海军陆战队员高温强训中注射丹参液前后血液流变学与凝血功能的变化

目的:观察高温高湿环境下负荷30kg野外强训时,注射丹参液前后海军陆战队员血液流变学和凝血系统的变化。方法:于2005-08在南海海面和某岛屿上选择男性解放军海军陆战队员112名,经医院全面检查身体合格,18～32岁,军龄1～14年。成建制的随机抽取55名作为训练组,负重30kg参加高温高湿环境下各种强训,另外57名陆勤人员作为对照组。训练组注射丹参液前后高温强训各8h,训练末各取1次血标本,检查两组队员凝血功能5项指标及血液流变学14项指标。结果:112名海军陆战队员均进入结果分析。①未注射丹参液前训练组全血低切黏度、全血中切黏度、全血高切黏度、血浆黏度、红细胞比积、红细胞沉降率、全血低切还原黏度、全血中切还原黏度、全血高切还原黏度、血沉方程K值、红细胞聚集指数、红细胞刚性指数、红细胞变性指数、红细胞电泳指数均高于对照组[训练组:(9.4±0.3)mPa.s,(5.6±0.4)mPa.s,(4.7±0.3)mPa.s,(1.5±0.2)mPa.s,(47.4±3.1)%,(7.3±1.1)mm/h,(19.4±3.8)mPa,s,(9.7±1.5)mPa.s,(8.0±1.5)mPa.s,(61.4±10.5),(2.7±0.2),(5.5±1.0),(0.8±0.1),(6.4±1.3);对照组:(8.1±0.5)mPa.s,(4.7±0.3)mPa.s,(4.1±0.3)mPa.s,(1.3±0.2)mPa.s,(44.8±3.3)%,(8.5±1.4)mm/h,(15.5±2.4)mPa.s,(8.2±1.5)mPa.s,(7.4±1.6)mPa.s,(51.8±9.7),(2.4±0.4),(4.5±0.8),(0.7±0.3),(5.3±1.1),P<0.05,P<0.01]。注射丹参液后训练组全血低切黏度、红细胞比积高于对照组[(8.7±0.3)mPa.s,(46.5±3.1)%;(8.1±0.5)mPa.s,(44.8±3.3)%,P<0.05]。②注射丹参液前训练组凝血酶原时间、部分活化凝血酶原时间、纤维蛋白原、D-二聚体、血清纤维蛋白降解产物、血小板与对照组相比,差异显著[训练组:(10.2±0.2)s,(23.9±1.1)s,(3.8±0.2)g/L,(309.6±13.3)mg/L,(7.7±0.4)mg/L,(270.1±14.2)×109L-1;对照组:(11.6±0.3)s,(28.4±1.4)s,(2.9±0.4)g/L,(241.5±13.7)mg/L,(5.9±1.2)mg/L,(174.5±22.0)×109L-1,P<0.01]。注射丹参液后训练组血小板、D-二聚体、纤维蛋白降解产物高于对照组[训练组:(236.2±11.2)×109L-1,(279.3±13.8)mg/L,(8.2±0.6)mg/L;对照组:(174.5±22.0)×109L-1,(241.5±13.7)mg/L,(5.9±1.2)mg/L,P<0.05,P<0.01],凝血酶原时间、部分凝血活酶时间、纤维蛋白原与对照组相比无差异。结论:注射丹参液可增强红细胞的柔软性,改善红细胞变形和聚集能力,降低纤维蛋白原含量,防止血液高黏发生。     

