体外循环手术期间精氨酸对红细胞作用的研究

目的 探讨体外循环(CPB)手术期间L-精氨酸对患者红细胞的影响.方法 择期在CPB下行瓣膜置换的风湿性心脏病患者30例,随机分为L-精氨酸组(L组)和对照组(C组),每组15例.于麻醉后切皮前,L组L-精氨酸按200 mg/kg的剂量加入5%葡萄糖注射液50 ml中,经颈内静脉用注射泵以99 ml/h的速度泵人体内.C组用等量5%葡萄糖注射液,用法同L组.分别于麻醉诱导后切皮前(基线值T1)、转流30 min(T2)、转流结束时(T3)、手术结束时(T4)、术后24 h(T5)五个不同时段分别抽取静脉血,测定血中游离血红蛋白(FHb)、丙二醛(MDA)、红细胞C3b受体花环率(RBC-C3bRR)、红细胞免疫复合物花环率(RBC-ICR).结果 (1)MDA、FHb、RBC-ICR:转机前,L、C组间无差异,两组转机30 min后的各标本含量均明显高于转机前(P<0.01),于CPB结束时各含量开始下降,转机30 min后L组含量又明显低于C组,组间差异有统计学意义(P<0.01).(2)RBC-C3bRR:转机前两组RBC-C3bRR无明显差异,转机30 min后两组RBC-C3bRR均明显低于转机前(P<0.01),于CPB结束时各含量开始回升,但转机30 min后L组RBC-C3bRR又明显高于C组,组间差异有统计学意义(P<0.01).结论 心内直视手术时CPB可导致红细胞损伤及其免疫功能下降.围手术期应用精氨酸对CPB中RBC及其免疫功能有较好的保护作用,并可促进CPB术后患者红细胞免疫功能的恢复.     

肝硬化患者低氧血症对红细胞免疫功能的影响

目的:探讨肝硬化患者低氧血症对红细胞免疫功能的影响。方法:采用红细胞酵母菌混合花环法检测48例肝硬化患者红细胞C3b受体花环率(RBC-C3bRR)和红细胞免疫复合物花环率(RBC-ICR);同时测定动脉血氧分压(PaO2)水平。结果:肝硬化患者RBC-C3bRR明显降低(P<0.01),RBC-ICR明显升高(P<0.01);PaO2降低,在肝性脑病(HE)组更明显。在动脉低氧血症时,RBC-C3bRR降低,RBC-ICR升高更加显著(P均<0.05)。结论:肝硬化患者红细胞免疫功能低下,在低氧血症时加重红细胞C3b受体的损害。     

乌司他丁在食道癌根治术中肺保护作用的研究

目的探讨乌司他丁对食道癌根治术所致肺损伤的保护作用。方法 40例限期行食道癌根治术患者,随机分为乌司他丁组(U组,n=20)和对照组(C组,n=20)。于手术开始前U组按1万U/kg静脉给予乌司他丁,C组给予等容量的生理盐水。于单肺通气开始时(T0)、单肺通气1h(T1)、双肺通气15min(T2)、术后24h(T3)各监测时间点抽取静脉血5ml,并于T0、T1、T2时间点收集支气管肺泡灌洗液(BALF)3ml。用酶联免疫吸附法(ELISA)法测定血浆中TNF-α、IL-6、IL-8、IL-10及BALF中IL-8的浓度。结果血浆中,两组患者T1、T2、T3时点的TNF-α、IL-6、IL-10及T2、T3时点的IL-8浓度均较T0明显升高(P<0.05);乌司他丁组TNF-α、IL-6、IL-8增高幅度较C组小,而IL-10增高幅度大于C组,差异均T1有统计学意义(P<0.05)。BALF中,C组T1、T2时间点、U组T2时间点IL-8浓度较T0均升高(P<0.05)。U组患者在T1、T2时间点IL-8浓度较C组明显降低(P<0.05)。结论乌司他丁可抑制血浆中TNF-α、IL-6、IL-8及BALF中IL-8浓度,并增加血浆中IL-10的浓度,从而减轻食道癌根治术引发的肺损伤。     

