Search

Purpose: This study aims to address the hypothesis that the high-mobility group A2 (HMGA2), an oncofetal protein, relates to survivability and serves as a prognostic biomarker for colorectal cancer (CRC).Experimental Design: This is a retroprospective multiple center study. The HMGA2 expression level was determined by performing immunohistochemistry on surgical tissue samples of 89 CRCs from a training set and 191 CRCs from a validation set. The Kaplan–Meier analysis and COX proportional hazard model were employed to analyze the survivability.Results: Multivariate logistic analysis indicated that the expression of HMGA2 significantly correlates with distant metastasis in training set (odds ratio, OR = 3.53, 95% CI: 1.37–9.70) and validation set (OR = 6.38, 95% CI: 1.47–43.95). Survival analysis revealed that the overexpression of HMGA2 is significantly associated with poor survival of CRC patients (P < 0.05). The adjusted HRs for overall survival were 2.38 (95% CI: 1.30–4.34) and 2.14 (95% CI: 1.21–3.79) in training and validation sets, respectively. Further investigation revealed that HMGA2 delays the clearance of γ-H2AX in HCT-116 and SW480 cells post γ-irradiation, which supports our finding that CRC patients with HMAG2-positive staining in primary tumors had augmented the efficacy of adjuvant radiotherapy (HR = 0.18, 95% CI: 0.04–0.63).Conclusion: Overexpression of HMGA2 is associated with metastasis and unequivocally occurred in parallel with reduced survival rates of patients with CRC. Therefore, HMGA2 may potentially serve as a biomarker for predicting aggressive CRC with poor survivability and as an indicator for better response of radiotherapy. Clin Cancer Res; 17(8); 2570–80. ©2011 AACR.

Objective To investigate the relationship between single nucleotide polymorphisms (SNPs) in human papillomavirus (HPV) receptor gene and HPV susceptibility and the outcomes in Chinese women. Study design A cohort study. Setting Lishui, Zhejiang, China. Population 3066 women were recruited. Methods 29 SNP sites of HPV receptor gene on women with available cytology residual samples were detected. Main outcome measures Develop to cervical intraepithelial neoplasia 2 and worse (CIN2+). Results: 2938 women with sufficient cytology samples performed SNP sites detection. Rs16894821 [GG vs. AA: OR =1.71 (1.08-2.69)] and rs724236 [TT vs. AA: OR=1.73 (1.14-2.62)] in SDC2 increased the HPV susceptibility. TT genotype of rs2575712 in SDC2 was associated with an increased HPV 16/18 susceptibility [OR=2.78 (1.22-6.36)]. Four SNPs (rs1047057 and rs10510097, rs2575735, and rs878949) were significantly associated with HPV persistent infection. In addition, the frequencies of genotype of rs16894821 under recessive model [OR=2.40 (1.12-5.15)] in SDC2, and rs11199993 under dominant model [OR=1.64 (1.01-2.68)] in FGFR2 were significantly associated with the disease progression. Importantly, we found that HPV test in combination with SNPs with sensitivity of 0.51 (0.36-0.66) and specificity of 0.96 (0.96-0.97) in predicting CIN2+ on women with non-HPV16/18+, which had similar performance to HPV test combined with cytology test with sensitivity of 0.44 (0.30-0.60) and specificity of 0.98 (0.97-0.99). Conclusion: Gene variants in HPV receptor related gene may influence the HPV susceptibilities and the outcome of HPV infection. HPV combined with SNPs is a promising alternative to HPV test combined with cytology test in patients with non-HPV16/18 infection.

