Influence of antibody and complement components on phagocytosis and chemiluminescence of macrophages

Macrophages are known to release reactive oxygen species (O₂^?, ¹O₂, H₂O₂, OH·) in response to various membrane stimuliHowever, our studies show that phagocytic stimulation of macrophages is not necessarily accompanied by a stimulation of the oxidative burstWhereas IgG-opsonized erythrocytes were capable to induce phagocytosis and a chemiluminescence response, both being dependent on the number of IgG bound per erythrocyte, C3b-bearing erythrocytes were well ingested but failed to induce any chemiluminescence reactionFurthermore, stimulation of macrophages, via the Fc-receptors, seems to alter their functional state in regard to the activation of a receptor, which enables them to recognize membrane lesions on the target erythrocyteThe presence of IgG and membrane lesions, e.gthe C5b-9-complex of complement, induced a marked increase in chemiluminescence compared with stimulation by IgG-bearing particles aloneThe augmented response of macrophages was at least in part due to an additional release of H₂O₂, which was not liberated in response to IgG-bearing erythrocytesThis «Alesion recognizing receptor» in the macrophage membrane could not be activated by stimulation of C3b-receptors, indicating its functional linkage to the Fc-receptors.     

UriSed尿沉渣分析仪的临床应用评估

目的对UriSed尿沉渣分析仪的临床应用进行评估。方法随机收集患者新鲜中段尿液标本,使用UriSed尿沉渣分析仪对尿液有形成分进行分析,评价分析仪的图像合格率、标记率、标记正确率、重复性、线性、互染率,并比较其与UF500尿沉渣分析仪对低值细胞的检出符合率。结果图像合格率为98.23%。有形成分标记率为:病理管型(100%)、白细胞(82.5%)、红细胞(74.7%)、小圆上皮细胞(73.6%)、上皮细胞(64.1%)、真菌(54.1%)。有形成分的标记正确率为:小圆上皮细胞(100%)、白细胞(99.9%)、上皮细胞(99.5%)、真菌(98.1%)、红细胞(97.4%)、病理管型(1.5%)。高、中、低水平白细胞尿标本的变异系数依次为:4.68%、3.45%、4.48%。分析仪对低值白细胞的具有良好的线性关系,但对低值红细胞的检测线性欠佳。测定白细胞的互染率为0。UriSed与UF500对100/L白细胞的检出符合率为97.4%,对100/L红细胞的检出符合率为96.8%。结论除对低值红细胞检测的线性关系不佳之外,UriSed尿沉渣分析仪主要指标能满足临床应用要求。     

Flow‐cytometric analysis of erythrocytes and reticulocytes in congenital dyserythropoietic anaemia type II (CDA II): value in differential diagnosis with hereditary spherocytosis

Congenital dyserythropoietic anaemia type II (CDA II) is the most common congenital dyserythropoietic anaemia. CDA II is frequently misdiagnosed as Hereditary Spherocytosis (HS) due to the presence of mild chronic haemolytic anaemia with splenomegaly, increased osmotic fragility, and presence of microspherocytes. Accurate diagnosis of CDA II is important to prevent severe iron overload. Erythrocyte and reticulocyte indices were assessed in 10 patients from six families with CDA II, 18 patients from eight families with HS, and 50 normal controls. Characteristic increases in distribution width were present in CDA II for cell volume (RDW, anisocytosis) and in HS for cell haemoglobin concentration (HDW, anisochromia), resulting in an RDW/HDW ratio which was significantly greater in CDA than HS (P < 0.0002). A cut‐off value for RDW/HDW of 5.34 resulted in 89% sensitivity and 70% specificity in distinguishing CDA II from HS. Distribution width for cell haemoglobin content of reticulocytes (CHDWr) was characteristically increased in CDA II, resulting in a CHDW/CHDWr ratio significantly lower in CDA II than HS (P < 0.0002). A cut‐off value of 0.98 provided 89% sensitivity and 80% specificity in distinguishing CDA II from HS. These differences in distribution widths of flow‐cytometric parameters of reticulocytes and mature erythrocytes reflect the different pathogeneses of the two diseases and are helpful for the differential diagnosis of these two conditions.     

