Unique genomic profiles obtained from cerebrospinal fluid cell-free DNA of non-small cell lung cancer patients with leptomeningeal metastases

Background: Leptomeningeal metastases (LM), associated with poor prognosis, are frequent complications of advanced non-small cell lung cancer (NSCLC) patients, especially in patients with epidermal growth factor receptor (EGFR) mutations. Due to limited access to leptomeningeal lesions, the mutational landscape of LM has not been comprehensively investigated in large cohorts and the underlining biology of LM remains elusive. Some studies have explored the potential of cerebrospinal fluid (CSF) in reflecting the molecular profile of LM but with limited number of patients enrolled.

Methods: In this study, we performed capture-based targeted sequencing using a panel consisting of 168 lung cancer-related genes on matched CSF and plasma samples from 72 advanced NSCLC patients with confirmed LM to interrogate the potential of CSF as a source of liquid biopsy.

Results: We revealed a rate of detection of 81.5% and 62.5% for CSF and plasma, respectively (p = 0.008). The maximum allelic fraction (MaxAF) was also significantly higher in CSF (43.6% vs. 4.6%) (p < 0.001). CSF, harboring a unique genomic profile by having a significant number of CSF-specific mutations, primarily copy number variations, is superior to plasma in reflecting the mutational profile of LM. Further pathway enrichment analysis revealed that most of CSF-specific mutations participated in pathways relevant to the tumorigenesis and the development of metastases. Moreover, our data also revealed that TP53 loss of heterozygosity (LOH) predominantly existed in CSF (p < 0.001).

Conclusions: Collectively, we demonstrated that CSF provides a more comprehensive profile of LM than plasma in a large cohort, thus can be used as an alternative source of liquid biopsy for LM patients.     

维生素D受体在糖尿病肾病足细胞损伤及蛋白尿缓解中的作用

目的探讨维生素D受体(VDR)在糖尿病肾病(DKD)足细胞中的表达水平及在足细胞损伤及蛋白尿缓解中的作用。方法(1)本研究纳入了65例诊断患有2型糖尿病(伴或不伴蛋白尿)的患者,并纳入了25例年龄和性别相匹配的健康体检者为对照组。根据白蛋白/肌酐(ACR)的尿排泄比例对2型糖尿病患者进行分组,分别为无蛋白尿(ACR<30 mg/g,n=24)、微量白蛋白尿(ACR 30~300 mg/g,n=18)和临床蛋白尿(ACR>300 mg/g,n=23)。另选择25例经肾活检确诊的DKD患者作为DKD组。正常肾脏组织标本均取自泌尿外科同一时期肾脏肿瘤切除患者10例。将各组检测指标进行对比,同时采用实时定量PCR、ELISA法和免疫组化法检测VDR在各组患者的血液、尿液样本和肾脏组织中的表达情况,以及使用Pearson相关分析分析VDR与尿蛋白的相关性。(2)在2型糖尿病肾病小鼠模型中对上述结果进行验证,将遗传背景均为C57BLKs/J的雄性db/db小鼠及同窝出生的db/m小鼠,随机分为正常对照组(A组)、DKD对照组(B组)、DKD二甲基亚砜处理组(C组)、DKD帕立骨化醇(VDR激动剂)处理组(D组),C、D组连续腹腔注射处理8周,对照组不做任何处理。小鼠10周龄时开始连续干预8周,在小鼠22周龄(开始干预后12周)检测各组小鼠体重、血、尿生化指标对比;Western印迹法检测β⁃catenin、VDR的变化;免疫荧光观察足细胞标志蛋白podocin及足细胞损伤蛋白α⁃SMA的表达变化。结果(1)与正常健康对照组相比,无蛋白尿组、微量白蛋白尿组和临床蛋白尿组的糖尿病患者血浆中VDR的mRNA和蛋白水平均较低(均P<0.05);与无蛋白尿组的糖尿病患者相比,微量白蛋白尿组和临床蛋白尿组的糖尿病患者血浆中VDR的mRNA和蛋白水平均较低(均P<0.05)。(2)与正常健康对照组相比,无蛋白尿糖尿病组和DKD组患者血浆中VDR的mRNA和蛋白水平均较低(均P<0.05);与无蛋白尿糖尿病组患者相比,DKD组患者血浆中VDR的mRNA和蛋白水平亦较低(均P<0.05)。(3)免疫组化结果显示,DKD组肾组织中VDR的表达明显少于正常对照组。(4)DKD患者血浆中VDR mRNA相对水平与ACR呈负相关(r=-0.342,P<0.05)。(5)各组尿液上清液中VDR的水平与血浆中的水平呈相反趋势。(6)Western印迹结果显示,B组、C组肾小球足细胞β⁃catenin蛋白表达高于D组(均P<0.05),VDR蛋白的表达低于D组(均P<0.05);免疫荧光结果显示,B组、C组肾小球足细胞podocin的表达低于D组(均P<0.05),α⁃SMA的表达高于D组(均P<0.05)。结论VDR高表达缓解DKD足细胞损伤及蛋白尿。     

