Elevated expression of complement C3 protein in chemically induced hepatotumorogenesis in Wistar rats: A correlative proteomics and histopathological study

Liver cancer remains the leading cause of cancer-related mortality worldwide. Early detection of liver cancer is problematic due to the lack of a marker with high diagnosis sensitivity and specificity. The present study was designed to determine the differently expressed proteins at early stage in the serum of animals with liver cancer vis-à-vis controls and figure out the function of the proteins. One-dimensional electrophoresis (1D), two-dimensional electrophoresis (2DE) and liquid chromatography mass spectrometry (LC–MS/MS) were used to screen the serum proteins of liver cancer induced in animals by diethyl nitrosamine (DEN) + 2-acetyl amino fluorine (2-AAF). From optimized 2DE image and computer assisted PD Quest analysis were found to be differentially expressed spots when the serum from normal and treated animals were compared. Among these, one spot was selected whose expression level was higher in DEN + 2-AAF treated animal sera than in adjacent normal animal sera. The target spot was excised from the 2D gel of liver cancer sera and the peptide mass fingerprinting as obtained LC–MS/MS analysis after digesting the chosen protein spot. This was identified to be complement C3 protein. The changes in complement C3 expression level were validated by Western blot analysis. We reported that the changes in complement C3 concentration start at very early stage of tumorogenesis. The fully grown tumors were developed at 120 days and hepatotumorogenesis was confirmed by histopathological examination. This protein may therefore represent a powerful tool in search for candidate biomarkers for HCC.     

Cell-free plasma hypermethylated CASZ1, CDH13 and ING2 are promising biomarkers of esophageal cancer

Identifying sensitive and specific biomarkers for early detection of cancer is immensely imperative for early diagnosis and treatment and better clinical outcome of cancer patients. This study aimed to construct a specific DNA methylation pattern of cancer suppressor genes and explore the feasibility of applying cell-free DNA based methylation as a biomarker for early diagnosis of esophageal squamous cell carcinoma (ESCC). We recruited early stage ESCC patients from Yangzhong County, China. The Illumina Infinium 450K Methylation BeadChip was used to onstruct a genome-wide DNA methylation profile. Then, differentiated genes were selected for the validation study using the Sequenom MassARRAY platform. The frequency of methylation was compared between cancer tissues, matched cell-free DNAs and normal controls. The specific methylation profiles were constructed, and the sensitivity and specificity were calculated. Seven CG sites in three genes CASZ1, CDH13 and ING2 were significantly hypermethylated in ESCC as compared with normal controls. A significant correlation was found between the methylation of DNA extracted from cancer tissues and matched plasma cell-free DNA, either for individual CG site or for cumulative methylation analysis. The sensitivity and specificity reached 100% at an appropriate cut-point using these specific methylation biomarkers. This study revealed that aberrant DNA methylation is a promising biomarker for molecular diagnosis of esophageal cancer. Hypermethylation of CASZ1, CDH13 and ING2 detected in plasma cell-free DNA can be applied as a potential noninvasive biomarker for diagnosis of esophageal cancer.     

DNA Methylation and Nonsmall Cell Lung Cancer

Lung cancer is the leading cause of cancer‐related death in men and women worldwide. Owing to the scarcity of effective tools for early detection and therapy strategies, the 5‐year survival rate of lung cancer is very poor. Because the accumulation of multiple genetic and/or epigenetic changes, including DNA methylation, has been suggested to contribute to development and progression of human cancers, improved understanding of the relationship between DNA methylation and lung cancer will provide new insights for identifying promising biomarkers for diagnosis, prognosis, and treatment of lung cancer. Here, we present a relatively comprehensive review of DNA methylation and lung cancer, discuss DNA methylation changes in carcinogenesis and metastasis of lung cancer, and explore the association of microRNA with DNA methylation. Additionally, we outline the applications of DNA methylation in clinical practice, such as diagnosis, prognosis, and therapy of lung cancer. Anat Rec, 2011. © 2011 Wiley‐Liss, Inc.     

