脓毒症患者血清TNF-α与T淋巴细胞亚群、凝血功能及预后的关系研究

摘要 目的：探讨脓毒症患者血清肿瘤坏死因子-α（TNF-α）与T淋巴细胞亚群、凝血功能及预后的关系。方法：选取2017年4月~2019年4月期间我院收治的脓毒症患者93例作为脓毒症组,另选取同期来我院行健康体检的志愿者90例作为对照组,比较脓毒症组、对照组的TNF-α、T淋巴细胞亚群、凝血功能,根据28d后转归情况分为死亡组和存活组,比较死亡组和存活组TNF-α、T淋巴细胞亚群水平、凝血功能,采用Pearson相关分析分析TNF-α与T淋巴细胞亚群、凝血功能的相关性。结果：脓毒症组TNF-α高于对照组,凝血酶原时间（PT）、活化部分凝血活酶时间（APTT）长于对照组,而CD3⁺、CD4⁺、CD4⁺/CD8⁺均低于对照组（P<0.05）;脓毒症组、对照组CD8⁺比较差异无统计学意义（P>0.05）。死亡组TNF-α高于存活组,PT、APTT长于存活组,而CD3⁺、CD4⁺、CD4⁺/CD8⁺均低于存活组（P<0.05）;死亡组、存活组CD8⁺比较差异无统计学意义（P>0.05）。Pearson相关分析结果显示,脓毒症患者血清TNF-α与PT、APTT呈正相关,与CD3⁺、CD4⁺、CD4⁺/CD8⁺呈负相关（P<0.05）;TNF-α与CD8⁺无明显相关性（P>0.05）。结论：脓毒症患者血清TNF-α、T淋巴细胞亚群、凝血功能均存在异常变化,TNF-α与患者的T淋巴细胞亚群、凝血功能和预后有关,检测TNF-α有助于评估脓毒症患者的病情和预后。     

三十六荡坎蛤散干预哮喘缓解期小鼠肠组织蛋白质组学差异表达研究

摘要 目的：用蛋白质组学方法探究三十六荡坎蛤散对哮喘缓解期小鼠肠组织蛋白差异表达的影响。方法：将45只SPF级雌性C57BL/6小鼠随机分为正常组、模型组、三十六荡坎蛤散组,采用HE染色法观察小鼠肺组织的病理变化,采用ELISA法检测肺泡灌洗液（BALF）中白介素-1β（IL-1β）、白介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）水平,提取肠组织蛋白,利用非标记（Label-free）定量蛋白质组学技术研究蛋白表达差异。结果：与模型组比较,三十六荡坎蛤散组显著改善了哮喘缓解期小鼠肺组织的病理状态,减轻炎症反应,改善小鼠哮喘症状;BALF中IL-1β,IL-6和TNF-α水平降低（P<0.01）;蛋白质组学研究共鉴定到6634个蛋白,模型组和三十六荡坎蛤散组共有232个差异蛋白,模型组和正常组共有34个差异蛋白,模型组与正常组比较有差异而三十六荡坎蛤散使差异趋势逆转的蛋白有6个,即Hsd17b2,Plp2,Vnn1,Olfm4,Hsbp1,Gimd。GO分析显示差异表达蛋白主要参与新陈代谢、炎症反应、神经系统调节等生物过程,KEGG分析显示,三十六荡坎蛤散主要涉及新陈代谢、炎症反应和免疫调节等10条信号通路。结论：三十六荡坎蛤散可能通过逆转Hsd17b2,Plp2,Vnn1,Olfm4,Hsbp1,Gimd的表达,参与多个生物过程,作用多条信号通路实现干预哮喘。     

