严重烧伤早期切痂对可溶性脂多糖受体CD14表达和肝损害的影响

目的研究休克期切痂对严重烧伤后可溶性脂多糖受体CD14(sCD14)和肝损害的影响,并探索其可能的内在机制.方法收集休克期切痂组和非休克期切痂组的大面积烧伤病人血标本,检测内毒素(LPS)、sCD14和肿瘤坏死因子(TNF-α)含量,同时测定血清谷丙转氨酶(ALT)和谷草转氨酶(AST)的水平.结果休克期切痂组的上述指标均明显低于非休克期切痂组(P＜0.05),sCD14和各项指标呈正相关.结论严重烧伤后LPS、sCD14和TNF-α途径是并发肝功能损害的一条重要途径,休克期切痂可能通过该途径在一定程度上控制早期主要炎症介质的释放,降低烧伤后肝损害的程度.     

肺挫伤患者血清可溶性白细胞分化抗原14、可溶性E选择素、肿瘤坏死因子α水平的变化及意义

目的：观察肺挫伤患者血清可溶性白细胞分化抗原14（soluble CD14,sCD14）、可溶性E选择素（soluble E-selectin,sE-selectin）、肿瘤坏死因子α（tumor necrosis factor-alpha,TNF-α）水平的动态变化及其与伤后肺损伤发展的关系。方法：用酶联免疫吸附试验（ELISA）法检测35例肺挫伤患者伤后6h、12h、1d、3d、5d、7d血清sCD14、sE-selectin、TNF-α含量及30例健康成人血清中以上3种因子的含量,比较血清中sCD14、sE-selectin、TNF-α含量在肺挫伤患者和健康成人中的差异以及sCD14、sE-selectin、TNF-α与伤后急性肺损伤（ALI）和急性呼吸窘迫综合征（ARDS）的关系。结果：肺挫伤患者伤后6h血清sCD14、sE-selectin、TNF-α含量即显著升高,且sCD14s、E-selectin含量与ALI和ARDS的发生关系密切,伤后血清sCD14、sE-selectin、TNF-α含量呈显著正相关。结论：血清sCD14、sE-selectin、TNF-α与肺挫伤的发生发展关系密切。     

创伤性多器官功能不全综合征患者血浆sTNFR-P55和TNFα比值变化临床意义

目的探讨血浆可溶性肿瘤坏死因子受体P55(sTNFR-P55)与肿瘤坏死因子α(TNFα)浓度比值(sT-NFR-P55/TNFα)变化在创伤患者多器官功能不全综合征(MODS)中的诊断价值.方法测定正常对照组、创伤MODS生存组和MODS死亡组血浆sTNFR-P55和TNFα的浓度,观察血浆sTNFR-P55/TNFα比值变化及主要器官功能受损指标和预后的关系.结果创伤MODS各组患者sTNFR-P55/TNFα比值明显低于正常对照组(P＜0.01),且MODS死亡组低于MODS生存组(P＜0.01).创伤MODS患者sTNFR-P55/TNFα与主要器官功能受损指标呈明显负相关.结论创伤MODS患者血sTNFR-P55浓度升高幅度不如TNFα.sTNFR-P55/TNFα比值可作为判断MODS患者器官功能不全程度及预后转归的预警指标.     

创伤性多器官功能不全综合征患者血浆sTNFR-P55和TNFα比值变化临床意义

目的　探讨血浆可溶性肿瘤坏死因子受体P55(sTNFR P55)与肿瘤坏死因子α(TNFα)浓度比值(sT NFR P55/TNFα)变化在创伤患者多器官功能不全综合征(MODS)中的诊断价值。方法　测定正常对照组、创伤 MODS生存组和MODS死亡组血浆sTNFR P55和TNFα的浓度,观察血浆sTNFR P55/TNFα比值变化及主要器 官功能受损指标和预后的关系。结果　创伤MODS各组患者sTNFR P55/TNFα比值明显低于正常对照组(P< 0.01),且MODS死亡组低于MODS生存组(P<0.01)。创伤MODS患者sTNFR P55/TNFα与主要器官功能受 损指标呈明显负相关。结论　创伤MODS患者血sTNFR P55浓度升高幅度不如TNFα。sTNFR P55/TNFα比值 可作为判断MODS患者器官功能不全程度及预后转归的预警指标。     

