Prognosis Monitoring Research Articles

Integrating Cell Phone Imaging with Magnetic Levitation (i-LEV) for Label-Free Blood Analysis at the Point-of-Living.

There is an emerging need for portable, robust, inexpensive, and easy-to-use disease diagnosis and prognosis monitoring platforms to share health information at the point-of-living, including clinical and home settings. Recent advances in digital health technologies have improved early diagnosis, drug treatment, and personalized medicine. Smartphones with high-resolution cameras and high data processing power enable intriguing biomedical applications when integrated with diagnostic devices. Further, these devices have immense potential to contribute to public health in resource-limited settings where there is a particular need for portable, rapid, label-free, easy-to-use, and affordable biomedical devices to diagnose and continuously monitor patients for precision medicine, especially those suffering from rare diseases, such as sickle cell anemia, thalassemia, and chronic fatigue syndrome. Here, a magnetic levitation-based diagnosis system is presented in which different cell types (i.e., white and red blood cells) are levitated in a magnetic gradient and separated due to their unique densities. Moreover, an easy-to-use, smartphone incorporated levitation system for cell analysis is introduced. Using our portable imaging magnetic levitation (i-LEV) system, it is shown that white and red blood cells can be identified and cell numbers can be quantified without using any labels. In addition, cells levitated in i-LEV can be distinguished at single-cell resolution, potentially enabling diagnosis and monitoring, as well as clinical and research applications.

Research Progress of Biomakers Proteomics-based in Lung Cancer

蛋白组学技术可以应用于癌症研究来检测差异蛋白质表达以发现癌症生物标志物。肺癌的生物标志物在肺癌早期诊断、指导治疗和预后监测方面起着关键作用。因此，迫切需要确定新的早期诊断和预后指标以开辟新的治疗途径。本文简要介绍了基于蛋白质组学的肺癌生物标志物的最新研究报告。他包括作为诊断、预后和预测性的生物标志物，以及基于最近发表文献的基础上和我们所做的相关工作的总结。

PWE-215 An isolated abnormal chromogranin B is associated with pancreato-duodenal nets and the presence of liver metastases

Introduction Chromogranin B (CgB) is a soluble secretory protein like chromogranin A (CgA) that has been reported to be elevated in patients with neuroendocrine tumours (NETs) but is not routinely assessed. This study explores whether an abnormal CgB paired with a normal CgA provides any useful clinical information in NET patients. Method A prospective NET database (01/2007–11/2014) was interrogated for paired radioimmunoassay CgA (two assays; normal Results 1161 paired CgA and CgB results (479 patients) were retrieved. 51 patients had an isolated abnormal paired CgB (79 results; mean CgB result=2.68xULN); 2 patients were excluded as coding error. Primary sites were 24 pancreas, 9 small bowel, 4 rectal, 3 duodenal, 3 unknown, 2 duodenal-jejunal, 2 pre-sacral, 1 caecal and 1 lung. In 8/9 small bowel NETs, the CgB was raised in isolation on only one occasion on longitudinal testing. The tumour grades were 24 G1, 18 G2, 4 G3 and 3 unknown. Liver metastases were present in 38/49 patients. Conclusion An isolated abnormal paired CgB was observed in 11% of NET patients. In 59% of patients it was associated with a pancreatic or duodenal primary NET. In 78% liver metastases were present. Further assessment is required to establish the role of CgB when paired with CgA as a biomarker of clinical utility in the diagnosis, prognosis and therapeutic monitoring of NETs. Disclosure of interest None Declared.

Prognosis and Molecular Monitoring in Chronic Myeloid Leukemia.

Tyrosine kinase inhibitors with activity against BCR-ABL form the cornerstone of CML therapy, and are particularly effective in those with chronic-phase disease. Because some patients exhibit primary resistance or secondary failure to TKI therapy, it is recommended that continued monitoring of disease burden be performed. In this article, we review methods of detecting the Philadelphia chromosome and BCR-ABL transcript, and discuss the correlation of response with patient outcomes. Expert guidelines that incorporate definitions and milestones of response are referenced to aid in clinical decision-making.

