Urinary metabolomic study of non‐small cell lung carcinoma based on ultra high performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry

Metabolic profiles from human urine reveal the significant difference of carnitine and acylcarnitines levels between non‐small cell lung carcinoma patients and healthy controls. Urine samples from cancer patients and healthy individuals were assayed in this metabolomic study using ultra high performance liquid chromatography coupled to quadrupole time‐of‐flight mass spectrometry. The data were normalized by the sum of all intensities and creatinine calibration, respectively, before orthogonal partial least squares discriminant analysis. Twenty differential metabolites were identified based on standard compounds or tandem mass spectrometry fragments. Among them, some medium‐/long‐chain acylcarnitines, for example, cis‐3,4‐methylene heptanoylcarnitine, were found to be downregulated while carnitine was upregulated in urine samples from the cancer group compared to the control group. Receiver operating characteristic analysis of the two groups showed that the area under curve for the combination of carnitine and 11 selected acylcarnitines was 0.958. This study suggests that the developed carnitine and acylcarnitines profiling method has the potential to be used for screening non‐small cell lung carcinoma.     

Serum metabolomics of laryngeal cancer based on liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry

The discovery of new laryngeal cancer‐related metabolite biomarkers could help to facilitate early diagnosis. A serum metabolomics study from laryngeal cancer patients and healthy individuals was conducted using liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry. Univariate and multivariate statistics were used to discriminate laryngeal cancer patients and healthy individuals. 1‐Palmitoyl‐sn‐glycero‐3‐phosphocholine (LysoPC 16:0), 1‐o‐hexadecyl‐2‐acetyl‐sn‐glycero‐3‐phosphocholine (PAF) and 1,2‐dipalmitoyl‐sn‐glycero‐3‐phosphocholine were found to be significantly different between the laryngeal cancer group and the healthy group. They are mainly involved in phospholipids catabolism, linoleic acid metabolism, α‐linoleic acid metabolism and arachidonic acid metabolism. The area under the curve of the biomarker combined by two metabolites (LysoPC 16:0 and PAF) was 0.935, the sensitivity was 0.962 and the specificity was 0.825. LysoPC 16:0 and PAF may show diagnostic potential for laryngeal carcinoma.     

GC/MS-based metabolomic approach to validate the role of urinary sarcosine and target biomarkers for human prostate cancer by microwave-assisted derivatization

A recent study showed that sarcosine may be potentially useful for the diagnosis and prognosis of prostate cancer (PCa). The aim of this study was to validate diagnostic value of sarcosine for PCa, to evaluate urine metabolomic profiles in patients with PCa in comparison of non-cancerous control, and to further explore the other potential metabolic biomarkers for PCa. Isotope dilution gas chromatography/mass spectrometry (ID GC/MS) metabolomic approach was applied to evaluate sarcosine using [methyl-D₃]-sarcosine as an internal standard. Microwave-assisted derivatization (MAD) together with GC/MS was utilized to obtain the urinary metabolomic information in 20 PCa patients compared with eight patients with benign prostate hypertrophy and 20 healthy men. Acquired metabolomic data were analyzed using a two-sample t test. Diagnostic models for PCa were constructed using principal component analysis and were assessed with receiver–operating characteristic curves. Results showed that the urinary sarcosine level has no statistical difference between the PCa group and the control group. In addition, nine metabolomic markers between the PCa group and the healthy male group were selected, which constructed a diagnostic model with a high area under the curve value of 0.9425. We conclude that although urinary sarcosine value has limited potential in the diagnostic algorithm of PCa, urinary metabolomic panel based on GC/MS assay following MAD may potentially become a diagnostic tool for PCa.     

