Targeting and exploitation of tumor-associated neutrophils to enhance immunotherapy and drug delivery for cancer treatment

Neutrophils,the most abundant leukocytes in human blood,are essential fighter immune cells against microbial infection.Based on the finding that neutrophils can either restrict or promote cancer progression,tumor-associated neutrophils(TAN)are classified into anti-tumor N1 and pro-tumor N2 subsets.One of the major mechanisms underlying the tumor-promoting function of N2-TANs is suppression of adaptive immune cells,in particular,cytotoxic T lymphocytes.Currently,no established methodologies are available that can unequivocally distinguish immunosuppressive TANs and granulocytic/polymorphonuclear myeloid-derived suppressor cells(G/PMN-MDSC).In view of the critical role of PMN-MDSCs in immune evasion and resistance to cancer immunotherapy,as established from data obtained with diverse cancer models,therapeutic strategies targeting these cells have been actively developed to enhance the efficacy of immunotherapy.Here,we have reviewed the available literature on strategies targeting PMN-MDSCs and summarized the findings into four categories:(1)depletion of existing PMN-MDSCs,(2)blockade of the development of PMNMDSCs,(3)blockade of PMN-MDSC recruitment,(4)inhibition of immunosuppressive function.Owing to their high mobility to inflamed organs and ability to trespass the blood-brain barrier,neutrophils are outstanding candidate carriers in nanoparticle-based therapies.Another attractive application of neutrophils in cancer therapy is the use of neutrophil membrane-derived nanovesicles as a surrogate of extracellular vesicles for more efficient and scalable drug delivery.In the second part of the review,we have highlighted recent advances in the field of neutrophil-based cancer drug delivery.Overall,we believe that neutrophil-based therapeutics are a rapidly growing area of cancer therapy with significant potential benefits.

Neutrophil peptide 1 accelerates the clearance of degenerative axons during Wallerian degeneration by activating macrophages after peripheral nerve crush injury

Macrophages play an important role in peripheral nerve regeneration,but the specific mechanism of regeneration is still unclear.Our preliminary findings indicated that neutrophil peptide 1 is an innate immune peptide closely involved in peripheral nerve regeneration.However,the mechanism by which neutrophil peptide 1 enhances nerve regeneration remains unclear.This study was designed to investigate the relationship between neutrophil peptide 1 and macrophages in vivo and in vitro in peripheral nerve crush injury.The functions of RAW 264.7 cells we re elucidated by Cell Counting Kit-8 assay,flow cytometry,migration assays,phagocytosis assays,immunohistochemistry and enzyme-linked immunosorbent assay.Axonal debris phagocytosis was observed using the CUBIC(Clear,Unobstructed Brain/Body Imaging Cocktails and Computational analysis)optical clearing technique during Wallerian degeneration.Macrophage inflammatory factor expression in different polarization states was detected using a protein chip.The results showed that neutrophil peptide 1 promoted the prolife ration,migration and phagocytosis of macrophages,and CD206 expression on the surfa ce of macrophages,indicating M2 polarization.The axonal debris clearance rate during Wallerian degeneration was enhanced after neutrophil peptide 1 intervention.Neutrophil peptide 1 also downregulated inflammatory factors interleukin-1α,-6,-12,and tumor necrosis factor-αin invo and in vitro.Thus,the results suggest that neutrophil peptide 1 activates macrophages and accelerates Wallerian degeneration,which may be one mechanism by which neutrophil peptide 1 enhances peripheral nerve regeneration.

Correlation of neutrophil to lymphocyte ratio to severity of coronary artery disease and in-hospital clinical outcomes in patients with acute coronary syndrome: A prospective observational study

Objective:To explore correlation of neutrophil-to-lymphocyte ratio(NLR)to severity of coronary artery disease(CAD)and in-hospital clinical outcomes in patients with acute coronary syndrome(ACS).Methods:In this prospective and observational study,we recruited 500 patients with ACS.For all the eligible patients,demographic details were collected,and laboratory parameters were evaluated.The CAD severity was evaluated in terms of the number of involved vessels.The NLR was calculated based on neutrophils and lymphocytes and the correlation of various risk factors and severity and outcome of CAD was performed.Results:77.2%of Patients was male,and 52%of the patients aged between 55-70 years.Based on the type of ACS,396 out of 500 patients had ST-elevation myocardial infarction.An ascending trend in the white blood cell levels and NLR value was noted as the severity of the ACS increased and the highest white blood cell levels and NLR was noted among classⅣpatients.The mean NLR value among the non-survivors were higher compared to the survivors(9.52±5.72 vs.4.76±2.36;P<0.01).Receiver operating curve showed that the cut-off NLR value was 5.76 with a sensitivity of 75.0%and a specificity of 77.3%.Conclusions:The NLR can be used as an independent prognostic marker in ACS.An elevated NLR value serves as a reliable predictor for short-term complications,notably in-hospital mortality.

