肝硬化患者血中LPS、TNF—α、sCD14的变化及临床意义

目的：探讨肠源性内毒素血症在肝硬化发生发展中的临床意义。方法：应用基质显色法及酶联免疫吸附试验，检测肝硬化患血中内毒素，肿瘤坏死因子-α，可溶性CD14水平。结果：随着肝硬化程度的加重，血中内毒素，肿瘤坏死因子-α，可溶性CD14水平明显升高，死亡内毒素，肿瘤坏死因子-α，可溶性CD14水平明显高于存活。结论：肝硬化胖肠源性内毒素血症患体内存在高水平的内毒素和可溶性CD14，在内毒素存在的情况下，通过肿瘤坏死因子-α，加剧肝细胞损伤。     

静脉注射sCD14对严重烫伤内毒素血症小鼠血清C-反应蛋白和降钙素原以及血糖水平的影响

目的观察可溶性白细胞分化抗原14(soluble Cluster of Differentiation,sCD14)对严重烫伤内毒素血症模型小鼠血清C-反应蛋白和降钙素原以及血糖的影响。方法 60只健康昆明小鼠随机分为烫伤sCD14治疗组(n=20),烫伤治疗对照组(n=20)和正常对照组(n=20),观察3组小鼠血清C-反应蛋白和降钙素原以及血糖水平变化。结果烫伤sCD14治疗组小鼠血清C-反应蛋白和降钙素原以及血糖水平均明显低于烫伤治疗对照组(P<0.01),但仍高于正常对照组(P<0.01)。结论静脉sCD14治疗可降低严重烫伤内毒素血症机体炎症反应,改善高血糖,提示sCD14可作为改善严重创伤内毒素血症炎症反应和高血糖的有效药物。     

肠源性内毒素血症对阿尔茨海默病发病的影响

目的 探讨肠源性内毒素血症对阿尔茨海默病（AD）患者的作用及其可能机制.方法 通过神经心理学测验检测入组患者认知功能,分为AD组和健康对照者各40例,采用显色基质鲎试剂法检测脂多糖（LPS）水平,ELISA法检测TNF-α和Tau蛋白水平.结果 AD患者MMSE、ADAS-Cog分数与对照组比较,MMSE分数明显低于对照组（t=16.473,P＜0.001）,ADAS-Cog分数明显高于对照组（t=18.067,P＜0.001）;LPS、TNF-α及Tau蛋白水平明显高于对照组（t=5.317,P＜0.001;t =5.014,P＜0.001;t=17.393,P＜0.001）.结论 AD患者存在肠源性内毒素血症,并可能在AD发病过程中发挥重要作用.     

微生态制剂对肝硬化患者肠源性内毒素血症的治疗作用

目的:探讨微生态制剂对肝硬化患者肠源性内毒素血症的治疗作用。方法:将58例肝硬化具有肠源性内毒素血症的患者分为观察组和对照组,两组间在性别、年龄、病程、临床特点及血清内毒素水平方面无统计学差异。观察组在对照组的基础上加用微生态制剂整肠生治疗,治疗2周后再次检测两组的血清内毒素水平,并进行统计学分析。结果:治疗前两组血清内毒素水平分别为(69.33±27.12)pg/ml和(67.79±26.07)pg/ml(P0.05),其间无统计学差异;治疗后两组血清内毒素水平分别为(32.04±14.76)pg/ml和(58.97±25.11)pg/ml(P0.05),差异有统计学意义。结论:微生态制剂对肝硬化患者肠源性内毒素血症有治疗作用。     

口服大黄对肝硬化腹水患者血浆内毒素水平的影响及临床意义

[目的]探讨中药大黄对肝硬化腹水患者血浆内毒素水平的影响及其控制肠源性内毒素血症的临床疗效.[方法]对86例肝硬化腹水患者采用大黄(治疗组)和贝飞达(对照组)治疗,比较2组4周内血浆内毒素水平的变化情况.[结果]两组在降低血浆内毒素水平,控制肠源性内毒素血症上均有作用,但治疗组效果居优(P＜0.05).[结论]大黄在降低血浆内毒素水平,治疗肠源性内毒素血症上优于贝飞达,肠源性内毒素血症控制后腹水消退加快,并发症发生率降低.     

