfulcoronary stenting, monocyteplatelet conjugates and monocytefunction were assessed by flow cytometric detection of GPIIb/IIIa(CD41) on monocytes and by monocyte surface exposure of Mac-1(CD11b/ CD18) and L-selectin (CD62L). Twenty patients receivingcombined antiplatelet therapy (ticlopidine, aspirin) were comparedto 20 patients with standard anticoagulation (phenprocoumon,overlapping heparin, aspirin). Before stenting, monocyte-plateletconjugates and Mac-1 surface expression in both groups weresignificantly increased, while L-selectin was significantlydiminished. Anticoagulation did not change these variables significantlyduring the subsequent 12 days. In contrast, antiplatelet therapyreduced platelet-monocyte conjugates by 46±9·3%(mean±SEM, P=0·0019) within 4 days, which wasassociated with a decrease in Mac-1 expression (28±6·7%,P=0·0013) and an increase in L-selectin (56±15·0%,P=0·0061). CONCLUSION: After intracoronary stenting, combined antiplatelet therapy,but not anticoagulation, causes reduction of monocyte-plateletinteraction, which is associated with monocyte deactivation.This may contribute to a decreased risk for thrombotic events.     

Small high-density lipoprotein is associated with monocyte subsets in stable coronary artery disease

Objective: High-density lipoprotein (HDL) particles are heterogeneous in structure and function and the role of HDL subfractions in atherogenesis is not well understood. It has been suggested that small HDL may be dysfunctional in patients with coronary artery disease (CAD). Monocytes are considered to play a key role in atherosclerotic diseases. Circulating monocytes can be divided into three subtypes according to their surface expression of CD14 and CD16. Our aim was to examine whether monocyte subsets are associated with HDL subfractions in patients with atherosclerosis. Methods: We included 90 patients with angiographically stable CAD. Monocyte subsets were defined as classical monocytes (CD14++CD16-; CM), intermediate monocytes (CD14++CD16+; IM) and non-classical monocytes (CD14+CD16++; NCM). HDL subfractions were measured by electrophoresis on polyacrylamide gel. Results: Serum levels of small HDL correlated with circulating pro-inflammatory NCM and showed an inverse relationship to circulating CM independently from other lipid parameters, risk factors, inflammatory parameters or statin treatment regime, respectively. IM were not associated with small HDL. In particular, patients with small HDL levels in the highest tertile showed dramatically increased levels of NCM (14.7 ± 7% vs. 10.7 ± 5% and 10.8 ± 5%; p = 0.006) and a decreased proportion of CM (79.3 ± 7% vs. 83.7 ± 6% and 83.9 ± 6%; p = 0.004) compared to patients in the two lower tertiles. In contrast, intermediate HDL, large HDL and total HDL were not associated with monocyte subset distribution. Conclusion: Small HDL levels are associated with pro-inflammatory NCM and inversely correlated with CM. This may suggest that small HDL could have dysfunctional anti-inflammatory properties in patients with established CAD.     

心肌血流储备分数在老年冠心病患者介入治疗中的应用

目的探讨心肌血流储备分数在老年冠心病患者PCI中对指导冠状动脉病变是否需行支架置入治疗的作用。方法选择老年冠心病患者18例,冠状动脉造影证实冠状动脉狭窄>70%,试验组8例患者18处病变行血流储备分数测定,对心肌血流储备分数≤0.75的狭窄病变处置入药物涂层支架;对照组10例患者21处病变常规行PCI置入药物涂层支架。比较2组患者支架置入数量、造影剂用量、手术时间、出院6个月后心绞痛发生情况以及主要心脏不良事件的发生率。结果试验组支架置入数量、造影剂用量明显少于对照组[(1.5±0.5)个vs(2.1±0.6)个,(196±68)ml vs(262±53)ml,P<0.05]。结论通过测定冠状动脉病变血流储备分数可以减少支架置入的数量和造影剂的用量,对老年冠心病患者PCI起到了较好的指导作用。     

吸烟冠心病病人的冠脉造影分析

目的　初步探讨吸烟冠心病病人的冠脉造影血管形态特点。方法　选择 2 0 0 1年 7月至 2 0 0 3年 7月间我院冠脉造影确诊冠心病的病人 2 0 8例 (男 179例 ,女 2 9例 ,年龄 38～ 75岁 ) ,吸烟年限 5～ 4 8年 ,平均 (36 4 5± 7 18)年 ;对照组选取同期病人 2 0 8例从不吸烟的患者 ,按照性别、年龄以及是否有高血压、糖尿病等冠心病危险因素严格配对。结果　吸烟组 13 9%有冠脉瘤样扩张 ;而对照组仅有 1 9% (P<0 0 0 1)。另外 ,在行冠脉成形术 (PTCA)的患者中吸烟组有4 0例 (19 2 % )有PTCA相关血管PTCA后的慢血流现象。而对照组仅有 7例 (3 83% ) (P <0 0 0 1)。结论　吸烟患者冠脉病变特点为冠脉瘤样扩张或冠脉扩张症多发。吸烟患者PTCA术中相关血管慢血流现象的发生率较高。