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The detection of human papillomaviruses (HPV) is a well-recognized strategy in early screening and prevention of cervical cancer. However, it's hard to carry out in undeveloped area because the sophisticated equipment that required in traditional methods is usually unavailable. To overcome this situation, we aim to establish a loop-mediated isothermal amplification (LAMP) method, which is simple and reliable for on-site detection of HPV.At least 3 sets of LAMP primers for each of the 13 types of high risk HPV were designed. After preliminary validation, the candidate primers were used in the detection of clinical samples and the results were head-to-head compared with a clinically approved real-time PCR assay. The performance of the LAMP method was assessed by kappa concordance test.Cervical secretions samples from 1412 patients were included, with 224 samples were used in the preliminary screening of the LAMP primers and the other 1188 samples were used in the verification. Compared with real-time PCR method, the specificity of our LAMP method for each type of HPV were 100 %, and 11 of the 13 types had a sensitivity greater than 80 %. Among them, HPV 31 and 52 demonstrated the best performance, both with Kappa value of 0.913 (P 0.0001). Besides, HPV 18, 35 and 56 only achieved a Kappa value less than 0.7, indicating their primers or reaction conditions may need further optimization. In general, the sensitivity, specificity, positive predictive value, negative predictive value and agreement of the LAMP assay in all HPV types was 86.9 %, 100 %, 100 %, 71.4 %, and 90.2 %, respectively (Kappa = 0.766, P 0.0001).In present study, we preliminary established and validated a LAMP method for HPV detection. This method could combine with self-sampling, thermostatic device, and appropriate dyes to form a simple and effective assay in the future, which would has good prospect and practical value in cervical cancer prevention, especially in undeveloped area.

ObjectiveThe main goal of our study is to clarify the EEG characteristics of the stress response caused by vestibular endurance training under the real conditions.MethodsTen pilot trainees received a series of acute anti-vertigo training stimulations on the rotary ladder while recording electroencephalographic data (64 electrodes). Afterwards, the subject’s anti-vertigo ability was tested for the best performance after 1 month of training, and verifying whether it is relating to the EEG signals we collected before.Results(1) The absolute power ofαwaves in the C3 and C4 regions is same as the difference between 1 min before and 2 min after stimulation, and their activity is enhanced by stimulation. Otherwise, the activation of the C3 region after 5min of stimulation is still significant changed. (2) Discover a spearman rank correlation, theαwaves in the C3 and C4 the greater the power change, the better the performance of the subject in the proficient stage.ConclusionsC3 and C4 areas are specific brain regions of the stress response of anti-vertigo endurance training, and the absolute power of theαwave can be used as a parameter for identifying the degree of motion sickness (MS). The absolute power changes ofαwaves in the C3 and C4 areas are positively correlated with their anti-vertigo potential.SignificanceThe increasing of the absolute power ofαwave in the C3 and C4 is a manifestation of MS stress adaptability.

ObjectivePhysical inactivity inChinese youth students particularly in senior high schools, who participate inthe National Higher Education Entrance Examination (NCEE) is very common. Inorder to explore the beneficial effects from physical exercise and education afterNCEE, we performed a Physicalexercise Intervention Program in the Youth (PiPy) to evaluate the interaction with PPARγ genetic variants on cardiovascular and metabolicparameters.MethodsA total of 772 freshmen (males 610/females162) from high schools to university were recruited into the PiPy cohort, which was designedaccording to the National Student Health Standards in China. Anthropometric data were collected, whilephysical activities and body composition at the baseline of PiPy cohort weremeasured with SECAprotocols. Eighttagged single nucleotide polymorphisms (SNPs) in the PPARγ gene were genotyped with TaqMan allelicdiscrimination.ResultsAfter physical exercise intervention forthree months, in parallel with increased physical activities, BMI and skeletalmuscle content in all subjects was enhanced, while heart rate and bloodpressures were decreased. Furthermore, SNPs in 5’-UTR of the PPARγ gene, including rs2920502,rs9817428 and rs2972164, were found to be associated with the changes of BMI.Body weight in the subjects with BMI 2 were increased,while the obese subjects (BMI ≥24.0 kg/m2) decreased.ConclusionThe present study for the first timedemonstrated that the PiPy could improve cardio-metabolic parameters such asheart rate, blood pressures and BMI for Chinese youth students after NCEE, inwhich the genetic interactive effects of PPARγ should be included into obesityintervention.

