下一代诊断病理学

随着人类基因组测序、生物大数据信息分析、分子病理检测和人工智能辅助病理诊断等技术进步及其应用, 临床医学发展迈向精准诊疗时代。这一时代背景下, 传统诊断病理学迎来前所未有的历史机遇, 正在向"下一代诊断病理学(next-generation diagnostic pathology)"迈进。下一代诊断病理学以病理形态和临床信息为诊断基础, 以分子检测与生物信息分析、智慧制样与流程质控、智能诊断与远程会诊、病灶活体可视化与"无创"病理诊断等创新前沿交叉技术为主要特征, 以多组学和跨尺度整合诊断为病理报告内容, 实现对疾病的"最后诊断", 并预测疾病演进和结局、建议治疗方案和评估治疗反应, 形成新的疾病诊断"金标准"。未来, 需要激发病理学科创新活力, 加快下一代诊断病理学成熟和应用, 重塑病理学科理论和技术体系, 发挥诊断病理学在疾病"防、诊、治、养"等过程中的重要作用, 促进临床医学进一步发展, 服务健康中国战略。     

结核性脑膜炎临床诊断评分体系的建立及评价研究

背景 结核性脑膜炎（TM）是临床常见的中枢性感染的一种，其起病较慢，症状不典型，病原学诊断困难，误诊率高。目前有效的TM诊断工具较少。利用常见的临床症状、检查指标等建立诊断评分系体可提高诊断准确率，减少误诊。 目的 建立TM临床诊断评分体系（TMCDS），并对其应用价值进行初步评价。 方法 选取2011年11月至2021年9月在柳州市人民医院感染病科住院并诊断为脑膜炎的患者187例为研究对象，采用SPSS 21.0统计软件将患者随机分成建模组（147例）和验模组（40例）。根据是否为TM将建模组分为非TM亚组（76例）和TM亚组（71例）。收集患者的一般资料，主要包括性别、年龄、临床症状（发热、头痛、意识障碍、颈抵抗），实验室及影像学检查结果，包括人类免疫缺陷病毒（HIV）感染情况、CD4+ T淋巴细胞计数、C反应蛋白、颅内压、脑脊液常规生化检查（糖、氯、蛋白、细胞数）。建模组采用多因素Logistic回归分析探讨TM的影响因素；根据每个因素的β值所占比重设立相应分值，建立TMCDS；采用受试者工作特征曲线（ROC曲线）分析TMCDS诊断TM的价值。 结果 两亚组头痛、HIV感染、CD4+ T淋巴细胞计数<200/μl、C反应蛋白升高、颅内压>200 mm H2O（1 mm H2O=0.009 8 kPa）、脑脊液糖降低、脑脊液氯降低、脑脊液蛋白升高、脑脊液单核细胞升高情况比较，差异有统计学意义（P<0.05）。多因素Logistic回归分析结果显示，头痛、CD4+ T淋巴细胞<200/μl、C反应蛋白升高、脑脊液糖降低、脑脊液蛋白升高均是TM的影响因素（P<0.05）。将以上5个影响因素同时结合临床经验纳入脑脊液氯、脑脊液细胞数再次进行多因素Logistic回归分析，结果显示，头痛、CD4+ T淋巴细胞<200/μl、C反应蛋白升高、脑脊液糖降低、脑脊液蛋白升高均是TM的影响因素（P<0.05）。根据上述7个因素β值建立评分系统，将脑脊液氯降低β值设定为1分，其他因素β值与其的倍数即为该因素所对应的分值，因2个影响因素评分为负值，为方便临床，每个因素对应分值增加2.5分，最终建立TMCDS。TMCDS诊断建模组TM的ROC曲线下面积（AUC）为0.807〔95%CI（0.735，0.879），标准误=0.037，P<0.001〕，最佳诊断界值为21.50分。TMCDS诊断验模组TM的AUC为0.766〔95%CI（0.610，0.921），标准误=0.079，P=0.004〕，灵敏度为0.789，特异度为0.667。 结论 通过7个变量建立的TMCDS简单易行，对于早期TM具有较高的临床诊断价值。     

多模态超声对侵袭性前列腺癌的诊断价值及ROC曲线分析研究

目的：探讨多模态超声(MUS)对侵袭性前列腺癌(PCa)的诊断价值及受试者工作特征(ROC)曲线分析。方法：选择2018年01月至2021年02月于本院行手术切除并经病理证实的86例PCa患者作为研究对象,根据病理Gleason评分可分为两组,高侵袭组(Gleason＞4+3,共46例),低-中侵袭组(Gleason≤4+3,共40例)。所有患者术前均行MUS检查,包括经直肠常规超声(TRUS)、剪切波弹性成像(SWE)、超声造影(CEUS)。比较两组之间各参数的差异,采用ROC曲线分析其对高侵袭组PCa的诊断效能。结果：高侵袭组与低-中侵袭组的TRUS表现显著不同(P<0.05),其中高侵袭组中TRUS主要表现为弥漫性。高侵袭组SWE中SR比值(23.86±13.67)显著高于低-中侵袭组(12.82±11.95),差异具有统计学意义(P<0.05)。高侵袭组的CEUS参数中初始强度、峰值强度显著高于低-中侵袭组(P<0.05),而两组达峰时间、峰值减半时间无显著差异(P>0.05)。MUS对高侵袭组PCa诊断的ROC曲线下面积最大,敏感度、特异度也最高(P<0.05)。结论：MUS对高侵袭性PCa具有更高的诊断价值,有助于指导临床对治疗方案的选择。     