Prospects of electrochemical immunosensors for early diagnosis of preeclampsia

Preeclampsia is a vascular multisystem disorder that accounts for varying degree of morbidity and mortality of mother and the fetus. This can be significantly averted if diagnosed at an early (18‐20 weeks) stage of gestation, as there is no known way to prevent preeclampsia. In spite of extensive work on biomarker discovery, the existing method for its detection is mostly based on colorimetric immunoassays whose sensitivity is ranging in nanomolar range. Further, it has also been observed that change in the expression of a single biomarker is not sufficient to diagnose this condition. So, for early diagnosis (by 18‐20 weeks), an immuno‐diagnostic platform with detection limits in picomolar range and beyond along with the ability to do simultaneous detection of multiple analyte would be of great importance. A nano‐immunosensors with an electrochemical readout system can be a potential alternative that promises for the ultrasensitive detection of analyte with high specificity as well as suitability for on‐site analysis. Coupling the lateral flow technology with immunosensors would make it feasible to detect more than one biomarker simultaneously on a microchip. This review intends to summarize the potential preeclampsia biomarkers, limitations of existing diagnostic methods along with the recent advancements, and prospects to develop electrochemical immunosensors for early clinical diagnosis.     

Ultrafast Nanolaser Flow Device for Detecting Cancer in Single Cells

Currently, pathologists rely on labor-intensive microscopic examination of tumor cells using staining techniques originally devised in the 1880s that depend heavily on specimen preparation and that can give false readings. Emerging BioMicroNanotechnologies (Gourley, 2005) have the potential to provide accurate, realtime, high throughput screening of tumor cells without invasive chemical reagents. These techniques are critical to advancing early detection, diagnosis, and treatment of disease. Using a new technique to rapidly assess the properties of cells flown through a nanolaser semiconductor device, we discovered a method to rapidly assess the respiratory health of a single mammalian cell. The key discovery was the elucidation of biophotonic differences in normal and transformed (cancer) mouse liver cells by using intracellular mitochondria as biomarkers for disease. This technique holds promise for detecting cancer at a very early stage and could nearly eliminate delays in diagnosis and treatment.     

Clinical application of supersensitive and multiplex assay, MUSTag technology

Recently, various sets of protein biomarkers have been discovered in important diseases such as cancers, brain stroke, heart attack, diabetes, and so on. Many of these biomarkers are expected to be extremely valuable as targets for clinical diagnosis and drug development; however, the clinical validation is difficult and time-consuming by individual assays or due to very low concentration in an early stage of disease. For the super-sensitive and multiplex detection of target biomarkers, we have developed MUSTag (Multiple Simultaneous Tag) assay technology with innovative modification of the immuno-PCR method. In MUSTag technology, specific antibodies against several important biomarkers were linked to 100-300bp long oligonucleotides as detection tags. Each different oligo-tag simultaneously detects multiplex protein targets with extremely high sensitivity(more than 10 fg (10(-15) g)/ml) in a dose-dependent manner by qRT-PCR-based (maximum 3 plexes) or capillary electrophoretic amplification (over 30 plexes). Here we report our recent results of multiple cytokine assay or disease-specific biomarker assay using MUSTag technology, and further, clinical results from patients with cancers, ischemic brain or heart attack, who need prompt and predictive diagnosis for adequate treatment.     

肝癌相关肿瘤标志物研究新进展

肝细胞癌( hepatocellular carcinoma, HCC )是常见的恶性肿瘤之一,其发病隐匿,恶性程度高,病死率高,因此早期诊断对于提高患者的生存率至关重要.目前临床上主要运用甲胎蛋白(alpha-fetoprotein, AFP )结合影像学及病理学检查进行肝癌的早期诊断;但是AFP对于肝癌筛查的特异性...     

联合检测在肝癌预警中的作用

为了探讨联合检测在肝癌预警中的作用,将584例肝病患者分为研究组（349例）和对照组（235例）,通过对研究组的监测,研究联合检测的具体项目及检查顺序,确定肝癌高危人群的风险分级及筛查间隔时间,并探讨联合检测的临床意义。结果显示,进入预警系统的研究组筛查出的肝癌数及小肝癌数大于对照组,发现肝癌的临床分期早于对照组,且三年生存率提高,P值均小于0.01。结论：联合检测为肝癌的早发现、早诊断、早治疗提供了一条新出路,具有重要的临床意义。     

肿瘤早期诊断分子标志物研究进展

早期诊断是肿瘤研究的重要方向之一。虽然在宫颈癌、乳腺癌等一些肿瘤中建立了早期筛查方法,但对于大部分肿瘤而言,尚没有敏感、特异的早期诊断方法。高通量分子生物学技术的建立,使研究者可以在全基因组、转录组、蛋白质组等水平筛选分子标志物,大大提高了筛选效率。已有研究筛选出了大量的肿瘤早期分子改变和候选早期分子标志物,未来将有更多敏感、特异的早期诊断分子标志物被发现并用于肿瘤早期筛查和诊断。     

