Diagnostic and prognostic value of plasma cell-free DNA combined with VEGF-C in laryngeal squamous cell carcinoma

BackgroundCirculating cell-free DNA (cfDNA) and vascular endothelial growth factor-C (VEGF-C) can be utilized to detect cancer and predict its prognosis. However, their potential application in laryngeal squamous cell carcinoma (LSCC) is unclear.PurposeThis study aimed to identify the diagnostic and prognostic value of cfDNA and VEGF-C in LSCC patients.MethodsThe plasma cfDNA of 148 LSCC patients and 43 non-tumor patients were isolated. Quantitative real-time PCR (qRT-PCR) was performed to assess long and short DNA fragments in plasma by amplifying the ALU repeats. ALU-qPCR results (ALU247/ALU115) were used to calculate cfDNA integrity index. Vascular endothelial growth factor-C (VEGF-C) level was detected by ELISA assay. Correlation between cfDNA and clinical features was analyzed. For detecting the sensitivity and specificity of cfDNA and VEGF-C alone or in combination for diagnosing LSCC, receiver operator characteristic (ROC) was established. For evaluating the overall survival (OS) of LSCC, Kaplan-Meier curves were established.ResultsLSCC patients had significantly higher levels of plasma cfDNA (ALU115, ALU247, and cfDNA integrity index) and VEGF-C than those without cancer (p < 0.05), showing area under the curve (AUC) values of 0.79, 0.74, 0.62 and 0.80, when cutoff value was correspondingly defined at 2.14 ng/mL, 1.39 ng/mL, 0.73 and 412.90 pg/mL, respectively. The AUC for distinguishing LSCC patients from non-tumor patients by plasma cfDNA combined with VEGF-C was 0.89 (95% CI: 0.83–0.94). A significant correlation was found between plasma cfDNA levels and Ki-67, tumor size, pT stage, and smoking history (p < 0.05). Based on survival analysis, low VEGF-C concentration groups had longer OS than those with high VEGF-C concentration (p = 0.02).ConclusionIndicators such as plasma cfDNA and VEGF-C may be used to diagnose and monitor LSCC for its noninvasiveness and rapid accessibility.     

Urinary pseudouridine in patients with lymphoma: comparison with other clinical parameters

BACKGROUND: Some patients with malignant lymphoma do not manifest superficial lymphadenopathy. In such cases, clinical parameters that indicate the number of tumor cells are important for the assessment of tumor growth and choice of proper treatment. We evaluated urinary pseudouridine (U-PU) as an indicator of the growth of malignant lymphoma by comparing its levels with serum concentrations of other clinical parameters in patients with various lymphomas at various stages. METHODS: Urine was obtained from 67 patients with lymphoma. U-PU was assayed by recombinant Fab-based inhibition ELISA. Serum soluble IL2 receptor (sIL2R), serum deoxythymidine kinase (dTK), serum beta-2 microglobulin (beta2MG) and serum lactate dehydrogenase (LDH) were also assayed. RESULTS: U-PU concentrations showed good correlations with serum concentrations of beta2MG, LDH, sIL2R and dTK. The level of U-PU was higher in stage IV than in stages I (P=0.023), II (P=0.006) and III (P=0.036). CONCLUSION: U-PU concentration correlates with the clinical stage of lymphoma and is a useful tool to assess the growth of lymphoma.     

Evaluation of TERT promoter mutations in urinary cell-free DNA and sediment DNA for detection of bladder cancer

BackgroundCell-free DNA (cfDNA) is proposed to be a valuable source of biomarkers in liquid biopsies for various diseases as it is supposed to partially originate from tumor cells. However, data about the diagnostic implications of cfDNA in urine for the detection of bladder cancer (BCa) is sparse.MethodsWe evaluated the usability of urinary cfDNA for diagnostic purposes compared to urine sediment DNA (sDNA) in 53 BCa patients and 36 control subjects by analyzing two abundant point-mutations (C228T/C250T) in the TERT promoter using Next-Generation Sequencing.ResultsMutations were detected in 77% of the urinary sDNA compared to 63% of the cfDNA samples. Moreover, the TERT mutation allele frequencies (MAF) were highly correlated in cfDNA and sDNA. In comparison, the accuracy of the TERT assay was higher in sDNA (84%) compared to cfDNA or voided urine cytology (both 77%). Interestingly, MAFs from leukocyte-rich urines were higher in cfDNA than in sDNA, indicating a diagnostic advantage of cfDNA in such urines.ConclusionsUrine-based mutation detection has the ability to augment and surpass voided urine cytology as the current gold-standard for the non-invasive detection and surveillance of BCa. The analysis of cell-free DNA provides no general diagnostic advantage compared to urine sediment DNA.     