剪切波弹性成像定量参数对良恶性乳腺肿块的鉴别及对乳腺癌术后复发的预测价值

探讨剪切波弹性成像定量参数对乳腺癌诊断和预后预测的作用.以乳腺癌患者60例作为观察组,乳腺良性结节患者60例作为对照组,利用剪切波弹性成像行超声检查,收集最大弹性模量值(Emax).t检验分析两组Emax差异,并使用ROC曲线分析Emax诊断乳腺癌的价值.结果显示,与对照组比较,观察组AE-max、Shell1Emax...     

18F-FDG PET/CT显像联合肿瘤标志物检查对女性不明原因腹腔积液的诊断价值

本文探讨了18 F-脱氧葡萄糖(18 F-FDG)正电子发射计算机断层显像(PET/CT)联合肿瘤标志物检查对女性不明原因腹腔积液的鉴别诊断价值。选取95例不明原因腹腔积液女性患者的18 F-FDG PET/CT影像资料及肿瘤标志物资料进行回顾性分析,并与病理诊断结果进行对照。结果显示:95例患者中,经病理证实恶性腹腔积液74例,良性腹腔积液21例。PET/CT对良恶性腹腔积液的诊断灵敏度为98.65%,特异度为19.05%,准确度为81.05%。恶性腹腔积液的SUV max与良性腹腔积液比较,差异无统计学意义(P>0.05)。将患者按病因进一步细分后发现,卵巢癌组的SUV max明显高于其他良性原因组(P<0.05),但与结核组和其他恶性肿瘤组比较,差异无统计学意义(P>0.05)。肿瘤标志物中以CA125检测效率最高,其对卵巢癌及结核所致腹腔积液的诊断灵敏度为98.2%,特异度为7.7%,准确度为81.4%。PET/CT和CA125联合诊断对卵巢癌及结核所致腹腔积液的灵敏度、特异度及准确度与单一PET/CT或CA125诊断比较差异均无统计学意义(P>0.05)。本实验表明18 F-FDG PET/CT诊断女性不明原因腹腔积液的良恶性有较高的灵敏度和准确度,但特异性较差,联合肿瘤标志物检查并不能提高对卵巢癌及结核所致腹腔积液的诊断效能。     

Screening for Oral Cancer Using Electrochemical Telomerase Assay

Electrochemical telomerase assay (ECTA) developed by our group was evaluated in an oral cancer screening using exfoliated oral cells and tissues obtained from patients of oral cancer, mucosa associated disease, or healthy volunteers. Telomerase activity from ECTA is correlated with hTERT mRNA expression level using a real‐time PCR and was increasing in the following order: healthy volunteer group<mucosa associated disease group<oral cancer group. Sensitivity and specificity of ECTA were 88 % and 72 %, respectively when used 17 % of the threshold value based on the receiver operating characteristic curve in ECTA data.     

磁共振T2WI、DWI对直肠癌新辅助治疗疗效的评估价值

本研究探讨磁共振成像T2WI、弥散加权成像(DWI)在直肠癌新辅助治疗疗效的评估价值。选取直肠癌患者92例,给予新辅助治疗,比较治疗前后T2WI、DWI参数变化。治疗无效患者治疗后肿瘤绝对信号值(SIT)明显高于治疗有效患者(P<0.05),而肿瘤绝对信号值减低率(SITRR)、治疗后肿瘤相对信号值(SIT/M)、肿瘤相对信号值减低率(SIT/MRR)明显低于治疗有效患者(P<0.05);治疗无效患者治疗后表观扩散系数(ADC)值、ADC变化值明显低于治疗有效患者(P<0.05);治疗后SIT、SITRR、SIT/MRR预测治疗有效的ROC曲线下面积分别为0.671、0.950和0.879(P<0.05);治疗后ADC值、ADC变化值预测治疗有效的ROC曲线下面积分别为0.717和0.977(P<0.05)。磁共振T2WI图像参数、ADC在预测直肠癌新辅助治疗疗效中有较高的应用价值。     

Extracellular matrix alterations in low‐grade lung adenocarcinoma compared with normal lung tissue by imaging mass spectrometry

