Development and validation of a risk prediction score for patients with nasopharyngeal carcinoma.

BACKGROUND: We aimed to develop and validate a predictive model for the overall survival (OS) of patients with nasopharyngeal carcinoma (NPC). METHODS: Overall, 519 patients were retrospectively reviewed in this study. In addition, a random forest model was used to identify significant prognostic factors for OS among NPC patients. Then, calibration plot and concordance index (C-index) were utilized to evaluate the predictive accuracy of the nomogram model. RESULTS: We used a random forest model to select the three most important features, dNLR, HGB and EBV DNA, which were significantly associated with the OS of NPC patients. Furthermore, the C-index of our model for OS were 0.733 (95% CI 0.673 ~ 0.793) and 0.772 (95% CI 0.691 ~ 0.853) in the two cohorts, which was significantly higher than that of the TNM stage, treatment, and EBV DNA. Based on the model risk score, patients were divided into two groups, associated with low-risk and high-risk. Kaplan-Meier curves demonstrated that the two subgroups were significantly associated with OS in the primary cohort, as well as in the validation cohort. The nomogram for OS was established using the risk score, TNM stage and EBV DNA in the two cohorts. The nomogram achieved a higher C-index of 0.783 (95% CI 0.730 ~ 0.836) than that of the risk score model 0.733 (95% CI 0.673 ~ 0.793) in the primary cohort (P = 0.005). CONCLUSIONS: The established risk score model and nomogram resulted in more accurate prognostic prediction for individual patient with NPC.

Circulating tumor cells: a valuable marker of poor prognosis for advanced nasopharyngeal carcinoma.

PURPOSE: To evaluate the prognostic value of circulating tumor cells (CTCs) in nasopharyngeal carcinoma (NPC). METHODS: Cox's proportional hazards regression models were used to identify whether CTCs was a poor prognostic factor for NPC. Chi-square tests were used to analyze and compare the distribution characteristics of CTCs in NPC. ROC curve was used to estimate the cut-off point of CTCs. Kaplan-Meier survival analyses were used to observe the prognostic value of CTCs alone and in combined with Epstein-Barr Virus DNA (EBV-DNA). RESULTS: CTCs was confirmed to be an independent risk factor for poor prognosis of NPC by Cox's regression models that enrolled 370 NPC cases and took age, gender, EBV-DNA and CTCs as variables. The proportion of CTCs in stage IV NPC was statistically different from that in stage III; the cut-off point of CTCs between stage IV (288 cases) and stage III (70 cases) NPC estimated by ROC curve was 0.5. The prognosis of advanced NPC patients became worse with the increase of CTCs count. The combined detection of CTCs and EBV-DNA could better predict the prognosis of NPC compared with the single detection of EBV-DNA.

Study on the degradation rule of calcitonin in vitro in patients with medullary thyroid carcinoma.

BACKGROUND: It was found that calcitonin in vitro can degrade rapidly whether in refrigeration or at room temperature, which increases the difficulty of accurate detecting of calcitonin greatly. This study was designed to solve this problem. METHODS: Serum samples with increased calcitonin from patients with medullary thyroid carcinoma were collected and divided into refrigeration and room temperature group. Analyze the degradation rule of calcitonin in the two groups according to the time of blood draw (T0), the time of sample detection (Tx), and the concentration (y) measured at Tx. RESULTS: The degradation rate of calcitonin was significantly different between the two storage conditions. The degradation has plateau stage and its concentration is associated with the original concentration. The degradation rule of the two groups can be expressed by the formula: y = -aln(x) + b. In the formula, b/a are constants associated with storage condition and gender. "x" is the time interval between blood draw and detection, that is, x = Tx - T0. CONCLUSIONS: The significance of the presence of immunoreactive isoforms and fragments of calcitonin with different stability in patients with medullary thyroid carcinoma is unclear. To make the inspection report more accurate and reliable, maybe the inspection personnel should report both the instrumental and corrected result, and also indicate the correction formula on the inspection report.

One-step sensitive detection of Salmonella typhimurium by coupling magnetic capture and fluorescence identification with functional nanospheres.

Sensitive, rapid, and reliable detection of bacteria has always been pursued due to the great threat of the bacteria to human health. In this study, a convenient one-step strategy for detecting Salmonella typhimurium was developed. Immunomagnetic nanospheres (IMNS) and immunofluorescent nanospheres (IFNS) were used to specifically capture and recognize S. typhimurium simultaneously. After magnetic separation, the sandwich immune complexes (IMNS-bacteria-IFNS) were detected under a fluorescence microscope with a detection limit as low as ca. 10 CFU/mL. When they were detected by fluorescence spectrometer, a linear range was exhibited at the concentration from 10(5) to 10(7) CFU/mL with R(2) = 0.9994. Compared with the two-step detection strategy, in which the bacteria were first captured with the IMNS and subsequently identified with the IFNS, this one-step strategy simplified the detection process and improved the sensitivity. Escherichia coli and Shigella flexneri both showed negative results with this method, indicating that this method had excellent selectivity and specificity. Moreover, this method had strong anti-interference ability, and it had been successfully used to detect S. typhimurium in synthetic samples (milk, fetal bovine serum, and urine), showing the potential application in practice.