Correlation of 18F-FDG PET/CT SUVmax with clinical features,D-dimer and LDH in patients with primary intestinal lymphoma

ObjectiveTo investigate the characteristics of fluorine-18-deoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) maximum standardized uptake value (SUVmax) in primary intestinal lymphoma (PIL) and its correlation with D-dimer and lactate dehydrogenase (LDH).MethodsFifty-two patients diagnosed with PIL from June 2016 to December 2019 were analyzed. All patients underwent 18F-FDG PET/CT. The relationships between SUVmax and different pathological subtypes, clinical stages and risk grades were analyzed. The correlations between SUVmax and Ki-67, LDH and D-dimer were determined. Additionally, PET/CT imaging results were collected from 35 patients with primary intestinal cancer (PIC) and compared with the imaging features of PIL.ResultsSUVmax was significantly different between PIL and PIC groups and various PIL pathological subgroups. Patients in the high-risk PIL group had markedly higher SUVmax values than the intermediate-risk and low-risk groups. A significant positive correlation was observed between SUVmax and Ki-67 in patients with PIL. SUVmax was significantly different between the elevated and normal D-dimer groups. D-dimer showed a positive correlation with SUVmax.Conclusion18F-FDG PET/CT SUVmax reflects the aggressiveness of lymphoma to a certain degree, is correlated with Ki-67 and determines the risk grades of PIL. Moreover, it facilitates differential diagnosis, clinical staging and treatment based on D-dimer levels.     

Systemic immune inflammation index,ratio of lymphocytes to monocytes,lactate dehydrogenase and prognosis of diffuse large B-cell lymphoma patients

BACKGROUNDIn malignant tumors, inflammation plays a vital role in the development, invasion, and metastasis of cancer cells. Diffuse large B-cell lymphoma (DLBCL), the most common malignant proliferative disease of the lymphatic system, is commonly associated with inflammation. The international prognostic index (IPI), which includes age, lactate dehydrogenase (LDH), number of extranodal lesions, Ann Arbor score, and Eastern Cooperative Oncology Group (ECOG) score, can evaluate the prognosis of DLBCL. However, its use in accurately identifying high-risk patients and guiding treatment is poor. Therefore, it is important to find novel immune markers in predicting the prognosis of DLBCL patients.AIMTo determine the association between the systemic immune inflammation index (SII), ratio of lymphocytes to monocytes (LMR), ratio of LMR to LDH (LMR/LDH), and prognosis of patients with DLBCL.METHODSA total of 68 patients diagnosed with DLBCL, treated in our hospital between January 2016 and January 2020, were included. χ² test, Pearson’s R correlation, Kaplan Meier curves, and Cox proportional risk regression analysis were used. The differences in the SII, LMR, and LMR/LDH among patients with different clinicopathological features were analyzed. The differences in progression-free survival time among patients with different SII, LMR, and LMR/LDH expressions and influencing factors affecting the prognosis of DLBCL patients, were also analyzed.RESULTSThe LMR and LMR/LDH in patients with Ann Arbor stage III–IV, ECOG score ≥ 2, and SII, IPI score 2–5 were significantly higher than those of patients with Ann Arbor stage I-II and ECOG score < 2 (P < 0.05). Patients with high SII, LMR, and LMR/LDH had progression-free survival times of 34 mo (95%CI: 32.52–38.50), 35 mo (95%CI: 33.42–36.58) and 35 mo (95%CI: 33.49–36.51), respectively, which were significantly lower than those with low SII, LMR, and LMR/LDH (P < 0.05); the SII, LMR, and LMR/LDH were positively correlated (P < 0.05). Cox proportional risk regression analysis showed that the SII, LMR, and LMR/LDH were influencing factors for the prognosis of DLBCL patients (hazard ratio = 1.143, 1.665, and 1.704, respectively; P < 0.05).CONCLUSIONThe SII, LMR, and LMR/LDH are related to the clinicopathological features of DLCBL, and they also influence the prognosis of patients with the disease.     