蛋白质指纹图谱技术在实验诊断与临床医学中的研究进展

蛋白质指纹图谱技术是近五年发展起来的实验室诊断新技术,它具有操作较简便、多样本检测、检测快速、灵敏性和特异性高等优点,是实验室诊断技术革命性的进展。蛋白质指纹图谱技术在医学领域中,主要用于多种疾病,如肿瘤的早期检测,对肿瘤早期检测的敏感性和特异性均在80%左右。以蛋白质指纹图谱技术为基础的免疫组质谱检测是在一个抗体组上同时捕获多个生物标志,并对捕获的变异或修饰的生物标志进行质谱精确分析。蛋白质指纹图谱技术及免疫组质谱检测技术特别对评估传统肿瘤标记物阴性的恶性肿瘤有意义。而且具有在分子或基因水平早期诊断的特点。蛋白质指纹图谱技术是一项发展前景非常好的诊断技术,具有广阔的临床应用前景。     

Screening and identification of potential novel lipid biomarkers for non-small cell lung cancer using ultra-high performance liquid chromatography tandem mass spectrometry

Lung cancer is characterized by a high incidence rate and low survival rate. It is important to achieve early diagnosis of the disease. We applied ultra-high performance liquid chromatography tandem mass spectrometry to screen plasma lipid spectrum in non-small cell lung cancer (NSCLC) patients, healthy controls (HC), and community-acquired pneumonia (CAP) patients. Modeling employing orthogonal partial least squares-discriminant analysis combined with t-test was used to screen the differential lipids. Logistic regression analysis was used to establish the diagnostic model, while the accuracy was verified by 10-fold cross-validation. The results showed that the abnormal metabolism of lipid in NSCLC mainly comprised fatty acid metabolism, phospholipid metabolism, and glyceride metabolism. Four potential biomarkers, including LPC (14:0/0:0), LPI (14:1/0:0), DG (14:0/18:2/0:0), and LPC (16:1/0:0), were fitted by the receiver operating characteristic curve model with the area under curve (AUC) value of 0.856, and the specificity and sensitivity were 87.0 and 78.0%, respectively. The results of cross validation showed that the AUC value of the model was 0.812, the sensitivity was 72.9%, and the specificity was 82.6%. The positive rate of four potential lipid biomarkers in this study (>60.0%) was higher than that of existing tumor biomarkers in the clinical application. We investigated the plasma lipid profile of NSCLC patients and identified lipid biomarkers with potential diagnostic values. From the lipidomics perspective, our study may lay a foundation for the biomarker-based early diagnosis of lung cancer.     

超声检查在高尿酸血症患者多脏器评估中的临床应用价值

目的探讨与评估彩色多普勒超声在高尿酸血症患者多脏器评估中的临床应用价值。方法纳入2015年7月至2017年5月于北京协和医院就诊的50例初治的高尿酸血症患者,记录血清尿酸(sUA)测值,应用彩色多普勒超声对每例患者的双侧第一跖趾关节、膝关节、肾脏、颈动脉及肝脏进行检查,评估sUA水平与上述脏器病变的相关性;进一步将研究对象分为痛风性关节炎(GA)组和无症状高尿酸血症(AH)组,分析并比较两组多脏器病变的情况。统计学方法采用卡方检验及Spearman相关分析。结果本研究50例初治高尿酸血症患者中,超声显示关节病变的检出率56.0%,肾脏形态结构改变的检出率72.0%;非酒精性脂肪肝(NAFLD)的检出率64.0%,其中59.4%为中重度脂肪肝,sUA水平与NAFLD的检出率呈正相关(P<0.05);颈动脉粥样硬化的检出率36.0%,sUA水平与颈总动脉内中膜厚度呈正相关(P<0.05);关节、肾脏、肝脏、颈动脉中两个/两个以上脏器病变的检出率80.0%,sUA水平与多脏器病变的检出率呈正相关(P<0.05)。GA组与AH组之间关节病变及颈动脉粥样硬化检出率差异有统计学意义(P<0.05)。结论彩色多普勒超声对于高尿酸血症患者关节病变和肾损害、非酒精性脂肪肝以及颈动脉粥样硬化等多脏器病变的早期诊断、全面评估病情进展和随访治疗后疗效具有一定的临床应用价值。     