Despite the progress on the analysis of proteins either in vitro or in vivo, detection and imaging of low-abundance proteins in living cells still remains challenging. Herein, a novel biodegradable and cofactor self-sufficient DNAzyme nanoprobe has been deve-loped for catalytic imaging of protein in living cells with signal amplification capacity. This DNAzyme nanoprobe is constructed by assembling a DNAzyme subunit-containing aptamer hairpin (HP), another DNAzyme subunit strand (DS), and the molecular beacon (MB) substrate strand onto pH-sensitive ZnO@polydopamine nanorods (ZnO@PDA NRs) that work as DNAzyme cofactor suppliers. Such a nanoprobe can facilitate cellular uptake of DNA molecules and protection of them from nuclease degradation as well as release of them in cells by lysosomal acid-triggered dissolution of ZnO@PDA NRs into Zn

In this paper, the azeotropic mixture of CO2/R170 (0.78/0.22, mass fraction) is presented as a refrigerant to replace R134a in an ASHPWH system. On the basis of the first and second laws of thermodynamics, system performance and the exergy efficiency analysis are carried out for the system using both refrigerants. The results show that both the COPheat (coefficient of heating performance) and exergy efficiency of the system using the CO2/R170 mixture refrigerant are higher than that using the R134a, and the values can be increased by 31.3% and 30.6%, respectively. Moreover, the compressor discharge temperature for CO2/R170 is about 34.3–49.8 °C lower and the compression ratio of compressor also has a significant reduction (from 12.0417 to 3.2734), which leads to a higher lifetime for the compressors. So, the CO2/R170 mixture may be a good choice to substitute for R134a in ASHPWH systems. Also, a CO2/R170 ASHPWH system may be more suitable in cold region.

Simple, rapid and selective methods for simultaneous determination of ultralow amount of multiple protein biomarkers in complicated biological samples are in great demand for early-stage diagnostics and biomedical research. Herein, we present a novel polydopamine nanoparticle (PDANP)-based multicolor fluorescent proximity immunoassay for ultrasensitive and multiplexed analysis of proteins in a single solu- tion using isothermal quadratic amplification (IQA). The assay system consists of DNA tethered-antibody pairs for different protein targets, different fluorophore-labeled DNA hairpin probes, DNA polymerase, T7 exonuclease and PDANPs. The DNA tethered- antibodies and fluorophore-labeled hairpin probes can coexist stably, and fluorophore- labeled DNA hairpin probes are adsorbed onto the PDANPs, leading to the significant quenching of the fluorescence of all fluorophores. When target proteins enter into the system, each pair of DNA tethered-antibodies simultaneously binds to a specific pro- tein target and forms the proximate complexes via proximity-dependent DNA hybridi- zation. The formed proximate complexes can then unfold the hairpin structures of the respective probes, leading to DNA polymerase/T7 exonuclease-catalyzed recycling of both proximate complexes and DNA strands to achieve IQA. This IQA results in the di- gestion of many fluorophore-labeled hairpin probes, causing a substantial increase in the fluorescence of corresponding fluorophores for detection of target proteins. As a proof of concept, we demonstrate that a three-color proximity immunoassay can si- multaneously detect prostate-specific antigen (PSA), carcinoembryonic antigen (CEA) and α-fetoprotein (AFP) with ultralow attomolar detection limits, high specificity and 6-log dynamic range. Furthermore, the application of this assay for direct and simulta- neous determination of multiple proteins in clinical sera has also been demonstrated. Our newly proposed assay can be expected to provide a generic method platform for multiplexed analysis of a variety of low-abundance proteins.

We develop a novel peptide-based four-color fluorescent polydopamine nanoprobe for multiplexed sensing and imaging of tumor-related proteases in living cells. This nanoprobe responds rapidly and selectively, enabling simultaneous and high-contrast imaging of multiple proteases in living cells. Furthermore, it could detect the changes of protease expression in living cells.