Detection of circulating tumor cells (CTCs) in central venous blood (CVB) versus peripheral venous blood (PVB) in patients with metastatic cancers.

e22039 Background: CTCs are considered as useful indicators of tumor aggressiveness, patient prognosis, and treatment effectiveness monitoring. Detection of CTCs in blood is difficult due to the ex...

MGMT promoter methylation in serum and cerebrospinal fluid as a tumor-specific biomarker of glioma.

O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation is a conventional technique to predict the prognosis or individualized treatment of glioma in tumor tissue following surgery or biopsy. However, the technique cannot be applied in those glioma patients with concomitant neurological dysfunctions or advanced age. The present study aimed to find a new minimally invasive and efficient alternative method for the detection of MGMT promoter methylation. The expression of MGMT promoter methylation was assessed in peripheral blood and cerebrospinal fluid (CSF), and compared to the corresponding tumor tissue from glioma patients. The 89 patients in the study [32 World Health Organization (WHO) grade II, 19 WHO grade III and 38 WHO grade IV) were pathologically-diagnosed glioma and received radiation therapy following sample collection. The resected glioma tumor tissue (89), corresponding serum (89) and CSF (78) samples were collected for the detection of MGMT promoter methylation using methylation-specific polymerase chain reaction. The sensitivity and specificity of detecting MGMT promoter methylation in CSF and serum were compared. Among the tumor tissue samples, 51/89 (57.3%) showed MGMT promoter methylation. The specificity of the detection in the CSF and serum samples reached 100%. The sensitivity of MGMT promoter methylation detection in CSF and serum were 26/40 (65.0%) and 19/51 (37.3%), respectively (P<0.05). In the WHO II, III and IV subgroups, the sensitivities of MGMT promoter methylation detection using CSF were 8/12 (66.7%), 11/18 (61.1%) and 7/10 (70.0%), respectively, which were significantly higher than the sensitivities using serum (7/21, 33.3%; 7/19, 36.8%; and 5/11, 45.5%, respectively P<0.05). Among patients with residual postoperative tumors, the sensitivities of detecting MGMT promoter methylation using CSF and serum were 18/25 (72.0%) and 10/24 (41.7%), respectively, both of which were significantly higher than the corresponding values for patients without residual tumors (8/15, 53.3% and 6/19, 31.6%, respectively; P<0.05). The detection of MGMT promoter methylation in CSF specimens shows higher sensitivity compared to the serum for glioma patients. Assessment of MGMT promoter methylation in CSF may provide a promising clinical methodology for early diagnosis, individual treatment, monitoring of recurrence and prognosis for glioma patients.

Fluorescent porphyrin with an increased uptake in peripheral blood cell subpopulations from colon cancer patients.

The intrinsic fluorescence of synthetic or natural porphyrins is regarded as an attractive characteristic exploited for assisting early cancer diagnosis and/or tumor localization. Single tumor cells circulating in the blood stream can be considered a major step in depicting dissemination of primary tumors, an event of clinical relevance for prognosis, staging or therapy monitoring of cancer. The third leading cause of cancer death in men is colorectal cancer and the hematogenous spreading of primary tumor cells is one of the main events in metastasis of this type of cancer. Hidden in the myriad of circulating blood cells, tumor cells need both a sensitive and affordable detection technique. 5- (3-methoxy)-4-methoxycarbonylphenyl)-10, 15, 20-tris-(4- methoxycarbonylphenyl) - 21, 23-H porphyne is a synthetic porphyrin with a noticeable preference of accumulation in peripheral blood mononuclear cells isolated from cancer patients as assessed by flow cytometry analysis. In addition, we found distinct accumulation of porphyrin depending on cancer type (cutaneous melanoma versus colorectal cancer). These data lead to the possibility of identifying circulating cells based on preferential accumulation of this new porphyrin in circulating tumor cells because, even accumulated in low percentage of cells the registered intensity of fluorescence was high. Selecting the genetic markers for circulating tumor cells is an option, but high costs and high level of know-how can be somewhat a hurdle for a rapid evaluation. Thus our approach with a new porphyrin can be developed in an accurate and innovative fast tracking method for circulating cancer cells, at least in colorectal cancer patients.