Relationship between plasma cell-free DNA (cfDNA) and prognosis of TACE for primary hepatocellular carcinoma

BackgroundOur study aims to investigate changes in cell-free DNA (cfDNA) concentration and integrity in primary hepatocellular carcinoma (PHC) patients before and after transcatheter arterial chemoembolization (TACE) treatment and their influence on the evaluation of prognosis of the disease.MethodsA total of 84 PHC patients admitted to the Affiliated Hospital of Nanjing University of Chinese Medicine from December 2016 to December 2017 were included as the study group, while 55 healthy people served as the control group. Plasma cfDNA concentration and integrity were determined using qRT-PCR. The correlation between cfDNA concentration/integrity and clinical characteristics of PHC patients were analyzed. A ROC curve was used to investigate the sensitivity and specificity of cfDNA as detection indices. Univariate and multivariate analyses were used to analyze factors affecting recurrence in PHC patients and compare recurrence-free survival (RFS) of PHC patients with high cfDNA expression and low cfDNA expression.ResultsPlasma cfDNA concentration and integrity were significantly higher in PHC patients before TACE treatment than in healthy people and significantly lower after treatment than before (P<0.05). The cfDNA concentration was significantly correlated with tumor size, lymph node metastasis, TNM stage, and BCLC stage, while cfDNA integrity was significantly correlated with tumor size, TNM stage, and BCLC stage (P<0.05). ROC results showed that the area under the curve (AUC) value of cfDNA concentration was the largest, with an optimal cut-off of 10.51 ng/mL. Multivariate regression analysis for COX showed that the TNM stage, cfDNA concentration, and AFP were independent risk factors that affected PHC patients’ survival.ConclusionsPlasma cfDNA concentration in PHC patients is more sensitive and specific than any other tumor marker. It is an independent risk factor for PHC patients treated with TACE. Therefore, it is hypothesized cfDNA is a potential biomarker for prognostic evaluation of PHC patients treated with TACE.     

大黄酸通过抑制STAT3基因的表达调控骨肉瘤细胞的增殖和凋亡

目的:探讨大黄酸对骨肉瘤细胞增殖和凋亡的影响及潜在的作用机制。方法:以人骨肉瘤MG-63细胞为研究对象，利用CCK-8实验检测大黄酸对MG-63细胞增殖的影响，并计算大黄酸作用48 h的半数抑制浓度（IC50），采用该浓度进行后续的实验。转染STAT3 siRNA至MG-63细胞，qRT-PCR和Western blot检测分别在mRNA和蛋白水平上检测转染效果，采用CCK-8实验、克隆形成实验、流式细胞术验证大黄酸和STAT3 siRNA及其联合作用对MG-63细胞存活率、克隆形成、凋亡能力的影响，Western blot检测各组MG-63细胞中凋亡相关蛋白Bax、Bcl-2、Cleaved Caspase-3的表达情况。结果:大黄酸能够呈浓度依赖性的抑制骨肉瘤MG-63细胞增殖，干预48 h的IC50为46.05 μmol/L。大黄酸或STAT3 siRNA均能降低MG-63细胞中STAT3的表达，抑制MG-63细胞存活率，阻碍细胞克隆形成能力，促进细胞凋亡，上调细胞中Bax和Cleaved Caspase-3的表达，下调Bcl-2的表达；且二者联合效果更显著。结论:大黄酸通过抑制STAT3基因表达抑制骨肉瘤细胞增殖并促进细胞凋亡。     

新型冠状病毒肺炎疫情下肿瘤患者居家防护和饮食指导

新型冠状病毒肺炎疫情下，肿瘤患者免疫力低下，属于易感人群。营养不良是肿瘤患者最常见的营养问题，肿瘤患者作为特殊的人群值得被关注。本文从患者居家防护与饮食指导方面进行阐述，以期在特殊时期为肿瘤患者提供指导。     

Cytogenetic and molecular landscape and its potential clinical significance in Hispanic CMML patients from Puerto Rico