左旋精氨酸与褪黑激素对大鼠肾脏缺血再灌注损伤的保护作用

目的:诱导型一氧化氮合酶来源的一氧化氮与内皮型一氧化氮合酶来源的一氧化氮的平衡对维持血流动力学稳定及内环境动态平衡有重要作用.观察左旋精氨酸与褪黑激素对肾脏缺血再灌注损伤中诱导型一氧化氮合酶与内皮型一氧化氮合酶的影响.方法:实验于2006-07/2007-03在中山大学医学院动物实验中心完成.①实验分组:健康雄性清洁级SD大鼠60只,体质量170～210 g,随机数字表法分为缺血再灌注组、左旋精氨酸治疗组、褪黑激素治疗组、联合治疗组,每组15只.②实验方法:建立大鼠肾脏缺血再灌注损伤模型,术前1 h各组大鼠腹腔内注射生理盐水及相应药物,每隔24 h重复注射1次,连续5d.③实验评估:观察各组术前及术后1,3,5 d血肌酐、诱导型一氧化氮合酶和内皮型一氧化氮合酶的变化规律及肾脏病理组织学的改变,并采用Paller法对术后第3天肾小管进行评分.结果:纳入大鼠60只,均进入结果分析.①血肌酐的表达:术后第1天联合治疗组水平显著低于缺血再灌注组(P＜0.05);术后第3,5天左旋精氨酸治疗组、褪黑激素治疗组与联合治疗组显著低于缺血再灌注组(P＜0.05).②内皮型一氧化氮合酶的表达:术后第3天缺血再灌注组比术前降低(P＜0.05);术后第3天和第5天左旋精氨酸治疗组、联合治疗组高于缺血再灌注组(P＜0.05).③诱导型一氧化氮合酶的表达:术后第1天缺血再灌注组与术前相比升高,第3天达到高峰,第5天仍高于术前(P＜0.05);褪黑激素治疗组、联合治疗组术后第3天与第5天明显低于缺血再灌注组(P＜0.05).④血肌酐水平与诱导型一氧化氮合酶的表达呈正相关,r=0.57,P＜0.01,与内皮型一氧化氮合酶/诱导型一氧化氮合酶呈负相关,r=-0.61,P＜0.01.⑤肾小管损伤Palller法评分:依次为联合治疗组＜褪黑激素治疗组＜左旋精氨酸治疗组＜缺血再灌注组.结论:通过调高内皮型一氧化氮合酶的表达、抑制诱导型一氧化氮合酶的表达可以减轻肾脏缺血再灌注损伤,联合应用左型精氨酸与褪黑激素即通过以上途径更有效的起到对肾脏缺血再灌注损伤的保护作用.     

Relative value of ZAP-70, CD38, and immunoglobulin mutation status in predicting aggressive disease in chronic lymphocytic leukemia

Leukemia-cell expression of ZAP-70, CD38, or unmutated immunoglobulin heavy chain variable region genes (U-IGHV) each is associated with aggressive disease in patients with chronic lymphocytic leukemia (CLL). To assess the relative strength of each marker, we defined thresholds for designating a case as positive for CD38 or ZAP-70 in a test cohort of 307 patients and used these data-defined criteria to stratify patients in an independent cohort of 705 patients. Multivariable analysis revealed that ZAP-70 was the strongest risk factor. Knowledge of the IGHV mutation status or CD38 did not improve our ability to predict the time to first treatment except for ZAP-70-negative cases, which could be segregated into 2 groups of intermediate-risk or low-risk disease based on whether they expressed unmutated or mutated IGHV. ZAP-70 maintained its high relative prognostic value for the subset of patients with early-stage, asymptomatic disease, including patients evaluated within 1 year of diagnosis. Although it is premature to recommend therapy based on these risk factors, patients with ZAP-70-positive CLL cells should be monitored closely for disease progression as they have a median time from diagnosis to requiring initial therapy by standard criteria of approximately 3 years.     