Effects of Bifidobacterium infantis on cytokine-induced neutrophil chemoattractant and insulin-like growth factor-1 in the ileum of rats with endotoxin injury

BACKGROUND The digestive tract is the maximal immunizing tissue in the body, and mucosal integrity and functional status of the gut is very important to maintain a healthy organism. Severe infection is one of the most common causes of gastrointestinal dysfunction, and the pathogenesis is closely related to endotoxemia and intestinal barrier injury. Bifidobacterium is one of the main probiotics in the human body that is involved in digestion, absorption, metabolism, nutrition, and immunity.Bifidobacterium plays an important role in maintaining the intestinal mucosal barrier integrity. This study investigated the protective mechanism of Bifidobacterium during ileal injury in rats.AIM To investigate the effects of Bifidobacterium on cytokine-induced neutrophil chemoattractant(CINC) and insulin-like growth factor 1(IGF-1) in the ileum of rats with endotoxin injury.METHODS Preweaning rats were randomly divided into three groups: Control(group C),model(group E) and treatment(group T). Group E was intraperitoneally injected with lipopolysaccharide(LPS) to create an animal model of intestinal injury.Group T was intragastrically administered Bifidobacterium suspension 7 d before LPS. Group C was intraperitoneally injected with normal saline. The rats were killed at 2, 6 or 12 h after LPS or physiological saline injection to collect ilealtissue samples. The expression of ileal CINC mRNA was evaluated by reverse transcription-polymerase chain reaction(RT-PCR), and expression of ileal IGF-1 protein and mRNA was detected by immunohistochemistry and RT-PCR,respectively.RESULTS The ileum of rats in Group C did not express CINC mRNA, ileums from Group E expressed high levels, which was then significantly decreased in Group T(F =23.947, P < 0.05). There was no significant difference in CINC mRNA expression at different times(F = 0.665, P > 0.05). There was a high level of IGF-1 brown granules in ileal crypts and epithelial cells in Group C, sparse staining in Group E, and dark, dense brown staining in Group T. There was a significant difference between Groups C and E and Groups E and T(P < 0.05). There was no significant difference in IGF-1 protein expression at different times(F = 1.269, P > 0.05). IGF-1 mRNA expression was significantly different among the three groups(P < 0.05),though not at different times(F = 0.086, P > 0.05).CONCLUSION Expression of CINC mRNA increased in the ileum of preweaning rats with endotoxin injury, and exogenous administration of Bifidobacterium reduced CINC m RNA expression. IGF-1 protein and mRNA expression decreased in the ileum of preweaning rats with endotoxin injury, and exogenous administration of Bifidobacterium prevented the decrease in IGF-1 expression. Bifidobacterium may increase IGF-1 expression and enhance intestinal immune barrier function in rats with endotoxin injury.

Neutrophil-derived interleukin-17A participates in neuroinflammation induced by traumatic brain injury

After brain injury, infiltration and abnormal activation of neutrophils damages brain tissue and worsens inflammation, but the mediators that connect activated neutrophils with neuroinflammation have not yet been fully clarified. To identify regulators of neutrophil-mediated neuroinflammation after traumatic brain injury, a mouse model of traumatic brain injury was established by controlled cortical impact. At 7 days post-injury(sub-acute phase), genome-wide transcriptomic data showed that interleukin 17 A-associated signaling pathways were markedly upregulated, suggesting that interleukin 17 A may be involved in neuroinflammation. Double immunofluorescence staining showed that interleukin 17 A was largely secreted by neutrophils rather than by glial cells and neurons. Furthermore, nuclear factor-kappaB and Stat3, both of which are important effectors in interleukin 17 A-mediated proinflammatory responses, were significantly activated. Collectively, our findings suggest that neutrophil-derived interleukin 17 A participates in neutrophil-mediated neuroinflammation during the subacute phase of traumatic brain injury. Therefore, interleukin 17 A may be a promising therapeutic target for traumatic brain injury.