Background: Over the last few decades, extra-thyroid 99mTc-pertechnetate uptake has rarely been reported. The clinical characteristics of extra-thyroid 99mTc-pertechnetate uptake were retrospectively analysed to explore the effect of the phenomenon on RAI therapy for DTC and its clinical significance. Methods: In this study, we retrospectively analysed 4930 RAI-treated DTC patients who had undergone 99mTc-pertechnetate scanning. Thirty-eight cases with extra-thyroid 99mTc-pertechnetate uptake were selected. The clinical features, location, location count and extra-thyroid 99mTc-pertechnetate uptake distribution were analysed, combined with the uptake rate, stimulated thyroglobulin (sTg) level, post-therapy whole-body scan and curative effect. Results: The results showed that sixty-five extra-thyroid 99mTc-pertechnetate foci were detected in 38 patients. The proportions of patients with abnormal uptake in the lymph nodes, lungs and bones were 68.4%, 10.5% and 10.5%, respectively. The corresponding uptake rates were 0.2%, 0.2% and 0.8%. The uptake rate was significantly lower in the lymph nodes than in the bones (Z = -2.722, p = 0.019). The uptake rate and sTg were positively correlated (r = 0.36, p = 0.027). 131I uptake was found in 36 cases at the technetium uptake site, and the number of iodine uptake foci was significantly higher than that of 99mTc-pertechnetate uptake foci. The sTg value and pathological staging significantly differed between the excellent and nonexcellent response groups (Z = 2.947, p = 0.003 and Z = 2.348, p = 0.019, respectively). Conclude: Extra-thyroid 99mTc-pertechnetate uptake mostly indicated metastases with specific clinical features, which may have prognostic value for the judgment of iodine uptake function and the RAI therapy plan.

It is universally acknowledged that the accumulation of frost on the evaporator surface will reduce the cooling capacity and performance of the cold storage. Therefore defrosting must be periodically and effectively carried out to avoid the evaporator blockage. The electric heater defrosting (EHD) and reversed cycle defrosting (RCD) methods are usually used in cold storages. However, no matter what method is adopted, there will be extra thermal load distributed to the compartments which lead to an unfavorable storage temperature fluctuation and then cause quality loss of the stored food. In order to solve the above-mentioned problems, a novel cold storage defrosting device based on EHD and RCD was proposed in this paper. Experiment was performed on the device to investigate its effect on several aspects including energy saving, reduction of cold storage temperature fluctuation and improvement of compressor operational stability etc. The results showed that this novel device could remarkably increase energy utilization and reduce temperature fluctuation of the cold storage, which was beneficial for long-term storage of products in the cold storage. Moreover, it was able to increase the operational stability of the compressor.

The extraordinary fluorescence quenching capability of graphene oxide (GO) was coupled to the specific recognition capability of aptamers to design a four-color fluorescent nanoprobe for multiplexed detection and imaging of tumor-associated proteins in living cells. Specifically, alpha-fetoprotein (AFP), vascular endothelial growth factor-165 (VEGF165), carcinoembryonic antigen (CEA), and human epidermal growth factor receptor 2 (HER2) were detected. Due to strong π interaction, the fluorescence of labeled aptamers is quenched by GO. Four fluorophore-labeled aptamers that bind the tumor-associated proteins were adsorbed on GO to form the four-color nanoprobe with quenched fluorescence. The nanoprobes were internalized into cells via endocytosis, where the aptamer/GO nanoprobes bind the intracellular tumor-associated proteins. The aptamer-protein complexes thus formed detach from GO, and fluorescence recovers. Each analyte has its typical color (AFP: blue; VEGF165: green; CEA: yellow; HER2: red). As a result, simultaneous detection and imaging of multiple tumor-associated proteins in living cells were achieved. This nanoprobe has a fast response and is highly specific and biocompatible. The linear ranges for AFP, VEGF165, CEA, and HER2 are 0.8 nM–160 nM, 0.5 nM–100 nM, 1.0 nM–200 nM, and 1.2 nM–240 nM, respectively. Detection limits were 0.45 nM for AFP, 0.30 nM for VEGF165, 0.62 nM for CEA, and 0.96 nM for HER2. The probe allows for a fast distinction between tumor cells and normal cells via imaging.

High mobility group AT-hook 2 (HMGA2) is involved in a wide spectrum of biological processes and is upregulated in several tumors. Here, we collected 273 breast cancer (BC) specimens as a training set and 310 specimens as a validation set to examine the expression of HMGA2 by immunohistochemical staining. It was found that HMGA2 expression was significantly positively correlated with advanced tumor grade and poor survival. Subgroup analysis indicated that high level of HMGA2 was significantly correlated with poor prognosis, especially in the subgroups of stage II-III, low pathological grade and non-triple negative breast cancer cases. Gene set enrichment analysis (GSEA) demonstrated a significant positive correlation between HMGA2 level and the gene expression signature of metaplastic and mesenchymal phenotype. Importantly, we also observed that ectopic expression of HMGA2 promoted the migration and invasion of breast cancer cells, and protected cancer cells against genotoxic stress from agents stimulating P53 (Ser15) phosphorylation. As a conclusion, expression of HMGA2 might indicate more advanced malignancy of breast cancer. Thus we believe HMGA2 could serve as a biomarker of poor prognosis and a novel target in treating BC tumors.

