纤维蛋白胶丁胺卡那霉素复合体防治深部创口感染

目的 观察纤维蛋白胶丁胺卡那霉素复合体治疗深部创口后白细胞计数和中性粒细胞分类变化,探讨该复合体控制感染的有效性.方法 采用临床病例对照研究方法进行研究.采用计算机随机摇号分组,将试验组100例患者与对照组100例患者按创口部位、大小、受伤到手术时间、合并损伤、全身应用抗生素配对分组,对比两组清创术后不同时间点白细胞计数和中性粒细胞分类计数变化,观察试验组有无与使用纤维蛋白胶丁胺卡那霉素复合体相关的毒副作用.结果 (1)试验组和对照组在性别分布、平均年龄、受伤机制方面基本相同(P>0.05),两组具有可比性.(2)试验组组内不同时间点白细胞计数和中性粒细胞分类计数差异有统计学意义(P<0.05),术后1 d白细胞计数和中性粒细胞分类计数最高,以后随着治疗时间延长而逐渐下降,术后24 d或出院前1 d最低.(3)试验组和对照组白细胞计数除术后1 d差异无统计学意义(P>0.05)外,其余时间点试验组均比对照组低(P<0.05).试验组和对照组术后不同时间点中性粒细胞分类计数比较,在术后1,2,12 d两组差异无统计学意义(P>0.05),其余时间点两组差异均有统计学意义(P<0.05),试验组不同时间点中性粒细胞分类计数均比对照组低.结论 纤维蛋白胶丁胺卡那霉素复合体治疗深部创口感染有较好的临床效果,安全性高,没有不良反应和毒副作用,值得推广使用.     

Simultaneous quantitative analysis of the expression of CD64 and CD35 on neutrophils as markers to differentiate between bacterial and viral infections in patients with rheumatoid arthritis

Abstract

Objectives The expression level of CD64 on neutrophils can be used to differentiate between an infection and a disease flare in rheumatoid arthritis (RA) patients. However, the CD64 expression is elevated by both bacteria and viruses, so it cannot be used to distinguish the type of infection. We herein investigated the results of a simultaneous quantitative analysis of the expression of CD64 and CD35 on neutrophils to determine whether these molecules can be used to distinguish between bacterial and viral infections in RA patients.

Methods We collected blood from 22 RA patients with pathogen-proven infections (15 bacterial and 7 viral infections). Blood samples were stained with QuantiBRITE CD64PE/CD45PerCP and CD35PE, and the mean fluorescence intensities were assessed by a flow cytometer. The mean numbers of molecules were calculated using QuantiBrite PE beads.

Results We calculated the ratio of CD64 to the CD35 level (CD35/CD64), and used a cut-off value of 2.8 for the CD35/CD64 ratio. At this value, the sensitivity for diagnosing a bacterial infection was 87%, and the specificity was 86%.

Conclusions Simultaneous quantitative analysis of CD64 and CD35 expression on neutrophils might be useful to distinguish between bacterial and viral infections in RA patients.     

Neutrophil function in hyperosmotic NaCl is preserved by phosphoenol pyruvate

Summary Hyperosmolority in the urinary tract inhibits the host defense against bacterial infection. NaCl contributes most to osmolority in the renal medulla and urine. Therefore, we studied the effect of hyperosmolar NaCl on neutrophil function. When osmolarity was increased by NaCl, polymorphonuclear leukocytes (PMNL) became defective in phagocytosis, intracellular killing of bacteria, chemotactic activity, and superoxide production. This coincided with a decrease in the intracellular adenosine triphosphate (ATP) content of PMNL. Both the inhibition of superoxide production and the reduction in ATP content did not occur at 4°C. By increasing ATP content, phosphoenol pyruvic acid (PEP) protected against the decrease in superoxide production. These results suggest that leukocyte function is inhibited by high concentrations of NaCl due to the activation of the Na⁺–K⁺ pump. PEP, an ATP precursor, can protect PMNL against osmotic stress by raising the intracellular concentration of ATP.     

