Whole-Blood Assay to Measure Oxidative Burst and Degranulation of Neutrophils for Monitoring Trauma Patients

AbstractBackground and Purpose: Polymorphonuclear neutrophils (PMNs) protect the host from invading microorganisms, but excessive PMN activation after trauma causes tissue injury. Rapid monitoring of PMN function is critical for the assessment of the inflammatory state of trauma patients. Here, the authors adapted two simple and rapid methods to measure oxidative burst and degranulation of human PMNs in whole blood to avoid potential interference of cell isolation procedures with the assessment of PMN function.Material and Methods: Heparinized blood was drawn from healthy volunteers or trauma patients, preincubated at 37 °C for 5 min, and stimulated with N-formyl-methionyl-leucyl-phenylalanine (fMLP). Four assays for oxidative burst were tested: (1) cytochrome C; (2) homovanillic acid (HVA); (3) Amplex^® Red; and (4) flow cytometry with dihydrorhodamine 123 (DHR). PMN degranulation was assessed with flow cytometry using antibodies to: (1) CD11b/Mac-1 (CD18); (2) CD63; and (3) CD66b (CD67).Results: With the exception of the DHR method, all methods to measure oxidative burst were found to be unsuitable in whole blood due to interference of plasma proteins and hemoglobin with the fluorimetric or photometric readouts. By contrast, all degranulation methods were suitable for whole-blood studies. However, for the assessment of formyl peptide-induced degranulation, anti-antibodies to CD11b/Mac-1 and CD66b were up to five times more sensitive than antibodies to CD63. Thus, the degranulation and DHR methods were optimized for increased sensitivity, speed, and specificity and their usefulness to measure PMN function in trauma patients was tested.Conclusion: The whole-blood methods based on flow cytometry with DHR, anti-CD11b/Mac-1, and anti- CD66b are rapid, simple, and reliable techniques to assess PMN function for trauma research.     

The relationship between the Stiles–Crawford effect of the first kind (SCE-I) and myopia

The Stiles-Crawford effect of the first kind (SCE-I) was measured on both emmetropic and myopic subjects at six different retinal locations. The results revealed a number of significant discrepancies in receptor alignment between the groups of different refractive errors. In myopic subjects, the receptors in the nasal retina (i.e. between the fovea and the optic nerve head) were found to be aligned nasally towards the optic nerve head, whereas the receptors in the temporal retina were aligned towards the centre of the exit pupil. In emmetropic subjects, the receptors across the retina were finely tuned towards the centre of the exit pupil. The magnitude of the receptor displacement in myopic subjects was found to be directly associated with the length of the eyeball.     

Review of long-term adverse effects associated with the use of chemically-modified animal and nonanimal source hyaluronic acid dermal fillers

Although only recently introduced, chemically-modified hyaluronic acid dermal fillers have gained widespread acceptance as “redefining” dermal fillers in the fields of dermatology and cosmetic facial surgery. Although hyaluronic acid-based dermal fillers have a low overall incidence of long term side effects, occasional adverse outcomes, ranging from chronic lymphoplasmacytic inflammatory reactions to classic foreign body-type granulomatous reactions have been documented. These long-term adverse events are reviewed.     

头低位模拟失重状态对前庭功能的影响

为研究头低位模拟失重对运动病症状、垂直视动眼震（ＶＯＫＮ）及体液重新分配的影响，在头低位－１０°的模拟失重状态下，采用大视野的垂直视动刺激，观察１８名正常人的运动病症状、ＶＯＫＮ、激素（ＡＶＰ、ＶＩＰ、ＣＯＲＴ、ＡＬＤＯ）的反应特点。结果表明，头低位－１０°状态下的大视野垂直视动刺激可以诱发出明显的运动病症状，头低位－１０°的垂直视动刺激比坐位更容易诱发运动病。坐位状态ＶＯＫＮ慢相速度有明显的方向性不对称，敏感组ＶＯＫＮ方向性不对称有显著差异（Ｐ＜０．０５）。头低位－１０°时ＶＯＫＮ的不对称现象不明显，向下方向运动的ＶＯＫＮ慢相速度显著增加。分析指出，头低位－１０°状态下垂直视动刺激比坐位和秋千刺激的贡献率大。尿中ＣＯＲＴ（皮质醇）在秋千和头低位的垂直视动刺激前后有显著性增加。提示：大视野的垂直视动刺激与头低位－１０°两种刺激的结合可能成为预测空间运动病的方法之一．     

火器伤后胸心异物存留

目的：探讨火器伤后胸心异物存留摘除的指征、时机、方法及结果。方法：回顾性分析了３９例火器伤后胸心异物存留摘除者的一般资料、临床表现、不同处理方法的选择及效果。结果：１９例患者早期因凝固性血胸及肺持续漏气伴异物存留或异物存留伴胸腔及心包腔化脓性感染，在开胸清创时做了异物摘除；另２０例患者为胸伤伤情恢复后单纯异物存留，因异物位于肺门、心脏及大血管旁或远期并发症等，而后期开胸异物摘除。结论：火器性胸部穿透伤后，因异物本身或其他原因所致早期并发症者，应在开胸清创的同时摘除异物；胸伤伤情恢复而单纯胸心异物存留，应选择性摘除或因并发远期并发症而摘除。     