Chronic myelomonocytic leukemia (CMML) is a clonal hematopoietic neoplasm that exhibits myelodysplastic and myeloproliferative characteristics with heterogeneous clinical and pathological features. There are limited publications on the ethnic and racial disparity of cytogenetics and genomics in CMML patients. This study aims to define the cytogenetic and molecular landscape in Hispanic CMML patients from Puerto Rico and explore its possible clinical significance. One hundred and eleven (111) Hispanic CMML patients from Puerto Rico were diagnosed in our institute from 2009 to 2018. Karyotypes were available in one hundred and seven (107) patients. Seventeen (17) patients had abnormal karyotypes (17/107, 16%). Compared to previously published data, Hispanic CMML patients in this study had significantly lower rates of overall cytogenetic abnormalities (16% vs 27–28%, p < 0.05) and trisomy 8 (2% vs 7%, p < 0.05). Among one hundred and eleven (111) Hispanic CMML patients, 40-gene myeloid molecular profile tests were performed in fifty-six (56) CMML patients. Gene mutations were identified in fifty-four (54) patients (96%). The most frequent mutated genes were: TET2, SRSF2, ASXL1, ZRSR2, DNMT3A, NRAS, CBL, and RUNX1. Twenty-nine (29) out of fifty-six (56) patients (29/56, 52%) had mutated TET2/wild type ASXL1 (muTET2/wtASXL1). Previous studies indicated that mutated ASXL1, DNMT3A, NRAS, RUNX1, and SETBP1 may associate with an unfavorable prognosis and muTET2/wtASXL1 may associate with a favorable prognosis in CMML patients. Compared to previously published data, Hispanic CMML patients from Puerto Rico in this study had significantly lower mutation rates in ASXL1 and SETBP1, and a higher rate of muTET2/wtASXL1. The findings raise the possibility of a favorable prognosis in Hispanic CMML patients.     

营养途径对食管癌围术期患者营养状况的影响

目的探讨肠内营养与肠外营养对食管癌围术期患者的营养状况和并发症的影响。方法回顾性分析淮安市第二人民医院2017年1月至2018年6月食管癌手术治疗患者73例，根据营养干预途径不同分为全肠外营养组（TPN）23例和全肠内营养组（TEN）50例。TPN组给予静脉营养治疗，TEN组给予经鼻饲管（经鼻至空肠起始位置）肠内营养治疗。比较两组患者营养指标和免疫指标、术后肠道功能和安全性指标。结果两组患者就诊前后体重丢失、入院时血清白蛋白、淋巴细胞数组间比较，差异无统计学意义（P>0.05）。TPN组血清白蛋白和淋巴细胞数不同时间点整体比较，差异均具有统计学意义（P<0.05）。TEN组淋巴细胞数不同时间点整体比较，差异均具有统计学意义（P<0.05）。两组手术后组间比较，TEN组体重丢失幅度小于TPN组；TEN组术后72h、术后120h血清白蛋白均短于TPN组，TEN组术后120h淋巴细胞数高于TPN组；TEN组肛门排气时间和肠鸣音恢复时间均低于TPN组，差异均具有统计学意义（P<0.05）。两组不良反应和不良事件比较，差异无统计学意义（P>0.05）。结论对食管癌围术期患者而言，肠内营养的疗效总体优于肠外营养，且安全性无明显差异。     

Enhanced generation of reactive oxygen species and photocatalytic activity by Pt-based metallic nanostructures: the composition matters

The modification of semiconductor nanostructures with metallic nanocomponents can promote the separation of electron/hole from photoexited semiconductors by forming heterojunctions, thus exhibit enhanced photocatalytic activities and potential applications. In this study, Pt-based NPs, including Pt, PtCu, and PtCuCo are employed as model co-catalysts to comparatively study their capability to enhance the photocatalytic activity of TiO₂ nanosheets. It was found that each of Pt, PtCu, and PtCuCo can greatly enhance the photocatalytic activity of TiO₂ toward degradation of organic dyes. Using electron spin resonance spectroscopy, we demonstrated that deposition of Pt-based NPs resulted in more production of reactive oxygen species including hydroxyl radicals, superoxide, and singlet oxygen. The enhancing effects of Pt-based NPs on generation of ROS and photocatalytic activity showed same trend: PtCuCo?>?PtCu?>?Pt. The mechanism underlying the enhancement differences in Pt-based NPs may be mainly related to electronic structure change of Pt in alloying with Cu and Co. These results are valuable for designing hybrid nanomaterials with high photocatalytic efficiency for applications in water purification and antibacterial products.     

我国延续性护理的概念分析

目的 通过文献对延续性护理的发展、概念应用、属性、案例、前因、后果等进行系统分析，从护理实践的角度厘清我国延续性护理的概念内涵。方法 收集2009~2019年中国知网、万方、维普、中国生物医学数据库、PubMed、CINAHL、Embase、PsycINFO等数据库中延续性护理相关文献，采用Walker和Avant的概念分析法进行分析。 结果 通过分析相关文献并结合我国护理发展新形势特点，归纳出我国延续性护理的概念属性：患者发生不同医疗机构转移或不同照护层次转变；提供连续性、协调性、整体性的照护服务；目的是维护患者健康或满足其健康需求；存在信息、关系和管理三方面的延续。结论 我国延续性护理的概念属性从护理实践角度出发，结合新形势下护理发展特点，可为我国具体开展延续性护理服务实践或研究提供清晰的概念支持，并为各级医院和社区开展相关延续性护理工作提供一定的理论和实践指导。