血清sP-selectin、sICAM-1和免疫球蛋白检测对乙肝肝硬化预后评估的意义

目的：旨在探讨血清sP-selectin、sICAM-1和免疫球蛋白检测对乙肝肝硬化预后评估的意义。方法：选取池州市人民医院于2014年1月~2016年12月收治的162例乙肝肝硬化患者作为研究对象,分别测定患者血清可溶性P-选择素(sP-selectin)、可溶性细胞间黏附分子-1(sICAM-1)、免疫球蛋白和相关血清指标。比较不同肝硬化患者和肝癌患者血清指标差异性,Pearson分析血清可溶性P-选择素(sP-selectin)、可溶性细胞间黏附分子-1(sICAM-1)、免疫球蛋白和相关血清指标相关性,logistic多因素回归分析肝硬化转肝癌危险因素,受试者工作特征（ROC）曲线评价OR>1参数对肝硬化转肝癌预测价值。结果：原发性肝癌患者sP-selectin、sICAM-1、IgM、IgG、IgA显著高于代偿期肝硬化和失代偿期肝硬化患者（P<0.05）。失代偿期肝硬化患者sP-selectin、sICAM-1、IgM、IgG、IgA显著高于代偿期肝硬化患者（P<0.05）。不同Child-Pugh分级肝硬化患者中,C级患者sP-selectin、sICAM-1、IgM、IgG、IgA显著高于B级和A级患者（P<0.05）。B级患者sP-selectin、sICAM-1、IgM、IgG、IgA显著高于A级患者（P<0.05）。多发癌灶患者和单发癌灶原发性肝癌患者sP-selectin、sICAM-1和免疫球蛋白表达差异无统计学意义（P>0.05）。Ⅱ期原发性肝癌患者sP-selectin、sICAM-1和免疫球蛋白显著高于Ⅰ期患者（P<0.05）。肝硬化患者sP-selectin、sICAM-1、免疫球蛋白之间均互为正相关性（P<0.05）。以肝硬化是否转原发性肝癌为因变量进行多因素Logistic回归分析,结果显示,ALT、AST、sP-selectin、sICAM-1、IgM、IgG、IgA均是肝硬化转原发性肝癌的危险因素。ROC曲线分析可得,sICAM-1和sP-selectin诊断肝硬化转肝癌敏感性和特异性均高于其他因素。结论：血清sP-selectin、sICAM-1对乙肝肝硬化预后评估具有较高临床诊断精度,对肝硬化转原发性肝癌的诊断特异度、敏感度高于单纯免疫球蛋白检测,故其联合检测诊断肝硬化预后值得临床推广。     

肝癌患者循环肿瘤细胞的检测及其临床应用研究进展

原发性肝癌发病率和死亡率均较高,其术后转移与复发已经成为影响患者预后的重要因素,临床上迫切需要能够有效预测和防治肝癌转移的检测手段。循环肿瘤细胞（circulating tumor cell,CTC）,作为近些年来新出现的肿瘤检测指标,因其能够实时、无创地监测肿瘤患者的病情状态,在早期诊断、早期治疗,判断预后和制定个体化治疗方案等方面具有重要意义而备受关注。CTC在乳腺癌、前列腺癌、结直肠癌、肺癌等疾病的应用已初步得到临床认可。最新的研究表明,肝癌患者病情状态与血液中CTC的数量也显示出极强的相关性。目前各种CTC检测体系主要包括富集纯化和鉴定两方面;但由于检测方法繁多,不同方法的敏感性、特异性不同限制了其临床应用。近年来,新的CTC检测方法层见叠出,涌现出了很多新技术用于肝癌CTC的临床研究。本文就其相关研究进展做一综述。     

血清GGT、ALP及ADA在肝癌诊断中的临床价值

目的探讨血清R-谷氨酰转移酶(GGT)、碱性磷酸酶(ALP)和腺苷脱氨酶(ADA)检测对肝癌诊疗的临床意义。方法随机选取肝癌患者、良性肝病患者及健康体检者(正常对照组)各45例,分别检测其血清GGT、ALP及ADA含量,统计分析各组间结果差异,并比较三种指标单独或联合检测对肝癌诊断的敏感性和特异性。结果血清GGT、ALP及ADA含量在肝癌患者组明显高于良性肝病患者组及正常对照组,差异有统计学意义(P0.05);对肝癌的诊断:单项检测GGT、ALP及ADA的敏感性分别为84%、69%、78%,特异性分别为76%、89%、91%;三项联合检测的敏感性为91%,特异性为69%。结论三项联合检测提高了对肝癌诊断的敏感性,有利于肝癌的早期诊断,降低了对肝癌的漏诊率。     