干化学法与沉渣镜检法对比检测尿中红细胞和白细胞

目的探讨迪瑞H-300尿液分析仪干化学法与尿沉渣镜法检测红细胞和白细胞的一致性。方法对3678例尿液分析仪干化学测定全部为阴性、619例测定为隐血±至＋＋同时白细胞阴性的尿液标本做尿沉渣镜检。结果3678例阴性标本尿沉渣镜检检出白细胞每高倍视野5～30个以上206例,假阴性率占5．6％;红细胞每高倍视野3～15个115例,假阴性率为3．1％。在619例隐血阳性同时白细胞阴性的尿标本中,红细胞假阳性率为3．1％,白细胞假阴性率为5．5％。结论临床上使用干化学法做尿常规出现全阴性时,为了避免漏、误诊,必须进行显微镜沉渣检查。     

缺氧缺血性脑病患儿血常规结果分析及其临床意义

目的探讨27项血常规参数在新生儿缺氧缺血性脑病（HIE）中的变化及其意义。方法利用全自动血细胞分析仪对29例HIE患儿和30例健康新生儿进行白细胞、红细胞及血小板等参数检测,分析其结果和临床意义。结果H1E组与对照组比较,（1）白细胞参数：淋巴细胞比率、单核细胞比率、中性粒细胞比率差异有统计学意义（P〈0．05）,而白细胞计数、嗜酸性粒细胞比率、嗜碱性粒细胞比率差异无统计学意义（P〉0．05）;（2）红细胞参数：血细胞比容、红细胞平均体积、平均血红蛋白浓度（MCHC）、红细胞分布宽度CV、红细胞分布宽度SD、网织红细胞计数、网织红细胞百分比、未成熟网织红细胞、低荧光网织红细胞比率、高荧光网织红细胞比率、有核红细胞计数、有核红细胞百分比等12项红细胞参数差异有统计学意义（P〈0．05）,而红细胞计数、血红蛋白、平均血红蛋白含量、中荧光强度网织红细胞比率等参数差异无统计学意义（P〉0．05）;（3）血小板参数：血小板计数、血小’板：压积差异有统计学意义（P〈0．05）,而平均血小板体积、血小板分布宽度及大型血小板比率等参数差异无统计学意义（P〉0．05）。结论利用全自动血细胞分析仪对HIE患儿外周血进行常规筛查和动态监测,计数快速、准确,能很好地满足临床应用,协助临床诊断HIE。     

Blood micronutrient, oxidative stress, and viral load in patients with chronic hepatitis C

AIM: To assess the extent of micronutrient and oxidative stress in blood and to examine their linkages with viral loads in chronic hepatitis C patients. METHODS: Hepatitis C virus (HCV)-RNA levels were quantified in the serum from 37 previously untreated patients with chronic hepatitis C. The plasma and erythrocyte micronutrients (zinc, selenium, copper, and iron) were estimated, and malondialdehyde (MDA) contents were determined as a marker to detect oxidative stress. Antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione reductase (GR) activities in blood were also measured. The control group contained 31 healthy volunteers. RESULTS: The contents of zinc (Zn), and selenium (Se) in plasma and erythrocytes were significantly lower in hepatitis C patients than in the controls. On the contrary, copper (Cu) levels were significantly higher. Furthermore, plasma and erythrocyte MDA levels, and the SOD and GR activities in erythrocytes significantly increased in hepatitis C patients compared to the controls. However, the plasma GPX activity in patients was markedly lower. Plasma Se (R=-0.730, P<0.05), Cu (r=0.635), and GPX (r= -0.675) demonstrated correlations with HCV-RNA loads. Significant correlation coefficients were also observed between HCV-RNA levels and erythrocyte Zn (r=-0.403), Se (r = -0.544), Cu (r= 0.701) and MDA (P=0.629) and GR (r =0.441). CONCLUSION: The levels of Zn, Se, Cu, and oxidative stress (MDA), as well as related anti-oxidative enzymes (GR and GPX) in blood have important impact on the viral factors in chronic hepatitis C. The distribution of these parameters might be significant biomarkers for HCV.     

长春新碱载体红细胞体内代谢分布的初步研究

目的：初步探讨长春新碱载体红细胞在小鼠体内的分布代谢情况。方法i昆明种小鼠腋窝皮下接种S180肉瘤细胞建立荷瘤小鼠模型,将荷瘤小鼠随机分为长春新碱、长春新碱载药红细胞2组,经尾静脉分别注射长春新碱200ug及长春新碱载药红细胞（浓度1cg／L）。注射后0、1、2、3、4、24、48及72h取小鼠血液、肝脏及肿瘤组织,用高效液相色谱法测定其中药物浓度,计算半衰期。结果：长春新碱进入体内后血浆浓度迅速增高,其代谢速率也很快,48h后完全测不出,半衰期1．53h。长春新碱载药红细胞在血浆中浓度稳定而持久,72h仍可测出,药物半衰期达4．1h,是前者的2．68倍。载药红细胞在肝脏及肿瘤中的浓度和稳定性均高于游离药物,且随着时间延长,这种优势越来越大。结论：载药红细胞延缓药物释放,延长药物作用时间;增强了药物向肝脏及肿瘤的靶向聚集,减少不良反应。为临床肿瘤治疗提供新思路和可行方法。     