乌司他丁对婴幼儿体外循环炎性反应的影响

目的:探讨乌司他丁对婴幼儿体外循环围手术期炎性介质的影响。方法:先天性心脏病患者30例,随机分为乌司他丁组(U组)和对照组(C组),各15例。U组患者预充液中加入乌司他丁,对照组行常规体外循环(CPB),取不同时间点检测血清中细胞因子浓度。结果:在T2、T3、T4时点U组TNF-α、IL-2、IL-6增值较C组低,IL-10增值较C组高。结论:乌司他丁应用于婴幼儿先天性心脏病手术可减轻CPB炎性反应。     

促红细胞生成素对脐血红细胞C3bRR及ICR表达的影响

目的探讨促红细胞生成素(EPO)对脐血与成人外周血红细胞免疫功能表达的影响。方法无菌采集正常孕妇分娩时胎儿脐血及献血者外周血各28份。应用ELISA法检测EPO;应用红细胞C3b受体花环试验和红细胞免疫复合物花环试验检测红细胞C3b受体活性率(RBC-C3bRR)和红细胞粘附免疫复合物率(RBC-ICR)。同时考查不同浓度的EPO对脐血体外培养红细胞C3bRR、ICR表达的影响。结果脐血EPO、RBC-C3bRR及RBC-ICR与成人外周血比较差异有显著性意义(P<0.01);脐血体外培养,随着EPO浓度的增加,RBC-C3bRR活性逐渐增强,对RBC-ICR产生的抑制率逐渐减弱;当EPO浓度达40mIU/ml时,脐血体外培养各时间段的RBC-C3bRR、RBC-ICR与EPO为0mIU/ml时比较差异有显著性意义(P<0.01)。结论脐血EPO及红细胞C3bRRI、CR高于成人外周血。细胞因子EPO在脐血体外培养中,促进RBC-C3bRR的生成,抑制RBC-ICR的表达。     

慢性再生障碍性贫血患者T细胞及红细胞免疫功能的改变及意义

目的观察慢性再生障碍性贫血（CAA）患者的T淋巴细胞亚群（CD4，CD8，CD4／CD8）、γ-干扰素（IFN-γ）、肿瘤坏死因子-α（TNF-α）、白细胞介素-2（IL-2）以及红细胞C3b受体花环率（RBC-C3bRR）和红细胞免疫复合物（RBC-CIC）的变化，探讨机体免疫功能和CAA发病的关系。方法T淋巴细胞亚群（CD4，CD8）采用流式细胞仪进行检测，血清IFN-γ、TNF-α、IL-2的测定采用酶联免疫吸附法（ELISA），RBC-C3bRR和RBC-CIC测定采用红细胞酵母花环法。结果与对照组相比。CD4、CD4／CD8以及RBC-C3bRR显著降低（P〈0．05和P〈0．01）。而CD8和IFN-γ、TNF-α、IL-2水平则显著升高（P〈0．05和P〈0．01）。结论CAA患者表现多种免疫功能改变。监测CAA患者的白细胞及红细胞免疫活性有助于了解病情变化。     

慢性肾病患者红细胞免疫功能与脂质过氧化的关系

目的 研究慢性肾病患者红细胞免疫功能的变化及其与脂质过氧化的关系.方法 应用红细胞酵母花环法测定33例慢性肾病患者红细胞免疫功能和化学比色法测定血浆丙二醛(MDA)、超氧化物岐化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)含量,并与35例健康人比较.结果 慢性肾病患者红细胞免疫复合物(RBC-ICR)花环百分率和MDA水平明显升高(P＜0.01),而红细胞C3b受体(RBC-C3bRR)、SOD、GSH-PX明显低于正常(P＜0.01).相关分析显示,RBC-C3bRR 花环与MDA呈显著负相关(r=-0.4812,P＜0.05).RBC-ICR与MDA 呈正相关(r=0.5128,P＜0.05).结论 慢性肾病患者红细胞免疫功能的降低,与活性氧代谢紊乱密切相关.     

