肺癌伴感染性发热患者外周血单个核细胞 TLR4及TLR2基因表达的检测价值

目的 分析肺癌伴感染性发热患者外周血单个核细胞(PBMC)Toll样受体4(TLR4)、Toll样受体2(TLR2)基因表达变化。方法 选择2019年1月-2021年1月医院收治的肺癌患者900例为研究对象,根据是否合并感染性发热,将患者分为感染性发热组(n=163)和非感染性发热组(n=737)。分析肺癌伴感染性发热患者的病原菌检出情况。采用实时荧光定量聚合酶链式反应法检测患者PBMC中TLR4、TLR2基因表达水平。采用酶联免疫吸附试验法检测患者血清炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6和IL-8水平。结果 163例肺癌伴感染性发热患者共检出病原菌172株,其中革兰阴性菌113株(65.70%),革兰阳性菌59株(34.30%);感染性发热组患者PBMC中TLR4、TLR2 mRNA表达水平以及血清TNF-α、IL-1β、IL-6和IL-8水平均高于非感染性发热组患者(P<0.05);感染性发热患者治疗后PBMC中TLR4、TLR2 mRNA表达水平以及血清炎性因子TNF-α、IL-1β、IL-6和IL-8水平均低于治疗前(P<0.0...     

慢性乙型肝炎患者外周单个核细胞分泌细胞因子的实验研究

目的探讨慢性乙肝患者外周血单个核细胞(PBMC)分泌肿瘤坏死因子α(TNF-α)、干扰素γ(IFN-γ)、白细胞介素6(IL-6)水平及与病毒载量的关系。方法采用ELISA法测定60例正常人和138例不同病程慢性乙肝患者PBMC中经植物血凝素(PHA)刺激前后TNF-α、IFNγ-、IL-6在上清液中的表达水平,并用荧光定量PCR法检测PBMC中HBV DNA水平。结果正常对照组及各种慢性乙肝患者PBMC上清液中TNF-α、IFNγ-、IL-6均有表达,HBV感染者三种细胞因子表达水平高于对照组,随着病情程度加重,表达水平逐渐增高(P<0.05);经PHA刺激后对照组及患者组TNFα-、IFN-γ、IL-6水平均有明显增高(P<0.01),但患者组升高水平均低于对照组(P<0.05);随着HBV DNA水平增高,PBMC中TNF-α、IFN-γ表达水平逐渐减低,而IL-6活性逐渐增高(P<0.05)。结论慢性乙肝患者PBMC可分泌TNF-α、IFNγ-、IL-6,其表达水平与宿主病毒载量有关。Th1/Th2细胞因子失衡在PBMC抗病毒免疫受抑,乙型肝炎慢性化进程中起着重要作用。     

IL-21诱导类风湿关节炎外周血单个核淋巴细胞促炎症细胞因子分泌研究

[目的]IL-21受体在类风湿关节炎(RA)患者外周血单个核淋巴细胞(PBMC)表面表达升高,本文分析IL-21对RA患者PBMC的免疫调节作用,并探讨其在疾病发生过程中的免疫病理作用.[方法]收集24例RA患者和20例健康成人对照组的外周血标本,分离PBMC,体外细胞培养,使用抗CD3+单克隆抗体和人重组IL-21刺激.48 h后收集培养上清液,使用ELISA检测促炎症细胞因子的分泌,并探讨IL-21在疾病发展过程中的作用.[结果]体外培养48 h后,检测上清液中促炎症细胞因子水平,主要包括TNF-α、IFN-γ和IL-2,结果发现RA患者外周血PBMC分泌TNF-α、IFN-γ和IL-2水平比健康者比较显著升高(P<0.005).RA患者接受有效药物治疗后,发现PBMC分分泌上述促炎症细胞因子水平显著下降(P<0.05).[结论]IL-21可显著诱导RA患者PBMC激活,并分泌大量促炎症细 胞因子,提示IL-21在关节炎症病变过程中起着重要作用,阻断体内IL-21的生物学活性可以用于治疗RA的发生.     

