Ultraviolet light A irradiation induces immunosuppression associated with the generation of reactive oxygen species in human neutrophils

Ultraviolet blood irradiation has been used as a physical therapy to treat many nonspeci¯c diseases in clinics;however,the underlying mechanisms remain largely unclear.Neutrophils,the first line of host defense,play a crucial role in a variety of in°ammatory responses.In the present work,we investigated the effects of ultraviolet light A(UVA)on the immune functions of human neutrophils at the single-cell level by using an inverted°uorescence microscope.N-Formylmethionyl-leucyl-phenylalanine(FMLP),a classic physiological chemotactic peptide,was used to induce a series of immune responses in neutrophils in vitro.FMLP-induced calcium mobilization,migration,and phagocytosis in human neutrophils was significantly blocked after treatment with 365 nm UVA irradiation,demonstrating the immunosuppressive effects of UVA irradiation on neutrophils.Similar responses were also observed when the cells were pretreated with H2O2,a type of reactive oxygen species(ROS).Furthermore,UVA irradiation resulted in an increase in NAD(P)H,a member of host oxidative stress in cells.Taken together,our data indicate that UVA irradiation results in immunosuppression associated with the production of ROS in human neutrophils.

The in vitro inhibitory effect of human neutrophil peptide-1 on human immunodeficiency virus type 1

In order to clarify, the mechanism of inhibition of human neutrophil peptide-1 （ HNP-1 ） on hu- man immunodeficiency vires type 1 （HIV-1 ）, CD4^ ＋ cells were used as the target cells for acute infection with HIV-1, and experiments were peffomed separately with the interaction of different concentrations of HNP-1 with free vires particles, un-infected and infected CD4^＋ cells. The activity of reverse transcriptase （RT） in the supematant of cell cultures of different lots of experiments were then assayed accordingly, and the toxicity effect on human lymphocytic cells MT4 was measured by MTT assay. The experimental results showed that pre-incubation of HNP-1 with the concentrated stock of vires could block the binding of vires to target cells with EC50 of 2.49 μg/ml, while pre-treatment of CD4^＋ cells with HNP- 1 prior to inoculation could reduce the ability of cells to bind vires with EC50 of 20.7 μg/ml. In addition, When culturing the infected CD4^＋ cells in the continuous presence of various concentrations of HNP-1 added immediately after infection, HNP-1 exhibited modest inhibitory effect on viral replication with reduced RT activities in comparison with those of the control group （ P 〈 0.05 at 100 μg/ml of the highest concentration） . No cytotoxieity effect of HNP-1 was observed as demonstrated by MTT assay. These results indicate that HNP-1 exerts anti-HIV activity by at least two levels： direct inactivation of vires particles and effect on the ability of target cells to bind with viruses. The evaluation of two parameters, inhibitoty effect and the cytotoxicity renders HNP-1 an available candidate for anti-HIV therapeutic agent.

Increased human neutrophil lipocalin and its clinical relevance in adult-onset Still’s disease

Background:Human neutrophil lipocalin(HNL)has been used extensively to differentiate acute bacterial infection from febrile diseases as a biomarker to reflect the activation of the neutrophil.The serum HNL levels in the adult-onset Still’s disease(AOSD)patients with and without infection,as well as the healthy controls(HCs),were analyzed statistically in this study to evaluate the value of HNL for the diagnosis of AOSD.Methods:A total of 129 AOSD patients were enrolled,from whom blood samples were drawn and the AOSD diagnosis was confirmed through the review of the medical records,where the systemic score,demographic characteristics,clinical manifestations,and laboratory parameters were also collected for the patients;in addition,a total of 40 HCs were recruited among the blood donors from the healthcare center with the relevant information collected.The HNL test was done for the blood samples with the enzyme-linked immunosorbent assay and the analyses were done for the correlations of HNL with clinical manifestations and diagnostic effectiveness.Results:The serum HNL increased significantly in the patients with only AOSD as compared with that in the HCs(139.76±8.99 ng/mL vs.55.92±6.12 ng/mL;P<0.001).The serum HNL level was correlated with the white blood cell(WBC)count(r=0.335,P<0.001),neutrophil count(r=0.334,P<0.001),erythrocyte sedimentation rate(r=0.241,P=0.022),C-reactive protein(r=0.442,P<0.0001),and systemic score(r=0.343,P<0.0001)in the AOSD patients significantly.Patients with fever,leukocytosis≥15,000/mm^(3),and myalgia in the HNL-positive group were observed relatively more than those in the HNL-negative group(P=0.009,P=0.023,and P=0.007,respectively).HNL was a more sensitive indicator than ferritin and C-reactive protein(CRP)to differentiate the AOSD patients with bacterial infection from AOSD-only patients,and the Youden index was 0.6 for HNL and 0.29 for CRP.Conclusion:Serum HNL can be used as a biomarker for the diagnosis of the AOSD,and HNL is also observed to be associated with the disease activity.