不同剂量乌司他丁对体外循环患者炎症反应及呼吸功能的影响

目的 研究不同剂量乌司他丁在体外循环(CPB)心脏直视手术中对肺损伤的保护作用机制.方法 采用前瞻性随机对照研究方法,选择42例心脏瓣膜置换术患者,随机分为对照组、乌司他丁8 kU/kg组(U1组)及乌司他丁12 kU/kg组(U2组),3组分别于麻醉诱导后术前(T1)及停止CPB 1 h(T2)、4 h(T3)、24 h(T4)取静脉血,检测血浆肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-10的浓度;同时测定动脉血气,对比观察氧合指数(PaO_2/FiO_2)及呼吸指数(RI).结果 ①3组患者T1时血浆TNF-α、IL-6和IL-10比较差异均无统计学意义(P均>0.05),T2～4时则均显著升高(P<0.05或P<0.01).与对照组比较,U1、U2组T2～4时血浆TNF-α、IL-6浓度降低,IL-10浓度升高,且U2组变化较U1组明显(P<0.05或P<0.01).②3组患者T1时PaO_2/FiO_2、RI比较差异均无统计学意义(P均>0.05),T2～4时则出现PaO_2/FiO_2减少、RI值增大(P<0.05或P<0.01).与对照组比较,U1、U2组T2～4时PaO_2/FiO_2增大、RI值减小,且U2组变化较U1组更为显著(P<0.05或P<0.01).结论 在CPB过程中,乌司他丁能明显抑制促炎因子TNF-α、IL-6的产生,上调抗炎因子IL-10的释放,减轻肺损伤,改善CPB术后肺功能,并呈剂量-效应关系.     

体外循环心内直视手术围术期乌司他丁对血浆S100β蛋白及TNF-α表达的影响

目的观察体外循环心内直视手术围术期乌司他丁对S100β蛋白及肿瘤坏死因子-α(TNF-α)表达的影响。方法30例择期心内直视手术患者随机分成两组：乌司他丁组(U组)和生理盐水对照组(C组),每组15例。U组于麻醉诱导后,体外循环前用微量泵输注溶于50 ml生理盐水的乌司他丁(1万U/kg),体外循环(CPB)开始时再将乌司他丁(1万U/kg)加入体外循环机预充液中。C组用等体积的生理盐水。分别于CPB开始后15 min(T_1)、CPB结束时(T_2)、CPB结束后2h(T_3)、24h(T_4)时点从颈内静脉采血,分别测定S100β蛋白和TNF-α的水平。结果与C组比较,T_1和T_4时点无统计学意义(P＜0．05),U组S100β和TNF-α含量T_2时点明显降低(P＜0．01);T_3时点也降低(P＜0．05)。两组T_2、T_3、T_4时S100β水平均高于T_1点(P＜0．01)。C组T_2、T_3时TNF-α水平明显高于T_1时点(P＜0．01);U组各时点TNF-α浓度之间差别无统计学意义(P＞0．05)。结论体外循环心内直视手术围术期乌司他丁可减少TNF-α炎性因子的产生,并降低血浆中脑损伤的生化标志物S100β蛋白的表达水平。     

体外循环中盐酸戊乙奎醚对肺损伤的保护作用

目的:研究盐酸戊乙奎醚时体外循环(CPB)炎症因子的影响及对肺损伤的保护作用.方法:24例择期首次行心脏瓣膜置换术患者,随机分为盐酸戊乙奎醚组(A组)和对照组(C组),每组12例.分别于诱导切皮前(T1)、转流30 min(T2)、停CPB 1 h(T3)、4 h(T4)、24 h(T5)抽取动脉血,测定红细胞压积和动脉血气分析,计算呼吸指数RI,测定血浆TNF-α、IL-6浓度.结果:两组间血浆IL-6、TNF-α水平和RI值在CPB前T1时差异无统计学意义(t≤1.82,P＞0.05).两组T2-5各时点TNF-α、IL-6均高于T1(F≥6.21,P＜0.05),A组T2-5各时点TNF-α、IL-6浓度明显低于C组同时点值(t≥2.41,P＜0.05),两组T3-T4时点RI均高于T1时点(F≥5.87,P＜0.05),A组中T3,T4时点RI较C组同时点明显降低(t≥2.38,P＜0.05).结论:盐酸戊乙奎醚能有效地抑制CPB心脏直视手术围术期炎症因子的释放,改善肺功能.     