Blocking TREM-1 signaling prolongs survival of mice with Pseudomonas aeruginosa induced sepsis.

TREM-1 is a recently discovered receptor expressed on neutrophils and macrophages. Blocking of TREM-1 signaling improves the survival of mice with bacterial sepsis. However, the precise mechanism by which TREM-1 modulates the inflammatory responses is poorly defined. In this study, we investigated the role of TREM-1 in Pseudomonas aeruginosa-induced peritonitis. Our results showed that TREM-1 was not expressed on lymphocytes but emerged on the cell surface of neutrophils and peritoneal macrophages. Blockade of TREM-1 signaling significantly prolonged survival of mice with P. aeruginosa-induced peritonitis. However, blocking TREM-1 signaling had no effect on macrophage phagocytosis in vitro. Interestingly, the expression of the costimulatory molecules CD40 and CD86 on macrophages was significantly decreased after blocking TREM-1 signaling. Furthermore, interfering with TREM-1 engagement led to significant reduction of pro-inflammatory mediators such as IL-1, TNF-α, MCP-1 and IFN-γ. Therefore, our results showed that TREM-1 could be a potential therapeutic target for bacterial sepsis.

Identification of Pathway-Based Biomarkers with Crosstalk Analysis for Overall Survival Risk Prediction in Breast Cancer.

Recently, many studies have investigated the role of gene-signature on the prognostic assessment of breast cancer (BC), however, the tumor heterogeneity and sequencing noise have limited the clinical usage of these models. Pathway-based approaches are more stable to the perturbation of certain gene expression. In this study, we constructed a prognostic classifier based on survival-related pathway crosstalk analysis. We estimated pathway's deregulation scores (PDSs) for samples collected from public databases to select survival-related pathways. After pathway crosstalk analysis, we conducted K-means clustering analysis to cluster the patients into G1 and G2 subgroups. The survival outcome of the G2 subgroup was significantly worse than the G1 subgroup. Internal and external dataset exhibits high consistency with the training dataset. Significant differences were found between G2 and G1 subgroups on pathway activity, gene mutation, immune cell infiltration levels, and in particular immune cells/pathway's activities were significantly negatively associated with BC patient's outcomes. In conclusion, we established a novel classifier reflecting the overall survival risk of BC and successfully validated its clinical usage on multiple BC datasets, which could offer clinicians inspiration in formulating the clinical treatment plan.

Molecular Subclassification Based on Crosstalk Analysis Improves Prediction of Prognosis in Colorectal Cancer.

The poor performance of single-gene lists for prognostic predictions in independent cohorts has limited their clinical use. Here, we employed a pathway-based approach using embedded biological features to identify reproducible prognostic markers as an alternative. We used pathway activity score, sure independence screening, and K-means clustering analyses to identify and cluster colorectal cancer patients into two distinct subgroups, G2 (aggressive) and G1 (moderate). The differences between these two groups with respect to survival, somatic mutation, pathway activity, and tumor-infiltration by immunocytes were compared. These comparisons revealed that the survival rates in the G2 subgroup were significantly reduced compared to that in the G1 subgroup; further, the mutational burden rates in several oncogenes, including KRAS, DCLK1, and EPHA5, were significantly higher in the G2 subgroup than in the G1 subgroup. The enhanced activity of the critical pathways such as MYC and epithelial-mesenchymal transition may also lead to the progression of colorectal cancer. Taken together, we established a novel prognostic classification system that offers meritorious insights into the hallmarks of colorectal cancer.

Efficacy of Transnasal Endoscopic Fine-Needle Aspiration Biopsy in Diagnosing Submucosal Nasopharyngeal Carcinoma.