New progress in prostate cancer biomarker research

Prostate cancer is one of the major problems of men's health. It has more obvious cancer heterogeneity, affecting the diagnosis, treatment and prognosis monitoring when comparing with other cancer. Thus, cancer related biomarkers are urgently needed to guide biopsy, treatment selection and follow-up. In the aspect of diagnosis, though the emerging of prostate specific antigen can raise the rate of diagnosis to some extent, it brings more unnecessary biopsies because of its low specificity. Cancer related long non-coding RNAs, single nucleotide polymorphism and fusion genes are proved to increase the diagnostic efficiency through high-throughput sequencing techniques. In the aspect of molecular classification, traditional Gleason score can discriminate different risk level of the cancer, but it is limited by the technicians. It is reported that copy number variations from DNA level and small RNAs, long non-coding RNAs from RNA level are related with the progress of cancer, indicating the potential role in cancer molecular classification. Although we are still on the way of the initial stage in exploring prostate cancer biomarkers, more useful and promising biomarkers will be found by high-throughput sequencing and molecular biotechniques.

Sensitive electrochemical immunosensor based on three-dimensional nanostructure gold electrode.

A sensitive electrochemical immunosensor was developed for detection of alpha-fetoprotein (AFP) based on a three-dimensional nanostructure gold electrode using a facile, rapid, “green” square-wave oxidation-reduction cycle technique. The resulting three-dimensional gold nanocomposites were characterized by scanning electron microscopy and cyclic voltammetry. A “sandwich-type” detection strategy using an electrochemical immunosensor was employed. Under optimal conditions, a good linear relationship between the current response signal and the AFP concentrations was observed in the range of 10–50 ng/mL with a detection limit of 3 pg/mL. This new immunosensor showed a fast amperometric response and high sensitivity and selectivity. It was successfully used to determine AFP in a human serum sample with a relative standard deviation of <5% (n=5). The proposed immunosensor represents a significant step toward practical application in clinical diagnosis and monitoring of prognosis.

Quantitation of Lan antigen in Lan+, Lan+(w) and Lan- phenotypes.

The Lan antigen (LAN1) is a clinically significant, high-frequency red blood cell (RBC) antigen. Lan was originally described in 1961 and assigned to the LAN blood group system in 20121,2. Lan is currently the only antigen within the LAN blood group system and Lan+, Lan−, Lan+w and Lan+w/− phenotypes have been defined1–4. The Lan− phenotype is very rare worldwide with a frequency of less than 1% in all populations tested to date2,5,6. For example, a Japanese screening study identified 14 Lan− individuals among 713,384 blood donors, giving a frequency of 0.002%2. Lan− individuals are usually identified due to the detection of anti-Lan during investigations into haemolytic disease of the foetus and newborn7,8 or when serological testing is performed to find compatible blood units for a patient5,9. Transfusion support for Lan− individuals is highly challenging due to the scarcity of both compatible blood and suitable anti-Lan reagents for screening for compatible blood. The first monoclonal anti-Lan antibody (OSK43) was produced in 2012 by Helias et al. and this antibody has proven to be of huge benefit as a reliable reagent for screening for Lan− individuals2. The carrier of the Lan antigen is the ABCB6 protein encoded by the ABCB6 gene at chromosome 2q36, containing 19 exons2. Numerous genetic variants have been identified as encoding for the Lan− phenotype2,4,10–12. The initial sequencing study identified ten novel alleles, including frameshift, nonsense and splice-site mutations, within 12 unrelated Lan− individuals and found that each individual was heterozygous for two ABCB6 null alleles2. Subsequently, Saison et al. characterised yet another ABCB6 null allele, a single nucleotide variant missense mutation (c.574C>T)10. To date this single nucleotide variant is the most common mutation causing the Lan− phenotype4,10. It has been suggested that the frequency of ABCB6 null alleles differs between populations as sequencing of 27 Japanese Lan− individuals identified a further ten novel alleles and revealed that none of the donors carried the c.574C>T variant12. The Lan+w phenotype has been described in individuals heterozygous for an ABCB6 null allele and a wild-type allele, and these individuals typically express 50% of the normal level of Lan antigen4,10. The fourth Lan phenotype is referred to as Lan+w/− as cells serologically type as either Lan+w or Lan− depending on the anti-Lan utilised3. A comprehensive study involving serological and molecular characterisation of Lan phenotypes was recently performed by Reid et al. and for the first time alleles encoding Lan+w/− phenotypes were defined4. Lan+w/− individuals are heterozygous for an ABCB6 null allele and a variant allele encoding for weakened Lan expression. Antigens can be quantified by flow cytometry by converting the fluorescent intensity of staining into an antibody-binding capacity (ABC), relating to the number of monoclonal antibody molecules bound to a cell. This is performed utilising populations of calibrated microspheres coated with defined amounts of a capture antibody, to create a calibration curve used for quantitation of ABC. The usefulness of determining ABC values has been well established, particularly for lymphocyte antigens13. In previous studies, calibrated microsphere-based assays have been validated to aid in the diagnosis, prognosis, and treatment monitoring of diseases including chronic lymphocytic leukaemia14,15 and human immunodeficiency virus infection16. In the context of RBCs, glycophorin A and RhD antigen expression has been investigated utilising traditional flow cytometric approaches17,18 and, more recently, calibrated microspheres19,20. The variability of Lan antigen expression has never been quantitatively investigated. In this study, we investigated the expression of Lan antigen by developing a novel indirect staining protocol capable of quantitating the number of Lan sites per RBC in samples reported as Lan+, Lan+w, and Lan−.