外周血细胞因子水平与颈椎病病情严重程度的相关研究

目的：分析白细胞介素（IL）1、IL-6、IL-10、肿瘤坏死因子-α（TNF-α）、人转化生长因子（TGF）-β1、一氧化氮（NO）在颈椎病的表达及与病情严重程度的关系。方法选择2009年2月至2013年3月江苏省沭阳县人民医院治疗的96例颈椎病患者,根据MRI结果将患者分为颈椎间盘退行性改变组（48例）和颈椎间盘突出组（48例）,选取同期治疗的颈椎外伤患者48例为对照组。对3组患者IL-1、IL-10、IL-6、TNF-α、TGF-β1、NO在颈椎病的表达、与JOA评分关系和与膝跳反射之间的关系进行研究。结果椎间盘突出组6项检测水平均明显高于其他两组,差异有统计学意义（P＜0．05）;JOA评分重度患者的6项检测结果明显高于中度和轻度患者（P＜0．05）;膝跳反射亢进患者的6项检测明显高于膝跳反射正常患者（P＜0．05）。结论IL-1、IL-6、TNF-α、TGF-β1和NO可有效显示颈椎病的严重程度,值得在临床中推广使用。     

Peripheral blood stem cell mobilization by cytokines

Hematopoietic stem cells circulate in the peripheral blood. These cells can be collected by apheresis techniques either in the unperturbed state, after mobilization following the administration of cytokines like G-CSF or GM-CSF, or during the phase of early blood count recovery following chemotherapy-induced myelosuppression. The number of cells collected following mobilization is greater than that obtained after apheresis in the unperturbed state. There are, however, qualitative differences between unperturbed and mobilized cells. Chemotherapy related mobilization can be potentially dangerous in that severe myelosuppression necessary to achieve mobilization can have serious consequences. There are no controlled studies that evaluate the relative merits of each method of collection. Regardless of the techniques employed peripheral blood stem cells can reliably accelerate hematologic recovery after potentially myeloblative therapy and provide an alternative to bone marrow support. © 1992 Wiley-Liss, Inc.     

外周血炎性细胞因子与类风湿关节炎

背景：由滑膜细胞、单核/巨噬细胞以及淋巴细胞等产生的炎性细胞因子在类风湿关节炎的发病中发挥着重要作用。白细胞介素17是近年来发现的一个炎性细胞因子,与很多自身免疫性疾病有关。目的：观察类风湿关节炎患者血清中辅助性T细胞17相关的白细胞介素17水平及其与红细胞沉降率、C-反应蛋白、疾病活动性的相关性。方法：纳入2008年1月至2009年12月在解放军成都军区总医院就诊的符合1987年美国风湿病协会修订的类风湿关节炎分类标准的类风湿关节炎患者79例作为病例组,同时选取同时期性别、年龄与病例组相匹配的50名健康体检者作为对照组。根据28处关节疾病活动性评分指数将病例组分为活动组49例和稳定组30例。结果与结论：ELISA检测结果显示,类风湿关节炎患者血清白细胞介素17水平明显高于对照组（P〈0.01）,且活动期患者的白细胞介素17水平高于稳定期患者（P〈0.01）。Pearson相关分析结果显示,活动期类风湿关节炎患者血清中白细胞介素17水平与红细胞沉降率、C-反应蛋白、28处关节疾病活动性评分指数均呈正相关（r=0.459,0.379,0.455;P〈0.05）。说明白细胞介素17参与了类风湿关节炎的炎症反应,与红细胞沉降率、C-反应蛋白、28处关节疾病活动性评分指数等同样能反映病情的活动性。     