Lung adenocarcinoma (LUAD) is the second most common cancer, affecting both men and women. Fibrosis is a hallmark of LUAD occurring throughout progression with excess production of extracellular matrix (ECM) components that lead to metastatic cell processes. Understanding the ECM cues that drive LUAD progression has been limited due to a lack of tools that can access and report on ECM components within the complex tumor microenvironment. Here, we test whether low‐grade LUAD can be distinguished from normal lung tissue using a novel ECM imaging mass spectrometry (ECM IMS) approach. ECM IMS analysis of a tissue microarray with 20 low‐grade LUAD tissues and 20 normal lung samples from 10 patients revealed 25 peptides that could discriminate between normal and low‐grade LUAD using area under the receiver‐operating curve (AUC) ≥0.7, P value ≤.001. Principal component analysis demonstrated that 62.4% of the variance could be explained by sample origin from normal or low‐grade tumor tissue. Additional work performed on a wedge resection with moderately differentiated LUAD demonstrated that the ECM IMS analytical approach could distinguish LUAD spectral features from spectral features of normal adjacent lung tissue. Conventional liquid chromatography with tandem mass spectrometry (LC‐MS/MS) proteomics demonstrated that specific sites of hydroxylation of proline (HYP) were a main collagen post translational modification that was readily detected in LUAD. A distinct peptide from collagen 3A1 modified by HYP was increased 3.5 fold in low‐grade LUAD compared with normal lung tissue (AUC 0.914, P value <.001). This suggests that regulation of collagen proline hydroxylation could be an important process during early LUAD fibrotic deposition. ECM IMS is a useful tool that may be used to define fibrotic deposition in low‐grade LUAD.     

二维超声联合超声造影TIC参数在乳腺导管内病变良/恶性诊断中的应用价值

本研究分析了二维超声(2DUS)联合超声造影(CEUS)时间-强度曲线(TIC)参数在乳腺导管内病变良/恶性鉴别诊断中的应用价值.选取乳腺导管内病变患者81例(共93个病变),均行2DUS及CEUS检查.结果显示,2DUS检出恶性病变17个,良性病变48个,诊断准确度69.89％(65/93)、敏感度53.13％(17...     

UPLC‐MS/MS based diagnostics for epithelial ovarian cancer using fully sialylated C4‐binding protein

Serum levels of fully sialylated C4‐binding protein (FS‐C4BP) are remarkably elevated in patients with epithelial ovarian cancer (EOC) and can be used as a marker to distinguish ovarian clear cell carcinoma from endometrioma. This study aimed to develop a stable, robust and reliable liquid chromatography–hybrid mass spectrometry (UPLC‐MS/MS) based diagnostic method that would generalize FS‐C4BP as a clinical EOC biomarker. Glycopeptides derived from 20 μL of trypsin‐digested serum glycoprotein were analyzed via UPLC equipped with an electrospray ionization time‐of‐flight mass spectrometer. This UPLC‐MS/MS‐based diagnostic method was optimized for FS‐C4BP and validated using sera from 119 patients with EOC and 127 women without cancer. A1958 (C4BP peptide with two fully sialylated biantennary glycans) was selected as a marker of FS‐C4BP because its level in serum was highest among FS‐C4BP family members. Preparation and UPLC‐MS/MS were optimized for A1958, and performance and robustness were significantly improved relative to our previous method. An area under the curve analysis of the FS‐C4BP index receiver operating characteristic curve revealed that the ratio between A1958 and A1813 (C4BP peptide with two partially sialylated biantennary glycans) reached 85%. A combination of the FS‐C4BP index and carbohydrate antigen‐125 levels further enhanced the sensitivity and specificity.     