A proteomic approach to better understand the role of human neutrophil peptides in the NSCLC microenvironment

Aim:Although treatment of lung cancer,one of the deadliest diseases worldwide,with immune checkpoint inhibitors(ICIs)has shown promising outcomes,these survival outcomes are only observed in a relatively small subset of lung cancer patients.In a previous study,we elucidated that the presence of human neutrophil peptide(HNP)1,2 and 3 in non-small cell lung cancerous(NSCLC)biopsies is associated with a clinical response towards treatment with PD-1/PD-L1 immune checkpoint inhibitors.Furthermore,HNP1 has shown in vitro an immune-activated function towards lung cancer cells,but the specific role of HNP1 in(lung)cancer is still unknown.The aim of this study was to provide a better understanding of HNP1 in an NSCLC microenvironment.Methods:To gain better insights into the role of HNP1 on cancer growth and to unravel immune responses,in vitro(SILAC-labelled)A549/PBMC(from three healthy donors)cocultures were set up and treated/not treated with HNP1.After 5 days,both secretome and cellular analysis using mass spectrometry were performed on these cocultures.After protein identification in all different tested conditions,pathway analyses(MetaCoreTM)were performed to investigate the biological significance of HNP1 stimulation on the A549/PBMC cocultures.The proteomic outcomes were confirmed by multiplex ELISA for a proinflammatory cytokine panel(TNF-α,IL-1β,IL-2,IL-4,IL-6,IL-12p70 and IL-18).Results:A number of biological pathways and process networks were observed to be upregulated after treatment of the coculture with HNP1.HNP1 stimulation leads to an increase in pathways and proteins stimulating chemotaxis(including plasmin signaling,leucocyte recruitment,CCL2 and CXCL8 expression),proinflammatory cytokine secretion(including IL-1β,IL-6 and TNF-α),dendritic cell(DC)maturation,phagocytosis and antigen presentation,leading to a more efficient adaptive anti-tumoral immunity.Conclusion:These results enhance our understanding of the role of HNP1 in the tumor microenvironment and suggest that HNP1 may be able to induce tumor necrosis by inducing prostimulatory immune responses.

Effect of everolimus on the immunomodulation of the human neutrophil inflammatory response and activation

The primary cause of mortality at 5 years following a cardiac transplantation is the development of atherosclerosis, termed coronary allograft vasculopathy （CAV）. This pathology is characterized by diffused intimal hyperplasia and emanates from coronary arterial injuries caused by immune inflammatory cells. Neutrophils play an important role in this inflammatory process; however, their potential participation in the pathogenesis of CAV is poorly understood. Despite their essential contribution to the prevention of graft rejection, immunosuppressive drugs could have detrimental effects owing to their pro-inflammatory activities. Thus, we investigated the impact of different immunosuppressive drugs on the inflammatory response of neutrophils isolated from the blood of healthy volunteers. Under basal conditions, mammalian target of rapamycin （mTOR） inhibitors （sirolimus and everolimus） had the most potent anti-inflammatory effect, decreasing both IL-8 release （≈-80%） and vascular endothelial growth factor （VEGF） release （≈-65%） and preserving the release of the anti-inflammatory cytokine interleukin-1 receptor antagonist （IL-1RA）. In TNF-α-treated neutrophils, pre-incubation with everolimus provided the most potent effect, simultaneously reducing the release of both VEGF and IL-8 while doubling the release of IL-1RA. This latter effect of everolimus was maintained even when administered in combination with other immunosuppressive drugs. Sirolimus and everolimus decreased the tumor necrosis factor （TNF）-α-induced adhesion of neutrophils to human endothelial cells and human extracellular matrix. This effect was largely dependent on the ability of these compounds to alter P2-integrin/CD18 activation. Our results suggest a potential mechanism for the beneficial effect of everolimus in the prevention of CAV in heart transplant recipients.

