烧伤延迟复苏对CD14+单核细胞HLA-DR表达率和外周血淋巴细胞凋亡率的影响

目的:探讨烧伤延迟复苏对CD14 单核细胞人类白细胞组织相容性抗原-DR(HLA-DR)表达率和外周血淋巴细胞凋亡率的影响及其机制。方法:50例烧伤面积大于30%的烧伤患者,按伤后是否及时有效复苏分为延迟复苏组和非延迟复苏组,于伤后1、3、7、14、28d取外周血。流式细胞术检测CD14 单核细胞HLA-DR表达率和外周血淋巴细胞凋亡率。20例健康体检者外周血检测上述指标作为对照。结果:烧伤患者外周血CD14 单核细胞HLA-DR表达率明显低于正常对照组(P均<0.01),延迟复苏患者明显低于非延迟复苏患者(P<0.05或P<0.01)。烧伤患者外周血淋巴细胞凋亡率明显高于正常对照组(P<0.05),延迟复苏患者明显高于非延迟复苏患者,伤后7和28d差异有显著性(P<0.05)。结论:烧伤患者免疫功能低下,动态检测患者的免疫功能状态对患者预后具有指导意义。     

严重烧伤患者人白细胞抗原DR定量表达的临床意义

目的探讨严重烧伤患者人白细胞抗原DR（HLA—DR）定量表达的变化规律及其临床意义。方法采集77例烧伤大于体表总面积30％的患者的血样,通过流式细胞技术对患者烧伤后不同时段CD14^＋单核细胞表面HLA—DR结合量进行动态的定量分析。结果严重烧伤患者伤后第1天开始CD14^＋单核细胞表面HLA—DR结合量明显低于正常对照组（P〈0．05）,其表达均值与烧伤面积呈显著负相关（r=-0．7232,P〈0．05）。并发多脏器功能障碍综合征（MODS）者其CD14^＋单核细胞表面的HLA—DR表达量持续下降,其中伤后第3、14、21、28天显著低于非MODS组（P〈0．05）。随着CD14^＋单核细胞HLA—DR表达水平的下降,MODS发生频率增加,患者预后不良。结论大面积烧伤可导致机体CD14^＋单核细胞HLA—DR表达严重受损和免疫功能障碍,动态观察其表达水平有助于烧伤后MODS的病程监测及患者预后判断。     

体外循环对单核细胞HLA-DR表达的影响

目的研究体外循环对单核细胞表面HLA-DR表达水平的影响.方法16例风湿性心脏病瓣膜置换术患者为试验组(心内直视术组),10例肺叶切除术患者为对照组(非心内直视术组).采用流式细胞术动态观察麻醉诱导前、气管插管后、体外循环心肺转流前、体外循环心肺转流后、手术后第1d、第3d及第5d淋巴细胞和单核细胞表面HLA-DR的表达水平,同时观察APACHEⅡ评分及手术后恢复情况.结果心内直视术组心肺转流后、术后第1d、第3dHLA-DR表达阳性的单核细胞百分率显著下降,且心肺转流后单核细胞数显著下降,单核细胞表面的HLA-DR分子数显著下降(P＜0.05).肺叶切除术组手术后HLA-DR表达阳性的单核细胞百分率明显下降,但单核细胞占外周血白细胞比例无显著性差异.HLA-DR表达阳性的淋巴细胞百分率在各个时间点均无统计学差异(P＞0.05);两组相比,手术后HIA-DR表达阳性的单核细胞百分率心内直视组显著低于肺叶切除组.2例心内直视术后并发感染患者于术后第6dHLA-DR表达阳性的单核细胞百分率仍低于40%.结论心肺转流后机体免疫功能的障碍与单核细胞的功能紧密相关.动态监测单核细胞表面HLA-DR的表达水平有助于预测心内直视术后感染的发展及预后.     