PD1/PD-L1、中性粒细胞弹性蛋白酶、赖氨酰氧化酶在膀胱癌中的表达及其与膀胱癌预后的关系

目的探讨膀胱癌组织中程序性细胞死亡受体1(PD1)、程序性细胞死亡配体1(PD-L1)、中性粒细胞弹性蛋白酶(NE)、赖氨酰氧化酶(LOX)的表达水平及其表达差异与膀胱癌患者临床病理特征和预后的关系。方法选取2018年1月至2020年12月江西省人民医院收治的56例膀胱癌患者的癌组织作为研究对象,并记录患者的复发和死亡情况进行分析研究。采用免疫组织化学染色法检测PD1、PD-L1、NE、LOX的表达情况,分析PD1、PD-L1、NE、LOX与患者的病理特征和预后的关系。结果免疫组化结果显示PD1在肿瘤相关免疫细胞中存在表达(46/56);TNM分期T2-T3期、高级别肿瘤和出现淋巴结转移的患者PD1和PDL-1的表达阳性率均明显升高,差异有统计学意义(P<0.05)。NE与PD1、PDL-1的表达呈正相关(P<0.05);LOX与PD-L1、PDL-1的表达呈正相关(P<0.05)。PD1阳性表达患者3年无进展生存率为58.70%(27/46),PD-L1阳性表达患者3年无进展生存率为43.48%(10/23),不同PD1、PD-L1表达患者的3年无进展生存率比较差异有统计学意义(P<0.05)。结论膀胱癌组织中PD1、PD-L1表达增加可作为膀胱癌患者潜在的预后预测指标,有助于筛选术后复发的高危患者,并为膀胱癌的免疫学治疗提供新靶点。

脓毒症患者血清HMGB1、sTLT-1、NLR变化及其对并发急性肺损伤的预测价值

目的探讨脓毒症患者血清高迁移率族蛋白B1(HMGB1)、可溶性髓样细胞触发受体样转录因子-1(sTLT-1)、中性粒细胞/淋巴细胞比值(NLR)变化及其对并发急性肺损伤(ALI)的预测价值。方法回顾性分析2022年1月至2023年12月新乡医学院第一附属医院收治的120例脓毒症患者的临床资料,根据住院期间是否发生ALI分为ALI组35例、非ALI组85例。比较ALI组与非ALI组、ALI组不同病情程度患者血清HMGB1、sTLT-1、NLR水平,并采用受试者工作(ROC)曲线分析血清HMGB1、sTLT-1、NLR对脓毒症并发ALI的预测价值。结果ALI组患者的血清HMGB1、sTLT-1、NLR水平分别为(74.21±11.70)pg/mL、(593.06±82.45)pg/mL、5.13±1.16,明显高于非ALI组的(60.15±7.95)pg/mL、(525.73±54.84)pg/mL、4.08±0.64,差异均有统计学意义(P<0.05);ALI组中,高危组患者的血清HMGB1、sTLT-1、NLR水平分别为(86.43±5.90)pg/mL、(686.31±23.91)pg/mL、(6.49±1.11),明显高于中危组的(76.64±11.44)pg/mL、(599.28±88.40)pg/mL、5.27±1.00及低危组的(64.48±5.06)pg/mL、(539.93±35.90)pg/mL、4.27±0.71,且中危组患者均高于低危组,差异均有统计学意义(P<0.05);血清HMGB1、sTLT-1、NLR联合预测脓毒症并发ALI的曲线下面积(AUC)、敏感度、特异度分别为0.890、90.60%、92.10%,HMGB1单独检测分别为0.848、84.70%、77.10%,sTLT-1单独检测分别为0.732、71.40%、68.20%,NLR单独检测分别为0.785、62.90%、84.20%,联合检测高于各指标单独检测(P<0.05)。结论脓毒症并发ALI患者血清HMGB1、sTLT-1、NLR明显升高,且三者联合检测对并发ALI有较高的预测价值。

复发鼻息肉组织各类炎性细胞变化的独立因素分析

目的探讨影响复发鼻息肉组织各类炎性细胞变化的独立影响因素。方法选择2007年12月~2021年12月原发鼻息肉与复发鼻息肉均在烟台毓璜顶医院耳鼻咽喉头颈外科行鼻内镜手术治疗的31例鼻息肉患者临床资料,进行逐步回归Logistic分析。结果相较于原发鼻息肉,复发鼻息肉各类组织浸润炎性细胞数量均发生变化,其中嗜中性粒细胞计数和百分比均显著升高(P<0.05)。Logistic分析发现吸烟是复发鼻息肉嗜酸性粒细胞增多的独立危险因素,年龄增大是促进嗜中性粒细胞降低的独立因素,合并变应性鼻炎和复发时间间隔分别是促进淋巴细胞降低和增多的独立因素。结论复发鼻息肉组织各类浸润细胞数量均较原发鼻息肉发生变化。吸烟是复发鼻息肉嗜酸性粒细胞增多的独立危险因素。