血清HNL、PCT、sCD163及IL-6联合检测在老年患者脓毒症诊断中的应用价值

目的探讨中性粒细胞载脂蛋白(HNL)联合其他感染相关生物标志物对老年脓毒症患者的诊断价值。方法选取2019年5月至2022年11月我院收治的115例老年患者为研究对象,根据“脓毒症-3”共识标准、临床病史和实验室检查将其分为脓毒症组(n=35)、急性局部细菌感染组(n=50)、病毒感染组(n=30)和健康对照组(n=50)。收集患者的静脉血样,分析全血细胞计数、血培养、微生物培养结果,HNL、降钙素原(PCT)、白介素-6(IL-6)和可溶性血红蛋白清道夫受体(sCD163)检测水平。结果在老年脓毒症患者诊断中,HNL和PCT联合检测的灵敏度为91.4%,特异度为98%,显著优于使用1.025 ng/ml作为PCT的最佳截止值所达到的灵敏度80%以及使用98.91 ng/ml作为HNL的最佳截止值所达到的特异度84%;在老年患者局部细菌感染诊断中,HNL和PCT联合检测的灵敏度为88%,特异度为90%,显著优于PCT和HNL单独检测的诊断效能。结论HNL和PCT联合检测有助于老年脓毒症以及局部细菌感染的早期诊断。

RO-heparin Inhibits L-Selectin-mediated Neutrophils Adhesion to Vascular Endothelium Under Flow Conditions

Selectins are carbohydrate-binding cell adhesion molecules that play a major role in the initiation of inflammatory responses. Accumulaed evidence has suggested that heparin＇s anti-inflammatory effects are mainly mediated by blocking L-or P-selectin-initiated cell adhesion. Recently, we have reported that periodate-oxidized, borohydridereduced heparin （RO-heparln） can inhibit P-selectin-mediated acute inflammation. Here we further examined the effect of RO-heparin on the adhesion of L-selectin-mediated leukocytes to vascular endothelium under flow conditions in vivo and in vitro. The results show that RO-heparin with a low anticoagulant activity can effectively reduce leucocyte roiling on thioglycoUate-induced rat mesenterlc venules and L-selectin-metadiated neutrophil roiling on TNF-α-induced human umbilical vein endothelial cells（HUVECs） under flow conditions. Our findings suggest that the effect of RO-heparin on inflammatory responses is mainly a result of its inhibiting the interaction between P- or L-selectin and its ligands. The findings also suggest that RO-heparin may be useful in preventing inflammation diseases.

大鼠脓毒症肺损伤与中性粒细胞载脂蛋白的相关性研究

目的探究大鼠脓毒症肺损伤与中性粒细胞载脂蛋白(HNL)的相关性。方法45只大鼠按照随机方法分为对照组、sham组及模型组,模型组建立脓毒症肺损伤大鼠模型,sham组开腹后仅游离盲肠后关腹,不进行结扎和穿刺,对照组为健康大鼠。使用试剂盒检测各组大鼠血清白细胞介素(IL)-6、IL-1β、肿瘤坏死因子-α(TNF-α)及HNL;血气分析仪检测氧合指数;计算肺组织湿/干比;HE染色观察肺组织病理形态,计算肺损伤评分。结果对照组及sham组12、24、36 h的IL-6、IL-1β、TNF-α、HNL水平、氧合指数、肺组织湿/干比、肺损伤评分比较,差异均无统计学意义(均P>0.05);与sham组相比,模型组12、24、36 h的IL-6、IL-1β、TNF-α、HNL水平、氧合指数、肺组织湿/干比、肺损伤评分均升高,差异均有统计学意义(均P<0.05)。对照组大鼠12、24、36 h肺组织结构正常,且未见水肿,sham组大鼠12、24、36 h肺组织仅有少量炎症细胞,模型组大鼠12、24、36 h肺组织结构严重破坏,肺泡萎陷,炎症细胞严重浸润,但随着时间延长病理改善。脓毒症肺损伤大鼠中,HNL与IL-6、IL-1β、TNF-α、肺部湿/干比、肺损伤评分呈现正相关性(均P<0.05),与氧合指数呈现负相关性(P<0.05)。结论脓毒症肺损伤大鼠中HNL显著升高,炎症反应严重,肺组织湿/干比、肺损伤加重,氧合指数降低,且HNL水平与IL-6、IL-1β、TNF-α水平、肺组织湿/干比、肺损伤呈正相关性,与氧合指数呈负相关性。