Tumor markers for diagnosis, monitoring of recurrence and prognosis in patients with upper gastrointestinal tract cancer.

To evaluate the value of combined detection of serum CEA, CA19-9, CA24-2, AFP, CA72-4, SCC, TPA and TPS for the clinical diagnosis of upper gastrointestinal tract (GIT) cancer and to analyze the efficacy of these tumor markers (TMs) in evaluating curative effects and prognosis. A total of 573 patients with upper GIT cancer between January 2004 and December 2007 were enrolled in this study. Serum levels of CEA, CA19-9, CA24-2, AFP, CA72-4, SCC, TPA and TPS were examined preoperatively and every 3 months postoperatively by ELISA. The sensitivity of CEA, CA19-9, CA24-2, AFP, CA72-4, SCC, TPA and TPS were 26.8%, 36.2%, 42.9%, 2.84%, 25.4%, 34.6%, 34.2% and 30.9%, respectively. The combined detection of CEA+CA199+CA242+CA724 had higher sensitivity and specificity in gastric cancer (GC) and cardiac cancer, while CEA+CA199+CA242+SCC was the best combination of diagnosis for esophageal cancer (EC). Elevation of preoperative CEA, CA19-9 and CA24-2, SCC and CA72-4 was significantly associated with pathological types (p<0.05) and TNM staging (p<0.05). Correlation analysis showed that CA24-2 was significantly correlated with CA19-9 (r=0.810, p<0.001). The levels of CEA, CA19-9, CA24-2, CA72-4 and SCC decreased obviously 3 months after operations. When metastasis and recurrence occurred, the levels of TMs significantly increased. On multivariate analysis, high preoperative CA72-4, CA24-2 and SCC served as prognostic factors for cardiac carcinoma, GC and EC, respectively. combined detection of CEA+CA199+CA242+SCC proved to be the most economic and practical strategy in diagnosis of EC; CEA+CA199+CA242+CA724 proved to be a better evaluation indicator for cardiac cancer and GC. CEA and CA19-9, CA24-2, CA72-4 and SCC, examined postoperatively during follow-up, were useful to find early tumor recurrence and metastasis, and evaluate prognosis. AFP, TPA and TPS have no significant value in diagnosis of patients with upper GIT cancer.