热应激对肝硬化内毒素血症大鼠HSP72表达及致炎细胞因子分泌的影响

目的:探讨热应激诱导肝硬化内毒素血症大鼠热休克蛋白72(HSP72)表达及对致炎细胞因子TNF-α分泌的影响.方法:CCl<,4>诱导的肝硬化SD大鼠及正常饮食的SD大鼠先后给予热应激处理及腹膜腔内注入脂多糖(LPS),另外两组肝硬化SD大鼠及正常饮食SD大鼠仅腹腔内注入LPS作为对照.EUSA检测并比较各组大鼠血浆内毒素、HSP72、TNF-α的含量,RT-PCR检测肝组织HS72 mRNA表达,评价HSP72表达对TNF-α分泌的影响.结果:外源给予LPS能加重肝硬化大鼠内毒素血症.热处理及LPS注入后,肝硬化大鼠及正常大鼠血浆及肝组织HSP72表达均较未热处理组增强,而未热处理的两组大鼠LPS注入后血浆TNF-α含量明显高于热处理组.结论:热应激能促进肝硬化内毒素血症大鼠肝组织及血浆HSP72表达,而抑制TNF-α分泌.     

原发性硬化性胆管炎患者血清LBP和sCD14水平及与预后的关系

[目的]探讨原发性硬化性胆管炎(PSC)患者血清脂多糖结合蛋白(LBP)和可溶性CD14(sCD14)水平及与预后的关系。[方法]选取2009年1月至2015年1月本院收治的87例PSC患者作为研究对象,采用酶联法检测患者血清LBP及sCD14水平,分析两者与PSC患者预后的关系。[结果]87例PSC患者预后不良发生率为40.23%(35/87).35例预后不良患者中肝衰竭11例(12.64%),进展为肝胆管癌8例(9.20%),继发性胆汁性肝硬化12例(13.79%),死亡4例(4.60%)。预后不良组患者血清LBP及sCD14水平均明显高于预后良好组。且差异均有显著性(P<0.01)。Logistic多因素回归分析显示血清白蛋白(ALB)、总胆红素(TBIL)、血清白介素-8(IL-8)、LBP及sCD14与PSC患者预后密切相关(P<0.05)。模型C评估PSC患者预后的AUC高于模型A及模型B,且差异均有显著性(P<0.05)。[结论]血清LBP及sCD14与PSC患者预后相关,检测血清LBP及sCD14有助于评估PSC患者预后情况。     

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。     

感染性疾病患者血浆内毒素和血清hs-CRP、TNF-α水平检测的临床意义

目的探讨感染性疾病患者血浆内毒素、超敏C反应蛋白(hs-CRP)和肿瘤坏死因子(TNF-α)水平的变化及意义。方法应用动态浊度法、免疫比浊法和放射免疫分析法对174例感染性疾病患者进行了血浆内毒素和血清hs-CRP、TNF-α水平检测,并与30例健康人作比较。结果感染性疾病患者血浆内毒素和血清hs-CRP、TNF-α水平均高于健康对照组,差异有统计学意义(P<0.01),无论是革兰阴性杆菌,还是阳性杆菌,或是血培养阴性的结果,其血浆内毒素、hs-CRP、TNF-α水平均显著地高于健康对照者(P<0.01)。结论内毒素血症是感染性疾病发病的重要因素之一,联合检测hs-CRP、TNF-α有助于疾病的诊断和治疗,有十分重要的临床意义。     