麻杏石甘汤对铜绿假单胞菌肺炎大鼠细菌清除效应及中性粒细胞浸润和炎症因子的影响

[目的]观察麻杏石甘汤对铜绿假单胞菌(PA)肺炎大鼠细菌清除效应及炎症因子和中性粒细胞(PMNs)浸润的影响。[方法]选取雄性SD大鼠120只,随机分为5组(空白组、模型组、西药组、中药组、中西医结合组),每组24只,采用气管滴注法,建立肺炎模型,空白组和模型组予生理盐水灌胃,西药组肌注头孢他啶,中药组予麻杏石甘汤浓缩煎剂灌胃,中西医结合组采用肌注头孢他啶联合麻杏石甘汤浓缩煎剂灌胃,分4个时点(造模后0、12、24、72 h)处死大鼠,每组每个时点处死6只,取肺组织及腹主动脉血,观察肺组织病理改变、检测肺组织匀浆细菌计数、髓过氧化物酶(MPO)表达、外周血PMNs、血清肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、白细胞介素-10(IL-10)水平。[结果]1)造模后各组组内前后时点比较,模型组和药物干预组(西药组、中药组和中西医结合组)肺部炎症均逐渐改善,肺组织细菌计数均逐渐下降,差异具有统计学意义(P<0.05);72 h时点,药物干预组与模型组比较,肺部炎症均明显改善,肺组织细菌计数均降低,差异具有统计学意义(P<0.05)。2)外周血PMNs计数,模型组和西药组呈持续上升趋势,中药组和中西医结合组呈先上升后下降趋势,不同时点组间比较,差异无统计学意义(P>0.05)。3)肺组织MPO表达量,模型组呈先上升后下降趋势,药物干预组均呈下降趋势;72 h时点,药物干预组MPO表达量低于模型组,差异有统计学意义(P<0.05)。4)炎症因子比较,造模后72 h内,模型组和药物干预组血清IL-10水平均呈上升趋势,72 h时点,中药组和中西医结合组IL-10水平均高于模型组,差异有统计学意义(P<0.05);造模后0、12、24 h,模型组和药物干预组血清TNF-α、IL-8水平均呈上升趋势,24 h后,中药组和中西医结合组TNF-α呈下降趋势,药物干预组IL-8均呈下降趋势,72 h时点,药物干预组TNF-α水平较模型组均降低,差异具有统计学意义(P<0.05)。[结论]麻杏石甘汤可促进机体清除PA,减少肺组织PMNs浸润,调节血清炎症因子TNF-α、IL-8、IL-10水平,减轻肺部炎症损伤。