Analysis of CLLU1 expression levels before and after therapy in patients with chronic lymphocytic leukemia

Objective: Chronic lymphocytic leukemia (CLL) is incurable, but therapy leading to eradication of minimal residual disease (MRD) in CLL is associated with improved clinical outcomes. CLL upregulated gene 1 (CLLU1) is solely upregulated in CLL patient samples. We hypothesized that CLLU1 could be used to monitor for residual disease in CLL patient samples after therapy. Methods: We examined whether the CLLU1 real‐time quantitative PCR (RQ‐PCR) could detect small numbers of CLL cells in mixtures of normal peripheral blood mononuclear (PBMC) cells. We then performed a retrospective analysis on time‐matched cryo‐preserved specimens from patients who achieved MRD‐negative remissions that underwent serial marrow biopsies for evaluation of residual disease by 4‐color flow cytometry. RNA from PBMC samples collected at the time of the marrow assessments was analyzed for CLLU1. Nine patients underwent a total of 46 paired blood and marrow evaluations (median 5 assessments per patient). Results: CLLU1 RQ‐PCR on PBMCs of healthy donors reconstituted with varying amounts of CLL cells demonstrated leukemia cells could be reliably detected with high sensitivities depending on the CLLU1 expression level. Analysis of time‐matched samples assessed for CLLU1 levels in the blood by RQ‐PCR and residual disease of the marrow determined by 4‐color flow cytometry revealed a correlation coefficient of 0.96 (P < 0.0001). Conclusion: The CLLU1 RQ‐PCR is a sensitive and specific assay for detecting residual CLL cells after therapy. Assessment of blood CLLU1 levels can be used as a reliable marker of tumor burden and has the potential to complement currently used techniques for MRD monitoring in patients with CLL.     

miR-181b is a biomarker of disease progression in chronic lymphocytic leukemia

MicroRNAs play a crucial role in chronic lymphocytic leukemia. We investigated whether microRNAs can discriminate patients with a progressive disease from patients with a stable disease. We analyzed microRNA expression on leukemic cells isolated from 358 sequential samples of 114 patients with either stable or progressive disease. We found that during the course of the disease the expression values of miR-181b, the most dysregulated microRNA, decreased in samples of patients with a progressive (P < .001, training and validation sets) but not in samples of patients with a stable disease (P = .3, training set; P = .2, validation set) over time. A drop of ≥ 50% between sequential samples and/or a miR-181b value ≤ 0.005 at the starting time point were significant to differentiate progressive from stable disease (P = .004, training set; P < .001, validation set). These parameters were associated with high risk of requiring treatment (risk ratio, 5.8; 95% confidence interval, 2.5-14.9). We also observed that miR-181b targets Mcl-1 protein and that the decrease of its expression inversely correlated with increased protein levels of MCL1 and BCL2 target genes. We conclude that parameters defined on the basis of the miR-181b expression values specify disease progression in chronic lymphocytic leukemia and are associated with clinical outcome.     

ZAP-70 enhances IgM signaling independent of its kinase activity in chronic lymphocytic leukemia

We transduced chronic lymphocytic leukemia (CLL) cells lacking ZAP-70 with vectors encoding ZAP-70 or various mutant forms of ZAP-70 and monitored the response of transduced CLL cells to treatment with F(ab)(2) anti-IgM (anti-mu). CLL cells made to express ZAP-70, a kinase-defective ZAP-70 (ZAP-70-KA(369)), or a ZAP-70 unable to bind c-Cbl (ZAP-YF(292)) experienced greater intracellular calcium flux and had greater increases in the levels of phosphorylated p72(Syk), B-cell linker protein (BLNK), and phospholipase C-gamma, and greater activation of the Ig accessory molecule CD79b in response to treatment with anti-mu than did mock-transfected CLL cells lacking ZAP-70. Transfection of CLL cells with vectors encoding truncated forms of ZAP-70 revealed that the SH2 domain, but not the SH1 domain, was necessary to enhance intracellular calcium flux in response to treatment with anti-mu. We conclude that ZAP-70 most likely acts as an adapter protein that facilitates B-cell receptor (BCR) signaling in CLL cells independent of its tyrosine kinase activity or its ability to interact with c-Cbl.     