恶性淋巴瘤患者凝血功能改变与国际预后指数间的相关性分析

本研究探讨淋巴瘤患者凝血功能改变的临床意义及其与国际预后指数（IPI）之间的相关性。应用磁珠法和速率法分别检测75例淋巴瘤患者和20例健康对照者的凝血4项即凝血酶原时间（PT）、部分活化凝血活酶时间（APTT）、凝血酶时间（TT）、纤维蛋白原（FIB）及血清乳酸脱氢酶（LDH）水平。结果表明：①淋巴瘤患者APTT、FIB较对照组明显增高（p〈0.05和〈0.01）,表现为高纤维蛋白原血症。不同年龄和结外病灶数之间凝血4项指标均改变不明显（p〉0.05）;②Ⅲ期和Ⅳ期患者APTT和FIB与Ⅱ期患者比较显著增高（p〈0.05和〈0.01）,Ⅳ期患者FIB较Ⅲ期患者增高（p〈0.05）;有全身症状患者FIB显著高于无症状者（p〈0.01）;③LDH升高患者APTT和FIB明显高于对照组（p〈0.05和〈0.01）,且FIB明显高于LDH正常患者（p〈0.05）。体力状态（PS）2-4分患者FIB明显高于PS0-1分患者（p〈0.01）;④低中危、高中危和高危患者FIB明显高于对照组（p〈0.01）,高中危和高危患者FIB较低危患者增高（p〈0.05）;⑤有全身症状、LDH升高、PS2-4分、Ⅲ-Ⅳ期淋巴瘤患者FIB升高病例数与对应无症状、LDH正常、PS0-1分、Ⅱ期患者比较明显增多（p〈0.05或0.01）。FIB与LDH水平、PS评分、Ann Arbor分期以及危险分级间存在正相关关系（p〈0.05或0.01）。结论：淋巴瘤患者常存在高纤维蛋白原血症,受Ann Arbor分期、全身症状、LDH水平和体力状态的影响,且FIB可能是反映淋巴瘤患者预后的有关指标。     

Cell-free DNA characteristics and chimerism analysis in patients after allogeneic cell transplantation

Cell-free DNA (cfDNA) isolated from plasma or serum has received increasing interest for diagnostic applications in pregnancy, solid tumors and solid organ transplantation. The reported clinical usefulness of cfDNA obtained from plasma or serum in patients undergoing allogeneic cell transplantation (alloHSCT) is scarce.ObjectiveTo analyze the potential clinical utility of cfDNA chimerism analysis after alloHSCT.Design and methodsA total of 196 samples obtained from 110 patients were investigated for their chimeric status both in peripheral blood and plasma using standard PCR for microsatellite amplification. Plasma DNA size distribution was analyzed using capillary electrophoresis.ResultsThe mean cfDNA concentration in the transplanted patients was 469 ng/ml (range: 50–10,700 ng/ml). The size range of almost 80% of the analyzed fragments was between 80 and 200 bp. In 41 out of the 110 patients included in the study a mixture of donor and recipient plasma cfDNA was detected. There was a statistically significant difference in the percentage of plasma mixed chimerism between the patients without transplant related complications and the patients with either GvHD (p < 0.05) or relapse (p < 0.01). In those patients who showed improvement of GvHD also displayed a decrease in the observable percentage of recipient cfDNA during GvHD treatment. In patients without improvement or even with worsening of acute GvHD, stable or increasing levels of recipient cfDNA were detected.ConclusionscfDNA in combination with peripheral blood and bone marrow cell chimerism analysis might improve its utility in the clinic in particular in those patients with clinical complications after alloHSCT.     

