C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 pathway as a therapeutic target and regulatory mechanism for spinal cord injury

Spinal cord injury involves non-reversible damage to the central nervous system that is characterized by limited regenerative capacity and secondary inflammatory damage.The expression of the C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 axis exhibits significant differences before and after injury.Recent studies have revealed that the C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 axis is closely associated with secondary inflammatory responses and the recruitment of immune cells following spinal cord injury,suggesting that this axis is a novel target and regulatory control point for treatment.This review comprehensively examines the therapeutic strategies targeting the C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 axis,along with the regenerative and repair mechanisms linking the axis to spinal cord injury.Additionally,we summarize the upstream and downstream inflammatory signaling pathways associated with spinal cord injury and the C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 axis.This review primarily elaborates on therapeutic strategies that target the C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 axis and the latest progress of research on antagonistic drugs,along with the approaches used to exploit new therapeutic targets within the C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 axis and the development of targeted drugs.Nevertheless,there are presently no clinical studies relating to spinal cord injury that are focusing on the C-C motif chemokine ligand 2/C-C motif chemokine receptor 2 axis.This review aims to provide new ideas and therapeutic strategies for the future treatment of spinal cord injury.

C–C motif chemokine ligand 16 inhibits the progression of liver cirrhosis via inactivating hepatic stellate cells

Background:Liver cirrhosis results from many forms of chronic damage,characterized by accumulation of extracellular matrix.The present study aimed to explore a potential non-invasive biomarker and its mechanism in the progression of liver cirrhosis.Methods:Gene Expression Omnibus(GEO)dataset(GSE15654,n=216)was analyzed to screen genes associated with progression of liver cirrhosis.A total of 181 plasma samples,including healthy control(HC,n=20),chronic hepatitis B(CHB,n=77)and HBV-related liver cirrhosis(LC,n=84),were enrolled for validation.In vitro and in vivo experiments were employed for the mechanistic investigation.Results:GEO dataset analysis showed that relatively low mRNA-expression of C–C motif chemokine ligand 16(CCL16)was associated with elevated Child-Pugh score(P=0.034)and worse prognosis(P=0.025).Plasma CCL16 level decreased in a stepwise pattern,with a median concentration of 10.29,6.57 and 4.47 ng/mL in the HC,CHB and LC groups,respectively(P<0.001).Low plasma CCL16 was significantly related to hepatic dysfunction both in the CHB and LC groups(P<0.05).Combination of CCL16 and ALT showed improved distinguishing capability for LC compared to either alone.In vitro,CCL16 expression was downregulated by lipopolysaccharide and hypoxia.Overexpression of CCL16 from human normal liver cell line(LO2)reduced the extracellular matrix associated proteins(Col1 and Col4)in human hepatic stellate cell line(LX-2).In vivo,the pathological feature of cirrhosis was alleviated by the hepatocytespecific expression of CCL16.Conclusions:CCL16 could be a feasible plasma marker to predict the occurrence and progression of liver cirrhosis.CCL16 might impact liver cirrhosis through inactivating hepatic stellate cells.

Macrophage migration inhibitory factor facilitates astrocytic production of the CCL2 chemokine following spinal cord injury

Spinal cord injury causes accumulation of a large number of leukocytes at the lesion site where they contribute to excessive inflammation.Overproduced chemokines are responsible for the migratory process of the leukocytes,but the regulatory mechanism underlying the production of chemokines from resident cells of the spinal cord has not been fully elucidated.We examined the protein levels of macrophage migration inhibitory factor and chemokine C-C motif chemokine ligand 2 in a spinal cord contusion model at different time points following spinal cord injury.The elevation of macrophage migration inhibitory factor at the lesion site coincided with the increase of chemokine C-C motif chemokine ligand 2 abundance in astrocytes.Stimulation of primary cultured astrocytes with different concentrations of macrophage migration inhibitory factor recombinant protein induced chemokine C-C motif chemokine ligand 2 production from the cells,and the macrophage migration inhibitory factor inhibitor 4-iodo-6-phenylpyrimidine attenuated the stimulatory effect.Further investigation into the underlying mechanism on macrophage migration inhibitory factor-mediated astrocytic production of chemokine C-C motif chemokine ligand 2 revealed that macrophage migration inhibitory factor activated intracellular JNK signaling through binding with CD74 receptor.Administration of the macrophage migration inhibitory factor inhibitor 4-iodo-6-phenylpyrimidine following spinal cord injury resulted in the reduction of chemokine C-C motif chemokine ligand 2-recruited microglia/macrophages at the lesion site and remarkably improved the hindlimb locomotor function of rats.Our results have provided insights into the functions of astrocyte-activated chemokines in the recruitment of leukocytes and may be beneficial to develop interventions targeting chemokine C-C motif chemokine ligand 2 for neuroinflammation after spinal cord injury.

