凉血活血方对脓毒症大鼠白细胞系列黏附分子表达的影响

目的:研究凉血活血方对脓毒症大鼠白细胞黏附分子表达的影响,探讨其药理作用机制。方法:Wistar大鼠90只,雌雄各半,随机分为假手术组(10只)、模型组(20只)、凉血活血方治疗组(分为大,中,小剂量组,每组20只)。采用盲肠结扎穿孔法制作脓毒症模型,造模同时分别进行灌胃治疗,给予大,中,小剂量的活血凉血方浸膏,观察72h动物死亡率;在造模72h后腹主动脉取血抗凝,溶血素处理后,分别加入不同荧光素标记的CD11b、CD31、CD54、CD62L抗体,经流式细胞仪检测白细胞表面4种黏附分子的表达情况;行气管插管进行肺泡灌洗,收集分离白细胞,流式细胞术检测4种黏附分子的表达情况;留取肝、肺及末段回肠组织,检测髓过氧化物酶(MPO)活性。结果:模型组大鼠血中及肺泡灌洗液中白细胞黏附分子CD11b、CD31、CD54、CD62L表达较假手术组显著增高;大,中剂量凉血活血方治疗可显著降低模型大鼠血中及肺泡灌洗液中白细胞CD11b、CD31、CD54、CD62L表达,显著降低肝、肺及末段回肠组织MPO活性;凉血活血方大剂量组动物死亡率显著降低。结论:脓毒症状态下白细胞黏附因子的异常高表达及其与血管内皮的黏附作用可介导多种脏器功能障碍,活血凉血中药可通过抑制血管内皮系列黏附分子高表达而发挥治疗作用。     

Haematological abnormalities in a 75-year-old population. Consequences for health-related reference intervals

A representative sample (n = 486) of a 75-year-old population was studied, and probands with defined laboratory aberrations were re-investigated. Anaemia was present in 6% of the men and 3% of the women; in 17/22 anaemic subjects a cause was found. The prevalence of plasma cobalamin concentrations less than 130 pmol/l was 6%, of iron deficiency approximately 6%. Divergences in white blood cell and platelet counts were rare. The observed haematological aberrations were almost always caused by disease. Reference intervals for haematological components were calculated in the total study group and two reference sample groups after exclusions based on anamnestic and/or laboratory screening criteria or anamnestic criteria and/or verified disease. The lower reference limits for B-Hb and P-B12 in a group obtained after exclusions based on anamnestic and screening data were considered to be minimum values for healthy subjects. The WHO criteria for anaemia were applicable.     

Performance of Cell Counters in Primary Health Care

The results are presented from two external quality control surveys of cell counters in primary health care. The precision of haemoglobin measurements in the normal range was better than that obtained by conventional haemometers. The analytical quality of leucocyte analyses was about the same as that performed by hospital instruments. The precision of cell counters with respect to thrombocytes was poorer than that obtained on hospital instruments, especially in the lower level. Celldyn and Sysmex instruments performed best among the instruments examined. Reliable cell counter results were associated with medical laboratory technicians as operators of the instruments.To improve the analytical quality of the laboratory in primary health care, it appears necessary to establish an external quality assurance programme, including a laboratory consultancy service.     

Toxicity evaluation of monodisperse PEGylated magnetic nanoparticles for nanomedicine