2型糖尿病肿瘤坏死因子-α及其可溶性受体P55水平变化

目的探讨2型糖尿病（T2DM）血浆肿瘤坏死因子α（TNF-α）及其可溶性肿瘤坏死因子受体P55（sTNFR—P55）水平变化特点及临床意义。方法严格选取95例T2DM患者及50例健康人为研究对象,血浆TNF—α和sTNFR—P55均采用酶联免疫吸附试验（ELISA）测定。结果T2DM患者血浆TNF—α及sTNFR-P55水平均显著高于健康对照组（P〈0．01）,且血浆TNF—α与sTNFR-P55水平具有相关性（r=0．7015、P〈0．01）。结论TNF-α及sTNFR—P55与T2DM患者的炎性反应有关。检测T2DM患者血浆TNF-α和sTNFRP55水平,有利于观察病情变化及指导治疗。     

2型糖尿病患者血浆肿瘤坏死因子-α及其可溶性受体P55水平变化

目的探讨2型糖尿病（T2DM）血浆肿瘤坏死因子-α（TNF-α）及其可溶性肿瘤坏死因子受体P55（sTNFR—P55）水平变化特点及临床意义。方法严格选取95例T2DM患者；K50名正常人为研究对象，血浆TNF—α和sTNFR—P55均采用酶联免疫吸附实验（ELISA）测定。结果T2DM患者血浆TNF—α及sTNFR-P55水平均显著高于正常对照组（P〈0．01），且血浆TNF—α与sTNFR—P55水平具有相关性（r=0．7015，P％0．01）。结论TNF—α及sTNFR—P55与T2DM患者的炎性反应有关。检测T2DM患者血浆TNF—α和sTNFR—P55水平，有利于观察病情变化及指导治疗。     

可溶性肿瘤坏死因子受体p75检测在X线照射大肠癌细胞株Lovo细胞中的意义

目的:研究X线照射在体外培养人大肠癌细胞株Lovo后,检测培养上清中可溶性肿瘤坏死因子受体p75(sTNFR-p75)水平变化情况.方法:ELISA法检测培养上清中sTNFR-p75水平,流式细胞仪分析细胞凋亡率,光镜及透射电镜观察细胞形态变化.结果:X线照射后大肠癌Lovo细胞上清sTNFR-p75水平显著低于照射前sTNFR-p75水平(P＜0.01);光镜观察发现细胞体积缩小,变圆,核缩小,透射电镜观察发现部分细胞核染色质边集,染色质发生固缩.部分细胞核内染色质凝聚,核膜孔消失,核膜呈波纹状皱缩,形成凋亡小体,内含有退变的细胞器.流式细胞仪分析细胞凋亡率显著增高(P＜0.05).结论:X线照射可减少体外培养的大肠癌细胞膜上的sTNFR-p75脱落到上清中,从而提高肿瘤坏死因子(TNF)与癌细胞的结合率.进而增强照射后TNF对肿瘤的诱导凋亡作用.     

创伤失血性多器官功能不全综合征患者血浆可溶性白细胞分化抗原14相对内毒素变化及其临床价值

多器官功能不全综合征(MODS)为同时或相继发生2个以上器官功能不全或衰竭。可溶性白细胞分化抗原14(sCD14 )是存在于人和动物血液和体液中的游离内毒素(LPS)缺陷受体,在严重创伤MODS引发全身炎症反应中,LPS和sCD14均明显升高[1] 。但国内外对二者间比值变化的临床意义的报道则鲜见。基于LPS和sCD14以量的关系结合,我们以创伤失血性MODS患者为研究对象,观察MODS生存组和死亡组sCD14与LPS浓度比值(sCD14 /LPS)变化及其与主要器官功能受损指标的相关性,旨在探讨sCD14与LPS变化在创伤MODS发生发展中的临床意义,为临床诊断和…     

还元注射液对出血性脑卒中患者肿瘤坏死因子－α 可溶性白介素－2受体的影响

目的观察还元注射液对急性出血性脑卒中患者肿瘤坏死因子－α(TNF－α)、可溶性白介素－2受体(sIL－2R)的影响。方法急性出血性脑卒中患者60例，随机分为还元液治疗组和甘露醇治疗对照组。观察给药前后临床神经功能缺损积分、CT、TNF－α和sIL－2R等参数的变化。结果出血性脑卒中患者自体脑出血模型动物血中TNF－α、sIL－2R明显上升，还元液可在一定程度上抑制这种改变。结论还元注射液对急性出血性脑卒中患者TNF－α、sIL－2R的影响可能是其治疗作用机理之一。     