新生儿持续肺动脉高压患儿血清CXCL8、CXCL12与一氧化氮吸入治疗临床转归的关系

目的探讨新生儿持续肺动脉高压(persistent pulmonary hypertension of newborn,PPHN)患儿血清人CXC型趋化因子配体8(C-X-C motif chemokine ligand 8,CXCL8)、CXCL12与一氧化氮吸入治疗临床转归的关系。方法选择2021-08/2023-05月作者医院收治并给予一氧化氮吸入治疗的PPHN患儿135例为研究对象。根据患儿出院时临床转归结局分为死亡组(n=32)和存活组(n=103)。比较两组PPHN患儿血清CXCL8、CXCL12水平。单因素及多因素Logistic回归模型分析接受一氧化氮吸入治疗PPHN患儿临床转归的影响因素。受试者工作特征(receiver operating characteristic,ROC)曲线分析血清CXCL8、CXCL12对接受一氧化氮吸入治疗PPHN患儿临床转归的预测价值。结果死亡组患儿血清CXCL8、CXCL12水平显著高于存活组(P均<0.05)。多因素Logsitic回归分析结果显示,血清CXCL8水平升高、血清CXCL12水平升高、早产、出生时Apgar评分0~3分、合并并发症是接受一氧化氮吸入治疗的PPHN患儿死亡的危险因素,肺表面活性物质应用、吸入一氧化氮早期反应则是保护因素(P<0.05)。ROC曲线分析结果显示,血清CXCL8、CXCL12联合检测对接受一氧化氮吸入治疗的PPHN患儿死亡预测的曲线下面积(area under the curve,AUC)为0.828,大于血清CXCL8、CXCL12单独检测(AUC分别为0.762、0.714)。结论PPHN患儿血清CXCL8、CXCL12水平升高与接受一氧化氮治疗的不良临床转归有关,且CXCL8、CXCL12水平升高是PPHN患儿死亡的危险因素。CXCL8、CXCL12联合检测对接受一氧化氮治疗PPHN患儿死亡具有较高的预测价值。

CXC趋化因子配体8、核转录因子κB、可溶性B7-H3在肺炎支原体肺炎患儿中的临床意义及预后评估价值

目的分析CXC趋化因子配体8(CXCL8)、核转录因子κB(NF-κB)、可溶性B7-H3(sB7-H3)在肺炎支原体肺炎(MPP)患儿中的临床意义及预后评估价值。方法选择在广州医科大学附属妇女儿童医疗中心增城院区小儿内科综合三区2022年10月至2024年3月就诊的160例MPP患儿作为MPP组,选择同期接受健康体检的100例儿童作为健康组。所有患儿均接受对症治疗,于治疗7d后随访,根据预后结局分为预后良好组(n=115)、预后不良组(n=45)。比较健康组、MPP组NF-κB、CXCL8、sB7-H3水平,比较不同预后情况MPP患儿的临床资料及NF-κB、CXCL8、sB7-H3动态变化情况;采用多因素logistic分析MPP患儿预后的影响因素,并使用受试者工作特征(ROC)曲线分析NF-κB、CXCL8、sB7-H3对MPP患儿预后的预测价值。结果MPP组NF-κB、CXCL8、sB7-H3水平高于健康组(P<0.05);预后不良组、预后良好组急性生理学与慢性健康状况评分系统Ⅱ(APACHEⅡ)评分比较有差异(P<0.05);预后不良组治疗前后NF-κB、CXCL8、sB7-H3水平均高于预后良好组(P<0.05);多因素logistic回归分析结果显示,APACHEⅡ评分、NF-κB、CXCL8、sB7-H3为MPP患儿预后的独立影响因素(P<0.05);ROC曲线分析结果显示,NF-κB、CXCL8、sB7-H3联合检测预测MPP患儿预后的曲线下面积(0.825)及灵敏度、特异度(81.37%、80.65%)均高于各项单一检测(P<0.05)。结论血清NF-κB、CXCL8、sB7-H3与MPP患儿预后存在密切关系,以上指标联合检测对预测MPP预后的价值较高。