肝硬化患者血清二胺氧化酶和内毒素检测及临床意义

目的:探讨肝硬化患者血清二胺氧化酶(diamine oxidase,DAO)和内毒素(endotoxin,ETX)水平的变化及临床意义.方法:52例肝硬化患者(肝硬化组),采用分光光度法和鲎试剂偶氮显色基质法检测治疗前、后外周血清中DAO和ETX水平,选择同期15名健康体检者作为对照组,并进行比较.结果:肝硬化组治疗前DAO水平高于时照组(P<0.01).肝硬化组Child-pugh A级与B级患者治疗前ETX水平比较差异无统计学意义(P>0.05),余各分级间DAO,ETX水平差异均有统计学意义(P<0.01);治疗后C级患者DAO水平升高(P<0.05),余各分级DAO,ETX水平均明显降低(P<0.01).合并腹腔积液患者DAO和ETX水平明显高于无腹腔积液患者(P<0.01);肝硬化患者DAO与ETX水平呈正相关(γ=0.28,P<0.05).结论:DAO可作为判断肠黏膜损伤的一项较敏感指标;肝硬化患者肠黏膜通透性升高,可能是导致内毒素血症发生的因素之一,且与肝硬化病情轻重有关.     

LBP and sCD14 patterns in total hip replacement surgery performed during combined spinal/epidural anaesthesia

Abstract

Background. Danger patterns and pattern recognition receptors have been targets in the investigation and treatment of systemic inflammatory response syndrome and sepsis. Lipopolysaccharide (LPS)-binding protein (LBP) presents LPS and gram-positive bacterial cell wall products to the receptors TLR4/MD-2 and TLR2, respectively. Low concentrations of LBP stimulate responses to LPS and peptidoglycan, whereas higher concentrations inhibit these responses. Soluble CD14 (sCD14) presents the LBP-LPS complex to CD14-negative cells, and it modulates the biological activity of circulating LPS. In this study, we aimed to elucidate the physiological reactions to LBP and sCD14 after total hip replacement surgery during spinal/epidural anaesthesia. Methods. Seven patients with coxarthrosis were operated upon with a total hip replacement, which is a defined trauma to bone and muscles in conjunction with a certain amount of blood loss. Venous blood samples were taken before the operation and at 1 h, 3 days and 6 days after surgery. LBP and sCD14 were measured by conventional ELISA. To correct for hemodilution, each parameter was adjusted for hematocrit. A panel of cytokines was measured using Luminex technology to evaluate the trauma reaction. Results. IL-6 levels peaked 24 h after the operation, whereas IL-1β and IL-10 levels remained unchanged. Systemic levels of LBP were increased 24 h after surgery, whereas sCD14 remained steady. However, the dilution-corrected sCD14 values increased significantly, and the levels of both LBP and sCD14 peaked at day 3 after surgery. Conclusion. Aseptic trauma primes the innate immune system for the posttraumatic release of LBP and sCD14.     

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.     

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.     

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.     

杀菌通透性增加蛋白体外抑制革兰阴性细菌脂多糖激活血小板的作用

本研究旨在探索杀菌通透性增加蛋白(bactericidal permeability-increasing protein,BPI)能否抑制G-细菌脂多糖(LPS)激活血小板的作用。取10例健康体检者全血制备富含血小板血浆(PRP,1×108/ml)。实验分为4组:正常血小板组:不作任何处理;LPS组:LPS(10μg/ml)刺激6 h;BPI组:BPI(100μg/ml)处理1 h;BPI+LPS组:BPI(100μg/ml)预孵育1 h后,再接受LPS(10μg/ml)刺激6 h。应用流式细胞术(FCM)检测各组血小板膜Toll样受体-4(TLR-4)的表达,酶联免疫吸附测定法(ELISA)测定PRP上清中细胞因子释放水平,包括肿瘤坏死因子-α(TNF-α)和白介素-6(IL-6)。结果表明,与正常血小板组相比,LPS刺激血小板后血小板膜TLR-4表达及上清中TNF-α和IL-6浓度均明显增高(P<0.001)。在接受BPI预处理后,LPS刺激血小板表达TLR-4及TNF-α和IL-6的作用明显下降,但仍高于正常血小板组。BPI单独刺激血小板不引起血小板TLR-4表达增高及细胞因子水平改变。结论:BPI能够抑制LPS诱导的血小板活化。     