妊娠期糖尿病与肿瘤坏死因子-α及白介素-6水平的相关性研究

目的:测定妊娠期糖尿病孕妇血清肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)的水平,探讨TNF-α及IL-2与妊娠期糖尿病的关系及其在发病中的作用。方法:测定67例妊娠期糖尿病孕妇(GDM)、38例妊娠期糖耐量受损孕妇(IGT)及42例正常妊娠妇女空腹血清TNF-α及IL-6水平。结果:与正常对照组相比,GDM组的血清TNF-α、IL-6水平明显升高,差异有显著性(P<0.01);与IGT组比较,GDM组的血清TNF-α、IL-6水平明显升高,差异有显著性(P<0.01);3组孕妇相比,血清TNF-α、IL-6水平呈逐渐上升趋势,GDM组血清TNF-α水平和血清IL-6水平显著高于其他两组(P<0.01)。结论:与正常妊娠妇女相比,妊娠期糖尿病孕妇血清TNF-α、IL-6水平明显升高,推测TNF-α、IL-6在妊娠期糖尿病的发病机制中可能起着重要作用。     

类视黄醇对呼吸道感染患儿细胞因子分泌的影响

目的 :　观察了视黄醇、视黄酸、维生素 A醋酸酯三种类视黄醇在体外对反复呼吸道感染 (RRI)患儿外周血单个核细胞 (PBMC)中的淋巴细胞分泌白细胞介素 2 (IL- 2 )及单核细胞分泌白细胞介素 1 (IL- 1 )的影响。方法 :　测定 RRI患儿 PBMC分泌 IL- 1、IL- 2活性以及在 1 0 -9mol/ L浓度时上述三种类视黄醇作用下分泌 IL- 1、IL- 2活性 ,与正常儿童相比较。结果 :　RRI患儿 PBMC分泌 IL- 1、IL- 2活性较正常儿童低 ,但经类视黄醇刺激后明显增高 ,可达到正常水平。三种类视黄醇均能明显促进 IL - 1、IL- 2分泌 ,其中视黄酸作用最强 ,VA醋酸酯次之 ,视黄醇最弱。结论 :　 RRI患儿 PBMC分泌 IL - 1、IL - 2活性经类视黄醇刺激后能明显增高 ,达到正常水平。     

肿瘤坏死因子-α、白细胞介素-10与新生儿先心病合并心力衰竭的关系分析

目的:探讨肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)水平及IL-10/TNF-α与新生儿先心病心力衰竭(CHF)严重程度之间的关系。方法:采用双抗体夹心酶联免疫吸附实验(ELISA)测定血清TNF-α、IL-10水平。结果:重度CHF患儿血清TNF-α为(859±221)ng/L,明显高于轻、中度CHF组及对照组(P0.001);血清IL-10水平各组间无显著性差异(P0.05);IL-10/TNF-α与新生儿CHF评分呈负相关(r=-0.36,P0.05)。结论:TNF-α、IL-10作为CHF发病机制的潜在介质,在新生儿CHF的发生、发展中起着重要的作用。     

白藜芦醇对脂多糖诱导小鼠腹腔巨噬细胞炎性因子生成的调控

目的探讨白藜芦醇(Res)对脂多糖(LPS)诱导小鼠腹腔巨噬细胞核因子-κB(NF-κB)活化及炎性细胞因子[肿瘤坏死因子(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)]基因表达的调节。方法分别用1mg/LLPS或25mmol/LRes+1mg/LLPS处理体外培养的小鼠巨噬细胞,采用电泳迁移率改变分析法(EMSA)检测细胞中NF-κB活性,逆转录-聚合酶链反应(RT-PCR)和酶联免疫吸附法(ELISA)检测细胞中TNF-α、IL-1β、IL-6mRNA和蛋白的表达。结果LPS组NF-κB活性和TNF-α、IL-1β、IL-6含量在刺激后6～12h明显高于正常对照组(P<0.001),而Res+LPS组NF-κB活性和TNF-α、IL-1β、IL-6含量均显著低于LPS组(P<0.005)。结论LPS可诱导巨噬细胞NF-κB活化,导致TNF-α、IL-1β、IL-6基因表达增强,而Res能抑制NF-κB活化而调节TNF-α、IL-1β、IL-6基因的表达。     

生物制剂对类风湿关节炎患者血清白细胞介素-2与肿瘤坏死因子-α及白细胞介素-13表达的影响

目的探讨生物制剂对类风湿关节炎(RA)患者血清白细胞介素-2(IL-2)、肿瘤坏死因子-α(TNF-α)及白细胞介素-13(IL-13)表达的影响。方法选择2018年1—12月医院收治的RA患者86例,按随机数字表法分为对照组和试验组,每组43例。对照组接受甲氨蝶呤治疗,试验组在对照组基础上加用重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)治疗,比较两组的临床疗效和IL-2、TNF-α、IL-13水平及不良反应发生情况。结果试验组治疗有效率为93.0%,高于对照组的74.4%,差异有统计学意义(P<0.05)。治疗后,试验组IL-2(312.54±84.43)pg/ml、TNF-α(6.82±2.07)pg/ml、IL-13(4.86±1.52)pg/ml,均低于对照组的(429.84±97.51)pg/ml、(12.72±3.74)pg/ml、(6.13±1.87)pg/ml,差异有统计学意义(P<0.05)。治疗期间仅对照组出现恶心/呕吐1例,未经对症处理自行恢复。结论RA患者接受rhTNFR:Fc治疗有助于下调血清IL-2、TNF-α及IL-13水平,抑制炎症反应,提升RA治疗效果,且药物不良反应较少。     