肺结核患者血清趋化因子CXCL8、RANTES与疗效的相关性

目的探究肺结核(pulmonary tuberculosis,PTB)患者血清CXC趋化因子配体8(C-X-C motif chemokine ligand 8,CXCL8)、调节活化正常T细胞表达和分泌因子(regulate upon activation normal T cell expressed and secreted,RANTES)水平与治疗效果的关系。方法选取2020年2月至2021年2月温州医科大学附属衢州医院收治的450例初诊涂阳的PTB患者为研究对象。所有患者均采用标准抗PTB治疗,根据治疗效果分为有效组(n=383)和无效组(n=67)。比较两组一般资料及血清CXCL8、RANTES水平,采用Logistic回归分析PTB患者治疗效果的影响因素以及CXCL8、RANTES对PTB患者治疗效果的预测价值。结果治疗后,450例PTB患者治疗有效率为85.11%。无效组的全程督导占比低于有效组,吸烟、外地户籍占比及治疗前血清CXCL8、RANTES水平高于有效组,差异均有统计学意义(P<0.05),性别、职业、年龄比较,差异无统计学意义(P>0.05)。多因素Logistic回归分析结果显示,吸烟、户籍类型、CXCL8及管理方式是PTB患者治疗无效的影响因素(P<0.05)。血清CXCL8、RANTES预测PTB患者治疗效果的曲线下面积(areas under the curve,AUC)分别为0.877、0.865,敏感度分别为77.6%、79.1%,特异性分别为88.3%、89.3%;CXCL8、RANTES联合预测PTB患者治疗效果的AUC为0.955,敏感度为94.0%,特异性为85.4%。结论血清CXCL8、RANTES高水平与PTB患者治疗不良密切相关,检测血清CXCL8、RANTES有利于评估PTB患者治疗效果,且二者联合预测效果更佳。

敲低CXCL8抑制口腔鳞癌的细胞增殖和转移

目的探究在CXC基序趋化因子配体8(CXC motif chemokine ligand 8,CXCL8)在口腔鳞癌(oral squamous cell carcinoma,OSCC)中的作用。方法实时荧光定量PCR(quantitative real-time PCR,RTqPCR)和Western印迹检测OSCC组织和细胞中CXCL8的表达;分析CXCL8表达与OSCC患者临床病理特征的相关性;细胞计数试剂盒8(cell counting kit-8,CCK-8)、克隆形成、流式细胞术、划痕实验、Transwell实验和Western印迹检测CXCL8敲低对细胞增殖、凋亡、迁移、侵袭和上皮间质转化(epithelial-mesenchymal transition,EMT)的影响。结果CXCL8在OSCC组织和细胞中的表达显著上调。CXCL8高水平与肿瘤大小、TNM分期和远处转移显著相关。CXCL8的敲低可以抑制SCC9细胞增殖、迁移、侵袭和EMT,促进细胞凋亡。结论CXCL8在OSCC中高表达,敲低其表达可抑制SCC9细胞的增殖和转移。

玉屏风颗粒联合阿奇霉素治疗肺炎支原体肺炎患儿的疗效及对CXCL8、P2X7受体的影响

目的探析玉屏风颗粒联合阿奇霉素治疗肺炎支原体肺炎(MPP)患儿的疗效及对外周血CXC趋化因子8(CXCL8)、血清腺嘌呤核苷酸离子通道型7(P2X7)受体的影响。方法纳入2019年1月至2022年6月南方医科大学附属珠海医院收治的100例MPP患儿,依据随机数字表法分为观察组和对照组,各50例。对照组给予阿奇霉素0.1 g/(kg·d),0.9%氯化钠注射液250 ml配制静脉滴注用药,每日1次;观察组在对照组的基础上联合玉屏风颗粒治疗,每次5 g,每日3次,两组均连续治疗2个疗程。比较两组的临床疗效,治疗前、治疗2个疗程时的免疫因子[免疫球蛋白(Ig)A、IgG、IgM、补体C3、C4]和炎症因子[降钙素原(PCT)、C反应蛋白(CRP)、CXCL8、P2X7受体]水平,以及不良反应发生情况。结果观察组治疗总有效率高于对照组[98.00%(49/50)比80.00%(40/50)](χ^(2)=8.274,P=0.004)。治疗2个疗程时,两组外周血IgA、IgG较治疗前升高(P<0.05),且观察组高于对照组[(1.41±0.20)g/L比(1.22±0.18)g/L、(9.41±2.23)g/L比(7.72±2.45)g/L](P<0.01);IgM、C3和C4较治疗前降低(P<0.05),且观察组低于对照组[(1.12±0.21)g/L比(1.39±0.27)g/L、(1.41±0.31)g/L比(1.89±0.45)g/L、(0.61±0.14)g/L比(0.85±0.18)g/L](P<0.01)。治疗2个疗程时,两组外周血PCT、CRP、CXCL8和P2X7受体均较治疗前降低(P<0.05),且观察组低于对照组[(3.15±0.63)μg/L比(4.72±0.89)μg/L、(3.11±0.52)mg/L比(5.16±0.73)mg/L、(192±18)ng/L比(229±20)ng/L、(80.4±14.3)μg/L比(99.8±15.3)μg/L](P<0.01)。两组患儿恶心呕吐、腹痛腹泻、头晕头痛、皮疹瘙痒的发生率比较差异无统计学意义(P>0.05)。结论玉屏风颗粒联合阿奇霉素治疗MPP患儿的疗效较好,患儿外周血CXCL8水平和P2X7受体表达明显降低,且联合用药的安全性良好。