脂多糖刺激前后小鼠肺肝脾组织中Toll样等受体基因表达情况

目的了解脂多糖(LPS)对肺、肝、脾组织中与LPS信号转导有关的受体的基因表达情况的影响。方法LPS刺激前后取小鼠肺、肝、脾组织总RNA，采用逆转录-聚合酶链反应(RT-PCR)半定量测定脂多糖结合蛋白(LBP)、CD14、Toll样受体2(TLR2)、TLR4和肿瘤坏死因子-α(TNF-α)等基因的表达情况。结果正常组织中，肺有TLR2的表达，肝有TLR2、CD14、LBP的表达，脾有TLR2的表达；注射LPS后的5h内，肺组织TLR2、TLR4、CD14和TNF-α，肝组织TLR2、CD14、LPB和TNF-α，脾组织TLR2、TLR4、CD14和TNF-α的基因表达均呈逐渐增加趋势。结论正常情况下TLR2等基因在不同组织中的表达有差异，LPS刺激5h内各基因的表达提高，从而提高机体对LPS等病原体模式识别分子的反应性。     

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。     

Polymorphisms in the lipopolysaccharide-binding protein and bactericidal/permeability-increasing protein in patients with myocardial infarction.

Gram-negative bacterial infection, namely Chlamydia pneumoniae has been recently discussed as a risk factor for myocardial infarction. The lipopolysaccharide-binding protein (LBP) and the bactericidal/permeability-increasing protein (BPI) play a role in the processes leading to recognition and neutralisation of the Chlamydia pneumoniae and their endotoxins lipopolysaccharides (LPS). LPS interact with plasma LBP, and LBP-LPS complex activates monocytes/macrophages, which can influence the atherosclerotic process. BPI is cytotoxic for Gram-negative bacteria and BPI-LPS complexes do not activate monocytes. We have analysed the polymorphisms in the LBP gene (Gly98-->Cys; Pro436-->Leu) and BPI gene (Lys216-->Glu; PstI polymorphism in intron-5; G545-->C) in 313 patients after myocardial infarction (MI) and in 302 control individuals. Genotype frequencies in the LBP gene and BPI gene did not differ between MI patients and control individuals. Our findings suggest that LBP and BPI polymorphisms do not influence the risk of MI.     

乳酸杆菌水平与糖尿病患者糖脂及炎症指标相关性分析

目的探讨乳酸杆菌水平与糖尿病患者糖脂代谢及炎症指标的相关性。方法于2014年8月至2015年8月,以50例2型糖尿病患者(糖尿病组)和50例体检健康者(对照组)作为研究对象,采用荧光定量聚合酶链反应检测粪便标本乳酸杆菌水平,同时检测外周血空腹血糖(FBG)、总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDLC)、糖化血红蛋白(HbA1c)、脂多糖结合蛋白(LBP)、肿瘤坏死因子α(TNF-α)、可溶性白细胞分化抗原14(sCD14)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、白细胞介素10(IL-10)水平。结果糖尿病组粪便标本乳酸杆菌拷贝数,以及FBG、TC、TG、HDLC、HbA1c、LBP、TNF-α、IL-1β、IL-6水平明显高于对照组(P0.05),糖尿病组sCD14、IL-10水平明显低于对照组(P0.05)。乳酸杆菌拷贝数与LDL-C水平呈负相关(P0.05),其对数值与TNF-α、IL-6水平呈正相关(P0.05)。结论糖尿病患者体内乳酸杆菌水平明显高于健康者,乳酸杆菌水平与糖尿病患者部分血脂及炎症指标存在相关性。