血清乳酸脱氢酶、β2微球蛋白及血管内皮生长因子水平检测对非霍奇金淋巴瘤的临床意义

本研究探讨血清乳酸脱氢酶(LDH)、β2微球蛋白(β2-MG)及血管内皮生长因子(VEGF)水平检测对非霍奇金淋巴瘤(NHL)的临床意义。选择我院2010年1月至2011年12月期间收治的初治NHL患者58例为研究组,另选取同期入院参加体检的58例健康体检者为对照组。采用生物化学方法测定血清LDH水平,采用酶联免疫吸附法测定血清β2-MG和VEGF水平,超过正常值范围判断为该指标阳性。结果表明,与正常对照组相比,NHL患者的LDH、β2-MG、VEGF水平均显著升高,差别均具有统计学意义(P<0.05)。在NHL组患者中,不同病理分型和临床分期患者LDH与VEGF水平变化显著。与低度恶性患者相比,中度恶性与高度恶性患者的LDH与VEGF水平明显提高,差异具有统计学意义(P<0.05)。与临床Ⅰ期患者相比,临床Ⅱ、Ⅲ及Ⅳ期患者血清LDH与VEGF水平明显提高,差异具有统计学意义(P<0.05)。在NHL组58例患者中,LDH阳性30例,占51.72%,β2-MG阳性39例,占67.24%,VEGF阳性29例,占50.00%。结论:血清LDH、β2-MG、VEGF水平检测对NHL具有一定的临床意义。     

原发性肾上腺淋巴瘤影像学表现及其病理相关性

目的 探讨原发性肾上腺淋巴瘤（PAL）的CT、MRI表现及其与病理学的相关性。方法 回顾性分析经病理证实的13例PAL的CT、MRI表现,均为弥漫大B细胞型非霍奇金淋巴瘤,其中3例经手术证实,10例经穿刺活检证实;其中8例乳酸脱氢酶（LDH）升高。11例（18个病灶）接受CT检查,6例（11个病灶）接受MR检查,4例同时接受CT和MR检查。结果 9例为双侧病灶,4例单侧病灶,共22个病灶。病灶最大径为2.60~13.40 cm,16个病灶呈类圆形、椭圆形,2个呈不规则形,4个基本保持肾上腺形状。18个病灶CT平扫呈软组织密度影,9个病灶MR T1WI呈低信号、T2WI呈稍高信号。CT或MR增强扫描中,病灶均呈轻中度渐进性均匀或不均强化,其中4个在MR增强静脉期或延迟期出现网格状强化。结论 PAL影像学表现具有一定特征性,结合LDH检查可提高术前正确诊断率。     

结外NK/T细胞淋巴瘤诊治进展

结外NK/T细胞淋巴瘤(ENKTCL)多见于亚洲和中南美洲,病因可能与EB病毒感染有关,病理学表现为多形态肿瘤组织侵犯血管。肿瘤细胞特异性表达CD2、CD56和胞浆CD3,但TCR基因重排阴性。确诊需病理组织学检查,影像学检查可对诊断提供帮助。化疗主要用于疾病播散、复发和放疗失败者。在传统的CHOP方案基础上加用足叶以甙、亚硝基脲类、左旋门冬酰胺酶等可能提高疗效,并可作为复发患者的挽救治疗。预后因素的研究显示分期、B组症状、LDH水平、初治疗效、血清nm23-H1水平与结外NK/T细胞淋巴瘤的预后有关。     

基于局部弥散一致性值观察终末期肾病继发性甲状旁腺功能亢进症患者脑白质改变及认知相关性

目的采用局部弥散一致性(LDH)探讨终末期肾病(ESRD)继发甲状旁腺功能亢进症(SHPT)患者脑白质改变。方法选取50例接受血液透析的ESRD患者,根据是否伴有SHPT分为SHPT组(25例)与非SHPT组(25例),利用蒙特利尔认知评估(MoCA)量表评估认知功能,并进行MR平扫及DTI数据采集,对全脑白质纤维LDH值进行基于体素的组间比较。结果相比非SHPT组,SHPT组出现双侧大脑半球弥漫性、对称分布的LDH减低区及右侧小脑半球局灶性LDH减低区(P均<0.01),LDH值升高区域集中在脑干及小脑(P均<0.01)。SHPT组MoCA总评分与全段甲状旁腺激素(iPTH)水平呈负相关(r=-0.525),与受教育水平呈正相关(r=0.413)、与左侧皮质脊髓束的LDH值呈正相关(r=0.500,P均<0.05);iPTH水平与差异脑区的LDH值均无明显相关(P均>0.05)。结论LDH值可提供ESRD伴SHPT患者脑白质LDH信息:脑白质LDH改变与认知及临床指标具有相关性。     