肺动脉高压新生儿外周血中TNF-α、ANG-1和IL-8细胞因子的变化水平

目的:探讨肺动脉高压新生儿外周血中肿瘤坏死因子-α(TNF-α)、血管生成素-1(ANG-1)和白细胞介素-8(IL-8)细胞因子的变化水平与疾病的关系。方法:以临床确诊的42例肺动脉高压患儿为研究对象,与16例正常新生儿进行比较,采用Elisa方法检测血清中TNF-α、ANG-1和IL-8蛋白含量。采用实时荧光定量PCR法检测TNF-α、ANG-1和IL-8mRNA水平。结果:与健康新生儿相比较,肺动脉高压患儿外周血ANG-1的蛋白含量及ANG-1mRNA表达水平明显降低(P<0.01);TNF-α、IL-8的蛋白含量及TNF-α、IL-8的mRNA表达水平明显升高(P<0.01)。结论:TNF-α、ANG-1和IL-8参与肺动脉高压的发病过程。     

胎膜早破患者血IL-6、IL-8及TNF-α变化的研究

目的:探讨白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子(TNF-α)对早产胎膜早破孕妇患绒毛膜羊膜炎的监测价值。方法:对68例早产胎膜早破孕妇血中IL-6、IL-8和TNF-α进行检测,并以正常孕妇30例作为对照组。结果:36例绒毛膜羊膜炎患者血清IL-6、IL-8均明显高于非绒毛膜羊膜炎者和对照组,差异有统计学意义(P<0.01);绒毛膜羊膜炎患者血清TNF-α水平也升高,但无统计学意义(P>0.05);绒毛膜羊膜炎患者血IL-8、TNF-α水平与IL-6含量呈正相关。结论:孕妇血中IL-6和IL-8联合测定,可作为监测绒毛膜羊膜炎的重要指标;血TNF-α的测定有一定的辅助作用。     

枸杞多糖抑制人白血病细胞生长的研究

用体外培养法研究枸杞多糖 (LBP X)对人白血病HL 6 0细胞生长的抑制作用并探讨其作用机制。结果表明 :LBP X(2 0、10 0、5 0 0和 10 0 0mg L)与HL 6 0细胞共培养 48h后 ,LBP X抑制HL 6 0细胞增殖呈剂量依赖性关系 ,且可降低HL 6 0细胞的膜流动性 ,HL 6 0细胞的DNA琼脂糖凝胶电泳可见明显的“DNAladder” ,TUNEL方法检测呈阳性。这提示诱导HL 6 0细胞凋亡是LBP X抗肿瘤的重要机制     

谷氨酰胺抑制单个核细胞细胞因子过度表达的研究

目的:探讨谷氨酰胺(Gln)抑制人外周血单个核细胞(PBMC)细胞因子过度表达的分子机制。方法:用Ficoll密度梯度离心法提取健康志愿者新鲜外周血单个核细胞,在培养液中加入Gln、P38MAPK通路阻滞剂(SB203580)和HSP70阻断剂(Quercetin),用内毒素刺激4 h后,留取细胞和上清液,ELISA法测定内毒素刺激下单个核细胞肿瘤坏死因子(TNF-α)和IL-10的表达,Western blot方法检测细胞内磷酸化P38的表达情况。结果:Gln能明显增加HSP70的表达,同时TNF-α和IL-10表达受抑;使用Quercetin和SB203580后,磷酸化P38减少的同时TNF-α和IL-10也减少了。结论:Gln下调LPS刺激下PBMC细胞因子的过度表达依赖于P38MAPK信号蛋白的抑制。     

枸杞多糖的分离纯化及其抗疲劳作用

采用萃取,离子交换层析及凝胶层析枸杞中获得枸杞多糖;将纯化的枸杞多糖（ＬＢＰ－Ｘ）按５,１０,２０,５０及１００ｍｇ．ｋｇ＾－１．ｄ＾－１分别灌喂小鼠,进行抗疲劳实验。结果表明,ＬＢＰ－Ｘ能显著地增加小鼠肌糖后,肝糖原储备量,提高运动前,后血液乳酸脱氢酶总活力;降低小鼠剧烈运动后血尿素氮增加量,加快运动后血尿素氮的清除速率,以ＬＢＰ－Ｘ １０ｍｇ．ｋｇ＾－１．ｄ＾－１剂量组效果最佳。     