微小RNA-223(miR-223)在脓毒症早期诊断、炎症程度评估中的价值研究

摘要 目的：分析微小RNA-223（miR-223）在脓毒症的早期诊断、炎症严重程度评估中的检测价值。方法：通过便利抽样法选定本院2020年5月至2022年5月住院的30例脓毒症患者作为观察组,选取同期门诊体检中心30例健康体检者作为对照组,检测、比较两组外周血miR-223、血清降钙素原（PCT）、C反应蛋白（CRP）,比较脓毒症患者中轻度组、严重组外周血miR-223、血清PCT、CRP,比较脓毒症患者中死亡组、存活组外周血miR-223、血清PCT、CRP,Spearman分析miR-223与PCT、CRP的相关性,绘制ROC曲线,计算曲线下面积（AUC）,分析miR-223与PCT、CRP对脓毒症的预测价值。结果：观察组外周血miR-223、血清PCT、CRP水平均高于对照组,P＜0.05（差异均具有统计学意义）。严重组外周血miR-223、血清PCT、CRP水平均高于轻度组,P＜0.05（差异均具有统计学意义）。死亡组外周血miR-223、血清PCT、CRP水平均高于存活组,P＜0.05（差异均具有统计学意义）。miR-223与PCT、CRP均呈正相关性,P＜0.05（差异均具有统计学意义）（r值=0.796、0.785）。ROC曲线结果显示：miR-223+PCT+CRP诊断脓毒症的灵敏度（89.54%）、特异度（86.52%）明显高于miR-223（82.37%、80.44%）、PCT（78.34%、75.34%）、CRP（75.12%、74.07%）,P＜0.05（差异均具有统计学意义）。结论：脓毒症患者机体外周血miR-223水平较高,且外周血miR-223与血清PCT、CRP呈正相关性,miR-223联合PCT、CRP可提高脓毒症诊断灵敏度、特异度。     

脓毒症患者短期预后的影响因素及YAP联合STAT3检测的预测价值研究

摘要 目的：研究脓毒症患者短期预后的影响因素及Yes相关蛋白（YAP）联合信号转导子和转录激活子3（STAT3） 信使RNA（mRNA）检测的预测价值。方法：选择我院2019年3月～2021年1月收治的131例脓毒症患者,将其按照28d预后情况的差异分成死亡组以及生存组。比较两组各项基线资料,外周血单个核细胞YAP、STAT3 mRNA表达水平。采用多因素Logistic回归分析脓毒症患者短期预后的影响因素。此外,通过受试者工作特征（ROC）曲线分析外周血单个核细胞YAP、STAT3 mRNA表达预测脓毒症患者短期预后的效能。结果：死亡组患者的外周血单个核细胞YAP、STAT3 mRNA相对表达量均高于生存组患者（P＜0.05）。死亡组年龄大于生存组,住院时间短于生存组,APACHEⅡ评分高于生存组,合并基础疾病、机械通气的患者比例高于生存组,差异均有统计学意义（P＜0.05）。多因素Logistic回归分析结果显示,年龄、APACHEⅡ评分、合并基础疾病、机械通气以及外周血单个核细胞YAP、STAT3 mRNA表达水平是脓毒症患者短期预后的影响因素（P＜0.05）。外周血单个核细胞YAP、STAT3 mRNA联合检测预测脓毒症患者短期预后的曲线下面积为0.845,高于两指标单独检测的0.623、0.687。结论：脓毒症患者短期预后的影响因素包括年龄、APACHEⅡ评分、机械通气以及合并基础疾病等,联合检测外周血单个核细胞YAP、STAT3 mRNA表达对脓毒症患者短期预后具有一定预测价值。     