纳米技术在癌症诊疗一体化中的应用

Cancer is one of the greatest killers and threats to human life in the worldwide. The early diagnosis and efficient drug treatment of cancer without side-effect were the most difficult bottleneck of cancer therapy. Recently, the rapid development of nanomedicine brought about new opportunities for cancer diagnosis and therapeutics. The researchers focused on building stable, efficient and safe nanocarriers loading anti-cancer drugs and cancer diagnostic probes. Multifunctional nanosystems integrating targeted therapy, <italic>in vivo</italic> tracing and prognosis monitoring, will be the future research trends of nanomedicine. Those will provide strong support to efficiently increase drug delivery and decrease drug side-effect. Herein, we highlighted the great challenges that facing human cancer therapy, and overviewed the latest development and the application in cancer theranostics of quantum dots, nanogold/nanosilver, carbon nanotube/graphene, magnetic nanoparticles, lipid/polymer nanoparticles, and mesoporous nanaparticles. The nanosystem integrated diagnosis and therapy is constantly improving to perfection. But more fundamental difficulties such as the impact of nanosystems to human cells/tissues/organs, quantified and standardized production, must be explored to provide safe, stable, effective, and non-toxic nanosystem to clinical theranostics.

Abstract 5588: Epithelial-to-mesenchymal markers of circulating tumor cells for detection of castration-resistant prostate cancer

Abstract Approximately 30% of prostate cancer (PCa) patients who receive androgen-deprivation therapy experience disease progression, including bone metastasis, in 18-36 months. Clinical diagnosis of this castration resistance is primarily based on a continuous rise in serum PSA levels during therapy. However, the underlying disease progression predates the clinical onset of castration resistance by many months. The clinical challenge is to distinguish indolent vs aggressive cancer for better disease management. During the prostate cancer progression, malignant circulating tumor cells (CTC) shed from the primary site into the bloodstream through epigenetic modifications and an epithelial-to-mesenchymal transition (EMT). Evidence suggests that increased CTCs are present in the blood of castration-resistant PCa patients and can be obtained through routine phlebotomy. EMT changes of CTCs present a great potential for disease prognosis, treatment stratification and subsequent disease monitoring. We have recently established the combined microfiltration-micromanipulation system (CM2S) to enrich prostate CTCs from blood as published in The Prostate. Our preliminary studies based on a relatively small sample size show that CTCs isolated from advanced PCa patients often lose the typical features of prostate epithelial cells and display incremental EMT-related gene expression signatures, higher elasticity and smoother membrane features. Furthermore, incremental expression of these genes and particular nanomechanical features in CTCs are associated with castration-resistant and metastatic PCa. In this study, we would like expand the sample size to clinically develop this methodology. CTCs were isolated from patients' blood (∼15 ml) using microfiltration system (ScreenCell). We analyzed castration-sensitive (CS) and castration-resistant (CR) PCa cells and CTCs using high throughput microfluidic single-cell RT-PCR. The data were subject to hierarchical clustering, violin plot and Ingenuity Pathway Analysis (IPA). Increased expression of EMT genes was found in CR PCa cells and CTCs as compared to CS counterparts. IPA indicated that these EMT genes are closely related to AKT, β-catenin, Myc and NFκB pathways. Some of CTCs isolated from patients were subject to nanomechanical and nanochemical analysis using PeakForce QNM Catalyst atomic force microscopy (AFM) (Bruker). CTCs of CR patients are about 3 fold more elastic and 7 fold more adherent than CTCs of CS patients. Additionally, CTCs of CR patients are about 3 fold more deformable than CTCs of CS patients. In conclusion, we confirmed our previous study with a larger patient sample size. The increased expression of EMT-related genes and nanomechanical and nanochemical phenotypes in CTCs are potential biomarkers for detection of the castration-resistant PCa, treatment stratification and subsequent disease monitoring. Citation Format: Chun-Liang Chen, Pawel Osmulski, Devalingam Mahalingam, Aaron M. Horning, Rohit R. Jadhav, Anna D. Louie, Chiou-Miin Wang, Tim H.-M. Huang. Epithelial-to-mesenchymal markers of circulating tumor cells for detection of castration-resistant prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5588. doi:10.1158/1538-7445.AM2014-5588

Abstract 4786: Circulating miR-21, miR-146a and miR-210 levels in plasma are associated with clinical outcome in breast cancer