Clinical management and susceptibility of primary hepatic lymphoma: A cases-based retrospective study

BACKGROUNDThe liver as a primary site of lymphoma is rarely seen, they are usually misdiagnosed as hepatocellular carcinoma, etc. In 2017, a review of primary hepatic lymphoma (PHL) was done in immunocompetent diffuse large B-cell lymphoma (DLBCL) patients. Yet questions that include treatment choosing or susceptibility of immunoincompetent patients remain disputable. AIMTo investigate the clinical characteristics of patients with PHL.METHODSWe collected PHL cases on PubMed, and extracted demographic and clinicopathological data to perform a systematic analysis. Survival analysis regarding age, lactate dehydrogenase (LDH), liver function abnormality (LFA), and treatment modalities were conducted. The Kaplan-Meier method and Cox regression were used to identify risk factors. RESULTSOf 116 PHL patients with DLBCL (62.1%) as the most common subtype. Biopsy methods before surgery produced a 97% positive rate. Progression-free survival (PFS) was significantly shortened in patients with elevated LDH [Hazard ratio (HR): 3.076, 95% confidence interval (CI): 1.207-7.840, P = 0.018] or LFA (HR: 2.909, 95%CI: 1.135-7.452, P = 0.026). Univariate Cox regression analysis suggesting that LDH, liver function, B symptom, hepatosplenomegaly, and lesion were significantly associated with PHL patients survival (P < 0.05). Heavy disease burden was observed in deceased patients. A few PHL patients (3.4%) have slightly higher tumor markers.CONCLUSIONPHL patients with elevated LDH and LFA tend to have shorter PFS. Biopsy before treatment in undecided patients with no tumor markers exceeds upper limits has the most essential clinical significance, especially in immunoincompetent patients.     

Beta-2 microglobulin as an immunological marker to assess the progression of human immunodeficiency virus infected patients on highly active antiretroviral therapy

BackgroundAcquired immune deficiency syndrome (AIDS) defines the end stage of Human immunodeficiency viral (HIV) infection before the introduction of highly active antiretroviral therapy (HAART). This study was carried out to assess the serum β-2 microglobulin (B2M) as a marker for progression of HIV infected patients undergoing HAART.MethodsBlood samples were collected from 50 subjects of HIV infected patients residing at semi urban area undergoing treatment at Chellam Hospital, Salem, India. Twenty five age-matched healthy subjects were taken as control group. Serum B2M level was measured by using enzyme immunoassay, absolute CD4 and CD8 counts were carried out by single platform (SP) technology using a flow cytometer. Viral RNA was measured by real time PCR. The serum LDH level and total WBC count were also measured.ResultsOur result showed a statistically high significant increase in B2M in HIV patients on HAART non complaint group whereas absolute CD4, CD8 count, CD4/CD8 ratio and WBC count were decreased significantly when compared to control and HAART complaint group. Statistically a significant negative correlation was observed between B2M and absolute CD4, CD8 count, CD4/CD8 ratio and WBC count. B2M showed a significantly positive correlation with viral RNA and LDH values.ConclusionsThe increase of B2M and reduced absolute CD4, CD8 count, CD4/CD8 ratio and WBC count in HIV patients on HAART non complaint group may have a contributory role in the immune progression of HIV with interruption of HAART. B2M plays an important role in the diagnosis of HIV and might indicate HIV progression.     