基于中性粒细胞胞外诱捕网探讨暖心康改善缺血性心力衰竭小鼠心功能的机制

目的探讨暖心康通过干预中性粒细胞胞外诱捕网形成,抑制炎症反应,改善心肌细胞内环境稳态,减少心肌细胞损伤,从而改善缺血性心衰小鼠心室重塑,提高心功能的作用机制。方法将30只成年健康雄性C57BL/6J小鼠随机分为3组:假手术组、模型组、暖心康组,模型组与暖心康组结扎冠状动脉左前降支建立缺血性心衰小鼠模型,假手术组仅开胸但不结扎冠状动脉。暖心康组给予暖心康灌胃,假手术组和模型组给予等量0.9%氯化钠溶液灌胃,1次/d,持续4周。对每组小鼠行心脏彩超检测左室射血分数(left ventricular ejection fraction,LVEF)、左室缩短分数(left ventricular fractional shortening,LVFS);苏木精-伊红染色法(Hematoxylin-eosin staining,HE)染色、天狼星红染色检测心肌结构及纤维化程度,RT-PCR检测各组心肌组织脑钠肽(Brain Natriuretic Peptide,BNP)、白细胞介素-6(Interleukin-6,IL-6)、白细胞介素-1β(Interleukin-1β,IL-1β)、肿瘤坏死因子α(Tumor Necrosis Factor-alpha,TNF-α)、髓过氧化物酶(myeloperoxidase,MPO)的表达;免疫组化染色(immunohistochemistry,IHC)观察MPO在心脏中的表达情况,酶联免疫吸附测定法(Enzyme linked immunosorbent assay,ELISA)检测小鼠外周血瓜氨酸化组蛋白3(citrullinated histone 3,CitH3)水平。结果与假手术组比较,模型组小鼠室壁运动幅度减弱,LVEF、LVFS显著降低(P<0.01),心脏组织炎性细胞浸润增多,心肌细胞肥大、排列紊乱,胶原纤维增多,心衰标志物BNP与炎症因子IL-6、IL-1β、TNF-α的mRNA表达水平升高(P<0.01),心脏组织中MPO的蛋白表达明显增多,mRNA表达水平显著升高(P<0.001),外周血CitH3含量显著升高(P<0.001);与模型组比较,暖心康组小鼠室壁运动幅度增大,左室射血分数、左室缩短分数升高(P<0.05),心脏组织炎性细胞浸润减少,心肌细胞结构与排列改善,心衰标志物BNP与炎症因子IL-6、IL-1β、TNF-α的mRNA表达水平降低(P<0.05),MPO的蛋白表达明显减少,mRNA表达水平显著下降(P<0.001),CitH3含量显著减少(P<0.001)。结论暖心康可改善缺血性心衰小鼠心功能,其作用机制可能是通过干预中性粒细胞胞外诱捕网形成,抑制炎症反应,改善心肌细胞内环境稳态,减少心肌细胞损伤,进而改善缺血性心衰小鼠心室重塑。

中性粒细胞明胶酶相关脂质运载蛋白和生长分化因子15在高血压肾损伤中的诊断价值

目的探讨血清中性粒细胞明胶酶相关脂质运载蛋白(NGAL)、生长分化因子15(GDF-15)联合检测在老年H型高血压患者肾损伤中的诊断价值。方法选取2021年4月至2022年6月承德医学院附属医院神经内科及全科医疗科收治的H型高血压患者177例,依据估算肾小球滤过率分为肾功能损伤组81例和肾功能正常组96例。收集2组患者一般资料、实验室检查指标,受试者工作特征曲线(ROC)分析血清NGAL、GDF-15联合检测对老年H型高血压患者肾损伤的诊断价值,Pearson相关性分析血清NGAL、GDF-15水平与老年H型高血压患者肾损伤的相关性。结果肾功能损伤组收缩压水平高于肾功能正常组[(148.53±14.62)mm Hg(1 mm Hg=0.133 kPa)vs(138.75±13.36)mm Hg,P<0.01],2组舒张压比较差异无统计学意义(P>0.05);肾功能损伤组肌酐、尿素、β_2微球蛋白(β_2-MG)、胱抑素C(Cys C)、尿微量白蛋白(MA)、NGAL及GDF-15水平高于肾功能正常组(P<0.01);ROC曲线分析显示,NGAL、GDF-15联合检测显著高于NGAL、GDF-15单独检测(P<0.01)。Pearson相关性分析显示,血清NGAL、GDF-15水平与肌酐、尿素、β_2-MG、Cys C、MA水平均呈正相关(P<0.01)。结论老年H型高血压患者出现肾损伤后血清NGAL、GDF-15水平升高,血清NGAL、GDF-15联合检测在老年H型高血压患者肾损伤诊断中具有较高的诊断价值。

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。

Toll样受体2对儿童肺炎支原体肺炎肺部炎症表现和肿瘤坏死因子-α的调节作用

目的 探讨Toll样受体2(TLR2)对肺炎支原体肺炎(MPP)病儿肺部炎症表现和肿瘤坏死因子-α(TNF-α)的调节作用。方法 选取2021年1月至2022年1月郑州大学附属儿童医院诊治的MPP病儿27例,其中难治性肺炎支原体肺炎(RMPP)病儿12例、MPP病儿15例。选取非感染病儿12例作为对照(NC)组。检测并比较不同病儿外周血TLR和TNF-α的表达,培养A549细胞并比较其与中性粒细胞肺炎支原体(MP)刺激的TNF-α表达,以及TLR2敲除后MP刺激的TNF-α表达。结果 MPP病儿血清TNF-α水平高于NC组,且RMPP组高于MPP组(P<0.05)。RMPP组外周血中性粒细胞TLR1和TLR2mRNA高于MPP组(P<0.05)。MPP组TLR2 mRNA高于NC组(P<0.05)。MP刺激组的TNF-α[A549细胞:(19.50±1.30)ng/L,中性粒细胞:(505.60±92.40)ng/L]和TLR2(A549细胞:3 760.17±772.06,中性粒细胞:4 224.00±711.12)表达显著高于NC组(P<0.05)。TLR2敲除后A549细胞后,与对照组相比,TLR2敲除的细胞MP刺激诱导的TNF-α表达受到显著抑制。蛋白质印迹法显示,MP刺激A549细胞中MyD88和NF-κB蛋白表达均明显增加(P<0.05)。阻断Myd88或NF-κB可明显减弱MP诱导的A549细胞TNF-α表达(P<0.05)。结论 TLR2可调节MPP通过TLR2-MyD88-NF-κB信号通路介导的肺部炎症和TNF-α释放。