Intestinal epithelial chemokine (C-C motif) ligand 7 overexpression protects against high fat diet-induced obesity and hepatic steatosis in mice

Background:We previously found that the intestinal epithelial chemokine(C-C motif)ligand 7(CCL7)plays an important role in the development of toxin-induced acute liver damage.The detailed effects of intestinal epithelial CCL7 on chronic diseases;however,are still unclear.Here,we aimed to investigate the impact of intestinal epithelial CCL7 overexpression on high-fat diet(HFD)-induced obesity and steatohepatitis in mice.Methods:Intestinal epithelial CCL7 overexpression(CCL7tgIEC)mice and their wild-type(WT)littermates were fed with normal chow or HFD for 16 weeks to induce obesity and non-alcoholic fatty liver disease.Body weight gain,as well as adipose tissue index were assessed.Liver injury was monitored by histological analysis and real time polymerase chain reaction.Gut microbial composition was analyzed by 16S rRNA gene sequencing.Results:We found that the CCL7tgIEC mice on a HFD had markedly decreased weight gain(8.9 vs.17.0 g,P<0.05)and a lower adipose tissue index that include mesenteric fat(1.0%vs.1.76%,P<0.05),gonadal fat(2.1%vs.6.1%,P<0.05),subcutaneous fat(1.0%vs.2.8%,P<0.05)compared to WT animals.HFD-induced glucose intolerance and insulin resistance were also significantly improved in CCL7tgIEC mice compared to WT.Furthermore,HFD-fed CCL7tgIEC mice displayed less hepatic lipid accumulation and lower expression of inflammatory factors than WT mice.16S rRNA gene sequencing demonstrated that CCL7 overexpression in intestinal epithelial cells improved HFD-induced gut microbial dysbiosis.Conclusions:Our study revealed that CCL7 overexpression in the intestinal epithelium protects mice against the progression of diet-induced obesity,hepatic steatosis,and enteric dysbiosis.

肺炎支原体肺炎患儿外周血CCL2、CCL4、CXCL8、CXCL9水平与心肌损伤的关系

目的探讨肺炎支原体肺炎(MPP)患儿外周血单核细胞趋化蛋白1(MCP-1/CCL2)、巨噬细胞炎性蛋白1β(MIP-1β/CCL4)、CXC趋化因子配体8(CXCL8)、CXC趋化因子配体9(CXCL9)水平变化及其与心肌损伤的关系。方法选取2018年3月—2019年5月收治的72例MPP患儿为研究对象,根据患儿疾病严重程度分为MPP轻症组54例与MPP重症组18例;选取同期门诊体检健康儿童35例为对照组。检测各组外周血CCL2、CCL4、CXCL8、CXCL9水平以及血清急性时相蛋白指标水平。根据MPP患儿是否发生心律失常分为心律失常组12例与非心律失常组60例并检测其心肌酶谱指标。采用Pearson法分析CCL2、CCL4、CXCL9、CXCL8与MPP患儿血清急性时相蛋白指标及其与心律失常患儿心肌酶谱指标的相关性。结果MPP轻症组和MPP重症组血清CCL2、CCL4、CXCL8、CXCL9、C-反应蛋白(CRP)、α1-酸性糖蛋白(α1-AGP)、触珠蛋白(HP)、铜蓝蛋白(CP)水平均显著高于对照组,且MPP重症组显著高于MPP轻症组(P<0.05)。CCL2、CCL4均与CRP、α1-AGP、HP、CP呈正相关(P<0.05或P<0.01),CXCL8、CXCL9均与CRP、α1-AGP呈正相关(P<0.05或P<0.01),CXCL9与CP呈正相关(P<0.05)。心律失常组血清CCL2、CCL4、CXCL8、CXCL9、乳酸脱氢酶(LDH)、α-羟丁酸脱氢酶(α-HBDB)、磷酸肌酸激酶(CK)、磷酸肌酸激酶同工酶(CK-MB)水平均显著高于非心律失常组(P<0.05或P<0.01)。CCL2、CXCL8、CXCL9均与LDH、α-HBDB、CK、CK-MB呈正相关(P<0.05或P<0.01),CCL4与LDH、α-HBDB、CK-MB呈正相关(P<0.05)。结论MPP患儿外周血CCL2、CCL4、CXCL8、CXCL9水平均明显升高,并与疾病严重程度及心肌损伤有关,对MPP病情评估及治疗方案的制定均具有重要意义。