MR-DWI成像预测乳腺癌腋窝淋巴转移的价值

本研究探讨磁共振弥散加权成像(MR-DWI)评估乳腺癌患者腋窝淋巴结转移的价值。回顾性选取乳腺癌患者68例(观察组),同时选取乳腺良性病变患者60例作为对照组,比较两组MR-DWI差异。观察组弥漫高信号、混杂高信号比例明显高于对照组(P<0.05);观察组病灶ADC值明显低于对照组(P<0.05);观察组Ⅲ期病灶ADC值明显低于Ⅰ~Ⅱ期(P<0.05);观察组腋窝淋巴结转移ADC值明显低于无淋巴结转移(P<0.05);弥漫高信号和其他信号组织ADC值比较差异无统计学意义(P>0.05);ADC值预测腋窝淋巴结转移的ROC曲线下面积为0.752,P<0.05。MR-DWI在乳腺癌腋窝淋巴结转移诊断应用价值较好,值得临床使用。     

Plasmonic Gold Chip for Multiplexed Detection of Ovarian Cancer Biomarker in Urine

Human epididymal protein 4(HE4), carbohydrate antigen 125 (CA125) and osteopontin(OPN) are three key biomarkers in detecting ovarian cancer. To explore the diagnosis value of combined detection of these three biomarkers for ovarian cancer, we developed a multiplexed assay on a plasmonic gold(pGOLD) platform for measuring HE4, CA125 and OPN in urine. The receiver operator characteristic(ROC) curve was drawn, and the diagnosis values of each biomarker alone or in combination for ovarian cancer were evaluated. In the analysis to distinguish ovarian cancer from other gynecological cancers, ovarian cysts and healthy people, the sensitivities of HE4, CA125 and OPN were 72.55%, 52.82% and 68.63%, the specificity values were 95.06%, 87.65% and 90.12%, while the areas under the curve(AUC) were 0.85, 0.75 and 0.77, respectively. The sensitivity and specificity for combination detection of the three markers were 90.20% and 80.25%. The detection methods of HE4, CA125 and OPN based on plasma fluorescence enhanced chip showed good analytic and diagnostic performance, and provided a non-invasive method for the diagnosis of ovarian cancer.     

DCE-MRI定量参数对子宫内膜癌的诊断价值及与临床病理因素的相关性

本文选取了子宫内膜癌患者108例作为观察组,另选取子宫内膜良性病变患者105例作为对照组,均行DCE-MRI检查后发现,观察组容量转移常数(Ktrans)、速率常数(Kep)、血管外细胞外间隙容积比值(Ve)高于对照组(P＜0.05);Ktrans、Kep、Ve值联合诊断子宫内膜癌的曲线下面积(AUC)为0.841;K...     

Pressurized CEC coupled with QTOF‐MS for urinary metabolomics

Pressurized CEC (pCEC) coupled with ESI‐QTOF‐MS using a sheathless interface was applied for metabolomics to develop an alternative analytical method for metabolic profiling of complex biofluid samples such as urine. The hyphenated system was investigated with mixed standards and pooled urine samples to evaluate its precision, repeatability, linearity, sensitivity, and selectivity. The applied voltage, mobile phase, and gradient elution were optimized and applied for the analysis of urinary metabolites. Multivariate data analysis was subsequently performed and used to distinguish lung cancer patients from healthy controls successfully. High separation efficiency has been achieved in pCEC due to the EOF. For metabolite identification, the pCEC‐MS separation mechnism was helpful for discriminating the fragment ions of glutamine conjugates from co‐eluted metabolites. Three glutamine conjugates, including phenylacetylglutamine, acylglutamine C8:1, and acylglutamine C6:1 were identified among 16 differential urinary metabolites of lung cancer. Receiver‐operating‐characteristic analysis of acylglutamine C8:1 resulted in an area‐under‐curve value of 0.882. Overall, this work suggests that this pCEC‐ESI‐QTOF‐MS method may provide a novel and useful platform for metabolomic studies due to its superior separation and identification.     