Pancreatic melatonin enhances anti-tumor immunity in pancreatic adenocarcinoma through regulating tumor-associated neutrophils infiltration and NETosis

Tumor microenvironment contributes to poor prognosis of pancreatic adenocarcinoma(PAAD)patients.Proper regulation could improve survival.Melatonin is an endogenous hormone that delivers multiple bioactivities.Here we showed that pancreatic melatonin level is associated with patients'survival.In PAAD mice models,melatonin supplementation suppressed tumor growth,while blockade of melatonin pathway exacerbated tumor progression.This anti-tumor effect was independent of cytotoxicity but associated with tumor-associated neutrophils(TANs),and TANs depletion reversed effects of melatonin.Melatonin induced TANs infiltration and activation,therefore induced cell apoptosis of PAAD cells.Cytokine arrays revealed that melatonin had minimal impact on neutrophils but induced secretion of Cxcl2 from tumor cells.Knockdown of Cxcl2 in tumor cells abolished neutrophil migration and activation.Melatonin-induced neutrophils presented an N1-like anti-tumor phenotype,with increased neutrophil extracellular traps(NETs)causing tumor cell apoptosis through cell-to-cell contact.Proteomics analysis revealed that this reactive oxygen species(ROS)-mediated inhibition was fueled by fatty acid oxidation(FAO)in neutrophils,while FAO inhibitor abolished the anti-tumor effect.Analysis of PAAD patient specimens revealed that CXCL2 expression was associated with neutrophil infiltration.CXCL2,or TANs,combined with NET marker,can better predict patients'prognosis.Collectively,we discovered an anti-tumor mechanism of melatonin through recruiting N1-neutrophils and beneficial NET formation.

扩大的血管周围间隙和中性粒细胞/淋巴细胞比值与视神经脊髓炎谱系疾病患者严重程度的关系

目的探讨扩大的血管周围间隙(enlarged perivascular space,EPVS)和中性粒细胞/淋巴细胞比值(neutrophil to lymphocyte ratio,NLR)与视神经脊髓炎谱系疾病(neuromyelitis optica spectrum disorders,NMOSD)患者疾病严重程度的关系及其在NMOSD患者发生重度神经功能缺损中的诊断价值。方法回顾性收集2019年11月至2023年4月郑州大学第一附属医院收治的NMOSD患者164例,根据扩展残疾状况量表评分(EDSS),将患者分为轻度组(EDSS<5分,独立行走不受限)和重度组(EDSS≥5分,独立行走受限),比较两组患者的临床资料差异。采用Spearman秩相关性分析EPVS、NLR与NMOSD患者EDSS评分的相关性,采用多因素Logistics回归分析NMOSD患者发生重度神经功能缺损的影响因素,采用受试者工作曲线(ROC)评价EPVS、NLR在识别NMOSD患者发生重度神经功能缺损中的诊断价值。结果重度组总EPVS个数及NLR水平均高于轻度组(P<0.05)。Spearman秩相关结果显示,总EPVS个数(r_(s)=0.374,P<0.01)、NLR(r_(s)=0.329,P<0.01)与NMOSD患者的EDSS评分呈正相关。多因素Logistic回归分析结果显示,年龄(OR=1.082,95%CI:1.046~1.120;P<0.01)、总EPVS个数(OR=1.083,95%CI:1.008~1.663;P<0.05)、NLR(OR=1.334,95%CI:1.077~1.653;P<0.01)、脊髓受累节段数(OR=1.168,95%CI:1.073~1.272;P<0.01)是NMOSD患者发生重度神经功能缺损的影响因素。EPVS联合NLR识别NMOSD患者发生重度神经功能缺损的ROC曲线下面积(AUC)为0.775(95%CI:0.696~0.854,P<0.01),其灵敏度和特异度分别为80.00%和71.43%。结论EPVS和NLR与NMOSD患者疾病严重程度呈正相关,两者在识别NMOSD患者发生重度神经功能缺损中具有一定临床价值,两者联合时其综合诊断效能更高。