人类白细胞抗原DR基因与狼疮性肾炎相关性的探讨

目的 从基因水平探讨人类白细胞DR抗原(HLA-DR)基因与中国北方汉人狼疮性肾炎(LN)的相关性。方法 采用特异性引物-聚合酶链式反应(PCR-SSP)方法,检测89例系统性红斑狼疮(SLE)患者及106例健康者HLA-DRB1基因类型,进行临床相关性分析。结果 SLE患者的HLA-DR2及DR9基因频率明显高于对照组(0.36/0.20,RR=2.36,P<0.001;0.27/0.18,RR=1.69,P<0.05);DR2及DR9同时阳性的35例患者中,LN的发病率明显高于其它患者(RR=4.93,P<0.005),而且抗dsDNA抗体的阳性率也较高(RR=2.66,P<0.05)。结论 中国汉人HLA-DR2及DR9基因可能与SLE的遗传易感性有关,两者有相加作用,DR2及DR9同时阳性的SLE患者易患LN。     

CD14基因多态性对严重烧伤后CD14表达的影响及其临床意义

目的探讨内毒素受体CD14C-159T基因多态性对烧伤后CD14合成与释放的影响及其与脓毒症易感性、患者预后的关系。方法26例烧伤面积大于30％的患者,采用聚合酶链反应及限制性内切酶HaeuI对PCR产物的消化作用检测CD14基因多态性。同时,对患者白细胞CD14、肿瘤坏死因子-α（TNF-α）mRNA表达,血清sCD14浓度与CD14-159位点基因型以及烧伤患者预后的关系进行了分析。结果Tr、TC、CC三种基因型患者白细胞CD14mRNA、TNF—amRNA表达,血清sCD14浓度存在明显差异。其中Tr、TC基因型CDl4mRNA表达均明显高于CC纯合子（P＜0．05或P＜0．01）,伤后第7天CC基因型血浆sCDl4水平显著低于TC基因型（P＜0．05）;同时,TT纯合子的TNF-αmRNA表达强度明显高于CC纯合子（P＜0．05）。此外,伤后第7、21天死亡组CDl4mRNA表达量显著高于存活组（P＜0．05）。结论CD14C-159T基因多态性可显著影响严重烧伤后CDl4的合成与释放,并与烧伤患者脓毒症易感性有关,T等位基因可能是患者预后不良的高危基因标志物。     

单核细胞表面人类白细胞抗原DR表达水平与慢性阻塞性肺疾病急性期感染的相关性研究

目的探讨人外周血单核细胞表面人类白细胞抗原DR(HLA-DR)表达水平与慢性阻塞性肺疾病(COPD)急性期感染的相关性。方法选取天津市南开医院慢性阻塞性肺疾病急性加重(AECOPD)患者和体检健康者共37例, 根据临床症状及感染指标分为对照组(15例)、急性期组(临床治疗干预前, 22例)和稳定期组(临床干预后, 16例)。采集患者外周静脉血, 用流式细胞仪分析CD14+单核细胞HLA-DR的表达水平, 并与患者C反应蛋白(CRP)、白细胞计数、中性粒细胞百分比、中性粒细胞计数、CD3+ T细胞百分比、CD3+CD4+ T细胞百分比、CD3+CD8+ T细胞百分比和CD4+/CD8+等进行相关性分析。结果与对照组比较, 急性期组患者外周静脉血CD14+单核细胞HLA-DR的表达水平降低(P<0.05)。与急性期组比较, 稳定期组患者外周静脉血CD14+单核细胞HLA-DR的表达水平升高(P<0.05), CRP的水平降低(P<0.05), 白细胞计数、中性粒细胞百分比、中性粒细胞计数下降(均P<0.05), CD3+CD4+ T细胞百分比、CD4+/CD8+增加...     

胸腺肽α1对烫伤延迟复苏大鼠单核细胞Ia抗原的影响

目的:研究烫伤延迟复苏后大鼠单核细胞表面Ia抗原变化规律及应用胸腺肽α1对单核细胞表面Ia抗原的影响。方法:将104只成年雄性Wistar大鼠随机分为正常对照组(8只)、单纯烫伤组(48只)和胸腺肽α1治疗组(48只)。正常对照组不作烫伤处理;其它组均造成30%TBSAⅢ度烧伤延迟复苏模型。分别于6h、1d、2d、3d、7d和14d处死动物,取血分离单核细胞,流式细胞仪检测Ia抗原表达率。结果:大鼠烫伤延迟复苏后单核细胞Ia抗原表达率降低,其中6h、和1d时间点Ia抗原表达率严重降低,3d、7d和14d时间点开始有所恢复,但明显低于正常大鼠(P均<0·01)。给予胸腺肽α1后3d、7d和14d时间点单核细胞Ia抗原表达率与单纯烫伤组比较明显提高(P均<0·01)。结论:严重烧伤延迟复苏后,单核细胞抗原递呈功能减弱,免疫功能障碍。胸腺肽α1作为一种免疫增强剂可明显改善烧伤延迟复苏后单核细胞抗原递呈功能。     