囊性颈动脉瘤动物模型建立与研究

目的：建立囊性颈动脉瘤的动物模型并研究动脉瘤的形成原因。方法：选择新西兰白兔随机分成3组，采用胰弹性蛋白酶血管壁外法建立囊性动脉瘤，并通过血流动力学改变来分析与动脉瘤形成的关系。结果：血管造影证实结扎对侧颈总动脉后，同侧颈总动脉上的成瘤率为100％，颈外动脉起点及颈外动脉分支起点处成瘤率为50％；不结扎对侧颈总动脉，同侧颈总动脉及颈外动脉起点上的成瘤率也为50％；其余成瘤率均为零。组织学检查显示实验侧动脉管壁破坏率为100％。结论：胰弹性蛋白酶血管壁外法是建立囊性颈动脉瘤的有效方法之一，动脉管壁的结构改变和血流动力学变化是形成囊性动脉瘤的两个主要因素。     

炎症反应中组织损伤机制的探讨:中性粒细胞化学发光和脱颗粒的研究

以大鼠脱精氨酸C5a(r-C5ai)、人工合成趋化因子N-甲酰-甲硫氨酰一亮氨酰-苯丙氨酸(fMet-Leu-phe,简称FMLP),血清调理的酵母多糖(serum treated zymosan,简称STZ)、热聚IgG(A-IgG)、钙离子载体A23187为刺激物,观察它们对中性粒细胞(PMN)化学发光和脱颗粒的影响,以便为以炎症反应为特征的疾病的发病学研究提供线索。结果表明,所用刺激物均显著提高PMN的化学发光,但不同刺激物对PMN脱颗粒时不同酶的释放影响不同。     

A simple assessment of human neutrophil adhesiveness

Summary The adhesiveness of human polymorphonuclear leukocytes was assessed in serum-coated polystyrene spectrophotometric cuvettes. Capped cuvettes, containing no more than 2×10⁶ resting or concanavalin A-treated (100 μg/ml) polymorphonuclear leukocytes, were laid horizontally and subjected to three 90° rotations on their major axis at fixed times. After incubation at room temperature, non-adherent cells remaining in suspension were counted on the Coulter counter STKS hematological analyzer. After a 16-min incubation (4 min each side of the cuvette) the adhesion of concanavalin A-activated neutrophils ranged from 98% to 100% and the adhesion of resting neutrophils from 30% to 35% (mean 32.4±2.2%,n=10). An 8-min incubation (2 min each side) led to approximately 50% adhesion of concanavalin A-activated neutrophils (mean 49.9±2.2%, range 46%–54%,n=16) whereas the adhesion of resting cells was about 21% (mean 21.4±1.6%, range 19%–24%,n=16). The variation in percentage adhesion in repeated assays did not exceed 4% using concanavalin A-activated cells and 7.5% with resting neutrophils. The procedure is very rapid, easy to perform and precise, and no special apparatus or glassware is necessary. The method also allows microscopic evaluation of shape changes of adherent neutrophils through the clear sides of the cuvettes.     

Sputum peroxidase activity correlates with the severity of lung disease in cystic fibrosis