Microarray kit analysis of cytokines in blood product units and segments

BACKGROUND: Cytokine concentrations in transfused blood components are of interest for some clinical trials. It is not always possible to process samples of transfused components quickly after their administration. Additionally, it is not practical to sample material in an acceptable manner from many bags of components before transfusion, and after transfusion, the only representative remaining fluid of the component may be that in the “segment,” because the bag may have been completely transfused. Multiplex array technology allows rapid simultaneous testing of multiple analytes in small‐volume samples. This technology was used to measure white blood cell (WBC) cytokine levels in blood products to determine 1) whether concentrations in segments correlate with those in the main bag and, thus, whether segments could be used for estimation of the concentrations in the transfused component and 2) whether concentrations after sample storage at 4°C for 24 hours do not differ from concentrations before storage, thus allowing for processing within 24 hours, rather than immediately after transfusion. STUDY DESIGN AND METHODS: WBC cytokines were measured in the supernatant from bags and segments of leukoreduced red blood cells (RBCs), nonleukoreduced whole blood, and leukoreduced plateletphereses using a human cytokine array kit (ProteoPlex, Novagen). RESULTS: Cytokine concentrations in RBCs and whole blood or plateletphereses stored at 4°C did not differ between bag and segment samples (all p > 0.05). There was no evidence of systematic differences between segment and bag concentrations. Cytokine concentrations in samples from plateletphereses did not change within 24 hours storage at 4°C. CONCLUSION: Samples from either bag or segment can be used to study cytokine concentrations in groups of blood products. Cytokine concentrations in plateletphereses appear to be stable for at least 24 hours of storage at 4°C and, thus, samples stored with those conditions may be used to estimate the cytokine concentrations of the component at the time of transfusion.     

植物凝集素刺激外周血单个核细胞增殖及分泌因子表达的变化

背景：植物凝集素可以刺激外周血单个核细胞分裂并引起免疫细胞的活化,是常用的细胞增殖模型。但是全血中的非外周血单个核细胞成分（血浆和无核细胞）在植物凝集素参与的体外培养过程中起到什么作用,以及内皮分泌因子的表达情况尚不明确。目的：在体外单独培养或全血培养外周血单个核细胞过程中,观察植物凝集素对其增殖和凋亡的影响,以及相关炎症因子以及内皮分泌因子的表达变化。方法：分离正常核型成人的外周血单个核细胞,应用无植物凝集素培养基和加入植物凝集素培养基分别进行单独培养,观察细胞形态学变化。收集全血培养或不同条件单独培养的外周血单个核细胞,提取mRNA,荧光定量RT-PCR检测细胞增殖、凋亡,以及炎症因子和内皮分泌因子的表达变化,并进行统计学分析。结果与结论：外周血单个核细胞单独培养和全血培养存在差异,植物凝集素会促进外周血单个核细胞Ki67、增殖细胞核抗原、蛋白水解酶3、γ-干扰素、肿瘤坏死因子β和白细胞介素6的基因表达,抑制蛋白C表达。提示植物凝集素促进体外培养外周血单个核细胞的增殖及凋亡,促使炎症因子表达上调,部分血液生物学相关的内皮分泌因子下调。     

G-CSF联合化疗动员外周血干细胞过程中某些生物因子的动态变化

目的　探讨IL 8、可溶性血管细胞粘附分子 1 (sVCAM 1 )及可溶性细胞间粘附分子 1(sICAM 1 )在自体造血干细胞动员过程中的变化及其意义。方法　采用酶联免疫吸附实验方法动态分析患者造血干细胞动员过程中血浆IL 8、sICAM 1及sVCAM 1水平的变化 ;通过免疫荧光标记和流式细胞仪检测CD3 4+细胞 ;体外半固体培养CFU GM集落及血细胞计数法观察白细胞和血小板数量变化。结果　①在动员过程中 ,外周血IL 8[(2 4 7.4± 84.2 ) μg L]、sICAM 1 [(530 .3± 2 86 .1 ) μg L]及sVCAM 1[(575 .3± 350 .4) μg L]含量均较动员前和正常人显著升高 (P <0 .0 1 ) ;②患者外周血中IL 8、sVCAM 1水平与CD3 4+细胞和CFU GM集落数呈正相关 (P <0 .0 0 1 )。结论　在自体干细胞动员过程中 ,血浆IL 8和sVCAM 1的含量与患者CD3 4+细胞数和CFU GM集落形成有显著的相关性 ,分析这些生物因子的变化具有临床实用价值     

Hemoglobin stimulates the release of proinflammatory cytokines from leukocytes in whole blood