尿液外泌体lncRNA PCAT-1评估非浸润性膀胱癌患者复发的价值

目的探究尿液外泌体长链非编码RNA前列腺癌相关转录本1（lncRNA PCAT-1）评估非浸润性膀胱癌患者复发的价值。方法随机选取2015年1月—2016年3月进行治疗的非浸润性膀胱癌患者50例作为非浸润性膀胱癌组，选取同期在进行健康体检的健康受检者50例作为对照组。测定两组尿液外泌体中lncRNA PCAT-1表达情况。分析尿液外泌体lncRNA PCAT-1表达与临床病理参数的相关性、尿液外泌体lncRNA PCAT-1与尿液脱落细胞学对非浸润性膀胱癌的诊断价值、影响非浸润性膀胱癌患者复发的相关因素，评估非浸润性膀胱癌复发的价值及尿液外泌体lncRNA PCAT-1表达与非浸润性膀胱癌患者预后的关系。结果与对照组比较，非浸润性膀胱癌组患者尿液外泌体lncRNA PCAT-1表达水平明显升高（P<0.01）。尿液外泌体lncRNA PCAT-1表达与病理分期具有明显相关性（P<0.01）。尿液外泌体lncRNA PCAT-1诊断非浸润性膀胱癌的曲线下面积明显高于尿液脱落细胞学。采用Cox比例风险因素回归模型分析，病理分期及lncRNA PCAT-1均为影响非浸润性膀胱癌患者复发的独立危险因素。以尿液外泌体lncRNA PCAT-1相对表达量5.14±1.04分为高表达组（23例）与低表达组（27例），Kaplan-Meier生存曲线分析结果显示，lncRNA PCAT-1低表达量组生存期明显高于lncRNA PCAT-1高表达组。结论外泌体lncRNA PCAT-1在非浸润性膀胱癌患者尿液中的表达水平较健康受检者明显升高，且其表达水平与患者病理分期明显相关。尿液外泌体lncRNA PCAT-1对评估非浸润性膀胱癌患者的复发具有重要作用。     

Validation of donor fraction cell-free DNA with biopsy-proven cardiac allograft rejection in children and adults

ObjectivesDonor-specific cell-free DNA shows promise as a noninvasive marker for allograft rejection, but as yet has not been validated in both adult and pediatric recipients. The study objective was to validate donor fraction cell-free DNA as a noninvasive test to assess for risk of acute cellular rejection and antibody-mediated rejection after heart transplantation in pediatric and adult recipients.MethodsPediatric and adult heart transplant recipients were enrolled from 7 participating sites and followed for 12 months or more with plasma samples collected immediately before all endomyocardial biopsies. Donor fraction cell-free DNA was extracted, and quantitative genotyping was performed. Blinded donor fraction cell-free DNA and clinical data were analyzed and compared with a previously determined threshold of 0.14%. Sensitivity, specificity, negative predictive value, positive predictive value, and receiver operating characteristic curves were calculated.ResultsA total of 987 samples from 144 subjects were collected. After applying predefined clinical and technical exclusions, 745 samples from 130 subjects produced 54 rejection samples associated with the composite outcome of acute cellular rejection grade 2R or greater and pathologic antibody-mediated rejection 2 or greater and 323 healthy samples. For all participants, donor fraction cell-free DNA at a threshold of 0.14% had a sensitivity of 67%, a specificity of 79%, a positive predictive value of 34%, and a negative predictive value of 94% with an area under the curve of 0.78 for detecting rejection. When analyzed independently, these results held true for both pediatric and adult cohorts at the same threshold of 0.14% (negative predictive value 92% and 95%, respectively).ConclusionsDonor fraction cell-free DNA at a threshold of 0.14% can be used to assess for risk of rejection after heart transplantation in both pediatric and adult patients with excellent negative predictive value.     

Performance of the new FilmArray Blood Culture Identification 2 panel and its potential impact on clinical use in patients with Gram-negative bacteremia

IntroductionRapid diagnostic tests have been developed recently for rapid species or resistance genes identification, offering the potential to improve the selection of appropriate antibiotics. The newly developed FilmArray Blood Culture Identification 2 (BCID2) panel, which can identify more species and resistance genes, such as extended-spectrum beta-lactamase, is expected to make an impact on antimicrobial practice.MethodsThe consecutive 50 inpatients with Gram-negative bacilli bacteremia were enrolled to this retrospective single-center study. In addition to the existing FilmArray Blood Culture Identification (BCID) panel, we have implemented BCID2 panel for positive blood culture. The sensitivity and specificity of BCID and BCID2 panel were respectively calculated, and a simulation study of time to effective, optimal and de-escalation therapy was performed based on BCID or BCID2 result.ResultsA total of 52 Gram-negative organisms in 50 patients were identified from blood cultures. Of these, 45 (87%) organisms were detected by BCID2 panel, which was more than BCID panel (41 organisms, 79%). BCID2 panel detected 5 CTX-M genes, which were concordant with conventional method. The time to effective therapy did not differ between BCID arm and BCID2 arm; however, the median time to optimal therapy (34 h in BCID arm and 26 h in BCID2 arm, P = 0.0007) and the median time to de-escalation therapy (42 h in BCID arm and 22 h in BCID2 arm, P = 0.0005) were significantly shortened.ConclusionsThis simulation study of BCID2 panel showed high sensitivity and specificity, and the potential impact on shortening the time to optimal and de-escalation therapy.