Analysis of cysteine glutathionylation in hemoglobin of gastric cancer patients using nanoflow liquid chromatography/triple‐stage mass spectrometry

Glutathione is an intracellular antioxidant capable of scavenging free radicals and detoxifying electrophiles from endogenous and exogenous sources via the free thiol group. Post‐translational glutathionylation at cysteine residues of proteins can affect the structure and cause a functional change of proteins. Protein glutathionylation has been proven to reflect the cellular redox status. Our previous report indicates that the levels of glutathionylation in hemoglobin from peripheral blood of smokers are significantly higher than in nonsmokers. In this study, a nanoflow liquid chromatography/nanospray ionization triple‐stage mass spectrometric (nanoLC/NSI‐MS³) method with a linear ion trap mass spectrometer was employed to quantify glutathionylated peptides in the trypsin digests of hemoglobin from gastric cancer patients. We compare the extent of glutathionylation in hemoglobin from nonsmoking gastric cancer patients with that from nonsmoking healthy adults. Using a carboxymethylated peptide as the reference peptide, the relative quantification of each glutathionylated peptide was measured as the peak area ratio of the modified peptide versus the sum of the peak areas of the modified and the carboxymethylated parent peptide in the selected reaction monitoring chromatograms. Using this method, we found that the extents of glutathionylation at Cys‐104 of the α‐globin and Cys‐93 of β‐globulin hemoglobin from 10 gastric cancer patients were significantly higher than those from 14 normal individuals with p values <0.0001. Our results suggest the possibility of using the extent of cysteine glutathionylation at β‐93 of hemoglobin as an oxidative stress biomarker candidate for gastric cancer.     

Mass spectrum analysis of serum biomarker proteins from patients with schizophrenia

Diagnosis of schizophrenia does not have a clear objective test at present, so we aimed to identify the potential biomarkers for the diagnosis of schizophrenia by comparison of serum protein profiling between first‐episode schizophrenia patients and healthy controls. The combination of a magnetic bead separation system with matrix‐assisted laser desorption/ionization time‐of‐flight tandem mass spectrometry (MALDI‐TOF/TOF‐MS) was used to analyze the serum protein spectra of 286 first‐episode patients with schizophrenia, 41 chronic disease patients and 304 healthy controls. FlexAnlysis 3.0 and ClinProTools^TM 2.1 software was used to establish a diagnostic model for schizophrenia. The results demonstrated that 10 fragmented peptides demonstrated an optimal discriminatory performance. Among these fragmented peptides, the peptide with m/z 1206.58 was identified as a fragment of fibrinopeptide A. Receiver operating characteristic analysis for m/z 1206.58 showed that the area under the curve was 0.981 for schizophrenia vs healthy controls, and 0.999 for schizophrenia vs other chronic disease controls. From our result, we consider that the analysis of serum protein spectrum using the magnetic bead separation system and MALDI‐TOF/TOF‐MS is an objective diagnostic tool. We conclude that fibrinopeptide A has the potential to be a biomarker for diagnosis of schizophrenia. This protein may also help to elucidate schizophrenia disease pathogenesis. Copyright © 2013 John Wiley & Sons, Ltd.     

Plasma Exosomes of Patients with Breast and Ovarian Tumors Contain an Inactive 20S Proteasome