Alu cell-free DNA concentration,Alu index,and LINE-1 hypomethylation as a cancer predictor

IntroductionThe liquid biopsy approach, a less-invasive diagnostic tool, enables the detection of disease-specific genetic and epigenetic aberrations. Approximately 66–69% of the human genome may be composed of transposable repetitive elements, including Alu and LINE-1. This study aimed to investigate whether Alu-derived cell-free DNA (cfDNA) concentrations, Alu index, and LINE-1 methylation could be used to distinguish patients with cancers from healthy individuals.MethodsTwo sets of primers, shorter and longer Alu fragments, were used to amplify Alu elements, followed by the quantitation of Alu DNA concentration and its integrity index. LINE-1 methylation status was then analyzed with quantitative PCR using methylation- and unmethylation-specific TaqMan probes.ResultsBoth Alu index and LINE-1 methylation level were significantly different in comparison between patients with lung or breast cancer and the healthy controls. The area under the ROC curve of the Alu index and LINE-1 hypomethylation was 0.742 and 0.848 for lung cancer, respectively, and 0.724 and 0.890 for breast cancer, respectively. However, Alu longer fragment DNA concentration was significantly correlated with Alu index in comparison to LINE-1 hypomethylation. Regression analysis suggested that the LINE-1 methylation level, rather than the Alu index, was a good discriminator for lung and breast cancers.ConclusionsThis study investigated the genome-wide Alu index and LINE-1 methylation status; their associations with cancers suggested that these combinatory panels could be implemented as a triage test to discriminate cancer patients from healthy individuals.     

The role of ctDNA detection and the potential of the liquid biopsy for breast cancer monitoring

Introduction: Recent advances in deep amplicon sequencing have enabled rapid assessment of somatic mutations and structural changes in multiple cancer genes in DNA isolated from tumour tissues and circulating cell-free DNA (cfDNA). This cfDNA is under investigation as a ‘liquid biopsy’ for the real time monitoring of patients with cancer in a growing number of research studies and clinical trials.

Areas covered: Here we will provide a brief overview of the potential clinical utility of cfDNA profiling for detection and monitoring of patients with breast cancer. The review was conducted in English using PubMed and search terms including ‘breast cancer’, ‘plasma DNA’, ‘circulating cell free DNA’ and ‘circulating tumour DNA’.

Expert commentary: Liquid biopsies through circulating tumor DNA (ctDNA) enable monitoring of patients with breast cancer. The challenge ahead will be to incorporate cfDNA mutation profiling into routine clinical practice to provide patients with the most appropriate and timely treatment.     

VEGF在淋巴瘤骨髓、血浆中的表达意义及贫血对其的影响

目的探讨血管内皮生长因子(VEGF)在淋巴瘤骨髓上清、血浆中的表达与意义及贫血因素对其的影响。方法对血液肿瘤科2007年7月至2008年7月收治的无骨髓浸润的非何杰金淋巴瘤患者30例(淋巴瘤组)、良性贫血患者8例(贫血组)及正常体检者5例(对照组),采用ELISA方法检测骨髓上清、血浆VEGF的表达水平。结果血浆、骨髓VEGF水平在淋巴瘤Ⅰ～Ⅳ期及贫血组均明显高于对照组(P均<0.05);在贫血组与淋巴瘤组差异无统计学意义(P均>0.05)。淋巴瘤仅IV期患者血浆VEGF水平明显高于Ⅰ、Ⅱ、Ⅲ期及贫血组(P均<0.05),但骨髓VEGF水平差异无统计学意义(P>0.05)。血浆、骨髓VEGF表达水平在淋巴瘤组(r=0.07813,P>0.05)、贫血组(r=0.08910,P>0.05)均无相关性。结论无论骨髓或血浆VEGF表达水平均可作为淋巴瘤的诊断依据,但Ⅰ～Ⅲ期需注意有无贫血因素的影响,Ⅳ期血浆表达水平未发现贫血因素的影响。骨髓VEGF表达水平对淋巴瘤的分期无指导意义。血浆VEGF表达水平对淋巴瘤分期有无指导意义有待进一步研究,但对评估预后有一定指导意义。骨髓、血浆VEGF表达水平无相关性。贫血对血浆、骨髓VEGF的表达有一定影响。     

Cross‐platform comparison of next‐generation sequencing and matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry for detecting KRAS/NRAS/BRAF/PIK3CA mutations in cfDNA from metastatic colorectal cancer patients