葛根素对大鼠成骨细胞增殖和分化的影响

目的:课题以往的研究已证实,葛根素在体外能够抑制破骨细胞的骨吸收活性,促进骨生长,实验将进一步验证葛根素对大鼠成骨细胞增殖和分化的影响,并评价其对骨形成的作用。方法:实验于2003-03/2004-03在武装警察部队医学院药物化学教研室药物活性筛选室进行。①实验材料:葛根素(Puerarin)购自中国药品生物制品检定所,雌酚酮购自浙江仙居制药厂。大鼠由解放军军事医学科学院实验动物中心提供。②实验方法:实验分5组:雌酚酮阳性对照组、空白对照组(不加药,只加等量细胞悬液和培养基)、0.01,0.1,1μmol/L葛根素组。由新生24hSD大鼠颅盖骨分离培养成骨细胞,调整成骨活细胞密度为0.9×107L-1,接种于24孔培养板,每组12个复孔,分别于第1,3,5天每组取3孔计数,绘制细胞生长曲线。③实验评估:分别于第1,3,5天采用四唑盐法测定吸光度,计算平均增殖率,采用硝基苯基质动力学法测定各组细胞内外碱性磷酸酶活性。结果:①与空白对照组相比,0.1,1μmol/L葛根素干预组与雌酚酮阳性对照组第3,5天细胞增殖速度均加快,差异有显著性意义(P<0.01)。②经1μmol/L葛根素处理的第1,3,5天,细胞内、外碱性磷酸酶活性均明显高于空白对照组(P<0.01)。经0.1μmol/L葛根素处理第3,5天,细胞内、外碱性磷酸酶活性明显高于空白对照组(P<0.05,P<0.01)。经0.01μmol/L葛根素处理后第5天,细胞外碱性磷酸酶活性明显高于空白对照组(P<0.01)。结论:葛根素对骨形成的作用:①通过影响碱磷酸酶活性,而促进成骨细胞分化。②通过雌激素受体介导促进成骨细胞的骨形成效应。     

服刑人员132例人格障碍筛查问卷调查

目的:了解人格障碍筛查问卷的初步诊断情况。方法:于2005-09/10选择广东省三水劳教所的服刑人员154人为调查对象,将人格障碍筛查问卷的软件中的106个检查条目制成“是”或“否”问卷进行调查。结果:发放问卷154份,回收问卷154份,有效问卷132份。132名服刑人员中未提示任何人格障碍和品行障碍的有10名(7.58%)。73名提示18岁前有品行障碍,提示有人格障碍的116名,99名存在2项以上人格障碍。结论:问卷结果提示的人格障碍和品行障碍有可能存在诊断过度的问题。对调查阳性者进行人格障碍临床半定式检测工具测试是下一步要进行的工作。     

An anti-B cell autoantibody from Wiskott-Aldrich syndrome which recognizes i blood group specificity on normal human B cells.

We previously identified IgM autoantibodies in the sera of patients with Wiskott-Aldrich syndrome (WAS) that react with a subset of normal human B lymphocytes and induce B cell differentiation in vitro. From splenocytes of a patient with WAS we generated heterohybridomas (HY18 and HY21) and a lymphoblastoid cell line (LWA10) that produce human IgM lambda or IgM kappa anti-B lymphocyte autoantibodies, respectively. Immunohistochemical and multiparameter flow cytometric analyses demonstrate that these autoantibodies are specific for lymphocytes of the B lineage and preferentially stain B cells that reside in the mantle zone of secondary follicles and that constitutively co-express the CD5 surface antigen and most major autoantibody-associated cross-reactive idiotypes; in addition, these antibodies stain most pre-B cells in adult bone marrow. Molecular studies show that these anti-B lymphocyte autoantibodies are encoded by a highly conserved VH4 gene, designated VH4.21. The gene encodes a number of autoantibodies, especially anti-i and anti-I IgM cold agglutinins. Hemagglutination and surface labeling studies reveal that HY18 and LWA10 recognize the "i" carbohydrate antigenic determinant(s) which is classically found on human cord red blood cells and, as shown now by this study, on a subpopulation of human B cells which expresses it early in B cell development. These studies raise the possibility that the gene product encoded by this highly conserved germ-line VH4 gene may play a physiological role in B cell development and/or differentiation.