脓毒症患儿血清SAA、PCT、CRP水平与预后的关系及其诊断价值分析

摘要 目的：探讨脓毒症患儿血清淀粉样蛋白A（SAA）、降钙素原（PCT）、C反应蛋白（CRP）与预后的关系,并分析三者对脓毒症的诊断价值。方法：纳入我院于2016年8月～2020年6月期间收治的脓毒症患儿60例开展回顾性研究,作为脓毒症组,选取同期于我院进行体检的健康儿童40例作为对照组,比较两组血清SAA、PCT、CRP水平。根据脓毒症患儿1个月内的生存、死亡情况,分成生存组（n=42）、死亡组（n=18）,比较两组临床资料及血清SAA、PCT、CRP水平,经COX回归模型分析脓毒症患儿死亡的危险因素。绘制受试者工作特征（ROC）曲线分析血清SAA、PCT、CRP对脓毒症的诊断价值。结果：脓毒症组血清SAA、PCT、CRP水平显著高于对照组（P＜0.05）。死亡组器官障碍数量＞2个、脓毒性休克患儿占比分别为55.56%、44.44%,显著高于生存组的19.05%、9.52%（P＜0.05）;死亡组入院后1 h内使用抗菌治疗患儿占比为38.89%,显著低于生存组的69.05%（P＜0.05）;死亡组血清SAA、PCT、CRP水平高于生存组（P＜0.05）。COX多因素分析结果显示,器官障碍数量＞2、脓毒性休克及血清SAA、PCT、CRP水平升高是脓毒症患儿死亡的危险因素（P＜0.05）,而入院后1 h内使用抗菌治疗是脓毒症患儿死亡风险的保护性因素（P＜0.05）。血清SAA、PCT、CRP单独及三者联合诊断脓毒症的曲线下面积（AUC）分别为0.808、0.780、0.761、0.912。结论：脓毒症患儿血清SAA、PCT、CRP明显升高,三者升高均为脓毒症患儿死亡的危险因素,且对脓毒症具有一定诊断价值。     

脓毒症患者血清H2S、GDF-15、PTX-3水平与凝血功能、炎症指标及病情评分的相关性分析

摘要 目的：探讨脓毒症患者血清硫化氢（H₂S）、生长分化因子-15（GDF-15）、穿透素-3（PTX-3）水平与其凝血功能、炎症指标及病情评分的相关性。方法：随机选取我院2018年2月～2020年2月收治的脓毒症患者52例作为脓毒症组,另选取我院同期收治的脓毒症休克患者46例作为休克组以及同期于我院进行体检的健康者50例作为对照组。检测三组血清H₂S、GDF-15、PTX-3水平以及凝血功能、炎症指标,其中凝血功能指标包括血小板计数（PLT）、部分凝血活酶时间（APTT）、凝血酶原时间（PT）、纤维蛋白原（FIB）。炎症指标包括降钙素原（PCT）、C反应蛋白（CRP）。采用急性生理学和慢性健康状况评分系统Ⅱ（APACHEⅡ）、快速序贯器官功能（qSOFA）评分对脓毒症组、休克组患者病情进行评分。分析血清H2S、GDF-15、PTX-3与患者凝血功能、炎症指标及病情评分的相关性。结果：脓毒症组、休克组的血清H₂S、PLT均低于对照组,且休克组低于脓毒症组（P＜0.05）。脓毒症组、休克组的血清GDF-15、PTX-3、APTT、PT、FIB、PCT、CRP均高于对照组,且休克组高于脓毒症组（P＜0.05）。脓毒症组APACHEⅡ、qSOFA评分均显著低于休克组（P＜0.05）。血清H2S与PLT呈正相关（P＜0.05）,与APTT、PT、FIB、PCT、CRP、APACHEⅡ评分、qSOFA评分呈负相关（P＜0.05）。血清GDF-15、PTX-3与PLT呈负相关（P＜0.05）,与APTT、PT、FIB、PCT、CRP、APACHEⅡ评分、qSOFA评分呈正相关（P＜0.05）。结论：脓毒症患者的血清H₂S明显下降,而血清GDF-15、PTX-3增高,三者与凝血功能、炎症以及病情评分均存在密切关联,这可能是影响脓毒症进展的重要原因之一。     