肿瘤坏死因子及其可溶性受体对狼疮活动的价值

目的 探讨肿瘤坏死因子(TNFα)和可溶性肿瘤坏死因子受体(sTNFR)对系统性红斑狼疮(SLE)疾病活动的价值。方法 分别采用狼疮活动标准记分(LACC)和SLE疾病活动指数(SLEDAI)判断狼疮活动和活动程度，采用双抗夹心ELISA法分别检测35例活动、25例稳定SLE和20例对照血浆TNFα、sTNFRl和sTNFR2水平。结果 SLE患者血浆sTNFR水平显著高于正常对照组。狼疮活动、稳定及对照sTNFR水平差异显著；SLE活动期sTNFRl、sTNFR2升高者显著多于稳定期。sTNF2水平与SLEDAI显著正相关(r＝0．40，P＜0．01)；sTNFR1与sTNFR2显著正相关(r＝0．62，P＜0．001)。结论 SLE患者血浆sTNFRl、sTNFR2均升高，狼疮活动时sTNFRl、sTNFR2升高的患者明显增多，可作为判断狼疮活动的指标。     

sCD14和sTNFR-P55治疗严重烫伤小鼠胰岛素抵抗和胰岛素分泌障碍实验研究

目的观察可溶性白细胞分化抗原14(sCD14)和可溶性肿瘤坏死因子受体-p55(sTNFR-P55)对严重烫伤小鼠胰岛素抵抗、胰岛素分泌以及糖代谢障碍的治疗效果。方法单项和联合静脉注射sCD14和sTNFR-P55给严重烫伤小鼠,观察不同治疗组及其对照组的死亡率、稳态模式胰岛素抵抗指数(HOMA-IR)和胰岛素分泌指数(HOMA-β)以及空腹血糖和乳酸含量变化。结果烫伤小鼠各治疗组HOMA-IR指数、空腹血糖和乳酸含量明显低于对照组,且联合治疗组低于单项治疗组。烫伤小鼠各治疗组HOMA-β指数明显高于对照组(P<0.01),且联合治疗组高于单项治疗组(P<0.01)。sCD14和sTNFR-P55联合治疗组烫伤小鼠死亡率明显低于治疗对照组(P<0.05),但其它治疗组与治疗对照组无显著差异(P>0.05)。结论静脉注射sCD14和sTNFR-P55能够有效改善严重烫伤小鼠胰岛素抵抗和胰岛素相对血糖分泌功能下降以及糖代谢障碍,且联合治疗效果优于单项治疗。     

Soluble CD14 participates in the response of cells to lipopolysaccharide

CD14 is a 55-kD protein found both as a glycosylphosphatidyl inositol-linked protein on the surface of mononuclear phagocytes and as a soluble protein in the blood. CD14 on the cell membrane (mCD14) has been shown to serve as a receptor for complexes of lipopolysaccharide (LPS) with LPS binding protein, but a function for soluble CD14 (sCD14) has not been described. Here we show that sCD14 enables responses to LPS by cells that do not express CD14. We have examined induction of endothelial-leukocyte adhesion molecule 1 expression by human umbilical vein endothelial cells, interleukin 6 secretion by U373 astrocytoma cells, and cytotoxicity of bovine endothelial cells. None of these cell types express mCD14, yet all respond to LPS in a serum-dependent fashion, and all responses are completely blocked by anti-CD14 antibodies. Immunodepletion of sCD14 from serum prevents responses to LPS, and the responses are restored by addition of sCD14. These studies suggest that a surface anchor is not needed for the function of CD14 and further imply that sCD14 must bind to additional proteins on the cell surface to associate with the cell and transduce a signal. They also indicate that sCD14 may have an important role in potentiating responses to LPS in cells lacking mCD14.     

Lipopolysaccharide binding protein and soluble CD14 catalyze exchange of phospholipids.