新型及传统炎症指标对血液疾病患者细菌感染的评估

目的探究新型及传统炎症指标中性粒细胞载脂蛋白(human neutrophil apolipoprotein,HNL)、中性粒细胞CD64(neutrophtl CD64,nCD64)、降钙素原(procalcitonin,PCT)、白细胞(white blood cell,WBC)计数对血液疾病细菌感染的诊断价值。方法回顾性分析85例细菌感染血液疾病患者临床资料,设为感染组,另外选取同期非细菌感染血液疾病患者62例为非感染组。比较2组患者一般资料、外周血HNL、nCD64、PCT、WBC计数水平差异,采用受试者操作特征(receiver operating characteristic,ROC)曲线描述上述指标应用于血液疾病细菌感染患者的诊断效能。结果感染组外周血PCT、HNL、nCD64、WBC计数水平均高于非感染组,差异有统计学意义(P<0.001),PCT、HNL、nCD64、WBC计数水平及联合预测诊断血液疾病细菌性感染的敏感度分别为81.18%、82.35%、78.82%、77.65%及96.47%,特异度分别为85.88%、89.41%、88.68%、88.24%及78.82%。ROC曲线结果显示,PCT、HNL、nCD64、WBC计数水平及联合预测诊断血液疾病细菌性感染的曲线下面积分别为0.871、0.907、0.891、0.788及0.984。结论外周血PCT、HNL、nCD64、WBC计数水平对诊断血液疾病细菌感染均有评估价值,其中以联合诊断价值最大。

CD4^(+)/CD8^(+)、NGAL和SAA与老年肾衰竭血液透析患者细菌感染的相关性及诊断价值

目的 研究外周血CD4^(+)/CD8^(+)、人中性粒细胞明胶酶相关脂质运载蛋白(neutrophil gelatinase associated lipocalin,NGAL)和淀粉样蛋白A(serum amyloid A protein,SAA)与接受维持性血液透析(maintenance hemodialysis,MHD)治疗的老年慢性肾衰竭(chronic renal failure,CRF)患者细菌感染的相关性及其诊断价值。方法 选取2019年10月—2021年10月在海南省中医院接受MHD治疗的老年CRF患者合并细菌感染者61例(感染组)和未合并感染者104例(非感染组)为样本开展横断面研究,检测2组CD4^(+)/CD8^(+)、NGAL和SAA表达水平并进行比较,建立Logistics回归模型分析各指标与老年CRF患者MHD治疗期间细菌感染的关系并作受试者工作特征(ROC)曲线分析其诊断价值。结果 感染组CD8^(+)(t=2.573,P=0.011)、NGAL(t=6.670,P<0.001)和SAA(t=10.116,P<0.001)表达水平高于非感染组,CD4^(+)(t=8.514,P<0.001)、CD4^(+)/CD8^(+)(t=3.904,P<0.001)表达水平低于非感染组。校正年龄、性别和透析龄等基本资料后进行Logistics回归分析(因变量赋值感染组=1,非感染组=0)显示CD4^(+)/CD8^(+)(OR=0.586,95%CI:0.440~0.780,P<0.001)、NGAL(OR=1.324,95%CI:1.149~1.525,P<0.001)和SAA(OR=1.419,95%CI:1.251~1.610,P<0.001)均与老年CRF患者MHD治疗期间细菌感染存在密切联系;ROC曲线分析显示CD4^(+)/CD8^(+)、NGAL和SAA用于老年MHD患者细菌感染诊断的曲线下面积(AUC)分别为0.576、0.738和0.838,敏感度分别为54.10%、62.30%和77.05%,特异度分别为63.46%、83.65%和77.88%,将3项指标建立Logistics回归模型进行综合判断,结果显示AUC为0.886,敏感度为85.25%,特异度为79.81%。结论 接受MHD治疗的老年CRF患者外周血CD4^(+)/CD8^(+)、NGAL和SAA与细菌感染存在密切联系,检测CD4^(+)/CD8^(+)、NGAL和SAA表达水平变化可为细菌感染早期准确诊断提供参考依据。

人中性粒细胞载脂蛋白、降钙素原、白介素-6对危重患者发生导管相关性血流感染的早期诊断价值

目的探讨人中性粒细胞载脂蛋白(HNL)、降钙素原(PCT)、白介素-6(IL-6)对长期留置中心静脉导管的危重患者发生导管相关性血流感染(CRBSI)的早期诊断价值。方法选取2022年1月至2023年9月山东第一医科大学第二附属医院重症医学科收治的189例留置中心静脉导管患者为研究对象,按照是否发生CRBSI分为CRBSI组(n=63)和非CRBSI组(n=126)。收集患者临床特征资料及留取血培养当天的HNL、PCT、IL-6结果。采用logistic回归分析建立HNL、PCT和IL-6的联合检测模型。采用受试者工作特征(ROC)曲线评价各项指标对危重患者发生CRBSI的早期诊断价值。结果CRBSI组HNL、PCT、IL-6水平均高于非CRBSI组,差异有统计学意义(P<0.05)。ROC曲线分析结果显示,HNL、PCT、IL-6及联合检测模型诊断CRBSI的ROC曲线下面积(AUC)分别为0.950、0.901、0.828、0.964,差异有统计学意义(P<0.05)。HNL、PCT、IL-6诊断CRBSI的最佳临界值分别为248.51 ng/ml、0.62 ng/ml、32.68 pg/ml,灵敏度分别为81.0%、79.4%、84.1%,特异度分别为99.2%、86.4%、69.6%。一致性检测结果显示,联合检测模型结果与预测诊断CRBSI存在较好的一致性(Kappa=0.810,P<0.05),HNL≥248.51 ng/ml与预测诊断CRBSI存在较好的一致性(Kappa=0.804,P<0.05)。结论相比于PCT、IL-6,HNL对危重患者发生导管相关性血流感染有较好的早期诊断价值。