Primary pyridylcarboxamides are prevalent parent structures in bioactive molecules and have the apparent advantages over N-protected derivatives as synthetic building blocks. However, no practical methods have been developed for direct synthesis of this compound class from unfunctionalized pyridines. We herein present a general, safe, concise, acid-free, and highly selective method for the C2-carba­moylation of pyridines with unprotected formamide and N-methyl formamide through the cleavage of two C–H bonds.

A novel gold nanoparticle-based four-color fluorescence proximity immunoassay is developed for one-step, multiplexed detection of protein biomarkers using ribonuclease H signal amplification. This assay could detect proteins with detection limits down to pg mL−1 and a 5-log dynamic range. Furthermore, it is suitable for rapid, direct quantification of multiple proteins in biological samples.

e14650 Background: Gene fusions, resulting from chromosomal breakage and rearrangement, play a critical role during oncogenesis. Recently, a growing number of gene fusions are being considered as important therapeutic targets in oncology, for instance, Anaplastic lymphoma receptor tyrosine kinase (ALK) fusion, which confer exquisite sensitivity to ALK tyrosine kinase inhibitors (TKIs). However, available methods are inadequate for unbiased identification of potentially targetable fusions. Hence, we developed a novel fusion gene analyzing method named AcornFusion by DNA-seq data. Methods: The method is purely based on the mapping results of soft-clipped reads and the distribution of the positions where the candidate breakpoints locate, which is significantly different from the traditional methods with assembly of short reads and efficient way to do the detection. 309 patients with advanced non-small cell lung cancer (NSCLC) were enrolled. Formalin-fixed, paraffin embedded (FFPE) samples were used to DNA extracting and analyzed by AcornFusion method. Immunohistochemistry (IHC) and Reverse transcriptase polymerase chain reaction (RT-PCR) platform were also applied. Results: Of 309 pts, 17 patients were identified as fusion-positive cases by AcornFusion. 15 patients were EML4-ALK fusion positive and 1 patient was HIP-ALK fusion positive, which were all confirmed by IHC. 1 patient was ROS1 rearrangement positive, which was validated by Reverse transcriptase polymerase chain reaction (RT-PCR) method. Conclusions: With this advantage of high consistency with IHC and RT-PCR, AcornFusion can be a useful tool for investigating gene fusion events in clinical settings.

Primary pyridylcarboxamides are prevalent parent structures in bioactive molecules and have the apparent advantages over N-protected derivatives as synthetic building blocks. However, no practical methods have been developed for direct synthesis of this compound class from unfunctionalized pyridines. We herein present a general, safe, concise, acid-free, and highly selective method for the C2-carba­moylation of pyridines with unprotected formamide and N-methyl formamide through the cleavage of two C–H bonds.

The high mobility group A1 (HMGA1) protein is associated with poor prognosis in patients with a wide range of cancers. However, the affect of HMGA1 on the risk of mortality from breast cancer (BC) has not been fully characterized. In the present retrospective multiple center study, the HMGA1 expression level was determined by performing immunohistochemistry on surgical tissue samples of 273 BC specimens from the Second Affiliated Hospital of Zhejiang University (Zhejiang, China) and 310 BCs from the National Engineering Center for Biochip (Shanghai, China). Kaplan-Meier analysis and Cox proportional hazard model were employed to analyze the survivability. HMGA1 expression was significantly associated with tumor histological degree and body mass index (BMI). However, HMGA1 expression showed no prognostic value in patients with BC. Combined evaluation of HMGA1 expression and high BMI (≥24 kg/m2) predicted worse overall survival of BC. Therefore, HMGA1 and BMI were considered to serve synergistic roles in the development and progression of BC, and combined evaluation of HMGA1 expression and high BMI may be an effective marker in predicting poor prognosis of BC patients.