Abstract

Innovative nanotechnology aims to develop particles that are small, monodisperse, smart, and do not cause unintentional side effects. Uniform magnetic Fe₃O₄ nanoparticles (12?nm in size) were prepared by thermal decomposition of iron(III) oleate. To make them colloidally stable and dispersible in water and cell culture medium, they were modified with phosphonic acid- (PA) and hydroxamic acid (HA)-terminated poly(ethylene glycol) yielding PA-PEG@Fe₃O₄ and HA-PEG@Fe₃O₄ nanoparticles; conventional γ-Fe₂O₃ particles were prepared as a control. Advanced techniques were used to evaluate the properties and safety of the particles. Completeness of the nanoparticle coating was tested by real-time polymerase chain reaction. Interaction of the particles with primary human peripheral blood cells, cellular uptake, cytotoxicity, and immunotoxicity were also investigated. Amount of internalized iron in peripheral blood mononuclear cells was 72, 38, and 25?pg Fe/cell for HA-PEG@Fe₃O₄, γ-Fe₂O₃, and PA-PEG@Fe₃O₄, respectively. Nanoparticles were localized within the cytoplasm and in the extracellular space. No cytotoxic effect of both PEGylated nanoparticles was observed (0.12–75?μg/cm²) after 24 and 72-h incubation. Moreover, no suppressive effect was found on the proliferative activity of T-lymphocytes and T-dependent B-cell response, phagocytic activity of monocytes and granulocytes, and respiratory burst of phagocytes. Similarly, no cytotoxic effect of γ-Fe₂O₃ particles was observed. However, they suppressed the proliferative activity of T-lymphocytes (75?μg/cm², 72?h) and also decreased the phagocytic activity of monocytes (15?μg/cm², 24?h; 3–75?μg/cm², 72?h). We thus show that newly developed particles have great potential especially in cancer diagnostics and therapy.     

尿沉渣Wright染色观察狼疮肾炎治疗前后白细胞尿的变化及其意义

目的对狼疮肾炎患者治疗前后进行尿沉渣Wright染色，探讨狼疮肾炎白细胞尿的变化及其临床意义。方法对76例尿中出现白细胞的狼疮肾炎患者，分别在狼疮治疗前后留取新鲜中段尿进行细菌定量培养，应用相差显微镜进行尿沉渣细胞学检查及1h尿白细胞计数，观察狼疮肾炎治疗前后的尿液中白细胞的变化。结果7例已婚狼疮肾炎患者清洁中段尿培养为大肠杆菌，其余69例尿细菌培养为阴性。经激素及CTX治疗以后狼疮肾炎病情缓解，尿中白细胞明显减少（P〈0．01）。结论狼疮肾炎活动期可以出现非感染性白细胞尿，尿中白细胞数的变化，可以作为观察狼疮肾炎疗效的一个客观指标。     

糖尿病视网膜病变粘附分子表达与白细胞粘附的干预性研究进展

糖尿病视网膜病变是糖尿病的重要微血管并发症之一,其发病机制复杂,尚未完全阐明。近年来研究发现白细胞和粘附分子在糖尿病性视网膜病变的病理发生过程中起重要作用。本研究就糖尿病时白细胞和粘附分子表达异常、对视网膜的损害及干预性药物治疗等方面进行了综述。     

Biochemical, genetic and ultrastructural study of a family with the sea-blue histiocyte syndrome/chronic non-neuronopathic Niemann- Pick disease

Abstract. Deficient leucocyte sphingomyelinase activity has been demonstrated in a patient with the sea-blue histiocyte syndrome. Family studies revealed that two other cases previously diagnosed on clinical and histo-chemical criteria also had a pronounced diminution of sphingomyelinase activity. Both parents of the affected individuals were carriers of the disease as indicated by sphingomyelinase activity intermediate between normal and diseased subjects. Additional heterozygous carriers were found among the siblings and other relatives of the patients. This family study supports further the hypothesis that the sea-blue histiocyte syndrome and chronic Niemann-Pick (Type B) disease are the same.     

Macrophage accumulation and renal fibrosis are independent of macrophage migration inhibitory factor in mouse obstructive nephropathy