Brain-computer interface (BCI) has been widely used in sports training and rehabilitation training. It is primarily based on action simulation, including movement imagery (MI) and movement observation (MO). However, the development of BCI technology is limited due to the challenge of getting an in-depth understanding of brain networks involved in MI, MO, and movement execution (ME). To better understand the brain activity changes and the communications across various brain regions under MO, ME, and MI, this study conducted the fist experiment under MO, ME, and MI. We recorded 64-channel electroencephalography (EEG) from 39 healthy subjects (25 males, 14 females, all right-handed) during fist tasks, obtained intensities and locations of sources using EEG source imaging (ESI), computed source activation modes, and finally investigated the brain networks using spectral Granger causality (GC). The brain regions involved in the three motor conditions are similar, but the degree of participation of each brain region and the network connections among the brain regions are different. MO, ME, and MI did not recruit shared brain connectivity networks. In addition, both source activation modes and brain network connectivity had lateralization advantages.

Background Droplet digital polymerase chain reaction (ddPCR) is a promising method for analyzing minor amounts of nucleic acid. However, its application has not been reported in pleural fluid, which is an ideal sample source for epidermal growth factor receptor (EGFR) mutation analysis in non-small-cell lung cancer (NSCLC) patients. Methods The extracted DNA from supernatants of pleural fluid was selected from our sample bank and re-analyzed by our previously established ddPCR assay. The results were compared with the former outcomes detected by direct sequencing or the amplification-refractory mutation system (ARMS). Results A total of 95 samples were selected, and 64 and 31 of them had been performed with direct sequencing and ARMS tests, respectively. The EGFR mutation detection rate of ddPCR was significantly elevated, compared with both direct sequencing (75.4% vs. 43.8%, P P = 0.016). Compared with ARMS, Fisher's exact test showed that EGFR-positive patients who were redefined by ddPCR had higher objective response rates (ORRs): 57.9% vs. 16.7%, P = 0.032. Compared with direct sequencing results, Kaplan-Meier curves demonstrated that EGFR-positive patients who were redefined by ddPCR had longer progression-free survival (PFS): 8.0 vs. 2.0 months, P = 0.0001. Conclusion We have demonstrated the clinical value of ddPCR in pleural fluid samples. The experience obtained from the present study is practical and favorable for the proper application of this new assay.

Collecting circulating tumor cells (CTCs) shed from solid tumor through a minimally invasive approach provides an opportunity to solve a long-standing oncology problem, the real-time monitoring of tumor state and analysis of tumor heterogeneity. However, efficient capture and detection of CTCs with diverse phenotypes is still challenging. In this work, a microfluidic assay is developed using the rationally-designed aptamer cocktails with synergistic effect. Enhanced and differential capture of CTCs for nonsmall cell lung cancer (NSCLC) patients is achieved. It is also demonstrated that the overall consideration of CTC counts obtained by multiple aptamer combinations can provide more comprehensive information in treatment monitoring.

We investigated the clinical significance of serum bone-specific alkaline phosphatase (BAP), tartrate-resistant acid phosphatase isoform 5b (TRACP 5b) and type I collagen carboxyterminal telopeptide (ICTP) as bone metabolic markers for bone metastasis (BM) screening in lung cancer patients.Newly diagnosed advanced lung cancer patients with (N = 130) and without (N = 135) BM were enrolled in the study. Serum BAP, TRACP 5b and ICTP were measured before the treatment.BAP, TRACP 5b and ICTP values were higher in patients with BM compared with patients without BM (all P0.0001). Area under ROC curve (AUC) of BAP, TRACP 5b and ICTP was 0.760, 0.753 and 0.835 (all P0.0001), respectively. The cut-off values for BAP, TRACP 5b and ICTP were 21.8 μg/l, 7.8 U/l and 8.8 μg/l, respectively. When TRACP 5b and ICTP were combined, AUC was elevated to 0.895 (P0.0001), and the cut-off values were TRACP 5b 7.6 U/l and ICTP 8.4 μg/l.We conclude that serum BAP, TRACP 5b and ICTP may serve as useful tools for BM screening in lung cancer patients.