3种中药多糖对人外周血单核细胞活性的影响

摘要:目的　比较3种中药多糖对人外周血中单核细胞活性的影响。方法　cong分离出人外周血分离出单核细胞,实验分为:正常对照组、枸杞多糖组、黄芪多糖组和当归多糖组。在体外培养72 h后,采用中性红实验分别检测单核细胞的吞噬能力;用酶细胞化学染色法检测单核细胞酸性磷酸酶的活性;ELISA 法检测培养液中肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的表达水平。结果　与正常对照组相比,3种中药多糖均能明显增强单核细胞的吞噬能力,增加单核细胞内酸性磷酸酶的活性,并使培养上清液中的TNF-α和IL-6的含量增加,其差异有统计学意义。其中以枸杞多糖的作用最为显著。结论　3种中药多糖均能增强人外周血单核细胞的活性,其中以枸杞多糖的作用最为显著。     

枸杞多糖粗品抗突变作用的研究

目的研究枸杞多糖粗品（LBPc）对遗传损伤的保护作用。方法采用微核试验、Ames试验和DNA修复试验方法评价不同浓度的枸杞糖粗品的抗突变作用。结果不同浓度的LBPc可使醋酸铅诱导的小鼠骨髓多染红细胞微核率明显降低，3个试验组的微核率分别为4．5‰、3．7‰、1．9‰，LBPc高剂量组（20mg/kg）与阳性对照组（5．3‰）相比，差异有非常显著性（P〈0．01），并呈剂量-反应关系；Ames试验结果表明，加入不同浓度的LBPc后，其抗突变作用由弱变强；职业人群服用LBPc1周后，其DNA修复速度明显提高（P〈0．01）。结论LBPe具有明显的抗突变活性。     

Recollections of pioneers in nutrition: from starvation to cholesterol.

Objective: The aim of this work was to determine the bioavailability of herbs and spices after human consumption by measuring the ability to protect lymphocytes from an oxidative injury and by examining the impact on inflammatory biomarkers in activated THP-1 cells.

Methods: Ten to 12 subjects in each of 13 groups consumed a defined amount of herb or spice for 7 days. Blood was drawn from subjects before consumption and 1 hour after taking the final herb or spice capsules. Subject serum and various extractions of the herbs and spices were analyzed for antioxidant capacity by oxygen radical absorbance capacity (ORAC) analysis or by 1,1-diphenyl-2-picrylhydrzyl (DPPH). Subject peripheral blood mononuclear cells (PBMCs) in medium with10% autologous serum were incubated with hydrogen peroxide to induce DNA strand breaks. Subject serum was also used to treat activated THP-1 cells to determine relative quantities of 3 inflammatory cytokine (tumor necrosis factor-α [TNF-α], interleukin-1α [IL-1α], and IL-6) mRNAs.

Results: Herbs and spices that protected PBMCs against DNA strand breaks were paprika, rosemary, ginger, heat-treated turmeric, sage, and cumin. Paprika also appeared to protect cells from normal apoptotic processes. Of the 3 cytokine mRNAs studied (TNF-α, IL-1α, and IL-6), TNF-α was the most sensitive responder to oxidized LDL-treated macrophages. Clove, ginger, rosemary, and turmeric were able to significantly reduce oxidized LDL-induced expression of TNF-α. Serum from those consuming ginger reduced all three inflammatory biomarkers. Ginger, rosemary, and turmeric showed protective capacity by both oxidative protection and inflammation measures.

Conclusions: DNA strand breaks and inflammatory biomarkers are a good functional measure of a food's bioavailability.     

Effects of natural eggshell membrane (NEM) on cytokine production in cultures of peripheral blood mononuclear cells: increased suppression of tumor necrosis factor-α levels after in vitro digestion