七氟醚通过影响外周血miR-340水平逆转脓毒症大鼠巨噬细胞吞噬功能抑制状态的机制

摘要 目的：探究七氟醚通过影响外周血miR-340水平逆转脓毒症大鼠巨噬细胞吞噬功能抑制状态的机制。方法：60只雄性Sprague-Dawley大鼠根据研究目的将大鼠分为对照组、脓毒症组和七氟醚组。通过RT-PCR分析各组大鼠外周血miR-340以及促炎因子IL-1β、IL-6和TNF-α的mRNA表达水平。统计各实验组大鼠的存活率。通过血琼脂平板对各组大鼠腹腔液和血液中的细菌进行计数。通过使用荧光显微镜测量吞噬率和吞噬指数。通过蛋白印迹分析p65和NF-κB的蛋白表达。结果：脓毒症组miR-340水平较对照组升高（P<0.05）,七氟醚组miR-340水平较脓毒症组降低（P<0.05）。三组不同时间点存活率相比差异无统计学意义（P＞0.05）;第4 d、8 d脓毒症组大鼠存活率较对照组大鼠降低（P<0.05）,七氟醚组大鼠存活率较脓毒症组升高（P<0.05）。脓毒症组IL-1β、IL-6和TNF-α的mRNA表达较对照组升高（P<0.05）,七氟醚组IL-1β、IL-6和TNF-α的mRNA表达较脓毒症组降低（P<0.05）。脓毒症组腹腔液和血液中的细菌数量较对照组升高（P<0.05）,七氟醚组腹腔液和血液中的细菌数量较脓毒症组减少（P<0.05）。血琼脂平板对各组大鼠腹腔液和血液中的细菌进行计数,脓毒症组腹腔液和血液中的细菌数量较对照组升高（P<0.05）,七氟醚组腹腔液和血液中的细菌数量较脓毒症组减少（P<0.05）。脓毒症组p65和NF-κB的蛋白表达较对照组升高（P<0.05）,七氟醚组p65和NF-κB的蛋白表达较脓毒症组降低（P<0.05）。结论：miR-340参与了脓毒症大鼠巨噬细胞吞噬功能调节,七氟醚通过降低外周血miR-340水平减少内毒素诱导的大鼠促炎细胞因子的释放,并恢复巨噬细胞吞噬功能。     

脓毒症患者发生凝血功能紊乱的临床特征及预后影响因素分析

摘要 目的：探究脓毒症患者发生凝血功能紊乱的临床特征及预后影响因素。方法：选择2019年9月~2023年1月本院收治的80例脓毒症患者为本次研究对象,对所有患者开展凝血功能检验,并依据检验结果,将患者分为凝血功能异常组（n=60）及正常组（n=20）,分析凝血功能障碍异常及正常患者临床特征、凝血功能障碍异常及正常患者病情严重程度,并依据脓毒症患者预后,将其分为存活组（n=64）及死亡组（n=16）脓毒症患者预后的单因素与多因素Logistic回归分析。结果：凝血功能正常组及异常组平均动脉压、体温、白细胞计数（WBC）、纤维蛋白原（FIB）、凝血酶时间（TT）指标水平无较大差异（P＞0.05）,与凝血功能正常组比较,凝血功能异常组患者呼吸、脉率、降钙素原（PCT）、C反应蛋白（CRP）指标水平相对较高,部分凝血活酶时间（APTT）、凝血酶原时间（PT）指标水平相对较长,PLT指标水平相对较高（P＜0.05）;与凝血功能正常的脓毒症患者相比较,凝血功能异常患者急性肾损伤（AKI）及急性呼吸窘迫综合征（ARDS）发生率、急性生理与慢性健康评分（APACHE Ⅱ）及急性生理功能评分（SOFA）评分更高,住ICU时间均相对较长（P＜0.05）,机械通气及住院时间比较（P＞0.05）;脓毒症患者预后影响因素分析中,结果显示,年龄、体质量指数（BMI）、性别、糖尿病史、高血压病史均未对脓毒症患者预后产生较大影响（P＞0.05）,PT、PCT、APTT、CRP、SOFA评分及APACHE Ⅱ评分对脓毒症预后产生严重影响,死亡组患者各指标水平均高于存活组（P＜0.05）;将影响脓毒症患者预后的单因素予以赋值,纳入Logistic回归分析,以PCT、PT、APTT、CRP、SOFA、APACHE Ⅱ 评分为自变量,结果显示,PCT、PT、CRP、SOFA、APACHE Ⅱ 评分是影响脓毒症患者预后的独立危险因素（P＜0.05）。结论：脓毒症患者发生凝血功能紊乱会对临床相关指标产生影响,从而增加疾病严重程度,依据脓毒症患者预后,研究结果显示,PT、PCT、CRP、SOFA及APACHE Ⅱ评分均会对脓毒症患者预后产生不良影响,检验上述指标水平,可为临床评估脓毒症预后提供一定参考价值。     