Low HLA-DR expression on CD14+ monocytes of burn victims with sepsis, and the effect of carbachol in vitro

This study aimed to investigate changes in the expression of human leukocyte antigen-DR (HLA-DR) on CD14⁺ monocytes in the peripheral blood of burn victims with delayed resuscitation in relation to the development of sepsis, and the effect of carbachol in vitro. The study population comprised 25 people with burns of at least 30% of total body surface area and delayed resuscitation, and 20 healthy volunteers as controls. Peripheral blood was collected on post-burn days 1, 3, 7, 14 and 28. When 7 participants developed sepsis, their peripheral blood was drawn on 2 consecutive days. Expression of HLA-DR on CD14⁺ monocytes in peripheral blood of burned participants was lower than that of controls, and fell further with the development of sepsis, when the rate and concentration of tumour necrosis factor-α (TNF-α) rose above those of controls and burned participants without sepsis. Expression of HLA-DR on CD14⁺ monocytes was negatively correlated with interleukin-10 (IL-10) levels on post-burn days 1, 7 and 28. In vitro, HLA-DR expression on monocytes also decreased with lipopolysaccharide (LPS) stimulation, but after treatment with carbachol, rose in a concentration-dependent manner. Thus expression of HLA-DR on CD14⁺ monocytes may be a useful parameter for monitoring the immune function of burn victims with and without sepsis. Carbachol significantly inhibited LPS-induced immunosuppression in human monocytes in vitro.     

高容量血液滤过对多脏器功能障碍综合征犬CD14+单核细胞白细胞-DR抗原表达的影响

目的 观察高容量血液滤过(HVHF)对多脏器功能障碍综合征(MODS)犬CD14+单核细胞犬白细胞DR抗原(DLA-DR)表达的影响.方法 通过2次打击建立犬MODS模型,随机分HVHF组和MODS组,HVHF组建模后给予HVHF治疗24h,MODS组不给HVHF治疗.监测CD14+单核细胞DLA-DR表达及主要器官功能指标变化.结果 与实验前(6.52±1.47)比较,MODS组CD14+单核细胞DLA-DR明显降低(P＜0.01).在T5、T7、T8、T9时间点HVHF组CD14+单核细胞DLA-DR水平显著高于MODS组(P＜0.05).HVHF组主要器官功能明显改善.结论 HVHF能改善器官功能,提高CD14+单核细胞DLA-DR的表达,有助于重建机体内稳态.     

高原严重烫伤大鼠延迟复苏肾组织细胞凋亡与c-fos蛋白表达的变化

目的：探讨高原不同海拔地区大鼠严重烫伤延迟复苏后肾脏组织c-fos蛋白的表达规律及其与肾组织细胞凋亡的关系.方法：雄性Wistar大鼠240只,分别在1517 m和3 848 m两个海拔高度进行实验,每个海拔高度的大鼠（n=120）均随机分为3组：①即时复苏组（IFR组,n=60）：伤后即刻腹腔注射等渗盐水4 ml/kg;②延迟复苏组（DFR组,n=50）：伤后6 h开始按4 ml/kg补液;③假伤组（n=10）：大鼠背部置于37 ℃水浴中20 s模拟烫伤,不补液.IFR组和DFR组大鼠将背部浸于90 ℃热水中20s建立30% TBSA Ⅲ度高原烫伤模型,分别于伤后1、6、12、24、72和168 h取肾组织,假伤组大鼠于模拟烫伤后1 h取肾组织,采用原位末端标记法（TUNEL）检测肾组织细胞的凋亡,计算凋亡指数;免疫组织化学染色与图像分析技术检测肾组织c-fos蛋白的表达.结果：肾组织细胞凋亡指数和c-fos蛋白表达水平在相同海拔高度DFR组和IFR组均高于假伤组,且DFR组均高于IFR组,高海拔地区高于低海拔地区（P〈0.001）;细胞凋亡指数与c-fos蛋白表达水平呈明显的正相关（r=0.968,P〈0.001）.结论：高原严重烫伤可导致 c-fos蛋白的表达增强,同时伴肾组织细胞凋亡加剧,延迟复苏较即时复苏加重细胞凋亡;c-fos蛋白的表达与高原严重烫伤大鼠肾组织细胞凋亡相关.     