BackgroundExamining tumor KRAS/NRAS/BRAF/PIK3CA status in metastatic colorectal cancer (mCRC) is essential for treatment selection and prognosis evaluation. Cell‐free DNA (cfDNA) in plasma is a feasible source for tumor gene analysis.MethodsIn this study, we recruited mCRC patients and analyzed their KRAS/NRAS/BRAF/PIK3CA status in cfDNA using two platforms, next‐generation sequencing (NGS) and matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF). The performance between the two platforms and the concordance rate between cfDNA and tissue were analyzed. The relationship between cfDNA‐related variables and clinical variables was also assessed. Tumor mutations in cfDNA from patients receiving continuous treatments were monitored in the follow‐ups.ResultsNext‐generation sequencing and MALDI‐TOF had similar specificity (100.0% vs. 99.3%) and negative predictive value (99.9% vs. 99.4%), whereas NGS had higher sensitivity (97.1% vs. 85.3% of MALDI‐TOF) and positive predictive value (100% vs. 82.9% of MALDI‐TOF). The overall concordance rate of NGS and MALDI‐TOF was 98.6%. For the reportable types of mutations in both cfDNA and tissue, the concordance rate was 96.1%. Among 28 tissue‐positive patients, the allele frequencies of tumor mutations in cfDNA were higher in patients with primary tumor burden (p = 0.0141). Both CEA and CA 19‐9 were positively correlated with cfDNA concentration (r = 0.3278 and r = 0.3992). The allele frequencies of tumor mutations changed with disease progression.ConclusionsNext‐generation sequencing showed slightly better performance in detecting cfDNA mutations and was more suitable for clinical practice. cfDNA‐related variables reflected the tumor status and showed a promising potential in monitoring disease progression.     

超声引导下穿刺活检对获得性免疫缺陷综合征患者合并淋巴结肿大的病因诊断价值

目的： 经超声引导下穿刺活检分析获得性免疫缺陷综合征（acquired immunodeficiency syndrome，AIDS）患者合并淋巴结肿大的病因。方法： 对2017年4月至2020年12月在上海市公共卫生临床中心行超声引导下穿刺活检的130例AIDS合并淋巴结肿大患者的病理诊断及实验室指标等进行回顾性分析，进而分析超声引导下穿刺活检对该类患者淋巴结肿大病因的诊断价值。结果： 130例患者中，最终诊断良性病变82例（63.1%），包括淋巴结核56例，非结核分枝杆菌感染7例，马尔尼菲篮状菌感染6例，坏死性淋巴结炎5例，隐球菌感染2例，巨细胞病毒感染2例，金黄色葡萄球菌感染、淋巴组织增生、淋巴囊肿、EB病毒感染各1例；恶性肿瘤36例（27.7%），包括淋巴瘤17例（其中弥漫大B细胞淋巴瘤、Burkitt淋巴瘤各5例，浆母细胞淋巴瘤、小淋巴细胞性淋巴瘤及间变大细胞淋巴瘤各1例，另4例未分型），转移性肿瘤17例，卡波西肉瘤2例；病因类型不明确12例，其中病理检查提示11例为良性病变、1例为肿瘤性坏死。130例患者淋巴结肿大病因总诊断率达90.8%。所有患者无并发症。结论： 超声引导下穿刺活检对AIDS合并淋巴结肿大患者淋巴结肿大病因诊断率高，且微创安全，对于该类患者淋巴结肿大临床早期诊断及精准治疗具有较高价值，值得临床推广应用。     

Detection of EGFR-SEPT14 fusion in cell-free DNA of a patient with advanced gastric cancer: A case report