Lipopolysaccharide binding protein (LBP) is a plasma protein known to facilitate the diffusion of bacterial LPS (endotoxin). LBP catalyzes movement of LPS monomers from LPS aggregates to HDL particles, to phospholipid bilayers, and to a binding site on a second plasma protein, soluble CD14 (sCD14). sCD14 can hasten transfer by receiving an LPS monomer from an LPS aggregate, and then surrendering it to an HDL particle, thus acting as a soluble "shuttle" for an insoluble lipid. Here we show that LBP and sCD14 shuttle not only LPS, but also phospholipids. Phosphatidylinositol (PI), phosphatidylcholine, and a fluorescently labeled derivative of phosphatidylethanolamine (R-PE) are each transferred by LBP from membranes to HDL particles. The transfer could be observed using recombinant LBP and sCD14 or whole human plasma, and the plasma-mediated transfer of PI could be blocked by anti-LBP and partially inhibited by anti-CD14. sCD14 appears to act as a soluble shuttle for phospholipids since direct binding of PI and R-PE to sCD14 was observed and because addition of sCD14 accelerated transfer of these lipids. These studies define a new function for LBP and sCD14 and describe a novel mechanism for the transfer of phospholipids in blood. In further studies, we show evidence suggesting that LBP transfers LPS and phospholipids by reciprocal exchange: LBP-catalyzed binding of R-PE to LPS x sCD14 complexes was accompanied by the exit of LPS from sCD14, and LBP-catalyzed binding of R-PE to sCD14 was accelerated by prior binding of LPS to sCD14. Binding of one lipid is thus functionally coupled with the release of a second. These results suggest that LBP acts as a lipid exchange protein.     

冠心病患者外周血中sCD14水平的研究

目的分析冠心病患者外周血中sCD14的水平及其与单核细胞数量的相关关系,探讨sCD14和动脉粥样硬化之间的关系。方法ELISA法测定269例冠心病患者和24例健康人血清中的sCD14水平,血细胞分析仪检测受试者抗凝全血中单核细胞的数目和比例。结果冠心病患者与健康对照组sCD14分别为:(3 011.4±1431.4)ng/L和(1 943.8±510.2)ng/L,二者结果有显著差异(t=3.7259,P=0.002);冠心病患者血清sCD14水平(Y)与血单核细胞数量(X)和比例(X)均无相关关系,直线回归方程分别为:Y=2 969.8-95.6X,r=-0.026,P=0.668;Y=2 763.6+1581.8X,r=0.060,P=0.329。结论冠心病患者血清sCD14水平显著性增高与血单核细胞数量和比例均无相关关系,可能与单核细胞的质量改变有关。     

Soluble CD14 acts as a shuttle in the neutralization of lipopolysaccharide (LPS) by LPS-binding protein and reconstituted high density lipoprotein

We have recently shown that lipopolysaccharide (LPS)-binding protein (LBP) is a lipid transfer protein that catalyzes two distinct reactions: movement of bacterial LPS (endotoxin) from LPS micelles to soluble CD14 (sCD14) and movement of LPS from micelles to reconstituted high density lipoprotein (R-HDL) particles. Here we show that LBP facilitates a third lipid transfer reaction: movement of LPS from LPS- sCD14 complexes to R-HDL particles. This action of LBP is catalytic, with one molecule of LBP enabling the movement of multiple LPS molecules into R-HDL. LBP-catalyzed movement of LPS from LPS-sCD14 complexes to R-HDL neutralizes the capacity of LPS to stimulate polymorphonuclear leukocytes. Our findings show that LPS may be transferred to R-HDL either by the direct action of LBP or by a two- step reaction in which LPS is first transferred to sCD14 and subsequently to R-HDL. We have observed that the two-step pathway of LPS transfer to R-HDL is strongly favored over direct transfer. Neutralization of LPS by LBP and R-HDL was accelerated more than 30- fold by addition of sCD14. Several observations suggest that sCD14 accelerates this reaction by serving as a shuttle for LPS: addition of LBP and sCD14 to LPS micelles resulted in LPS-sCD14 complexes that could diffuse through a 100-kD cutoff filter; LPS-sCD14 complexes appeared transiently during movement of LPS to R-HDL facilitated by purified LBP; and sCD14 could facilitate transfer of LPS to R-HDL without becoming part of the final LPS-R-HDL complex. Complexes of LPS and sCD14 were formed transiently when LPS was incubated in plasma, suggesting that these complexes may play a role as intermediates in the neutralization of LPS under physiological conditions. These findings detail a new activity for sCD14 and suggest a novel mechanism for lipid transfer by LBP.     

Human hepatocytes secrete soluble CD14, a process not directly influenced by HBV and HCV infection

BACKGROUND: Chronic hepatitis B (HBV) and hepatitis C (HCV) patients have elevated plasma levels of soluble CD14 (sCD14). We examined whether human hepatocytes produce sCD14 in vivo, and whether HBV or HCV infections influence this chimeric production. METHODS: uPA-SCID mice were transplanted with primary human hepatocytes and some animals were subsequently infected with HBV or HCV. Plasma from these mice was analyzed for the presence of human sCD14. The liver was examined via immunohistochemistry. RESULTS: A soluble form of human CD14 could be detected in the plasma from successfully transplanted mice, while it was completely absent in non-transplanted control animals. The isoform of this human sCD14 corresponded with the most abundant isoform found in human plasma. CD14 levels in circulation were not significantly different between non-infected, HBV infected and HCV infected animals. CONCLUSIONS: Our data indicate that human hepatocytes produce sCD14 in vivo, and that liver cells might be the major source of sCD14 in normal human plasma. In addition we demonstrate that HBV and HCV infections have no direct influence on the production of sCD14 by human hepatocytes in this chimeric model.     