Influence of CD14 polymorphism on CD14 expression in patients with extensive burns

We sought to investigate the association of CD14 genotype with the risk of mortality after burn, and we also attempted to evaluate whether CD14-159 C/T polymorphism affects the kinetics and extent of CD14 expression as well as its release, and TNF-alpha expression in burned patients. The study involved 64 patients in Chinese Han population incurring burns covering more than 30% of the total body surface area. CD14 polymorphism was determined by polymerase chain reaction (PCR) and subsequent restriction fragment length polymorphism (RFLP) analysis. Meanwhile, leukocyte CD14 mRNA expression and soluble CD14 (sCD14) levels were measured during a 28-day observation period. TNF-alpha mRNA and protein levels were also determined in patients with different genotypes of CD14. On day 21 after burn, CD14 mRNA expression and sCD14 levels were significantly higher in TT homozygotes than in CC genotypes (1.33+/-0.36 microg/ml vs. 0.75+/-0.28 microg/ml and 16.1+/-4.6 microg/ml vs. 9.7+/-3.4 microg/ml, P<0.05), and these values were also higher in non-survivors than in survivors (1.32+/-0.40 microg/ml vs. 0.87+/-0.32 microg/ml and 14.8+/-4.5 microg/ml vs. 11.1+/-4.8 microg/ml, P<0.05). In addition, TNF-alpha mRNA and protein levels were significantly lower in both CC homozygotes and survivors than in TT genotypes or non-survivors during the 28-day observation period (P<0.05). However, TT genotype did not impart an increased risk for burn mortality in this small study. In conclusion, CD14-159 C/T polymorphism might be associated with the kinetics and extent of CD14 expression as well as its release, and it was also related to TNF-alpha expression. However, this study did not confirm CD14-159 C/T polymorphism was associated with the outcome of extensive burns.     

Changes in CD14 expression on monocytes and changes in serum soluble CD14 level during hemodialysis

Background. CD14 is a receptor of lipopolysaccharide (LPS) and LPS binding protein (LBP) complex expressed on monocytes, and changes in the cell surface CD14 expression are thought to be a marker of activation of these cells. CD14 is shed from the cell surface when monocytes are activated. In this study, we assessed the influence of dialyzer membrane material and dialysate purity on monocyte CD14 expression and serum soluble CD14 (sCD14) levels in chronic hemodialysis (HD) patients in vivo during HD. Methods. We measured LPS concentrations in dialysate at two institutions by limulus assay. From one institution where LPS was undetectable in dialysate over a 2-year period, we selected seven patients. In the first period of the study, they were treated with a regenerated cellulose (RC) dialyzer, and then they were treated with a polysulfone (PS) dialyzer. We named them the "RC group" and the "PS group", respectively. From the other institution, where dialysate was contaminated with LPS, we selected eight patients. They were treated with a PS dialyzer, and were named the "PS + LPS group". CD14 expression on monocytes and serum sCD14 concentrations were measured by flow cytometry analysis and enzyme-liked immunosorbent assay, respectively. Results. During HD, in the RC group, upregulation of CD14 expression across the dialyzer was greater than in the PS group. There was no significant variation in serum sCD14 levels during HD in the RC and PS groups, while in the PS + LPS group, serum sCD14 level on the venous side of the dialyzer was significantly increased at 30 and 180 min after the initiation of HD compared with the predialysis value, and at 30 and 180 min compared with the level on the arterial side of the dialyzer. These results suggest that the changes in CD14 expression reflected the effect of dialyzer membrane material, while the changes in serum sCD14 levels reflected the effect of LPS influx from the dialysate. Conclusion. Dialysate purity may be an important factor in preventing monocyte activation during HD. Received: April 13, 1999 / Accepted: July 16, 1999     