钼靶X线联合高频彩超在乳腺癌早期筛查中的应用价值

目的 探讨钼靶X线联合高频彩超在乳腺癌早期筛查中的应用价值。方法 选取2016年1月~2018年12月于什邡市人民医院就诊的126例早期乳腺癌患者（直径＜2 cm）作为研究对象。所有患者均被序贯予以钼靶X线和高频彩超检查,比较钼靶X线、高频彩超及联合检测早期乳腺癌筛查的血流信号检出率、微钙化灶检出率及诊断准确率。结果 高频彩超血流信号检出率为80.16%,高于钼靶X线的21.43%;高频彩超微细钙化检出率为33.33%,低于钼靶X线的59.52%,差异有统计学意义(P＜0.05)。高频超声诊断准确率为82.54%,高于钼靶X线的80.16%,差异无统计学意义(P＞0.05);而联合检查诊断准确率为96.83%,高于单用高频超声和单用钼靶X线,差异有统计学意义(P＜0.05)。结论 钼靶X线、高频彩超应用于乳腺癌的早期筛查各有优劣,联合应用能提高诊断乳腺癌的准确率,具有较高的临床价值。     

Hypermethylated FAM5C and MYLK in serum as diagnosis and pre-warning markers for gastric cancer

Most cases of gastric cancer (GC) are not diagnosed at early stage which can be curable, so it is necessary to identify effective biomarkers for its diagnosis and pre-warning. We have used methylated DNA immunoprecipitation (MeDIP) to identify genes that are frequently methylated in gastric cancer cell lines. Promoter regions hypermethylation of candidate genes were tested by methylation-specific polymerase chain reaction (MSP) in serum samples, including GC (n=58), gastric precancerous lesions (GPL, n=46), and normal controls (NC, n=30). Eighty two hypermethylated genes were acquired by array analysis and 5 genes (BCAS4, CHRM2, FAM5C, PRAC and MYLK) were selected as the candidate genes. Three genes (CHRM2, FAM5C and MYLK) were further confirmed to show methylation rates increased with progression from NC to GPL, then to GC. There was obvious decrease in detection of FAM5C and MYLK hypermethylation, but not CHRM2, from preoperative to postoperative evaluation (P< 0.001). Combined detection of FAM5C and MYLK hypermethylation had a higher sensitivity in GC diagnosis (77.6%,45/58) and pre-warning (30.4%,14/46) than one single gene detection and also had a high specificity of 90%. The combined hypermethylated status of FAM5C and MYLK correlated with tumor size (P<0.001), tumor invasion depth (P=0.001) and tumor-node-metastasis (TNM) stage (P=0.003). Hypermethylated FAM5C and MYLK can be used as potential biomarkers for diagnosis and pre-warning of GC.     

Contribution of (1,3)-Beta-d-Glucan to Diagnosis of Invasive Candidiasis after Liver Transplantation

Invasive candidiasis (IC) causes high morbidity and mortality rates after liver transplantation, in part due to delayed diagnosis. The fungal cell wall component (1,3)-beta-d-glucan (BG) could be an early biomarker of IC. This preliminary prospective study was designed to evaluate the contribution of BG measurements to the diagnosis of IC after liver transplantation. All consecutive patients who underwent liver transplantation at Henri Mondor Hospital in France between January and June 2013 were enrolled prospectively in the study. They were monitored weekly for colonization by Candida, and colonization index values were calculated. Serum samples were tested for BG (Fungitell; Cape Cod Inc.) at least weekly between liver transplantation and discharge from the hospital. A total of 52 patients (including 39 male patients) were enrolled, with a median age of 55 years (range, 31 to 69 years). The median Model for End-Stage Liver Disease (MELD) score was 27 (range, 6 to 40). Cultures from 42 patients (81%) yielded Candida spp., with the most common Candida species isolated being Candida glabrata (47%). Six cases of documented IC were found for four of the 52 patients. On the day the clinical diagnosis of IC was made, analysis based on combining two sequential BG-positive samples (>146 pg/ml) and a colonization index of ≥0.5 revealed sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) results of 83%, 89%, 50%, and 97.6%, respectively. The detection of BG associated with Candida colonization may be a promising tool based on a high NPV that can rule out IC among high-risk patients.