Ionic liquid-enhanced silica aerogels for the specific extraction and detection of aflatoxin B1 coupled with a smartphone-based colorimetric biosensor.

The polymer ionic liquid (1-allyl-3-butylimidazolium bromide) enhanced silica aerogel was modified onto the surface of stainless-steel mesh to immobilize aptamer-1 for the specific recognition of AFB1. The porous channels of silica aerogel could prevent the interference of macromolecules in food samples. Enzyme kinetic analysis showed that the MoS2/Au was an effective peroxidase mimic with a relatively low Michaelis constant (Km) value of 0.17 mM and a high catalytic rate of 3.87 × 10-8 mol (L·s)-1, which exhibited obvious superiority compared with horseradish peroxidase. The established "sandwich-structure" biosensor was coupled with the smartphone "Color Picker" application was used to detect AFB1 with a wide linear range (1-100 ng mL-1) and low detection limit (0.25 ng mL-1). The anti-interference ability of the established biosensor was evaluated by adding different concentrations of standards in corn, peanut, and wheat and matrix effects were 90.84-106.11 %. The results showed that this method demonstrated high specificity, sensitivity, rapidity and low interference in food samples.

KLF5 and p53 comprise an incoherent feed-forward loop directing cell-fate decisions following stress.

In response to stress, cells make a critical decision to arrest or undergo apoptosis, mediated in large part by the tumor suppressor p53. Yet the mechanisms of these cell fate decisions remain largely unknown, particularly in normal cells. Here, we define an incoherent feed-forward loop in non-transformed human squamous epithelial cells involving p53 and the zinc-finger transcription factor KLF5 that dictates responses to differing levels of cellular stress from UV irradiation or oxidative stress. In normal unstressed human squamous epithelial cells, KLF5 complexes with SIN3A and HDAC2 repress TP53, allowing cells to proliferate. With moderate stress, this complex is disrupted, and TP53 is induced; KLF5 then acts as a molecular switch for p53 function by transactivating AKT1 and AKT3, which direct cells toward survival. By contrast, severe stress results in KLF5 loss, such that AKT1 and AKT3 are not induced, and cells preferentially undergo apoptosis. Thus, in human squamous epithelial cells, KLF5 gates the response to UV or oxidative stress to determine the p53 output of growth arrest or apoptosis.

Integration of Metallic Nanomaterials and Recognition Elements for the Specifically Monitoring of Pesticides in Electrochemical Sensing.

Although all countries have been controlling the excessive use of pesticides, incidents of pesticide residues still existed. Electrochemical biosensors are extensively applied detection techniques to monitor pesticides with the help of different types of biorecognition components mainly including, antibodies, aptamers, enzymes (i.e., acetylcholinesterase, organophosphorus hydrolase, etc.), and synthetic molecularly imprinted polymers. Besides, the electrode materials mainly affected the sensitivity of electrochemical biosensors. Metallic nanomaterials with various structures and excellent electrical conductivity were desirable choice to construct electrochemical platforms to achieve the detection with high sensitivity and good specificity toward the target. This work reviewed the developed metallic materials including monometallic nanoparticles, bimetallic nanomaterials, metal atoms, metal oxides, metal molybdates, metal-organic frameworks, MXene, etc. Integration of recognition elements endowed the electrode materials with higher specificity toward the target pesticide. Besides, future challenges of metallic nanomaterials-based electrochemical biosensors for the detection of pesticides are also discussed and described.

Post-esophagectomy chylothorax refractory to mass ligation of thoracic duct above diaphragm: a case report.