Isolated mononuclear leukocytes, when incubated with purified hemoglobin Ao (HbAo), release the proinflammatory cytokines interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha). In this study we examined whether leukocytes in whole blood, when incubated with HbAo, release IL-8, TNF-alpha, and IL-6. Leukocytes in whole blood incubated with HbAo for 4 hours at 37 degrees C, 5% CO2, and 95% humidity released 187, 1313, and 50 pg/mL of IL-6, IL-8, and TNF-alpha, respectively, as compared with 6, 192, and 2 pg/mL released by leukocytes in blood incubated with human serum albumin (HSA). Furthermore, plasma from blood incubated with HbAo exhibited chemotactic activity and stimulated human umbilical vein endothelial cells to become adherent to neutrophils. These activities were 3.3 and 2.6 times those measured in plasma from blood incubated with HSA. Hydrocortisone (0.05 micromol/L to 50 micromol/L) inhibited cytokine release in a dose-dependent manner with ED50 values of 0.23 micromol/L, 0.19 micromol/L, and 0.10 micromol/L for IL-6, IL-8, and TNF-alpha, respectively. The release of proinflammatory cytokines in whole blood after exposure to hemoglobin solutions is consistent with the possibility that an inflammatory reaction could develop on infusion of hemoglobin, whereas inhibition of cytokine release by hydrocortisone suggests that the inclusion of anti-inflammatory compounds in hemoglobin solutions may prevent undesirable effects caused by inflammation after infusion.     

红细胞在外周血单个核细胞活化过程中对细胞因子分泌的影响

目的 通过检测外周血单个核细胞（PBMC）培养液中细胞因子IFN-γ、TNF—α、IL-1β的浓度，观察和分析红细胞在抗CD3单克隆抗体（CD3McAb）诱导PBMC活化过程中对细胞因子分泌的影响，为临床在分离PBMC进行自体淋巴细胞免疫治疗的应用过程中取舍红细胞数量多少为宜提供实验依据。方法 获取健康成年人PBMC，采用人工方法加入不同数量的红细胞，模拟在制备PBMC过程中残余的红细胞数量，然后根据红细胞数量与PBMC中细胞数量之比的不同组成3个不同的实验组（RBC／PBMc=2：1、10：1、50：13组），同时附设单独PBMC为对照组。利用CD3McAb分别对以上4组细胞进行激活，然后检测各组细胞培养液中细胞因子IFN-γ、TNF-α、IL-1β的含量。结果 随着红细胞加入数量的增加，细胞培养液中各细胞因子的浓度有不同程度的上升，具有统计学意义，并且与加入的红细胞数量呈正相关关系。结论当RBC／PBMC≤50：1时，红细胞在CD3McAb诱导PBMC活化过程中对细胞因子IFN-γ，TNF—α、IL-1β的分泌具有一定的促进作用。     

TSC for hemorrhagic shock: effects on cytokines and blood pressure

Previous studies have shown that administering trans-sodium crocetinate (TSC) as a treatment of hemorrhagic shock leads to increased whole-body oxygen consumption and survival as well as protection of the liver and kidney. It has been suggested that TSC increases oxygen delivery by increasing the diffusivity of oxygen through plasma. However, as with any novel mechanism of action, there are always questions about whether the results could also be ascribed to other, previously described mechanisms of action. This study was designed to look at some aspects of that by examining the effect of different TSC dosing regimens on the blood pressure and the production of cytokines after hemorrhage because both responses have been reported with compounds that act via other mechanisms. In a constant-pressure rat model of hemorrhagic shock, it was seen that a singe bolus injection of TSC results in an immediate but transient increase in the arterial blood pressure. This is similar to the effect reported previously for using 100% oxygen. It was also found that if the TSC injections were repeated periodically over an hour, a sustained increase in the blood pressure would occur. Because inflammatory cytokines have been implicated in mortality and tissue damage, it has been suggested that TSC may affect the production of cytokines. Thus, the effect of TSC on the production of TNF-alpha and IL-10 was also examined. The data show that treatment with TSC results in lower concentrations of TNF-alpha in the liver and spleen as well as lower concentrations of IL-10 in the spleen. Again, similar effects on other cytokines have been seen with 100% oxygen. These results support the hypothesis that the effects of TSC on hemorrhagic shock are mediated via an effect on oxygen.