OBJECTIVES/HYPOTHESIS: The routine practices of examining submucosal lesions are not suitable for deep lesions. Therefore, we evaluated the efficacy of non-real-time image-guided transnasal endoscopic fine-needle aspiration biopsy (FNAB) in diagnosing nasopharyngeal carcinoma (NPC) with submucosal lesions. STUDY DESIGN: The effectiveness evaluation of diagnostic methods. METHODS: Fifty suspected NPC patients who failed in conventional biopsies were enrolled in this study. The efficacy, maneuverability, and safety of FNAB in diagnosing these intractable cases were evaluated. RESULTS: The definitive diagnostic results of these 50 patients were NPC (34/50, 68.0%), nasopharyngeal necrosis (1/50, 2.0%), nasopharyngeal mucositis (12/50, 24.0%), and other cancers (3/50, 6.0%), respectively. The results of the diagnostic efficacy of FNAB were sensitivity, 89.2%; specificity, 100.0%; positive predictive value, 100.0%; negative predictive value, 76.5%; and accuracy, 92.0%, respectively. The area under the receiver operating characteristic curves was 0.946 (95% confidence interval = 0.884-1.00, P < .001). No severe complications occurred after FNAB. CONCLUSIONS: FNAB can improve the diagnostic efficiency of NPC occurring in the submucosal space. It can be an additional option for routine nasopharyngeal biopsy and is worthy of clinical application. LEVEL OF EVIDENCE: 4 Laryngoscope, 131:1798-1804, 2021.

Plasma Macrophage Inhibitory Cytokine-1 as a Complement of Epstein-Barr Virus Related Markers in Identifying Nasopharyngeal Carcinoma.

BACKGROUND: We evaluated the diagnostic value of plasma Macrophage inhibitory cytokine-1 (MIC-1) in distinguishing patients with nasopharyngeal carcinoma (NPC) and explored its complementary role with widely used Epstein-Barr virus (EBV) related markers, EBV capsid antigen-specific IgA (VCA-IgA) and EBV copy number. METHODS: ELISA was used to analyze the plasma MIC-1 levels in 190 NPC patients, 72 VCA-IgA-positive healthy donors (VP), and 219 normal subjects with negative VCA-IgA (VN). 10 pairs of plasma samples before and after radiotherapy were also included. RESULTS: The plasma MIC-1 levels were significantly higher in NPC patients (Median: 678.39 ng/mL) than those in VN and VP (310.29 and 294.59, p < 0.001). Receiver operating characteristic (ROC) curves of the MIC-1 concentrations revealed that the area under the ROC curve (AUC) was 0.790 (95% confidence interval [CI]: 0.748-0.832), with a sensitivity of 63.7%, and a specificity of 85.9% respectively, for distinguishing NPC patients from the healthy donors. Similarly, between NPC and VP, ROC was 0.796 (0.738-0.853) with sensitivity of 63.7%, and specificity of 88.9%. In addition, between NPC and VN, ROC was 0.788(0.744-0.832) with sensitivity of 63.7%, and specificity of 84.9%. Further, we found that MIC-1 could complement VCA-IgA and EBV DNA markers, with a negative rate of 88.9% in VCA-IgA-positive healthy controls, and a positive rate of 59.0% in EBV DNA negative NPC patients, respectively. Also, the MIC-1 plasma concentration dropped significantly after radiotherapy (p = 0.027). CONCLUSIONS: MIC-1 can complement VCA-IgA titers and EBV DNA copy number tests in NPC detection, improve identification of EBV DNA-negative NPC patients, and distinguish NPC from VCA -IgA positive healthy controls.

Combined determination of Epstein-Barr virus-related antibodies and antigens for diagnosis of nasopharyngeal carcinoma.

BACKGROUND AND OBJECTIVE: With the development of molecular biology in recent years, many indexes for detecting Epstein-Barr virus (EBV) have been developed. This study was to evaluate the diagnostic value of combined determination of EBV-related antibodies and antigens, including VCA-IgA, EA-IgA, EBV-DNase antibody and EBV-DNA, in diagnosing nasopharyngeal carcinoma (NPC). METHODS: Serum and plasma samples from 160 untreated NPC patients and 76 healthy donors were collected. VCA-IgA and EA-IgA in the serum samples were detected by immunoenzyme staining method. Raji cells were stimulated by ortho-butanoic acid and croton oil to detect EBV-DNase antibody. The content of EBV-DNA in the plasma samples was detected by real-time fluorescence quantitative polymerase chain reaction (RQ-PCR). The diagnostic values of the indexes for NPC were evaluated. RESULTS: The sensitivity and specificity for diagnosing NPC were 90.0% and 89.5% for VCA-IgA, 75.0% and 94.7% for EA-IgA, 76.3% and 90.8% for EBV-DNase antibody, 68.8% and 88.2% for EBV-DNA, and 98.8% and 84.2% for combined determination. The positive rates of VCA-IgA and EA-IgA had no relationship with clinical stage of NPC (p > 0.05); nevertheless, the positive rates of EBV-DNase antibody and EBV-DNA were related with clinical stage (p < 0.05). CONCLUSIONS: The sensitivity of VCA-IgA and the specificity of EA-IgA are the highest while detecting solely. Combined determination could improve the diagnostic sensitivity and accuracy for NPC. EBV-DNase antibody and EBV-DNA could be helpful to evaluate the course of disease and classify the clinical stage of NPC.