血清新喋呤与烧伤病人内毒素血症及脓毒症的关系

为探讨血清新喋呤与大面积烧伤后内毒素血症及脓毒症的关系,对35例烧伤面积大于30%(30%～98%)患者的血清新喋呤、血浆内毒素的变化进行了动态观察。结果表明。烧伤后第3天大多数患者新喋呤升高(P<0.05),但与烧伤面积无显著相关(P>0.05)。严重烧伤第2周以后内毒素血症患者新喋呤水平显著高于无内毒素血症思者(P<0.05～0.01)。同时,伤后第14,21天内毒素血症患者的循环内毒素与新喋呤呈显著正相关(第14天:r=0.368,P<0.05;第21天:r=0.439,P<0.01)。在整个监测期间,脓毒症患者(15例)血清新喋呤水平持续升高,伤后第14,28天与非脓毒症患者相比差异有显著或非常显著意义(P<0.05～0.01)。提示烧伤可引起血清新喋呤升高,但与烧伤面积无关。烧伤后内毒素血症对新喋呤的不断产生可能有显著影响。血清新喋呤的持续升高与严重烧伤后脓毒症的发生与发展密切相关。     

血清新喋呤与烧伤病人内毒素血症及脓毒症的关系

为探讨血清新喋呤与大面积烧伤后内毒素血症及脓毒症的关系，对３５例烧伤面积大于３０％（３０％～９８％）患者的血清新喋呤、血浆内毒素的变化进行了动态观察。结果表明，烧伤后第３天大多数患者新喋呤升高（Ｐ＜０．０５），但与烧伤面积无显著相关（Ｐ＞０．０５）。严重烧伤第２周以后内毒素血症患者新喋呤水平显著高于无内毒素血症患者（Ｐ＜０．０５～０．０１）。同时，伤后第１４，２１天内毒素血症患者的循环内毒素与新喋呤呈显著正相关（第１４天：ｒ＝０．３６８，Ｐ＜０．０５；第２１天：ｒ＝０．４３９，Ｐ＜０．０１）。在整个监测期间，脓毒症患者（１５例）血清新喋呤水平持续升高，伤后第１４，２８天与非脓毒症患者相比差异有显著或非常显著意义（Ｐ＜０．０５～０．０１）。提示烧伤可引起血清新喋呤升高，但与烧伤面积无关。烧伤后内毒素血症对新喋呤的不断产生可能有显著影响。血清新喋呤的持续升高与严重烧伤后脓毒症的发生与发展密切相关。     

烫伤合并金葡菌感染大鼠组织CD14 mRNA的改变

目的　探讨细菌脂多糖受体CD14在烫伤合并金黄色葡萄球菌 (金葡菌 )感染中的变化规律及其意义。　方法　采用大鼠 2 0 %总体表面积Ⅲ度烫伤合并金葡菌攻击造成脓毒症模型 ,动态检测心、肝、肺、肾等重要器官中CD14mRNA表达的改变 ,同时观察内毒素在动物循环及主要脏器内的分布特点。　结果　烫伤合并金葡菌脓毒症早期 ,各脏器内毒素含量即明显高于正常对照组 ,并于2～ 6h达峰值 ,其中以肝、肺组织内毒素水平升高幅度最为显著 (P <0 .0 5 )。而血浆内毒素水平亦于伤后 2h显著高于正常对照组 (分别为 0 .30 5 6EU/ml和 0 .12 5 0EU/ml,P <0 .0 5 )。与此同时 ,小肠组织中二胺氧化酶的活性明显降低 (P <0 .0 5 )。烫伤合并金葡菌感染后 ,各组织CD14mRNA的表达亦呈不同程度升高 (P <0 .0 5 ) ,其中肺脏改变尤为显著 ,伤后 6、2 4h肺脏CD14mRNA表达分别为正常对照组的 1.80和 1.81倍。　结论　烫伤合并金葡菌攻击可导致内毒素移位和组织CD14mR NA表达不同程度升高 ,CD14基因表达的上调可能与移位内毒素的刺激作用有关。