Exosomes are directly involved in governing of physiological and pathological conditions of an organism through the transfer of information from producing to receiving cells. It can be assumed that exosomes are one of the key players of tumor dissemination since they are very stable and small enough to penetrate from various tissues into biological fluids and then back, thus interacting with tissue target cells. We evaluated the enzymatic activity and the level of 20S proteasome in tissue and exosomes of healthy females (n = 39) and patients with ovarian (n = 50) and breast (n = 108) tumors to reveal the critical role of exosomal cargo in the mediation of different types of metastases. Exosomes from plasma and ascites were isolated and characterized in according to International Society for Extracellular Vesicles guidelines. The level of 20S proteasome in tissue and exosomes was determined using Western blot analysis. Chymotrypsin- and caspase-like (ChTL and CL, respectively) peptidase activities of the proteasomes were determined using fluorogenic Suc-LLVY-AMC and Cbz-LLG-AMC substrates, respectively. We observed increased levels of 20S proteasome in ovarian cancer tissue and luminal B subtype breast cancer tissue as well as in plasma exosomes from cancer patients. Moreover, the level of the 20S proteasome in plasma exosomes and ascites exosomes in patients with ovarian tumors is comparable and higher in ovarian cancer patients with low volume ascites than in patients with moderate and high-volume ascites. We also found increased ChTL and CL activities in breast cancer and ovarian cancer tissues, as well as in peritoneal metastases in ovarian cancer, while proteasomal activity in exosomes from plasma of healthy females and all patients, as well as from ascites of ovarian tumor patients were lower than detection limit of assay. Thus, regardless of the type of tumor metastasis (lymphogenous or peritoneal), the exosomes of cancer patients were characterized by an increased level of 20S proteasome, which do not exhibit enzymatic activity.     

Design and Synthesis of Novel Triazolyl Benzoxazine Derivatives and Evaluation of Their Antiproliferative and Antibacterial Activity

A series of novel triazolyl benzoxazine derivatives have been synthesized via Cu(I)‐catalyzed ‘Click’ cycloaddition. All of the compounds were fully characterized from their spectral data, and their antiproliferative activity was evaluated against three selected human cancer cell lines: cervical cancer cells (HeLa), colorectal adenocarcinoma (HT‐29), and ovarian adenocarcinoma (SKOV‐3). A few representative compounds have also been evaluated for their antibacterial potential against two bacterial strains Pseudomonas aeruginosa and Bacillus subtilis.     

MRI-DWI技术诊断复杂性肛瘘的效能及与治疗效果的关联性分析

选取83例复杂性肛瘘患者作为观察组,选取同期80例健康体检者作为对照组,均接受磁共振弥散加权成像(MRI-DWI)技术检查,统计两组表观扩散系数(ADC)值、肛周疾病活动指数(PDAI)。结果发现,病灶ADC值与肛周异常排泄物评分、并发肛周疾病类型评分、肛周硬结评分呈负相关(P<0.05);ROC曲线显示,ADC诊断复杂性肛瘘价值的曲线下面积(AUC)值为0.826,诊断敏感度及特异度分别为85.54%和66.25%。由此可见,ADC值与复杂性肛瘘患者肛周疾病活动性、疗效及肛周功能具有一定关联性,早期采用MRI-DWI技术检查,可为复杂性肛瘘诊治、病情程度与疗效判断、预后评估提供辅助影像学手段。     

Searching for serum tumor markers for colorectal cancer using a 2‐D DIGE approach

Identification of specific protein markers for colorectal cancer (CRC) could provide a basis for its early diagnosis and detection, as well as clues to the molecular mechanisms governing cancer progression. In the present study, 2‐D DIGE coupled with MS was used to screen for biomarker candidates in the serum proteome of ten human CRC samples and ten healthy control samples. After pooling identical amounts of serum proteins (based on total protein concentration), albumin/IgG was depleted under partially denaturing conditions. Subsequently, the serum samples were labeled with three different CyDyes, and separated by 2‐D DIGE. After analysis with the biological variation analysis module of the DeCyder software, only three spots were found to be significantly elevated in all patient groups (with ratios from 1.52 to 9.08), whereas five spots were significantly down‐regulated in patients (with ratios from ?1.23 to ?10.21) (t‐test; p<0.05). Finally, two potential biomarkers, Transaldolase 1 and thyroid receptor interactor, were chosen for validation and analysis by ELISA with the serum of 30 CRC patients and 30 healthy controls. The serum levels of the two proteins correlated well with the 2‐D DIGE results. Thus, 2‐D DIGE approaches show great promise for biomarker discovery in CRC.