亚急性梭曼染毒豚鼠颈部脊髓差异表达蛋白分析

目的：利用蛋白质组学技术研究亚急性梭曼染毒豚鼠颈部脊髓中的差异表达蛋白,探讨梭曼中毒影响的主要生物学通路,为其中毒诊断、治疗及预后提供重要生物标志物。方法：以雄性成年豚鼠为研究对象,连续14天每天1次对豚鼠背部皮下注射梭曼(0.2×LD₅₀)进行亚急性染毒;末次染毒后取颈部脊髓组织,利用蛋白质组学技术分析梭曼染毒组与对照组之间的差异表达蛋白;利用KEGG数据库对差异表达蛋白进行功能注释与通路富集分析;对重要通路中的差异蛋白进行功能分析与讨论。结果：豚鼠颈部脊髓组织中共鉴定出通过KEGG库注释的蛋白质数目为3 563个;采用主成分分析和偏最小二乘法分析表明,梭曼染毒后共有407个差异表达蛋白,其中276个蛋白质表达上调,131个蛋白质表达下调,这些差异蛋白主要与代谢、遗传信息处理、环境信息处理、细胞过程、组织系统、人类疾病相关;上调蛋白质主要富集于27个通路(P<0.05),下调蛋白质主要富集于8个通路(P<0.05);上调蛋白质的主要富集通路包括吞噬体、紧密连接、细胞外基质-受体相互作用、PI3K-Akt信号通路、JAK-STAT信号通路、补体和凝血级...     

不同病原学结果血流感染脓毒症患者PT、NLR、PCT/ALB的变化及其与预后的关系研究

摘要 目的：探讨不同病原学结果血流感染（BSI）脓毒症患者凝血酶原时间（PT）、中性粒细胞/淋巴细胞比值（NLR）、降钙素原/白蛋白（PCT/ALB）的变化及其与预后的关系。方法：选取2021年1月～2023年3月复旦大学附属上海市第五人民医院收治的BSI脓毒症患者130例,根据不同病原学结果分为真菌组11例、革兰阳性菌（G⁺菌）组49例、革兰阴性菌（G^-菌）组70例。根据28d临床结局将BSI脓毒症患者分为死亡组36例和存活组94例。检测并计算PT、NLR、PCT/ALB。采用多因素Logistic回归分析影响BSI脓毒症患者预后的因素,采用受试者工作特征（ROC）曲线分析PT、NLR、PCT/ALB对BSI脓毒症患者死亡的预测价值。结果：G^-菌组PT长于真菌组、G⁺菌组,NLR、PCT/ALB高于真菌组、G⁺菌组（P＜0.05）,真菌组与G⁺菌组PT、NLR、PCT/ALB比较无差异（P＞0.05）。多因素Logistic回归分析显示脓毒性休克、PT延长,序贯器官衰竭评估（SOFA）评分升高、急性生理和慢性健康评估Ⅱ（APACHEⅡ）评分升高、NLR升高、PCT/ALB升高为影响BSI脓毒症患者预后的独立危险因素（P＜0.05）。ROC曲线分析显示PT、NLR、PCT/ALB联合预测BSI脓毒症患者死亡的曲线下面积为0.897,大于PT、NLR、PCT/ALB单独预测（P＜0.05）。结论：相比真菌和G⁺菌,G^-菌BSI脓毒症患者PT、NLR、PCT/ALB明显升高,且PT、NLR、PCT/ALB升高是影响BSI脓毒症患者预后的独立危险因素,三者联合对BSI脓毒症患者预后的预测价值较高。     