IL-8、CXCL14、IGFBP-2联合在特发性肺纤维化中的诊断价值

目的探讨白细胞介素(IL-8)、C-X-C Motif趋化因子配体14(CXCL14)、胰岛素样生长因子结合蛋白2(IGFBP-2)联合在特发性肺纤维化中的诊断价值。方法选取特发性肺纤维化患者30例为纤维组,细菌性肺炎患者30例为肺炎组,同时选取体检健康者30例为对照组,检测3组血清中IL-8、CXCL14、IGFBP-2的表达量。用受试者工作特征曲线(ROC曲线)下面积分析IL-8、CXCL14、IGFBP-2对特发性肺纤维化的诊断价值。结果特发性肺纤维化患者血清中IL-8、CXCL14和IGFBP-2的表达量均明显高于细菌性肺炎患者和体检健康者,差异有统计学意义(P<0.05)。诊断特发性肺纤维化患者与细菌性肺炎患者IL-8、CXCL14、IGFBP-2的灵敏度分别为80%、77%、70%,特异度为80%、87%、87%;诊断特发性肺纤维化患者与体检健康人群IL-8、CXCL14、IGFBP-2的灵敏度分别为83%、97%、83%,特异度为83%、87%、87%;IL-8、CXCL14、IGFBP-2联合对特发性肺纤维化患者诊断的灵敏度为86.67%,特异度为83.33%,准确度为84.44%。结论IL-8、CXCL14、IGFBP-2可作为联合诊断特发性肺纤维化的潜在标志物。

肿瘤坏死因子α刺激基因-6和CXC趋化因子配体8在瘢痕疙瘩不同部位中的表达及意义

目的探讨肿瘤坏死因子α刺激基因-6(TSG-6)和CXC趋化因子配体8(CXCL8)在瘢痕疙瘩不同部位中的表达及意义。方法选取2016年3月至2018年10月在河南大学淮河医院进行手术切除联合放射疗法治疗的瘢痕疙瘩病人25例,术中收集各病人浸润部、增生部和老化部瘢痕疙瘩组织,另外17例正常皮肤对照取自四肢损伤接受手术的病人,采用酶联免疫吸附法(ELISA)检测各皮肤组织标本中TSG-6蛋白、CXCL8蛋白以及细胞凋亡、胶原代谢相关蛋白的表达量并进行比较。结果对照组、老化部组、增生部组、浸润部组相比,TSG-6,第二个线粒体衍生的半胱天冬酶激活蛋白(second mitochondria-derived activator of caspase,Smac)、半胱氨酸蛋白酶(caspase-3)、胶原代谢相关蛋白IGF-1表达量均依次降低(P<0.05),CXCL8蛋白,凋亡相关蛋白B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、黑色素瘤凋亡抑制蛋白(Livin)、胶原代谢相关蛋白转化生长因子-β1(transforming growth factorβ1,TGF-β1)、I型胶原(Type I collagen,COL-I)、基质金属蛋白酶2(Matrix Metallo Proteinase 2,MMP-2)表达量均依次升高(P<0.05)。对照组、浸润部组、增生部组、老化部组TGS-6蛋白表达量分别为(1.81±0.19)、(0.34±0.07)、(0.64±0.12)、(1.37±0.14)ng/mL,CXCL8蛋白表达量分别为(37.54±5.61)、(248.65±40.62)、(174.01±26.25)、(66.43±10.23)ng/mL;Bcl-2蛋白表达量分别为(61.87±2.12)、(167.21±6.34)、(114.45±3.26)、(81.64±3.41)ng/mL,Smac蛋白表达量分别为(618.42±47.85)、(176.55±19.64)、(348.71±31.58)、(424.54±36.13)ng/mL,Caspase-3蛋白表达量分别为(17.67±2.64)、(5.48±1.74)、(8.87±1.54)、(15.25±2.67)ng/mL,Livin蛋白表达量分别为(1.25±0.28)、(3.94±0.35)、(2.26±0.31)、(1.51±0.24)ng/mL,P53蛋白表达量分别为(2.19±0.28)、(0.64±0.08)、(1.58±0.11)、(1.81±0.17)ng/mL;TGF-β1蛋白表达量分别为(275.54±76.16)、(421.57±56.01)、(361.55±52.15)、(324.16±74.72)ng/mL,COL-1蛋白表达量分别为(0.98±0.05)、(3.80±0.54)、(2.51±0.61)、(1.43±0.16)ng/mL,MMP-2蛋白表达量分别为(2.47±0.26)、(25.39±6.41)、(19.12±5.45)、(5.17±1.42)ng/mL,IGF-1蛋白表达量分别为(251.05±77.25)、(81.51±14.50)、(130.48±21.52)、(234.49±82.42)ng/mL;Pearson相关分析结果显示,瘢痕疙瘩病人TGS-6蛋白表达量与Bcl-2、Livin、TGF-β1、COL-1、MMP-2均呈负相关(P<0.05),与Smac、Caspase-3、p53、IGF-1均呈正相关(P<0.05);CXCL8蛋白表达量与Bcl-2、Livin、TGF-β1、COL-1、MMP-2均呈正相关(P<0.05),与Smac、Caspase-3、p53、IGF-1均呈负相关(P<0.05)。结论TSG-6和CXCL8在瘢痕疙瘩不同部位中表达有明显差异,可能通过调节细胞增殖、凋亡及胶原蛋白的合成、代谢,参与瘢痕疙瘩的发生与发展。