This study was initiated to evaluate the clinical significant of HER2 extracellular domain (ECD) in the real-time management of breast cancer patients.Five-hundred forty-six eligible breast cancer patients were divided according to their clinical background. The correlation between ECD, tissue HER2, and clinical outcome of the patients were analyzed.Receiver operating characteristic analysis revealed that ECD measured before receiving neoadjuvant therapy yielded the highest area under the curve (0.9185; P0.0001), indicating that ECD and tissue HER2 levels are consistent in untreated tumor-bearing patients. At cut-off of 15.0ng/ml, the prognostic value of ECD was demonstrated using univariate (HR=1.664, P0.0001) and multivariate (HR=1.547, P=0.011) Cox regression analysis. Kaplan-Meier survival curves revealed that patients with elevated ECD had shorter progression-free survival (PFS) (4.0 vs. 6.1months, P0.0001). Elevated ECD was also an adverse predictor for PFS in response to anti-HER2 therapy (4.3 vs. 10.2months, P=0.0155). In contrast, ≥20%, decreased ECD was associated with longer PFS in patients who received anti-HER2 therapy (10.9 vs. 2.4months, P=0.0164) and overall (10.7 vs. 2.8months, P=0.0034).A patient's clinical history can help determine whether ECD could provide added value for breast cancer management.

The intensity and percentage of HER2 expression on CTCs. (DOC 99Â kb)

Characteristic of the All Enrolled Patients. (DOC 56Â kb)

The rapid determination of metastasis in sentinel lymph nodes (SLNs) of breast cancer patients plays a significant role in the selection of a surgery strategy. Although a previous one-step nucleic acid amplification assay that uses reverse-transcription (RT) loop-mediated isothermal amplification (LAMP) has showed specific advantages over traditional pathological examination, its target marker requires optimisation. In addition to epithelial-specific CK19, the internal control gene PBGD and the breast-specific PIP were included in the new method. After the RT-LAMP primers were designed and verified using a cell line, the performance of our method was evaluated by comparing it with the corresponding result of the Food and Drug Administration approved breast lymph node (BLN) assay and routine pathological examination. One hundred and seventy-four valid SLN samples from 101 patients were collected from five hospitals. The threshold of reaction time for CK19, PIP and PBGD was defined as 16, 20 and 20 min, respectively. Compared with the BLN assay, the concordance rate of our method was 95.4 % (166/174). Statistical analysis revealed that the two methods are consistent (kappa = 0.890, P

Background This study was initiated to investigate the prognostic significance of circulating tumor cell (CTC) enumeration and the predictive value of CTC HER2 expression for efficient anti-HER2 therapy in HER2-positive metastatic breast cancer (MBC) patients. Methods Sixty HER2-positive MBC patients were enrolled in the present study. Before the initiation of systemic treatment, CTCs from 7.5 ml of blood were analyzed using the CellSearch system. The progression-free survival (PFS) of the patients was estimated using Kaplan-Meier survival curves. Results CTCs were detected in 45% (27/60) of the patients, who had shorter median PFS than those without CTCs (2.5 vs. 7.5 months, P = 0.0125). Furthermore, referring to the standard HER2 testing that uses immunohistochemistry (IHC), we proposed a CTC HER2-positive criterion, defined as >30% of CTCs over-expressing HER2. Among patients undergoing anti-HER2 therapy, those with HER2-positive CTCs had longer PFS (8.8 vs. 2.5 months, P = 0.002). Among patients with HER2-positive CTCs, the median PFS for those receiving anti-HER2 therapy was significantly longer than those who were not (8.8 vs. 1.5 months, P = 0.001). Notably, up to 52% (14/27) of the HER2-positive patients were CTC HER2-negative, and anti-HER2 therapy did not significantly improve the median PFS in these patients (2.5 vs. 0.9 months, P = 0.499). Conclusions Our findings underscore the necessity of a comprehensive CTC analysis, which may provide valuable prognostic and predictive information for optimizing individually tailored therapies in HER2-positive MBC patients. To test this idea, additional large cohort, multi-center and prospective clinical trials are needed.