应用GSEA和WGCNA方法分析细菌性败血症宿主关键差异基因及意义

【目的】采用生物信息学方法分析公共数据库来源的细菌性败血症患者全血转录组学表达谱,探讨细菌败血症相关的宿主关键差异基因及意义。【方法】基于GEO数据库中GSE80496和GSE72829全血转录组基因数据集,采用GEO2R、基因集富集分析(GSEA)联用加权基因共表达网络分析(WGCNA)筛选细菌性败血症患者相比健康人群显著改变的差异基因,通过R软件对交集基因进行GO功能分析和KEGG富集分析。同时,通过String 11.0和Cytoscape分析枢纽基因,验证枢纽基因在数据集GSE72809(Health组52例,Definedsepsis组52例)全血标本中的表达情况,并探讨婴儿性别、月(胎)龄、出生体重、是否接触抗生素等因素与靶基因表达谱间的关系。【结果】分析GSE80496和GSE72829数据集分别筛选得到932个基因和319个基因,联合WGCNA枢纽模块交集得到与细菌性败血症发病相关的10个枢纽基因(MMP9、ITGAM、CSTD、GAPDH、PGLYRP1、FOLR3、OSCAR、TLR5、IL1RN和TIMP1);GSEA分析获得关键通路(氨基酸糖类-核糖代谢、PPAR信号通路、聚糖生物合成通路、自噬调控通路、补体、凝血因子级联反应、尼古丁和烟酰胺代谢、不饱和脂肪酸生物合成和阿尔兹海默症通路)及生物学过程(类固醇激素分泌、腺苷酸环化酶的激活、细胞外基质降解和金属离子运输)。【结论】本项研究通过GEO2R、GSEA联用WGCNA分析,筛选出与细菌性败血症发病相关的2个枢纽模块、10个枢纽基因以及一些关键信号通路和生物学过程,可为后续深入研究细菌性败血症致病机制奠定理论依据。     

Quantitative proteomics analysis of human vitreous in rhegmatogenous retinal detachment associated with choroidal detachment by data-independent acquisition mass spectrometry

The prognosis of rhegmatogenous retinal detachment (RRD) with choroidal detachment (RRDCD) is often poor and complicated. This study focused on the identification of the characteristic proteins and signal pathways associated with the etiology of RRDCD and to provide guidance for diagnosis and treatment of RRDCD. In this study, vitreous humor samples were obtained from 16 RRDCD patients, 14 with RRD, 12 with idiopathic epiretinal macular membrane (IEMM), and 5 healthy controls from donated corpse eyes. Data-independent acquisition mass spectrometry and bioinformatics analysis were employed to identify differentially expressed proteins (DEPs). In the vitreous humor, 14,842 peptides were identified. Patients with RRDCD had 249 DEPs (93 upregulated and 156 downregulated), with 89 in patients with RRD and 61 in patients with IEMM. Enrichment analysis of the GO and Kyoto Encyclopedia of Genes and Genomes DEP databases indicated functional clusters related to inflammation and immunity, protein degradation and absorption, cell adhesion molecules (CAMs), the hedgehog signaling pathway, and lipid metabolism. Weighted gene co-expression network analysis showed that DEPs with positive co-expression of RRDCD participated in immune-related pathways led by the complement and coagulation cascade, whereas DEPs with negative co-expression of RRDCD participated in protein degradation and absorption, CAMs, and the hedgehog signaling pathway. In summary, our study provides important clues and the theoretical basis for exploring the pathogenesis, progression, and prognosis of ocular fundus disease.

     

Proteomics-based analysis of potential therapeutic targets in patients with peritoneal dialysis-associated peritonitis

BackgroundPeritoneal dialysis-associated peritonitis (PDAP) is the most common complication in peritoneal dialysis patients. We propose screening for characteristic expressed proteins in the dialysate of PDAP patients to provide clues for the diagnosis of PDAP and its therapeutic targets.MethodsDialysate samples were collected from patients with a first diagnosis of PDAP (n = 15) and from patients who had not experienced peritonitis (Control, n = 15). Data-independent acquisition (DIA) proteomic analysis was used to screen for differentially expressed proteins (DEPs). Co-expression networks were constructed via weighted gene co-expression network analysis (WGCNA) for detection of gene modules. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used for functional annotation of DEPs and gene modules. Hub proteins were validated using the parallel reaction monitoring (PRM) method.ResultsA total of 142 DEPs in the dialysate of PDAP patients were identified. 70 proteins were upregulated and 72 proteins were downregulated. GO and KEGG analysis showed that DEPs were mainly enriched in cell metabolism, glycolysis/glycogenesis and hypoxia-inducible factor-1 signaling pathway. Subsequently, a co-expression network was constructed and four gene modules were detected. Myeloperoxidase (MPO) and myeloperoxidase (HP) were the key proteins of the blue and turquoise modules, respectively. Additionally, PRM analysis showed that the expression of MPO and HP was significantly upregulated in the PDAP group compared to the non-peritonitis group, which was consistent with our proteomics data.ConclusionMPO and HP were differentially expressed in the dialysate of PDAP patients and may be potential diagnostic and therapeutic targets for PDAP.     