Neuroinflammation in mild respiratory COVID‑19:insights into cognitive impairment in milder cases

Severe acute respiratory syndrome coronavirus 2(SARSCoV-2)infection has been extensively shown to cause many neurological sequelae,and cognitive deficits(known as“brain fog”)may particularly and widely occur even in individuals with mild symptoms[1].Peripheral hyperinflammation as well as central nervous system(CNS)local immune responses may synergistically contribute to brain autoimmune injury.In addition to the direct neuroinvasion of SARS-CoV-2 and nonimmune effects such as severe systemic hypoxemia and vascular thrombosis,the central mechanism by which SARSCoV-2 accelerates cognitive-related symptoms may be related to immune effects[2].However,the precise neuroinflammatory mechanisms of SARS-CoV-2 infection have not been fully established.Fernández-Casta-da et al.[3]provided direct evidence and unique insights into the potential mechanism of cognitive impairment in mild respiratory coronavirus disease 2019(COVID-19)cases.

支原体肺炎患儿血清CXCL10和CCL8水平与疾病严重程度的关系

目的探究支原体肺炎(MPP)患儿血清CXC趋化因子配体10(CXCL10)、CC趋化因子配体8(CCL8)水平与病情严重程度的关系。方法选取海南医学院附属海南医院2020年11月-2021年11月收治的97例MPP患儿为MPP组,其中高危20例、中危35例、低危42例。另选取同期健康体检的95名健康儿童作为对照组。比较两组患儿血清CXCL10、CCL8水平。通过二元logistic回归分析MPP严重程度的独立危险因素。通过Pearson相关分析CXCL10、CCL8水平与CURB-65评分的相关性。结果MPP组血清CXCL10、CCL8水平分别为(533.27±161.96)pg/mL和(349.21±120.51)pg/mL,均高于对照组(122.35±52.15)pg/mL和(135.68±39.67)pg/mL,差异均有统计学意义(t=23.762、16.559,P均<0.05)。中、高危组患儿血清CXCL10、CCL8水平均较低危组升高,且高危组血清CXCL10、CCL8水平高于中危组,3组比较差异均有统计学意义(F=9.510、6.772,P均<0.05)。二元logistic回归分析显示,CXCL10、CCL8是MPP严重程度的独立影响因素(P均<0.05)。血清CXCL10、CCL8水平与CURB-65评分成正相关(r=7.926、7.849,P均<0.05)。受试者工作特征(ROC)曲线分析结果显示,血清CXCL10、CCL8均对MPP重症具有一定的诊断价值,且联合诊断的灵敏度和特异性优于单一指标诊断(Z=5.168、5.032,P均<0.05)。结论MPP患儿血清CXCL10、CCL8水平均升高。同时血清CXCL10、CCL8水平是影响MPP严重程度的主要因素,在预测MPP严重程度方面具有一定的指导价值。

A549肺癌细胞条件培养液促进人骨髓间充质干细胞的增殖和迁移

目的:探讨A549肺癌细胞对人骨髓间充质干细胞(hBMSCs)增殖和运动的影响。方法:以提取A549肺癌细胞和BEAS-2B正常人肺上皮细胞上清液制备条件培养基,培养的hBMSCs分别为A549组和BEAS-2B组;2组均添50 nmol/L趋化因子-8(CXCL-8),分别为A549 C组和BEAS-2B C组,若同时添加100 nmol/L triciribine(Akt抑制剂)分别为A549 CT组和BEAS-2B CT组;A549组加入100 nmol/L triciribine为A549 T组。正常条件下培养的hBMSCs为对照组。ELISA检测细胞上清液中CXCL-8含量和hBMSCs裂解液Akt、P-Akt蛋白的表达。MTT实验、流式细胞仪和transwell细胞小室实验分别检测各组hBMSCs吸光度值(A490值)、细胞凋亡率和细胞迁移数目。结果:与BEAS-2B上清液相比,A549上清液中CXCL-8含量明显升高。各组hBMSCs的A490值、细胞凋亡率、细胞迁移数目、Akt和P-Akt蛋白表达等均具有显著差异,尤以A549 C组hBMSCs最明显。与A549 C组相比,A549 CT组hBMSCs的A490值和细胞迁移数目、Akt和P-Akt蛋白表达均降低,细胞凋亡率升高较显著。结论:A549细胞表达的CXCL-8能激活Akt通路,促进hBMSCs增殖和运动。