Patients with cystic fibrosis (CF) of the same age differ significantly in their degree of pulmonary disease. Based on preliminary observations, we postulated that the activity of my-eloperoxidase would be significantly increased in patients with greater structural lung damage than in those with less lung damage. Acid extracts of weighed sputum samples were assayed for lactoferrin concentrations by ELISA. Activities of peroxidase, cathepsin G, and elastase (with and without proteinase 3) were determined by kinetic analysis using chromogenic substrates. The patients were divided into quartiles based on their Brasfield chest-radiograph score. Patients in the first quartile (least amount of structural lung abnormality) were compared to those in the fourth quartile. The concentration of lactoferrin, a specific (secondary) granule protein of neutrophils, did not differ between the two patient groups. However, the activities of the neutrophil primary granule proteins, peroxidase, elastase, and elastase plus proteinase 3, were significantly elevated in the group with the most structural lung abnormality. Sputum albumin concentration was used to estimate leakages of plasma proteins into the airways. Peroxidase activity, but not the activity of cathepsin G, of elastase, or of elastase plus proteinase 3, correlated significantly with albuminig sputum in both quartile groups. To confirm the association of sputum peroxidase activity with differences in lung structure and to test its correlation with lung function, spirometry was performed in a second group of patients during the week prior to the time of sputum sampling. In this second group, increased sputum peroxidase activity was associated with worse Brasfield scores and with decreased percent-predicted forced expiratory volume in 1 sec. Lactoferrin concentration again did not differ between the groups. Bacterial factors also did not differ. The results indicate that the degree of lung damage in patients with CF was more closely related to sputum peroxidase activity than to age, gender, CFTR genotype, or sputum bacterial parameters. Further study is required to determine whether or not peroxidase activity in the airway causes this lung damage. © 1995 Wiley-Liss, Inc.     

Chemotaxis of canine polymorphonuclear neutrophil granulocytes using the under-agarose method applied to glass microscope slides

The under-agarose chemotaxis assay method applied to glass microscope slides was optimised for canine polymorphonuclear granulocytes (PMN) with gelatin as protein supplement. Optimum chemotactic response occurred after approximately 135 min of incubation at 37 °C with 5 × 10⁵ cells/well and zymosan-activated serum (ZAS) as chemoattractant. Chemotactic (C) and random migration (R) were stated separately for each dog. Storage of blood samples for 90 min before isolation of PMN did not influence the C or R values (p > 0.05). The preanalyfcal variation was investigated and the proportion of variance, expressed as a percentage, due to each source for the C and R values, respectively, was determined: dog, 92%, 96%; blood sample, 0.4%, 0%; interslide, 3%, 2% and intraslide, 3%, 3%. The mean chemotactic responsiveness against ZAS and ZAS diluted 1:1 with RPMI 1640 (dZAS) was determined using 15 laboratory beagle dogs: C_ZAS = 0.67 mm (range: 0.25–2.33 mm), coefficient of variation (CV) = 24%, C_dZAS = 0.47 mm (range: 0.18–2.12 mm), CV = 22%. Also, the mean random migration was determined: 0.11 mm (range: 0.05–0.34 mm), CV = 25%. Neither age nor sex had any significant influence on chemotactic or random migration (p>0.05). The effect of heat-inactivated pooled canine serum (HI-serum) incorporated into agarose was examined and it was found that this had a significant stimulatory effect on both chemotactic and random migratory responses (p<0.05). Therefore, agarose with HI-serum is not useful when investigating the chemotactic function.     

Serum enzymatic changes modulated using trypsin: chymotrypsin preparation during burn wounds in humans

The levels of marker enzymes for liver function, namely transaminases (SGPT, SGOT), creatine phosphokinase (CPK), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) were estimated in the sera of burn patients by administering trypsinchymotrypsin preparation and comparing with an untreated group. Neutrophil proteolytic activity was also measured by assaying the lysosomal enzymes, namely neutrophil clastase and cathepsin D. Our earlier studies have already proved the efficacy of the above enzyme preparation to burn patients on the enhancement of vascular responses during the acute phase of the burn injury. These beneficial responses were brought about by the modulation of acute phase proteins expressed in the liver. Hence, it is of interest to study the changes in the above mentioned liver enzymes and certain lysosomal enzymes in the serum during the first 10 days of burn injury. The levels of liver and lysosomal enzymes markedly decreased in the treated group when compared with the untreated group. The enzyme studies clearly indicated that the initial rise in the liver enzymes was minimized in the treated group when compared with the untreated group and this helped in reducing the stress to the liver in the treated cases. The increase in the activity of ₁-antitrypsin and ₂-macroglobulin and decreased levels of C-reactive protein are atributed to the reduction of proteolytic enzyme levels in the treated group and minimizing the degradative changes during wound repair.