Quantitative plasma proteome analysis reveals aberrant level of blood coagulation-related proteins in nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC), one of the most common cancers in Southeast Asia, is not easily diagnosed until advanced stages. To discover potential biomarkers for improving NPC diagnosis, we herein identified the aberrant plasma proteins in NPC patients. We first removed the top-seven abundant proteins from plasma samples of healthy controls and NPC patients, and then labeled the samples with different fluorescent cyanine dyes. The labeled samples were then mixed equally and fractionated with ion-exchange chromatography followed by SDS-PAGE. Proteins showing altered levels in NPC patients were identified by in-gel tryptic digestion and LC-MS/MS. When the biological roles of the 45 identified proteins were assessed via MetaCore? analysis, the blood coagulation pathway emerged as the most significantly altered pathway in NPC plasma. Plasma kallikrein (KLKB1) and thrombin-antithrombin III complex (TAT) were chosen for evaluation as the candidate NPC biomarkers because of their involvement in blood coagulation. ELISAs confirmed the elevation of their plasma levels in NPC patients versus healthy controls. Western blot and activity assays further showed that the KLKB1 active form was significantly increased in NPC plasma. Collectively, our results identified the significant alteration of blood coagulation pathway in NPC patients, and KLKB1 and TAT may represent the potential NPC biomarkers.     

不同抗性苹果品种应答轮纹病菌胁迫的差异蛋白质组分析

为鉴定不同抗性苹果(Malus domestica)品种响应轮纹病菌胁迫的抗性相关蛋白表达差异, 以抗病品种华月及易感品种金冠为试材, 采用高通量同位素标记定量(IBT)技术结合液相色谱-串联质谱(LC-MS)鉴定技术, 对病原菌处理前后抗、感病品种叶片的蛋白质组差异表达进行分析, 共鉴定出171个差异表达蛋白(DEPs)。GO富集及KEGG通路分析表明, 在细胞组分、分子功能和生物过程3类中共注释到686个GO条目, 其中52个DEPs注释于KEGG通路的18个显著差异途径(P<0.05)。亚细胞定位预测分析表明, 171个DEPs中有170个分别定位于8类细胞器。蛋白功能注释分析表明, 46个DEPs注释于7类抗性相关蛋白, 包括类甜蛋白、过氧化物酶、多酚氧化酶、过敏原蛋白、几丁质酶、内切葡聚糖酶以及主乳胶蛋白。此外, 还对抗性相关蛋白的表达特点及基因定量结果进行了分析。该研究结果可为进一步解析抗、感病苹果品种应答轮纹病菌胁迫的抗性机制提供参考。     

Systematic proteomics analysis of lysine acetylation reveals critical features of placental proteins in pregnant women with preeclampsia

Preeclampsia (PE) is a dangerous hypertensive disorder that occurs during pregnancy. The specific aetiology and pathogenesis of PE have yet to be clarified. To better reveal the specific pathogenesis of PE, we characterized the proteome and acetyl proteome (acetylome) profile of placental tissue from PE and normal-term pregnancy by label-free quantification proteomics technology and PRM analysis. In this research, 373 differentially expressed proteins (DEPs) were identified by proteome analysis. Functional enrichment analysis revealed significant enrichment of DEPs related to angiogenesis and the immune system. COL12A1, C4BPA and F13A1 may be potential biomarkers for PE diagnosis and new therapeutic targets. Additionally, 700 Kac sites were identified on 585 differentially acetylated proteins (DAPs) by acetylome analyses. These DAPs may participate in the occurrence and development of PE by affecting the complement and coagulation cascades pathway, which may have important implications for better understand the pathogenesis of PE. In conclusion, this study systematically analysed the reveals critical features of placental proteins in pregnant women with PE, providing a resource for exploring the contribution of lysine acetylation modification to PE.     