中性粒细胞载脂蛋白、D-二聚体在儿童重症细菌性脓毒症早期诊断中的临床应用价值研究

目的探讨中性粒细胞载脂蛋白(HNL)、D-二聚体(D-D)在儿童重症细菌性脓毒症早期诊断中的应用价值。方法选取30例重症细菌性脓毒症患儿作为研究组,另选取同期30例普通细菌性脓毒症患儿作为对照A组,30例普通细菌感染患儿作为对照B组。三组患儿均在入院时检测HNL、D-D、C反应蛋白(CRP)、降钙素原(PCT)、白细胞计数(WBC)、天门冬氨酸氨基转移酶(AST)、谷丙氨转氨酶(ALT)、尿素氮(BUN)、血清肌酐(Cr)水平。比较三组HNL、D-D、CRP、PCT、WBC、AST、ALT、BUN、Cr水平。结果研究组患儿的HNL、D-D、CRP、PCT、WBC水平分别为(625.17±69.14)ng/ml、(7549.51±1532.75)ng/ml、(95.62±6.56)mg/L、(21.37±4.24)ng/ml、(14.83±1.79)×10^(9)/L,对照A组患儿别为(371.42±42.37)ng/ml、(1032.28±201.63)ng/ml、(21.41±3.43)mg/L、(10.41±2.26)ng/ml、(15.78±2.03)×10^(9)/L,对照B组患儿分别为(191.58±17.89)ng/ml、(382.16±56.58)ng/ml、(21.79±2.48)mg/L、(0.82±0.11)ng/ml、(15.37±1.44)×10^(9)/L。三组患儿的WBC水平比较无统计学差异(P>0.05);对照A组患儿的CRP水平与对照B组患儿比较无统计学差异(P>0.05);研究组患儿的HNL、D-D、CRP、PCT水平均高于对照A组与对照B组,对照A组患儿的HNL、D-D、PCT水平均高于对照B组,差异有统计学意义(P<0.05)。研究组患儿的AST、ALT、BUN、Cr水平分别为(93.82±8.56)U/L、(193.67±11.50)U/L、(15.75±4.06)mmol/L、(179.26±16.35)μmol/L,对照A组患儿分别为(42.58±6.91)U/L、(34.78±4.13)U/L、(4.48±1.47)mmol/L、(77.40±11.76)μmol/L,对照B组分别为(29.77±3.72)U/L、(32.39±4.89)U/L、(3.71±1.28)mmol/L、(75.57±6.24)μmol/L。研究组患儿的AST、ALT、BUN、Cr水平均高于对照A组与对照B组,对照A组患儿的AST水平高于对照B组,差异有统计学意义(P<0.05);对照A组患儿的ALT、BUN、Cr水平与对照B组患儿比较无统计学差异(P>0.05)。结论HNL、D-D有助于重症细菌性脓毒症、普通细菌性脓毒症、普通细菌感染患儿的早期鉴别诊断。

脓毒症相关生物标志物的研究进展

脓毒症是指由于宿主对感染反应失调而导致的危及生命的器官功能障碍,是危重患者死亡的主要原因之一。脓毒症的早期诊断与正确治疗是降低病死率的关键,但目前尚无诊断的金标准。理想的脓毒症生物标志物应该具有早期诊断和预测不良预后的能力,且具有较好的敏感性和特异性。脓毒症的候选生物标志物众多,本文主要综述中性粒细胞CD64(Neutrophil CD64,nCD64)、降钙素原(Procalcitonin,PCT)、C-反应蛋白(C-reactive protein,CRP)、人类白细胞抗原-DR(Human leukocyte antigen-DR,HLA-DR)、白细胞介素6(Interleukin-6,IL-6)等关注度较高的候选生物标志物的研究进展。