BACKGROUND AND AIM: The progression of renal injury, initiated by either an immune or non-immune insult, is closely associated with the accumulation of leucocytes and fibroblasts in the damaged kidney. Macrophage migration inhibitory factor (MIF) regulates leucocyte activation and fibroblast proliferation in vitro. Studies have identified a pathological role for MIF in immune-initiated renal injury in the rat. In this study, we examined the role of MIF in obstructive nephropathy, where renal injury is initiated by a non-immune insult. METHODS AND RESULTS: Unilateral ureteric ligation was performed on MIF wildtype (+/+) and MIF deficient (-/-) mice. Groups of five mice were killed at days 0, 1, 5 or 10 after obstruction, and kidneys were examined via immunohistochemistry and northern blotting. In MIF +/+ mice, expression of the MIF protein increased in obstructed kidneys compared to normal control kidneys. Interstitial macrophage and T cell accumulation was significantly increased in obstructed kidneys at day 5 and 10, but was unaffected by MIF deficiency. Osteopontin and macrophage colony stimulating factor (M-CSF) mRNA expression in obstructed kidneys were equally increased in both genotypes, indicating that expression of these chemokines is not influenced by MIF. No difference was detected in the development of renal fibrosis in obstructed MIF +/+ and MIF -/- kidneys, as assessed by myofibroblast accumulation and proliferation and expression of profibrotic molecules (transforming growth factor-beta 1(TGF-beta1) and collagen). CONCLUSION: These results demonstrate that MIF expression is increased in obstructive nephropathy without affecting kidney leucocyte accumulation or the development of renal fibrosis. This suggests that the progression of renal injury in obstructive nephropathy is independent of MIF.     

Retinol binding protein isolated from acute renal failure patients inhibits polymorphonuclear leucocyte functions

BACKGROUND: Protein factors accumulating in sera of patients with end-stage renal disease (ESRD) that interfere with the nonspecific immune response by inhibiting essential functions of polymorphonuclear leucocytes (PMNLs) have previously been described. No such factor has been isolated from acute renal failure (ARF) patients to date. MATERIALS AND METHODS: Using a three-step chromatographic procedure involving ion exchange, size exclusion and hydrophobic interaction chromatography we purified the apo- and holo-form of retinol binding protein (RBP) from high-flux dialyser (polyacrylonitrile; AN69) ultrafiltrates of patients with ARF. Their effect on the chemotaxis of PMNLs isolated from healthy donors was determined by the under-agarose method. Whole-blood assays applying flow cytometry were used to assess phagocytosis and the oxidative metabolism of PMNLs. Apoptosis was assessed by determining the DNA content using propidium iodide. RESULTS: Isolated apo- and holo-forms of RBP were truncated on their C-terminus as determined by mass spectrometry. All isolates significantly inhibited the chemotactic movement of PMNLs obtained from healthy donors and the PMNL oxidative metabolism stimulated by E. coli. These effects were concentration dependent. Retinol binding protein had no influence on the PMNL oxidative metabolism stimulated by PMA and on PMNL phagocytosis. Commercially available RBP isolated from urine influenced PMNL functions in the same way. Inhibition of p38 mitogen-activated protein kinase (MAPK) by SB203580 significantly attenuated the phagocytosis-induced respiratory burst and RBP did not lead to a further decrease. Polymorphonuclear leucocyte apoptosis was significantly inhibited by RBP. CONCLUSIONS: The apo- and holo-forms of RBP isolated from the ultrafiltrate of ARF patients inhibit PMNL chemotaxis, oxidative metabolism and apoptosis. Therefore, RBP may be considered a uraemic toxin contributing to a disturbed immune defence.     

Cellular and biochemical markers in semen indicating male accessory gland inflammation

Leucocytospermia is considered to be a sign of male accessory gland inflammation. The leucocytes in semen are mainly polymorphonuclear neutrophilic granulocytes. Leucocytospermia is not associated with the presence of bacteria and antibiotic treatment does not significantly lower the extent of leucocytospermia. A higher frequency of elevated herpes simplex antibodies titres were found in men with leucocytospermia. The concentration of inflammatory cytokines, interleukin-6 and -8, is closely correlated with the number of leucocytes. Their determination does not provide additional information. Reactive oxygen species (ROS) are generated at least in part by seminal leucocytes in response to stimulating factors. Purified leucocytes produce high levels of ROS. The determination of ROS appears to represent a parameter of functional activity of leucocytes. The role of chlamydiae in male accessory gland infection is unclear. Their determination in semen by DNA amplification and by immunological tests does not provide reliable results.