Motile hepatocellular carcinoma cells preferentially secret sugar metabolism regulatory proteins via exosomes

Exosomes are deliverers of critically functional proteins, capable of transforming target cells in numerous cancers, including hepatocellular carcinoma (HCC). We hypothesize that the motility of HCC cells can be featured by comparative proteome of exosomes. Hence, we performed the super‐SILAC‐based MS analysis on the exosomes secreted by three human HCC cell lines, including the non‐motile Hep3B cell, and the motile 97H and LM3 cells. More than 1400 exosomal proteins were confidently quantified in each MS analysis with highly biological reproducibility. We justified that 469 and 443 exosomal proteins represented differentially expressed proteins (DEPs) in the 97H/Hep3B and LM3/Hep3B comparisons, respectively. These DEPs focused on sugar metabolism‐centric canonical pathways per ingenuity pathway analysis, which was consistent with the gene ontology analysis on biological process enrichment. These pathways included glycolysis I, gluconeogenesis I and pentose phosphate pathways; and the DEPs enriched in these pathways could form a tightly connected network. By analyzing the relative abundance of proteins and translating mRNAs, we found significantly positive correlation between exosomes and cells. The involved exosomal proteins were again focusing on sugar metabolism. In conclusion, motile HCC cells tend to preferentially export more sugar metabolism‐associated proteins via exosomes that differentiate them from non‐motile HCC cells.     

Quantitative proteomic analysis of hepatic tissue in allotetraploid hybridized from red crucian carp and common carp identified crucial proteins and pathways associated with metabolism and growth rate

Allotetraploid is a new species produced by distant hybridization between red crucian carp (Carassius auratus red var., abbreviated as RCC) and common carp (Cyprinus carpio L., abbreviated as CC). There is a significant difference in growth rate between allotetraploid and its parents. However, the underlying molecular mechanism is largely unknown. In this study, to find direct evidence associated with metabolism and growth rate in protein level, we performed quantitative proteomics analysis on liver tissues between allotetraploid and its parents. A total of 2502 unique proteins were identified and quantified by SWATH-MS in our proteomics profiling. Subsequently, comprehensive bioinformatics analyses including gene ontology enrichment analysis, pathway and network analysis, and protein–protein interaction analysis (PPI) were conducted based on differentially expressed proteins (DEPs) between allotetraploid and its parents. The results revealed several significant DEPs involved in metabolism pathways in liver. More specifically, the integrative analysis highlighted that the DEPs ACSBG1, OAT, and LDHBA play vital roles in metabolism pathways including “pentose phosphate pathway,” “TCA cycle,” and “glycolysis and gluconeogenesis.” These could directly affect the growth rate in fresh water fishes by regulating the metabolism, utilization, and exchange of substance and energy. Since the liver is the central place for metabolism activity in animals, we firstly established the comprehensive and quantitative proteomics knowledge base for liver tissue from freshwater fishes, our study may serve as an irreplaceable reference for further studies regarding fishes’ culture and growth.     

三棱内酯B调控人冠状动脉内皮细胞基因表达谱的生物信息学分析

目的:分析三棱内酯B在人冠状动脉内皮细胞中的表达谱数据集,寻找三棱内酯B调控血管内皮功能的关键作用靶点。方法:基于GEO公共数据库,下载原始表达谱数据集(GSE44598),经过差异基因筛选,功能注释,通路富集,信号通路网络以及基因互作网络分析,找出三棱内酯B对人冠状动脉内皮细胞基因表达谱产生影响的关键基因和信号通路。结果:同对照组相比,三棱内酯B给药组共有5224个基因有显著性差异,包括2628个上调基因和2596个下调基因。基因功能注释和信号通路富集分析表明,差异基因主要参与了细胞周期过程。网络分析显示,MAPK信号通路、细胞周期通路以及PLCG2,PRKACA和ADCY4等为关键信号通路和基因。结论:三棱内酯B通过影响PLCG2,PRKACA,ADCY4等基因的表达,参与MAPK和细胞周期等信号通路,从而调节人冠状内皮细胞的功能。这些关键基因和信号通路是三棱内酯B在心血管疾病治疗应用中潜在的作用靶点。