动物实验性脑出血后血肿内红细胞管涌现象研究

目的探索脑出血后血肿内红细胞通过"管涌现象"的分布特征。方法 SD雄性大鼠60只,随机分为实验组30只和对照组30只,每组分为5个不同观察时间点:1h、6h、24h、3d、7d,每个时间点5只。采用异硫氰酸荧光素(FITC)标记血液中红细胞,建立大鼠自体动脉血脑出血模型,分别于不同时间点进行大鼠脑组织、颈深淋巴结组织病理切片,荧光显微镜下观察以及HE染色后显微镜下观察,对比分析红细胞的分布特征。结果实验组血肿形成1h后,病灶以及血肿周围可见到FITC-RBC和游离FITC分布,双侧大脑半球和脑干的维乔-罗宾间隙内也有游离FITC分布,双侧颈深淋巴结内均可见FITC-RBC和游离FITC分布。6h后血肿内开始出现含有黄色或黄绿色荧光颗粒的吞噬细胞,同时FITC-RBC和游离FITC逐渐减少。随时间延长,含有黄色荧光颗粒的吞噬细胞在各脑区内逐渐增多,淋巴结生发中心内可见成团分布的黄色荧光颗粒。对照组未见以上变化。结论血肿内红细胞通过"管涌现象"在脑出血后扩散范围比较局限,游离红细胞成分扩散范围比较广泛;"管涌现象"是血肿内有红细胞吸收途径之一。     

Role of FcγRI (CD64) in erythrocyte elimination and its up-regulation in thalassaemia

To examine any role of the high affinity Fcgamma class I receptor (FcgammaRI) (CD64) in erythrocyte elimination by mononuclear phagocytes (MP) in thalassaemia (thal), we investigated the in vitro interaction of beta-thalassaemic erythrocytes with monocytes (Mo) whose FcgammaR expression had been modulated by cytokines. Treatment of Mo with interferon (IFN)-gamma or interleukin (IL)-10 which up-regulate FcgammaRI, caused a dose-dependent increase in binding of beta-thalassaemic erythrocytes, whereas stimulation with IL-4 which down-regulates the receptor, reduced this interaction, in a dose-dependent manner, to that of normal erythrocytes. Binding of thalassaemic erythrocytes by IFN-gamma or IL-10-treated Mo was inhibited by FcgammaRI-specific reagents. In addition, Mo expression of FcgammaRI and HLA class II DR was determined by flow cytometry in Thai patients with HbH disease (alpha1/alpha2 or alpha1/Hb Constant Spring) (n = 15) or beta degrees -thal/HbE (n = 16). In both groups of patients FcgammaRI expression was increased as compared to normal controls (n = 14): mean fluorescence intensity (+/-SD) 124.79 +/- 38. 77 in HbH disease and 121.86 +/- 18.23 in beta degrees -thal/HbE versus 91.94 +/- 17.36 in normal controls (P < 0.01 and P < 0.001, respectively). In contrast, HLA class II DR expression was similar in patients and controls. The results suggest that, in thalassaemia, up-regulated FcgammaRI on mononuclear phagocytes plays a role in their interaction with erythrocytes and that this process can be modified by cytokines.     

Quantitative analysis of erythrocytes containing fetal hemoglobin (F cells) in children with sickle cell disease

Variation in the level of fetal hemoglobin (HbF) accounts for much of the clinical heterogeneity observed in patients with sickle cell disease (SCD). The HbF level has emerged as an important prognostic factor in both sickle cell pain and mortality, and a % HbF of 10–20% has been suggested as a threshold level for diminished clinical severity. The number of erythrocytes that contain HbF (termed F cells) may also be critically important, as F cells resist intravascular sickling and have preferential in vivo survival. Since F cells can be enumerated with high accuracy using flow cytometry methods, we prospectively studied a cohort of 242 children with SCD. Children with HbS and hereditary persistence of fetal hemoglobin (S/HPFH) had essentially 100% F cells. In contrast, children with homozygous sickle cell anemia (HbSS), HbS/β⁰ thalassemia, or HbS/β⁺ thalassemia had significantly lower mean % F cell values (55.9, 61.6, and 51.3%, respectively; P < 0.001), and children with HbSC had even fewer F cells (27.0%; P < 0.001). There was a highly significant correlation between the % F cells and the log (% HbF), which was observed for the total population of children (r = 0.95, P < 0.001), as well as for each of the individual subgroups of children with HbSS (r = 0.94, P < 0.001), HbSC (r = 0.89, P < 0.001), or HbS/β⁰ thalassemia and HbS/β⁺ thalassemia (r = 0.95, P <0.001). This logarithmic correlation between % F cells and % HbF has not been previously described and has important implications for the pharmacologic manipulation of HbF in patients with SCD. Am. J. Hematol. 54:40–46, 1997 © 1997 Wiley-Liss, Inc.