Tumor necrosis factor-α (TNF-α) plays an important role in inflammatory processes. This study examined the effects of natural eggshell membrane (NEM(?)) (ESM Technologies, LLC, Carthage, MO, USA) on interleukin (IL)-2, IL-4, IL-6, IL-10, interferon-γ (IFN-γ), and TNF-α cytokine production by 4-day peripheral blood mononuclear cell (PBMC) cultures exposed to serial dilutions of either an aqueous extract of natural eggshell membrane (NEM-AQ) or NEM subjected to in vitro digestion (NEM-IVD). The effects on cytokine production were also assessed in the presence of phytohemagglutinin (PHA) and pokeweed mitogen (PWM) where exposure to NEM-AQ resulted in reduced levels of proliferation and statistically significant effects on IL-6, IL-10, IFN-γ, and TNF-α cytokine production. NEM-AQ reduced levels of IL-6, IL-10, IFN-γ, and TNF-α in cultures exposed to PHA. In cultures containing PWM, NEM-AQ reduced production of IL-10 and at the highest dose tested increased IL-6 and decreased TNF-α cytokine levels. NEM-IVD, at the two lowest concentrations of product, significantly reduced TNF-α production by PBMC cultures exposed to PWM compared with the in vitro digest control or native NEM. Taken together, these results suggest that NEM-AQ can influence signaling events in response to the T cell-specific mitogen PHA as well as to the mitogen PWM that require cellular cross-talk and that these effects may be partially mediated through a reduction in level of the pro-inflammatory cytokine TNF-α. The suppression of TNF-α production in the presence of NEM-IVD is promising for the use of NEM as a consumable anti-inflammatory product.     

Are elite gymnasts really malnourished? Evaluation of diet,anthropometry and immunocompetence

The purpose of this study was to evaluate the in vitro cytokine production capacity by peripheral blood mononuclear cells (PBMC) in a group of female elite gymnasts. Methods: Study subjects were 13 young sportswomen from the Federación Española de Gimnasia, whose regular training accounted for a mean exercise performance of 48 h/wk. A control group of 20 healthy volunteers (<12 h/wk of exercise) were also studied. IL-2, IFN-γ, IL-1β, IL-6 and TNF-α production was measured in 48 h cultures of PHA stimulated PBMC by ELISA. Anthropometrical parameters and a 7-day dietary record questionnaire were also assessed. Results: Weight, BMI and calorie intake (1258 ± 133 vs 2034 ± 301 kcal/d) were significantly lower in gymnasts compared to the control group. IL-2 production showed lower values and IFN-γ and IL-1β showed higher values in gymnasts than in controls. No differences were found in IL-6 and TNF-α production between both groups. Cytokine production is altered in highly trained elite gymnasts in basal conditions reflecting a particular status of their immunocompetence that could be affected by the intensity and regularity of their exercise, the hormonal changes, stressful life conditions and very impaired nutrient intakes.     

外周血单核细胞及相关细胞因子检测在肺结核中的临床价值

目的回顾性分析初治活动性肺结核(APTB)患者与健康体检者外周血单核细胞及相关细胞因子的动态变化，探讨两者的相关性及临床应用价值。 方法使用迈瑞BC-6900全自动血细胞分析仪检测72例APTB患者(病例组)和79例健康体检者(正常对照组)的外周血血常规，记录单核细胞计数(MO#)和单核细胞百分率(MO%)；测定两组血清白介素-1β(IL-1β)、白介素-6(IL-6)以及肿瘤坏死因子-α (TNF-α)表达水平，然后进行组间指标比较分析，并对单核细胞参数和细胞因子浓度进行相关性分析，同时绘制受试者工作特征(ROC)曲线，评价各指标诊断APTB的敏感度和特异度。 结果病例组外周血MO%、MO#明显高于正常对照组(t值分别为5.135、5.510，P值均<0.05)，APTB患者血清IL-1β、IL-6以及TNF-α浓度与正常对照相比，均发生了明显升高，差异具有统计学意义(U=405.000、P<0.001，U=543.500、P<0.001，U=563.000、P<0.001)。APTB患者外周血MO#与血清IL-6表达水平呈弱负相关(r=-0.247、P<0.05)。项目间诊断性能比较分析显示，指标IL-1β、IL-6以及TNF-α的曲线下面积(AUC)分别为0.891、0.918和0.887均高于MO%和MO#(AUC分别为0.758、0.735)。当IL-1β临界值取9.017 ng/L时，其诊断APTB的敏感度为83.3%，特异度为81.2%；当IL-6临界值取3.835 ng/L时，诊断APTB的敏感度为86.1%，特异度为84.4%；当TNF-α临界值取138.545 ng/L时，其诊断APTB的敏感度为80.6%，特异度为81.2%，均优于MO%(65.3%、62.0%)和MO#(70.8%、64.6%)。此外，同时测量IL-1β，IL-6和TNF-α可获得的曲线下面积高达0.987，其诊断APTB的灵敏度和特异度分别为95.8%和92.8%。 结论结核分枝杆菌感染机体引起APTB，外周血单核细胞迁移至肺泡从而衍变为巨噬细胞(M?)，活化M?产生一系列细胞因子，在机体循环池单核细胞数目变化中可能产生了一定影响，可以作为区分APTB和正常人群的理想标志物。