Abstract Introduction: The discovery of novel non-invasive tumor biomarkers for early diagnosis, prognosis and therapy monitoring remains an undeniable imperative necessity. Circulating miRNAs have recently attracted a lot of interest as novel promising tumor biomarkers. In the present study, we investigated the expression levels of three cancer-associated miRNAs in plasma and evaluated their prognostic significance in breast cancer. Materials and Methods: We examined the expression levels of miR-21, miR-146a and miR-210 in plasma samples of 134 breast cancer patients (57 patients with verified metastasis and 77 patients with operable breast cancer) and 20 healthy individuals. Circulating miRNAs were isolated from 200μL of plasma using mirVana PARIS kit and in each sample 25fmol of the synthetic cel-miR-39 were spiked as exogenous control. For the quantification of plasma miRNAs, a stem-loop cDNA approach and TaqMan real time PCR (LightCycler 2, Roche Applied Science) were used. The normalization procedure was based on a combination of cel-miR-39 as exogenous control and hsa-miR-191 as endogenous control. Relative expression of circulating miRNAs in plasma was evaluated as previously described (Sourvinou et al., J. of Mol. Diagn, 2013). Results: Our results indicate that the expression levels of all three circulating miRNAs are significantly increased in plasma both in operable and metastasis verified breast cancer patients in comparison to those of healthy individuals (p&amp;lt;0.001). The expression levels of plasma miR-210 are slightly increased in patients with operable breast cancer in respect to patients with verified metastasis (p=0.034). In patients with verified metastasis, Kaplan-Meier survival analysis demonstrated an association between elevated levels of plasma miR-21 and overall survival (OS) (p= 0.001) and there was a trend with disease-free interval (DFI) (p=0.053). Concerning miR-146a, increased plasma expression levels were correlated with short OS (p=0.029), but there was no correlation with DFI (p=0.914). Finally, respecting miR-210, Kaplan-Meier survival analysis did not show any association with OS or DFI of these patients. Conclusions: Our results indicate that quantification of circulating miR-21 and miR-146a in plasma are promising prognostic biomarkers in breast cancer and should be further evaluated in a large number of patients. Citation Format: Ioanna S. Sourvinou, Athina Markou, Nikolaos Malamos, Vasilis Georgoulias, Εvi S. Lianidou. Circulating miR-21, miR-146a and miR-210 levels in plasma are associated with clinical outcome in breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4786. doi:10.1158/1538-7445.AM2014-4786

Characteristics and significance of atypical flow cytometric immunophenotype in plasma cell myeloma

Objective To investigate the characteristics and significance of atypical flow cytometric immunophenotype in plasma cell myeloma. Methods Using case-control study, 48 cases with atypical immunophenotype of plasma cell myeloma and 42 cases as control group were studied by flow cytometry, the former cases were classified into three groups according to the expression of CD38, CD138, CD19, CD56 : CD38+/CD138+/CD19+/CD56+ , CD38+/CD138+/CD19+/CD56- , CD38+/CD138+/CD19-/CD56-. And compare the three groups with intracellular immunoglobulin light chain (cKappa, cLambda) , bone marrow morphology and immunofixation. The positive rate was compared with chi-square test. Results Bone marrow plasma cells showed expression of particular antigens in the following proportion of the 48 cases: CD45 29.17%, CD38 100%, CD138 100%, CD19 95.83%, CD56 43.75%, cKappa 43.75% and cLambda 56.25%. And in the three groups, the expression of monoclonal immunoglobulin were 43.75%, 52.08% and 4.17%, which bone marrow morphology and immunofixation were 57.14%,80%,100% and 71.43%,88%,100%. The positive rate of flow cytometry, bone marrow morphology and serum immunofixation electrophoresis were 100%, 70.83% and 81.25%.While the expression of particular antigens in the control group were: CD45 47.62%, CD38 100%, CD138 100%, CD19 100%, CD56 0%, cKappa 100% and cLambda 100%. And no abnormalities were detected in bone marrow morphology and immunofixation. Conclusions Compared with the bone marrow morphology and immunofixation，multiparameter flow cytometry has more helpful to find out atypical immune phenotype of plasma cell myeloma, and differentiate malignant and benign plasma cell, contributes to the diagnosis of clinical plasma cell myeloma, prognosis and treatment monitoring.（Chin J Lab Med,2014,37:687-690） Key words: Multiple myeloma; Immunophenotyping; Flow cytometry

Circulating cell-free DNA in serum as a biomarker for diagnosis and prognostic prediction of colorectal cancer.