人脐带间充质干细胞来源外泌体驱动中性粒细胞向N2促血管生成表型转换的研究

背景人脐带间充质干细胞来源外泌体(human umbilical cord mesenchymal stem cell-derived exosomes,HucMSCExo)已成为再生医学领域中一种应用前景广阔的治疗手段,但人们对其作用的确切机制仍知之甚少。目的探索HucMSCExo能否驱动中性粒细胞向N2促血管生成亚型转换。方法采用超速离心法分离提纯HucMSC-Exo,通过透射电镜、粒径分析和Western blot对提取的外泌体进行鉴定。成功分离HucMSC-Exo后与中性粒细胞共培养,利用qRT-PCR检测中性粒细胞的表型相关分子表达情况。将接受HucMSC-Exo刺激的中性粒细胞与血管内皮细胞进行细胞共培养,通过CCK-8实验、Edu实验、划痕实验和Transwell实验观察中性粒细胞对血管内皮细胞的增殖、迁移和侵袭能力的影响,采用qRT-PCR分析中性粒细胞在接受HucMSC-Exo刺激后,血管生成相关基因的表达变化。结果提取物在透射电镜下显示为类圆形膜状结构,直径约100 nm,能够表达外泌体标记物CD9和Alix,表明提取物为HucMSC-Exo。与PBS处理组相比,HucMSC-Exo处理中性粒细胞后,qRT-PCR发现与N2表型相关的标志物Arg1、CD163和CD206的表达增多(P<0.05)。将HucMSCExo刺激的中性粒细胞与血管内皮细胞进行细胞共培养处理后,通过CCK-8实验、Edu实验、划痕实验和Transwell实验观察到血管内皮细胞的增殖、迁移和侵袭能力均增强(P<0.05)。与PBS处理组相比,中性粒细胞在接受HucMSC-Exo刺激后,促血管生成因子BV8、VEGFα表达升高(P<0.05)。结论HucMSC-Exo可使中性粒细胞向N2表型转换,这种N2型中性粒细胞可促进血管内皮细胞增殖、迁移和侵袭。

CD133、HER2、HIF-1α在胃癌组织中的表达及与患者Hp感染、淋巴结转移和预后的关系分析

目的探究中性粒细胞表面抗原(CD133)、人类表皮生长因子受体2(HER2)、缺氧诱导因子-1α(HIF-1α)在胃癌组织中的表达及与患者幽门螺杆菌(Hp)感染、淋巴结转移和预后的关系。方法回顾性选取2020年1月至2021年1月在黄石市中心医院/湖北理工学院附属医院进行治疗的100例胃癌患者作为研究对象。所有患者中,存在Hp感染75例,未发生Hp感染25例;存在淋巴结转移45例,未发生淋巴结转移55例。随访2年,56例患者发生复发或死亡,纳入预后不良组,其余44例纳入预后良好组。取患者的胃癌组织及癌旁组织,对组织中CD133、HER2、HIF-1α表达情况进行比较,并探究CD133、HER2、HIF-1α表达情况与患者Hp感染、淋巴结转移和预后的关系,并采用受试者工作特征(ROC)曲线分析CD133、HER2、HIF-1α表达对预后不良的预测价值。结果胃癌组织中CD133、HER2、HIF-1α表达阳性率分别为81.00%、25.00%、66.00%,均高于癌旁组织(18.00%、4.00%、44.00%),差异均有统计学意义(P<0.05)。发生Hp感染的患者中胃癌组织的CD133、HER2、HIF-1α表达阳性率分别为74.67%、30.67%、72.00%,均显著高于未发生Hp感染患者(48.00%、8.00%、48.00%),差异均有统计学意义(P<0.05)。淋巴结转移患者胃癌组织的CD133、HER2、HIF-1α表达阳性率分别为93.33%、48.89%、80.00%,均高于未发生淋巴结转移患者(70.91%、5.45%、54.55%),差异均有统计学意义(P<0.05)。预后不良组患者胃癌组织的CD133、HER2、HIF-1α表达阳性率分别为83.93%、37.50%、75.00%,均显著高于预后良好组患者(47.73%、9.09%、54.55%),差异均有统计学意义(P<0.05)。ROC曲线结果显示,CD133、HER2表达阳性率对胃癌预后不良的预测曲线下面积(AUC)值分别为0.781、0.762,HIF-1α表达阳性率对胃癌预后不良的预测AUC值为0.602。结论CD133、HER2、HIF-1α在胃癌组织及发生感染Hp、淋巴结转移患者中存在高表达,且CD133、HER2表达对患者预后有一定预测价值。