不同浓度A549细胞上清液对人骨髓间充质干细胞迁移的影响

目的:验证不同浓度A549细胞上清液对人骨髓间充质干细胞(hBMSC)迁移的影响.方法:提取A549细胞上清液,以5%和25%条件培养基(CM)培养hBMSCs为5%A549-CM组、25%A549-CM组,正常培养的hBMSCs为对照组.Transwell细胞迁移实验检测各组hBMSCs划痕面积闭合率;Western blot检测各组hBMSCs中CXCL-8蛋白含量;ELISA检测各组hBMSCs的Erk、p-Erk和PI3K、p-PI3K蛋白的表达.结果:对照组、5%A549-CM组、25%A549-CM组hBMSCs细胞划痕闭合率逐渐升高,差异有统计学意义(P<0.01).与对照组比较,5%A549-CM组、25%A549-CM组hBMSCs的Erk、p-Erk和PI3K、p-PI3K蛋白表达明显增高,差异均有统计学意义(P<0.01).与对照组比较,5%A549-CM组、25%A549-CM组hBMSCs的CXCL-8蛋白表达明显增高(P<0.01).结论:A549细胞上清液能够激活hBMSCs内PI3K-Erk信号通路促进hBMSCs迁移.

Transforming growth factor-β and toll-like receptor-4 polymorphisms are not associated with fibrosis in haemochromatosis

AIM:To investigate the role of genetic polymorphisms in the progression of hepatic fibrosis in hereditary haemochromatosis.METHODS:A cohort of 245 well-characterised C282Y homozygous patients with haemochromatosis was studied,with all subjects having liver biopsy data and DNA available for testing.This study assessed the association of eight single nucleotide polymorphisms(SNPs)in a total of six genes including toll-like receptor 4(TLR4),transforming growth factor-beta(TGF-β),oxoguanine DNA glycosylase,monocyte chemoattractant protein 1,chemokine C-C motif receptor 2 and interleukin-10 with liver disease severity.Genotyping was performed using high resolution melt analysis and sequencing.The results were analysed in relation to the stage of hepatic fibrosis in multivariate analysis incorporating other cofactors including alcohol consumption and hepatic iron concentration.RESULTS:There were significant associations between the cofactors of male gender(P=0.0001),increasing age(P=0.006),alcohol consumption(P=0.0001),steatosis(P=0.03),hepatic iron concentration(P<0.0001)and the presence of hepatic fibrosis.Of the candidate gene polymorphisms studied,none showed a significant association with hepatic fibrosis in univariate or multivariate analysis incorporating cofactors.We also specifically studied patients with hepatic iron loading above threshold levels for cirrhosis and compared the genetic polymorphisms between those with no fibrosis vs cirrhosis however there was no significant effect from any of the candidate genes studied.Importantly,in this large,well characterised cohort of patients there was no association between SNPs for TGF-βor TLR4and the presence of fibrosis,cirrhosis or increasing fibrosis stage in multivariate analysis.CONCLUSION:In our large,well characterised group of haemochromatosis subjects we did not demonstrate any relationship between candidate gene polymorphisms and hepatic fibrosis or cirrhosis.

CXCL8/CXCR1驱动FOXS1蛋白促进食管癌ECA109细胞增殖、侵袭及迁移

为了探讨CXC趋化因子配体8(CXC motif chemokine ligand 8,CXCL8)/CXC趋化因子受体1(CXC chemokine receptor 1,CXCR1)对体外人食管癌细胞ECA109增殖、凋亡及侵袭迁移的影响,并观察其与叉头框家族S1(forkhead box S1,FOXS1)蛋白的关系,将人食管癌细胞ECA109转染后分为对照组、CXCL8组、CXCL8+Con-siRNA组和CXCL8+CXCR1-siRNA组。CXCL8终浓度为10μg/mL。采用MTT试验检测细胞增殖情况,克隆试验检测克隆形成情况,流式细胞仪进行细胞凋亡测试,侵袭与迁移能力由Transwell试验和划痕试验测定,Western blotting检测细胞中CXCR1、FOXS1蛋白表达水平。结果显示,与对照组比较,CXCL8能够显著促进ECA109细胞增殖、克隆、侵袭和迁移(P<0.05),而干扰CXCR1后这些作用被阻断(P<0.05)。CXCL8组、CXCL8+Con-siRNA组细胞凋亡率与对照组比较,差异无统计学意义(P>0.05),CXCL8+CXCR1-siRNA组细胞凋亡率显著下降(P<0.05)。与对照组比较,CXCL8能够上调食管癌细胞CXCR1和FOXS1蛋白表达(P<0.05),而干扰CXCR1后CXCR1和FOXS1蛋白表达均出现显著下降(P<0.05)。该研究提示,CXCL8/CXCR1可促进食管癌细胞增殖、侵袭和迁移,其作用可能与驱动FOXS1蛋白有关。