Evaluation of a newly identified soluble CD14 subtype as a marker for sepsis

CD14, a high-affinity receptor for lipopolysaccharide (LPS), is a glycoprotein expressed on the surface membranes of monocytes/macrophages. We have identified a previously unknown form of soluble CD14, named soluble CD14 subtype (sCD14-ST), that is increased in patients with sepsis. To measure sCD14-ST concentrations in plasma, we prepared anti-sCD14-ST antibodies and developed an enzyme immunoassay (EIA) for this soluble form of CD14. With this assay, quantitative measurements are available within 4 h, and we compared the levels of sCD14-ST in plasma from normal subjects (healthy controls), patients with systemic inflammatory response syndrome (SIRS), and sepsis patients. The level of sCD14-ST in subjects with sepsis was much higher than the levels in subjects with SIRS and the healthy controls. Additionally, when a subject's sCD14-ST level was used as a diagnostic marker for sepsis, the area under the receiver operating characteristic (ROC) curve was 0.817, thereby demonstrating that elevated sCD14-ST levels were a better marker for sepsis than the other molecular markers we tested. sCD14-ST levels also correlated with procalcitonin (PCT) levels and with sequential organ failure assessment (SOFA) scores. Finally, changes in sCD14-ST concentration correlated with the severity of sepsis. Taken together, these results indicate that sCD14-ST is a useful marker for the rapid diagnosis of sepsis and for monitoring the severity of the disease.     

重症脓毒症患者血清脂多糖结合蛋白及其受体变化的临床研究

目的观察重症脓毒症患者血清脂多糖结合蛋白（LBP）及其受体CD14（CD14）的变化，探讨两者与重症脓毒症严重程度及预后的关系。方法采用酶联免疫吸附法（ELISA）测定41例重症脓毒症患者发病后不同时间血清LBP和可溶性CD14（sCD14）浓度以及发病后24h内血清前降钙素（PCT）和内毒索（LPS）浓度，并且进行相关性分析。结果重症脓毒症患者发病后各时间点血清LBP和sCD14水平较正常对照均显著升高（P均〈0．01）；死亡组患者各时间点血清LBP水平与存活组间差异均无显著性（P均〉0．05），而sCD14则均较存活组显著升高（P均〈0．05）。血清LBP水平与PCT、急性生理学与慢性健康状况评分Ⅱ（APACHEⅡ）无显著相关性，与LPS呈显著正相关；而sCD14与PCT、APACHEⅡ评分呈显著正相关，与LPS无显著相关性。结论重症脓毒症患者血清LBP和sCD14浓度均显著升高，LBP仅反映机体急性炎症反应，而不能作为判断疾病严重程度及预后的指标；而sCD14升高程度可提示预后，在一定程度上反映脓毒症的严重程度。     

Plasma CD14 decreases monocyte responses to LPS by transferring cell-bound LPS to plasma lipoproteins

CD14, a myeloid cell-surface receptor and soluble plasma protein, binds LPS and other microbial molecules and initiates the innate immune response to bacterial invasion. The blood concentration of soluble CD14 (sCD14) increases during the systemic response to infection. Although high sCD14 blood levels have correlated with increased risk of dying from severe sepsis, sCD14 can diminish cell responses to LPS. We show here that in human serum, sCD14 increases the rate at which cell-bound LPS is released from the monocyte surface and binds to plasma lipoproteins. This enhanced rate of LPS efflux is associated with a significant reduction in the ability of monocytes to produce cytokines in response to LPS. Serum from septic patients reduced the LPS-monocyte interaction by as much as tenfold, and depletion of sCD14 from the serum restored LPS-monocyte binding and release kinetics to near normal levels. In serum from septic patients, monocyte-bound LPS also moved more rapidly into lipoproteins, which completely neutralized the biologic activity of the LPS that bound to them. In human plasma, sCD14 thus diminishes monocyte responses to LPS by transferring cell-bound LPS to lipoproteins. Stress-related increases in plasma sCD14 levels may help prevent inflammatory responses within the blood.