NLR对PD-1抑制剂治疗HER2阴性晚期胃癌患者预后的预测价值分析

目的探索基线血清中性粒细胞/淋巴细胞(NLR)对细胞程序性死亡受体1(PD-1)抑制剂联合化疗治疗晚期胃癌患者的预后价值。方法选取2018年1月至2021年1月秦皇岛市第四医院使用免疫联合化疗治疗方案的晚期胃癌患者168例为研究对象,随访截止时间为2023年1月。收集患者资料,联合初次治疗前血液学及肿瘤相关标志物进行分析,利用X-tile软件确定NLR的最佳截断值。采用Kaplan-Meier生存曲线分析NLR表达对晚期胃癌患者生存率的影响,受试者工作曲线(ROC)分析NLR对晚期胃癌患者预后的预测价值,结合Cox比例风险模型筛选影响晚期胃癌患者疾病进展的相关因素。结果168例患者治疗前1周血清NLR的最佳截断值为2.41,将患者分为高NLR组(NLR>2.41,n=93)和低NLR组(NLR≤2.41,n=75),NLR水平与肿瘤分化、是否远处转移、细胞程序性死亡-配体1综合阳性评分、癌胚抗原、癌抗原125有关(P<0.05);低NLR组的免疫治疗联合化疗有效率明显高于高NLR组(P<0.05);低NLR组患者的中位无进展生存期(PES)和总生存期(OS)均较高NLR组延长(PFS:P=0.006;OS:P=0.023);ROC分析显示,NLR对晚期胃癌患者预后的曲线下面积为0.740,敏感度为81.50%,特异度为69.73%;在多变量分析中,除了治疗前NLR,肿瘤分化程度、是否远处转移也为晚期胃癌患者预后不良的独立预测因素(P<0.05)。结论在接受免疫联合化疗的晚期胃癌患者中,治疗前NLR与疾病控制及PFS/OS具有相关性,对预测晚期胃癌免疫治疗预后有较高的价值。

重症肺炎患者外周血CD64指数、NLR及CD14^(+)单核细胞HLA-DR的检测意义分析

目的:探讨重症肺炎患者外周血CD64指数、中性粒细胞/淋巴细胞比值(NLR)及CD14^(+)单核细胞人类白细胞DR抗原(HLA-DR)的检测意义。方法:收集2022年1月—2024年3月抚州市第一人民医院收治的80例重症肺炎患者的临床资料进行研究。根据28 d内患者预后情况分组,将死亡患者作为死亡组(n=32),存活患者作为存活组(n=48)。比较两组一般资料及CD64指数、NLR、CD14^(+)单核细胞HLA-DR水平,分析CD64指数、NLR、CD14^(+)单核细胞HLA-DR单独及联合预测重症肺炎预后的价值,以及重症肺炎患者病死的危险因素。结果:两组年龄、入院时急性生理学和慢性健康状况评价Ⅱ(APACHEⅡ)比较,差异均有统计学意义(P<0.05)。死亡组CD64指数、NLR均高于存活组,CD14^(+)单核细胞HLA-DR水平低于存活组(P<0.05)。受试者操作特征(ROC)曲线结果显示,CD64指数、NLR、CD14^(+)单核细胞HLA-DR单独及联合预测重症肺炎预后不良的曲线下面积(AUC)分别为0.812、0.769、0.728、0.893,均具有一定预测价值(P<0.05)。logistic回归分析显示,CD64指数>9.15、NLR>12.19、CD14^(+)单核细胞HLA-DR≤44.60%、年龄≥65岁、入院时APACHEⅡ≥21.69分是重症肺炎患者病死的独立危险因素(P<0.05)。结论:CD14^(+)单核细胞HLA-DR水平偏低,CD64指数、NLR偏高提示重症肺炎患者预后不佳,三者联合检测可为临床预测重症肺炎患者预后提供参考。