Measurement of biological activity of somatotropin in hypophysectomized rats

目的：建立测定生长激素（ＧＨ）在体生物活性的方法．方法：以去垂体大鼠体重增长（ＢＷＧ）和胫骨骺软骨板宽度（ＴＥＷ）为指标，观察动物性别、给药途径、次数和周期不同对效应的影响；同时进行４ｄＢＷＧ，６ｄＢＷＧ和６ｄＴＥＷ法，测定ＧＨ的效价（平行线３×３设计）．结果：♀和♂ｓｃ和ｉｍ给药以及每日给药１次和２次的ＢＷＧ和ＴＥＷ差异无显著意义．给药６ｄ比给药４ｄ引起较大的ＢＷＧ和ＴＥＷ（Ｐ＜００５）．４ｄＢＷＧ法和６ｄＢＷＧ法在００２０－０５００ＩＵ·ｄ－１有较好的λ值（００６６０和０１７４７）和ｒ值（０９０００和０９２３７）；４ｄＢＷＧ，６ｄＢＷＧ和６ｄＴＥＷ法测得ｒｈＧＨ的效价为４６１３２，３９８２９和４８０２３ＩＵ／ａｍｐ．６ｄＢＷＧ法有较小的λ值和较低的ＡＲＦＬ值．结论：可在同一组去垂体大鼠体内同时用４ｄＢＷＧ，６ｄＢＷＧ和６ｄＴＥＷ法测ＧＨ活性，以６ｄＢＷＧ法较好．     

CT对眼球异物的诊断及定位

本文比较了11例眼球异物CT与X线影像，并经手术摘出异物证实，CT眼球异物检出及定位准确性均优于X线，尤其对球壁异物的定位，而且能显示断层眼球壁轮廓，具有直观效果，CT异物影像比异物明显扩大，应警惕CT伪影可能使球壁异物定位发生误差，CT目前尚不能完全取代眼球异物常规X线检查。     

Collection of blood mononuclear cells by leukapheresis for transplantation in a yucatan miniature swine model

A large animal model is needed to evaluate new apheresis technologies. These technologies include novel methods of harvesting the blood mononuclear cell population which contains the hematopoietic stem cells needed to restore hematopoiesis in recipients of hematopoietically lethal therapy and the use of cytokines to provide a safe and predictable method of manipulating these circulating hematopoietic stem cells. We describe the methods used to collect mononuclear cells by leukapheresis from Yucatan miniature swine. These animals are of sufficient size to tolerate the procedures and have many physiologic and hematologic similarities to man. They are of good temperament and are easily trained. Long-term venous access was obtained using single lumen silicone rubber catheters placed in the inferior vena cava. The animals were apheresed while fully awake using a Haemonetics Model V50 machine and a modified lymphocyte collection protocol. The procedure was highly efficient for the collection of mononuclear cells and a 10 pass procedure yielded a product which contained 19.7 × 10⁹ mononuclear cells, 10.7 × 10⁹ granulocytes, and 17 ml of erythrocytes in a volume of approximately 100 ml. This product can be cryopreserved and used for subsequent transplantation. The content of four apheresis procedures provides hematopoietic reconstitution of lethally irradiated swine on a time scale equivalent to transplantation of optimal numbers of bone marrow cells. © 1992 Wiley-Liss, Inc.     

Central tumour necrosis factor-α mediates the early gastrointestinal motor disturbances induced by lipopolysaccharide in sheep

Cytokines are involved in fever and other symptoms of the acute phase response induced by endotoxins. The aim of this work was to study the involvement of central tumour necrosis factor-alpha (TNF-alpha) in the changes induced by lipopolysaccharide (LPS) on gastrointestinal (GI) motility in sheep. Body temperature and myoelectric activity of the antrum, duodenum and jejunum was recorded continuously. Intravenous (i.v.) administration of LPS (0.1 micro g kg-1)-induced hyperthermia, decreased gastrointestinal myoelectric activity and increased the frequency of the migrating motor complex (MMC). These effects started 40-50 min after LPS and lasted for 6-7 h. TNF-alpha (50 and 100 ng kg-1) mimicked these effects when injected intracerebroventricularly (i.c.v.) but not i.v. Pretreatment with soluble recombinant TNF receptor (TNFR:Fc, 10 micro g kg-1, i.c.v.) abolished the TNF-induced actions and reduced those evoked by LPS. Furthermore, the effects induced by either LPS or TNF were suppressed by prior i.c.v. injection of indomethacin (100 micro g kg-1). In contrast, the i.v. injections of TNFR:Fc or indomethacin were ineffective. Our data suggest that LPS disturbs GI motility in sheep through a central pathway that involves TNF-alpha and prostaglandins sequentially.     

咳喘患儿气管支气管异物的检出

气管支气管异物是引起小儿咳喘的重要原因之一。本文就X线诊断中的几点体会作了详细介绍,认为X线检查对气管支气管异物的诊断和治疗有极为重要的价值。