BACKGROUND: Post-esophagectomy chylothorax is a relatively rare but potentially lethal complication. The treatment strategy of post-esophagectomy chylothorax remains a subject of debate which mainly focuses on the indication and timing of surgical intervention. For cases in which the leakage site is not localized, a mass ligation of the thoracic duct above diaphragm is advocated as the surgical procedure is believed to ensure sealing all the accessory ducts that could be the source of the chylothorax. But in this paper, we report a case of post-esophagectomy chylothorax which was refractory to mass ligation of thoracic duct above diaphragm. CASE PRESENTATION: A 59-year old man suffered from high output chylothorax (> 1000 ml/24 h for more than 30 days) after esophagectomy through left thoracotomy. Considering the failure of lymphangiography, we performed mass ligation of thoracic duct above diaphragm. However, we failed to close the chylous leakage. Finally, we found that a rare variated tributary of thoracic duct was the resource of the chylous output. Both the variation of lymphatic system and the coincidence of injured site lead to the invalidness of reoperation. After definitely ligating the variated tributary, chylothorax was cured. CONCLUSION: This case supplies a direct evidence that mass ligation of thoracic duct is of no avail in some refractory chylothorax, which indicates the importance of chylous leakage localization.

Downregulation of fibulin-4 inhibits autophagy and promotes the sensitivity of esophageal squamous cell carcinoma cells to apatinib by activating the Akt-mTOR signaling pathway.

BACKGROUND: Fibulin-4, namely, EFEMP2, is an essential matricellular protein associated with a variety of malignancies. The aim of this study was to explore the role of fibulin-4 in the progression of esophageal squamous cell carcinoma (ESCC), as well as its effect on ESCC sensitivity to apatinib treatment. METHODS: The expression of fibulin-4 in ESCC tissues and cell lines was detected. Stably transfected ESCC cells were established by transducing lentiviral vectors for silencing or overexpressing the fibulin-4 gene into ESCC cells, and a subcutaneous xenograft tumor model of ESCC in mice was successfully established. IHC, RT-qPCR and western blotting were used to detect the expression of related genes and proteins. The CCK8 assay, EdU cell proliferation assay, wound healing assay, transwell assay and flow cytometry were used to evaluate the proliferation, invasion, migration and apoptosis of ESCC cells. After mice were sacrificed, the transplanted tumors were resected, and their volumes were measured. RESULTS: The expression of fibulin-4 was significantly increased in both ESCC tissues and cell lines, and the high expression was closely related to the poor clinicopathological features. Downregulation of fibulin-4 inhibited the proliferation, invasion and migration of ESCC cells in vitro and in vivo. Meanwhile, fibulin-4 knockdown inhibited autophagy of tumor cells by activating the Akt-mTOR signaling pathway and significantly promoted apatinib-induced apoptosis of ESCC cells. CONCLUSION: Our study showed that fibulin-4 is an oncogene that can promote ESCC progression and inhibit apoptosis. Downregulation of fibulin-4 enhances the sensitivity of ESCC cells to apatinib by inhibiting cellular protective autophagy through activating the Akt-mTOR signaling pathway.

[Corrigendum] STMN1, a prognostic predictor of esophageal squamous cell carcinoma, is a marker of the activation of the PI3K pathway.

Following the publication of the above article, an interested reader drew to the authors' attention that Fig. 10A, showing the migration and invasion capabilities of Eca109 and EC9706 cells transfected with STMN1 or normal control shRNA, contained three panels with apparently overlapping data, such that the images were likely to have been derived from the same original source. The authors examined their original data and realized that errors had been made in the compilation of the panels shown in this Figure. The authors were unable to guarantee the accuracy of the images obtained from their previous experiments, so these migration and invasion assay experiments were repeated, and the results obtained were found to be consistent with those presented in the original figure. The revised version of Fig. 10 is shown on the next page. The authors have confirmed that the errors associated with the original figure did not have any significant impact on either the results or the conclusions reported in this study, and are grateful to the Editor of Oncology Reports for allowing them the opportunity to publish this Corrigendum. Furthermore, they apologize to the readership of the Journal for any inconvenience caused. [the original article was published in Oncology Reports 39: 834-842, 2018; DOI: 10.3892/or.2017.6145].

RUNX3 inhibits the invasion and migration of esophageal squamous cell carcinoma by reversing the epithelial­mesenchymal transition through TGF­ß/Smad signaling.