BACKGROUNDGastric cancer is the fifth most diagnosed cancer worldwide and the third most common cause of cancer-related death. In recent decades, increasing application of next-generation sequencing has enabled detection of molecular aberrations, including fusions. In cases where tissue is difficult to obtain, cell-free DNA (cfDNA) is used for detecting mutations to identify the molecular profile of cancer. Here, we report a rare case of EGFR-SEPT14 fusion detected from cfDNA analysis in a patient with gastric cancer. CASE SUMMARYA 49-year-old female diagnosed with advanced gastric cancer in July 2019 received capecitabine and then combination chemotherapy of ramucirumab and paclitaxel, but ascites was detected. The therapy was switched to nivolumab, but disease progression was observed on a positron emission tomography/computed tomography scan in May 2020. Therapy was discontinued, and cfDNA next-generation sequencing was immediately evaluated. All genomic variants, including fusions, were analyzed from cfDNA. The following somatic alterations were detected from the patient’s cfDNA: an APC frameshift mutation (NM_000038.5:c.6579del, p.V2194fs) with variant allele frequency of 0.5%, an EGFR amplification with a copy number of 17.3, and an EGFR-SEPT14 fusion with variant allele frequency of 45.3%. The site of the fusion was exon 24 of EGFR fused to exon 10 of SEPT14. The fusion was in-frame and considered to be protooncogenic. Although the patient refused to continue therapy, we suggest that EGFR-targeted therapies be tried in such future cases. CONCLUSIONThe expanded applications of the cfDNA assay may open a new horizon in treatment of patients with advanced gastric cancer.     

High serum lactate dehydrogenase and dyspnea: Positive predictors of adverse outcome in critical COVID-19 patients in Yichang

BACKGROUNDCoronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak in China, constitutes a Public Health Emergency of International Concern. It is well known that COVID-19 patients may have increased serum lactate dehydrogenase (LDH) levels in the early stage. The clinical changes in LDH may have predictive value in disease evolution and prognosis in critically ill COVID-19 patients.AIMTo examine serum LDH and clinical characteristics in patients with COVID-19 and their predictive value for prognosis.METHODSThis retrospective study analyzed the clinical data of forty-seven critical COVID-19 patients in the intensive care unit of the Third People''s Hospital of Yichang City from January 27 to March 25, 2020 and divided them into survivors and non-survivors. The patients were diagnosed according to the World Health Organization interim guidance and critical cases met any one of the following criteria: Respiratory failure and required mechanical ventilation, the occurrence of shock, and the combined failure of other organs that required intensive care unit monitoring and treatments, according to the diagnostic criteria of critical COVID-19. Clinical data including symptoms, detection of SARS-CoV-2, chest computed tomography (CT) images, changes in serum LDH in different clinical phases, and prognosis were collected. Statistical analysis of the data was performed. Continuous variables were expressed as median (interquartile range) and compared with the Mann-Whitney U test. Categorical variables were compared with the Chi-square test. Survival data were analyzed using Kaplan-Meier survival curves and log-rank tests.RESULTSAccording to chest CT images, we observed the alveolitis and fibrosis stages in all critical patients in this study. Most non-survivors died in the fibrosis stage. Non-survivors had fewer days of hospitalization, shorter disease duration, shorter duration of alveolitis and fibrosis, and had dyspnea symptoms at disease onset (P = 0.05). Both first and lowest LDH values in the alveolitis stage were more pronounced in non-survivors than in survivors (449.0 U/L vs 288.0 U/L, P = 0.0243; 445.0 U/L vs 288.0 U/L, P = 0.0199, respectively), while the first, lowest and highest values of serum LDH in non-survivors were all significantly increased compared to survivors in the fibrosis phase (449.0 U/L vs 225.5 U/L, P = 0.0028; 432.0 U/L vs 191.0 U/L, P = 0.0007; 1303.0 U/L vs 263.5 U/L, P = 0.0001, respectively). The cut-off points of first LDH values in the alveolitis and fibrosis phase for distinction of non-survivors from survivors were 397.0 U/L and 263.0 U/L, respectively. In the fibrosis stage, non-survivors had more days with high LDH than survivors (7.0 d vs 0.0 d, P = 0.0002). Importantly, patients with high LDH had a significantly shorter median survival time than patients with low LDH in the alveolitis phase (22.0 d vs 36.5 d, P = 0.0002), while patients with high LDH also had a significantly shorter median survival time than patients with low LDH in the fibrosis phase (27.5 d vs 40.0 d, P = 0.0008). The proportion of non-survivors with detectable SARS-CoV-2 until death in the alveolitis stage was significantly increased compared with that in the fibrosis stage (100% vs 35.7%, P = 0.0220).CONCLUSIONHigh LDH and dyspnea symptoms were positive predictors of an adverse outcome in critical COVID-19. The rapid progressive fibrosis stage was more perilous than the alveolitis stage, even if SARS-CoV-2 is undetectable.