中性粒细胞百分比与白蛋白比值对吉兰-巴雷综合征患者短期预后的预测价值

目的探讨中性粒细胞百分比与白蛋白比值(NPAR)对吉兰-巴雷综合征(Guillain-Barrésyndrome,GBS)患者短期预后的预测价值。方法回顾性分析2018年1月至2022年12月南阳市中心医院收治的226例GBS患者的临床资料。根据患者出院时休斯功能分级量表(Hughes functional grading scale,HFGS)评分将患者分为短期预后良好组(HFGS<3分,n=119)和短期预后不良组(HFGS≥3分,n=107)。比较两组患者间性别构成、年龄、临床症状、体征、血液检查、脑脊液检查、免疫治疗等指标的差异。采用Spearman相关性分析NPAR与GBS患者短期预后的关系;采用多因素Logistic回归分析影响GBS患者短期预后的危险因素;绘制ROC曲线分析NPAR对GBS患者短期预后的预测价值。结果与短期预后良好组比较,短期预后不良组患者住院时间更长,肌张力低、腱反射减弱或消失、使用呼吸机、入住ICU、血浆置换患者比例高,白细胞计数、淋巴细胞计数、中性粒细胞百分比、脑脊液葡萄糖水平、NPAR高,白蛋白水平、预后营养指数更低(均P<0.05)。NPAR与GBS患者短期预后不良呈正相关(r=0.478,P<0.01)。多因素Logistic回归分析显示,NPAR、白蛋白、入住ICU、住院时间是影响GBS患者短期预后的独立危险因素(P<0.05)。ROC曲线结果显示,NPAR预测GBS患者短期预后的AUC为0.776(95%CI:0.716~0.836,P<0.01)。结论入院时NPAR与GBS患者短期预后不良相关,NPAR对评估GBS患者的短期预后具有较高的临床价值。

NLR对局晚期口腔鳞状细胞癌患者术前尼妥珠单抗联合新辅助化疗疗效预测价值研究

目的探讨尼妥珠单抗联合新辅助化疗前外周血中性粒细胞和淋巴细胞比值(neutrophil to lympho-cyte ratio,NLR)对局晚期口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)患者术前新辅助治疗近期疗效的预测价值,为临床提供参考。方法本研究获得伦理委员会审批和患者知情同意,收集2020年9月至2023年6月就诊于山西医科大学第一医院口腔颌面外科的Ⅲ、Ⅳ期OSCC患者59例,所有患者临床资料完整,都经病理学确诊为鳞状细胞癌,并接受术前尼妥珠单抗+TP(多西他赛+顺铂)新辅助化疗;分析其临床资料,收集治疗前及治疗后外周血中性粒细胞数值、淋巴细胞数值;计算比值NLR,使用受试者工作特征曲线(receiver operating characteristic curve,ROC)计算得到阈值,根据尼妥珠单抗联合TP新辅助化疗前NLR阈值将患者分为高NLR组和低NLR组;根据实体瘤疗效评价标准评估尼妥珠单抗联合TP新辅助化疗后临床疗效,分析NLR与近期疗效的相关性;使用免疫组织化学染色方法检测尼妥珠单抗联合TP新辅助化疗前后OSCC组织中表皮生长因子受体(epidermal growth factor receptor,EGFR)的表达,分析不同NLR组间EGFR表达差异。结果共收集59例晚期OSCC患者,根据ROC曲线得到NLR阈值为2.377,将患者分为<2.377组(低NLR组)24例,>2.377组(高NLR组)35例;低NLR组较高NLR组近期疗效好(P<0.05);低NLR组和高NLR组治疗后EGFR表达均下降,低NLR组较高NLR组下降幅度更大,差异有统计学意义(P<0.05)。结论尼妥珠单抗联合TP新辅助化疗前低NLR的患者有较好的疗效,此类患者更有可能在术前尼妥珠单抗联合新辅助化疗中受益。