Background:To verify whether the concentrations and integrity index of circulating cell-free DNA (ccf-DNA) in serum may be clinically useful for the diagnosis and progression monitoring of colorectal cancer (CRC) patients.Methods:Serum samples were collected from 104 with primary CRC, 85 with operated CRC, 16 with recurrent/metastatic CRC, 63 patients with intestinal polyps and 110 normal controls. Long (247 bp) and short (115 bp) DNA fragments in serum were detected by real-time quantitative PCR by amplifying the ALU repeats (ALU-qPCR). Serum carcinoembryonic antigen (CEA) level was detected by ARCHITECT assay.Results:The median absolute serum ALU115 and ALU247/115 in primary CRC group was significantly higher than those in intestinal polyp and normal control groups (both P<0.0001), in recurrent/metastatic CRC was significantly higher compared with primary CRC (P=0.0021, P=0.0018) or operated CRC (P<0.0001, respectively) and during follow-up, ALU115 and ALU247/115 were increased before surgery and decreased significantly after surgery.Conclusions:Combined detection of ALU115, ALU247/115 and CEA could improve the diagnostic efficiency for CRC. Serum DNA concentrations and integrity index may be valuable in early complementary diagnosis and monitoring of progression and prognosis of CRC.

New era in autoantibodies detection:quantitation

Autoantibodies are useful laboratory parameters for diagnosis of autoimmune diseases (AID). Methods for the detection of autoantibodies have achieved quantitative or semi-quantitative to some extent.Furthermore, with the development of detection technology, high-throughout and quantitative technology has been the trend in autoantibody measurement.Compared with qualitative results, the quantitative ones may provide more values for diagnosis, prediction, prognosis and therapeutic monitoring for AID.Therefore, although quantitative technology of autoantibodies is still faced a number of challenges, the detection for autoantibodies has come into the era of quantitation.（Chin J Lab Mes，2014,37： 561-563） Key words: Autoantibodies; Autoimmune diseases

Electrochemical biosensor for detection of BCR/ABL fusion gene based on isorhamnetin as hybridization indicator

A facile electrochemical DNA biosensor using isorhamnetin (ISO) indicator for detection of the sequence-specific detection of BCR/ABL fusion gene related to chronic myelogenous leukemia was developed. Results revealed that ISO bound to DNA via the intercalative binding mode with binding number of 2 and binding constant β=13.2. In the simulated interference background system, the block action of proteins in serum resulted to the increase of peak current difference between the probe modified GCE and after hybridization with target sequence. Under the optimal experimental condition, the proposed method could respond to target concentrations with a linear calibration range from 10pmolL−1 to 500nmolL−1, with a detection limit of 3pmolL−1. This biosensor exhibited high sensitivity and selectivity, and was successfully used in determining the BCR/ABL fusion which was added in simulated interference background system with a relative standard deviation (n=6) <5.5%. This new biosensor served as a significant step toward the practical application in clinical diagnosis and prognosis monitor.

Potential applications of concurrent transcranial magnetic stimulation and functional magnetic resonance imaging in acquired brain injury and disorders of consciousness

Background: Diagnostic assessment, prognosis and treatment monitoring in patients with disorders of consciousness (DoC) rest largely on behaviorally-based procedures. This approach can lead to misdiagnosis, inaccurate outcome prediction and inappropriate judgements regarding the effectiveness of treatment interventions. Concurrent transcranial magnetic stimulation (TMS) and functional magnetic resonance imaging (fMRI) may provide a biological measure of conscious awareness, aid clinicians in clinical decision-making and provide a treatment alternative for DoC.Study: This paper reviews the use of TMS and fMRI in the assessment of patients with DoC and suggests potential applications for concurrent use of these procedures.

Non-occlusion Monitoring of Arterial Pressure Dynamics from Pulsation Wave Propagation Time

A method and system for non-occlusion monitoring of human blood pressure and prognosis of hypertensive crisis is described. The method is based on a model of pulsation wave propagation. The structure of a developed hard-ware-software system for real-time monitoring and prediction of dangerous changes in arterial pressure is described. The system makes it possible to avoid hypertensive crisis and concomitant complications.