R406对人RASFs炎性因子分泌及生物学功能调节、关节炎小鼠关节炎症缓解作用观察

目的探讨R406(free base)对人类风湿关节炎关节滑膜成纤维细胞(RASFs)炎性因子分泌的抑制及生物学功能(侵袭、迁移和凋亡)的调节作用,观察R406对关节炎小鼠关节炎症的缓解作用。方法取3~7代生长的RASFs,分为实验组、阴性对照组和阳性对照组。实验组分别用1、10、50、100 ng/mL R406和10 ng/mL IL-1β干预,阴性对照组加入DMSO,阳性对照组加入DMSO和10 ng/mL IL-1β干预。干预24 h,采用qRT-PCR法检测各组炎性因子IL-6、IL-8、CCL2 mRNA。选择最适浓度(50 ng/mL)的R406作用于RASFs(实验组)不同时间(6、12、24、48 h),阴性对照组及阳性对照组处理方法同上,采用qRT-PCR检测各组IL-6、IL-8、CCL2 mRNA,ELISA法检测各组IL-6、IL-8、CCL2蛋白,Trans Well检测各组RASFs的侵袭能力;划痕法检测细胞迁移能力;流式细胞术检测细胞凋亡率。建立Ⅱ型胶原诱导型关节炎(CIA)模型鼠,将18只8周龄的DBA 1/J小鼠平均分为实验组、阴性对照组和阳性对照组。实验组诱发CIA后腹腔注射R406,200 mg/kg,每两天注射1次;阴性对照组不诱发CIA;阳性对照组诱发CIA后腹腔注射等量DMSO,每两天注射1次。R406治疗30 d后,ELISA法检测各组小鼠血清中炎性因子IL-6、IL-8、IL-1α、COMP。结果与阳性对照组比较,实验组中不同剂量R406作用后RASFs中炎性因子IL-6、IL-8、CCL2 mRNA表达被抑制,且在50 ng/mL时抑制效果最为显著(P均<0.01)。与阳性对照组比较,实验组中50 ng/mL R406作用于RASFs不同时间,细胞中炎性因子IL-6、IL-8、CCL2蛋白的表达均被抑制,且在24 h抑制效果最显著(P均<0.01);与阳性对照组比较,实验组RASFs上清液中炎性因子IL-6、IL-8、CCL2水平降低(P均<0.01)。与阳性对照组比较,实验组RASFs侵袭和迁移数均降低,凋亡率增加(P均<0.05)。与阳性对照组比较,实验组小鼠血清中炎性因子IL-6、IL-8、IL-1α、COMP水平降低(P均<0.01)。结论R406可以抑制RASFs中炎性因子的分泌,调节RASFs侵袭、迁移、凋亡能力,对CIA模型鼠关节炎症具有抑制作用。

NFATc2对卵巢癌细胞的生长调控及其机制研究

目的 探讨活化T细胞核因子c2(NFATc2)对人卵巢癌细胞增殖和凋亡的影响,并研究其可能的机制。方法 选取人上皮性卵巢癌细胞株A2780,分别构建NFATc2过表达(过表达组)和低表达[转染小干扰RNA(siRNA),NFATc2 siRNA组]的A2780细胞株,并设立对照组(不进行任何处理);采用噻唑蓝(MTT)法检测细胞的增殖水平,采用流式细胞仪检测细胞的凋亡水平;采用Western blot检测凋亡相关蛋白和CXC趋化因子配体8(CXCL8)表达水平。分别用正常的A2780细胞(空白组)、转染空白载体的A2780细胞(对照组)和低表达NFATc2的A2780细胞(NFATc2 siRNA组)建立裸鼠皮下移植瘤的卵巢癌模型,测量肿瘤的体积和质量,用Western blot检测肿瘤组织中CXCL8的表达水平。结果 与对照组比较,NFATc2 siRNA组细胞增殖率降低,细胞凋亡率升高,半胱氨酸蛋白酶3剪切体(cleaved caspase-3)表达水平增加,Bcl-2/Bax比值降低,CXCL8表达水平降低,差异均有统计学意义(P<0.01);过表达组细胞株则呈相反效应。NFATc2 siRNA组裸鼠肿瘤质量和体积、CXCL8表达水平均明显低于空白组和对照组(P<0.01)。结论 下调NFATc2可通过抑制A2780细胞中CXCL8的表达,减少细胞增殖,增加凋亡相关蛋白表达继而促进卵巢癌细胞凋亡。