Pu.1和cMyb在斑马鱼中性粒细胞发育中的相互作用

中性粒细胞发生是一个高度有序且被精密调控的过程,造血相关转录因子在其中起着关键作用。造血相关转录因子通过与其辅因子相互作用或转录因子之间相互作用形成复杂的调控网络,调控网络的异常可导致白血病的发生。目前参与该过程的转录因子有几十种,它们的结构与功能被广泛研究,但对转录因子之间调控关系的研究则相对缺乏。人PU.1和cMYB参与中性粒细胞发生的多个阶段且它们的异常通常与血液疾病相关,但目前对于在体情况下两者之间是否存在调控关系以及如何相互作用尚不明确。本研究利用cMyb过表达(cmybhyper)和Pu.1缺陷(pu.1^(G242D/G242D))的斑马鱼模型,通过整体原位杂交、qRT-PCR、荧光报告系统以及拯救实验检测中性粒细胞发育过程中Pu.1和cMyb的相互作用关系。结果显示,在pu.1^(G242D/G242D)突变体中,中性粒细胞的cmyb表达量显著升高,而在cmybhyper中,中性粒细胞的pu.1表达量则无明显变化。进一步在pu.1^(G242D/G242D)突变体中注射MO(morpholino)来降低cmyb的表达,并使用SB以及BrdU染色检测中性粒细胞的数量和增殖情况,发现cmyb的敲低可以拯救突变体中中性粒细胞异常增生的表型。这些结果表明,在中性粒细胞发育过程中Pu.1可以负调控cMyb的表达量。最后,本研究通过构建多位点突变质粒和荧光报告系统,证实了Pu.1能够直接结合在cmyb启动子+72 bp位点,从而负调控其表达。综上所述,本研究明确了Pu.1可以通过调控cmyb的表达参与中性粒细胞的发育,为理解两者之间调控关系及其在疾病中的作用提供了新的线索。

中老年2型糖尿病合并肌少症与NLR等炎症标志物的相关性

目的探讨中老年2型糖尿病(T2DM)合并肌少症与中性粒细胞计数/淋巴细胞计数(NLR)等炎症标志物的关系。方法选择2022年11月至2023年5月在华北理工大学附属医院内分泌科及全科医学科住院的2型糖尿病患者251例,依据合并肌少症情况将患者分为T2DM组和T2DM合并肌少症组。收集患者基本情况、临床指标等数据,计算NLR等炎症标志物。采用多因素Logistic回归分析肌少症与NLR等炎症标志物的关系。结果与T2DM组比较,T2DM合并肌少症组年龄、白细胞、尿酸、NLR、MHR水平均升高,HDL-C水平降低,差异有统计学意义(P<0.05)。多因素Logistic回归结果显示,NLR水平升高、增龄是T2DM合并肌少症的独立危险因素,HDL-C水平升高是T2DM合并肌少症的独立保护因素(P<0.05)。ROC曲线分析显示,研究对象NLR预测T2DM合并肌少症的AUC为0.737(95%CI:0.646~0.828)。结论在中老年T2DM患者中,NLR水平升高是肌少症的独立危险因素,NLR对预测T2DM合并肌少症有一定的价值。

F-NLR评分评估自发性基底节脑出血患者短期预后的临床研究

目的 探讨外周血纤维蛋白原和中性粒细胞/淋巴细胞比值联合评分(F-NLR)对自发性基底节区脑出血短期预后的预测作用。方法 回顾性分析2021年1月-2022年12月自发性基底节区脑出血患者154例,通过改良Rankin量表(mRS)评分系统进行评估,将患者分成预后良好组(n=50)和预后不良组(n=104)。应用单因素和多因素回归分析基底节区脑出血的风险因素,并利用ROC曲线评估F-NLR在基底节区脑出血预后中的应用价值。结果 2组患者的年龄、血肿体积、是否破入脑室、格拉斯哥昏迷评分(GCS评分)、白细胞(WBC)计数、中性粒细胞(ANC)计数、淋巴细胞(ALC)计数、中性粒细胞/淋巴细胞比值(NLR)、纤维蛋白原(FIB)、F-NLR水平差异有统计学意义(P<0.05)。多元logistic回归分析显示基底节区脑出血患者的血肿体积、GCS评分、高WBC、低ALC、高F-NLR评分是其短期预后不良的重要独立危险因素(P<0.05)。ROC曲线表明F-NLR评分预测基底节区脑出血预后不良的AUC值为0.799(95%CI:0.723~0.874),敏感度为66.3%,特异度为86.0%。结论 F-NLR评分对基底节区脑出血患者的近期预后具有重要指导意义,可作为进一步临床应用的依据。