微小RNA-138/NGAL对缺血再灌注诱导人肾小管上皮细胞的影响

目的探究微小RNA-138(microRNA-138,miR-138)调控中性粒细胞明胶酶相关脂质运载蛋白(neutrophil gelatinase-associated lipocalin,NGAL)对缺血/再灌注(ischemia/reperfusion,I/R)诱导人肾小管上皮(HK-2)细胞损伤的影响。方法HK-2细胞构建I/R模型,分别转染miR-138模拟物、miR-138抑制剂、NGAL、NGAL+miR-138模拟质粒,qRT-PCR测定不同细胞miR-138或NGAL mRNA表达鉴定转染结果,细胞活力检测(cell counting kit-8,CCK-8)法和流式细胞术测定细胞活性和凋亡。ELISA、Western blot法分别测定miR-138模拟物、miR-138抑制剂对I/R细胞白细胞介素(interleukin,IL)-6、IL-1β、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平,以及Toll样受体4(toll like receptor 4,TLR4)、核因子κB(nuclear factor kappa-B,NF-κB)、NF-κB抑制蛋白(inhibitor-NF-κB,IκBα)、磷酸IκBα(p-IκBα)、NGAL蛋白表达的影响。结果I/R细胞miR-138 mRNA表达、细胞活力降低,凋亡增加(P<0.01)。质粒转染成功,miR-138模拟物促进IR细胞活力,降低凋亡和NGAL mRNA表达(P<0.01);miR-138抑制剂、NGAL模拟降低IR细胞活力,增加凋亡和NGAL mRNA表达(P<0.01);NGAL模拟对IR细胞的损伤作用可被miR-138模拟逆转。miR-138抑制剂可增加I/R细胞IL-6、IL-1β、TNF-α水平和凋亡,上调TLR4、NF-κB、p-IκBα、NGAL蛋白表达,下调IκBα蛋白表达(P<0.05);miR-138模拟物可降低I/R细胞IL-6、IL-1β、TNF-α水平和凋亡,下调TLR4、NF-κB、p-IκBα、NGAL蛋白表达,上调IκBα蛋白表达(P<0.05)。结论miR-138通过调节NGAL和TLR4/NF-κB途径降低细胞凋亡和炎症因子水平,对I/R诱导的HK-2细胞发挥保护作用。

降钙素原、人中性粒细胞载脂蛋白和中性粒细胞CD64检测在细菌性感染早期诊断中的应用

目的研究细菌性感染患者血液中的降钙素原(PCT)、人中性粒细胞载脂蛋白(HNL)和中性粒细胞CD64(CD64)的表达水平,为临床细菌性感染早期诊疗提供科学依据。方法选取2013年2月—2017年5月就诊治疗确诊的210例感染患者,另选取80例同期健康体检人员为对照组,感染患者分为细菌组105例、病毒组105例,采用热景公司上转发光免疫分析仪检测PCT和HNL、BD FACS calibur流式细胞仪测定并计算CD64百分比、制定工作曲线选取PCT、HNL及CD64截断值,从而分析比较细菌组、病毒组和对照组的PCT、HNL及CD64表达水平和诊断细菌性感染疾病的特异度和灵敏度。结果细菌组与病毒组、对照组的PCT、HNL、CD64和WBC水平分别比较,差异均具有统计学意义(P<0.05)。病毒组与对照组的CD64、WBC水平比较,差异均具有统计学意义(P<0.05)。病毒组与对照组的PCT和HNL水平差异无统计学意义(P>0.05)。PCT的受试者操作特征(ROC)曲线下的面积(AUC)为0.855,AUC(HNL)为0.930,AUC(CD64)为0.928,AUC(WBC)为0.729。PCT、HNL和CD64诊断细菌感染性疾病的截断值分别为:>?0.79 ng/m L、>87.43 ng/m L、>9.01%。结论细菌感染性疾病导致PCT、HNL和CD64表达水平升高,PCT、HNL和CD64可用于细菌感染性疾病的早期诊断,其中HNL的诊断价值最高。

红茶提取物在炎性气道黏液高分泌中的作用

目的观察茶黄素(TF)各单体在人中性粒细胞弹性蛋白酶(HNE)诱导气道黏液高分泌中的作用。方法采用HNE刺激人肺腺癌细胞A549,导致黏液高分泌,以TF各单体(TF1、TF2、TF3)进行干预。采用MTT法测定各单体对A549细胞的活性影响,确定作用剂量后将A549细胞分为4组进行实验,分别为①对照组:无血清培养基培养;②HNE刺激组:HNE(50 nmol/L)刺激24 h;③TF单体干预组:分别用TF1、TF2、TF3(浓度分别为50、100、200μg/mL)作用24 h后,再予HNE刺激24 h;④AG1478干预组:先用表皮生长因子受体阻断剂AG1478(5μmol/L)处理30 min后,再予HNE刺激24 h。测定不同剂量TF1、TF2、TF3作用后黏蛋白(MUC)的变化及TF3(200μg/mL)作用时间不同(12、24、36 h)后MUC的变化。RT-PCR检测MUC5AC mRNA的变化;ELISA测定MUC5AC蛋白表达的变化。结果HNE刺激组的MUC5AC mRNA表达和MUC5AC蛋白含量显著高于对照组(P<0.01);而给予TF1、TF2、TF3预处理后,与HNE刺激组相比,两指标均明显下调(均P<0.01),且呈剂量依赖关系,其中以TF3作用最明显。TF3的作用时间也与MUC5AC的下调呈正相关;而AG1478干预组对黏液的下调作用较TF各组更强(P<0.01)。结论TF各单体虽不能完全抑制炎性黏液的分泌,但可明显降低炎性气道的黏液高分泌状态,以TF3作用最强。