Runt­related transcription factor 3 (RUNX3) is a candidate tumor suppressor, and its inactivation may play a crucial role in the carcinogenesis process of numerous cancer types, including esophageal squamous cell carcinoma (ESCC). We previously revealed that RUNX3 inactivation was correlated with lymph node metastasis (LNM) and ESCC recurrence. However, the exact mechanisms of this process are still under investigation. The aim of the present study was to examine the potential roles and underlying molecular mechanisms of RUNX3 in ESCC metastasis and the epithelial­mesenchymal transition (EMT). According to the results, RUNX3 expression in ESCC tissue was significantly reduced compared with that in adjacent normal tissue (0.50±0.20 vs. 0.83±0.16; P<0.001). In addition, statistical analysis revealed a close association between decreased RUNX3 expression and T status (P=0.027) and LNM (P=0.017) in ESCC patients. Pearson's correlation coefficient analysis was then used to evaluate correlations between RUNX3 and EMT­related marker expression. The results revealed that RUNX3 expression in ESCC tissues was negatively correlated with the expression of N­cadherin (r=­0.429; P<0.01) and Snail (r=­0.364; P<0.01) and positively correlated with the expression of E­cadherin (r=0.580; P<0.01). Moreover, Eca109 and EC9706 cell invasion, migration, MMP­9 expression and EMT were significantly inhibited by RUNX3 overexpression. Notably, further analysis revealed that RUNX3 overexpression markedly inhibited the phosphorylation of Smad2/3; RUNX3­overexpressing cells also displayed less sensitivity to TGF­ß1­induced EMT than control cells. Thus, RUNX3 may inhibit the invasion and migration of ESCC cells by reversing EMT through TGF­ß/Smad signaling and may be useful as a therapeutic target.

KLF4 activates NFκB signaling and esophageal epithelial inflammation via the Rho-related GTP-binding protein RHOF.

Understanding the regulatory mechanisms within esophageal epithelia is essential to gain insight into the pathogenesis of esophageal diseases, which are among the leading causes of morbidity and mortality throughout the world. The zinc-finger transcription factor Krüppel-like factor (KLF4) is implicated in a large number of cellular processes, such as proliferation, differentiation, and inflammation in esophageal epithelia. In murine esophageal epithelia, Klf4 overexpression causes chronic inflammation which is mediated by activation of NFκB signaling downstream of KLF4, and this esophageal inflammation produces epithelial hyperplasia and subsequent esophageal squamous cell cancer. Yet, while NFκB activation clearly promotes esophageal inflammation, the mechanisms by which NFκB signaling is activated in esophageal diseases are not well understood. Here, we demonstrate that the Rho-related GTP-binding protein RHOF is activated by KLF4 in esophageal keratinocytes, leading to the induction of NFκB signaling. Moreover, RHOF is required for NFκB activation by KLF4 in esophageal keratinocytes and is also important for esophageal keratinocyte proliferation and migration. Finally, we find that RHOF is upregulated in eosinophilic esophagitis, an important esophageal inflammatory disease in humans. Thus, RHOF activation of NFκB in esophageal keratinocytes provides a potentially important and clinically-relevant mechanism for esophageal inflammation and inflammation-mediated esophageal squamous cell cancer.

CEP55 promotes the proliferation, migration and invasion of esophageal squamous cell carcinoma via the PI3K/Akt pathway.

BACKGROUND: Centrosomal protein 55 (CEP55) is an important prognostic biomarker that plays an essential role in the proliferation, migration and invasion of multiple tumors. We aimed to investigate the prognostic value of CEP55 in pN0 esophageal squamous cell carcinoma (ESCC) and explore its biological function in ESCC cells. METHODS: We used immunohistochemistry and Western blot analysis to detect the expression of CEP55 in ESCC. Furthermore, both in vitro and in vivo assays were used to determine the effect of CEP55 on malignant behavior in ESCC cells. RESULTS: As expected, we found that CEP55 was overexpressed in ESCC. Univariate and multivariate analyses demonstrated that patients with CEP55 overexpression had a poor prognosis. Additionally, the abilities of proliferation, migration and invasion of cells, as well as the epithelial-mesenchymal transition markers, were all altered with the changed CEP55 expression levels in ESCC cells. Further study elucidated that CEP55 facilitated ESCC via the PI3K/Akt pathway. Blockade of this pathway markedly attenuated CEP55-mediated proliferation, migration, invasion and epithelial-mesenchymal transition of ESCC cells. CONCLUSION: Oncogenic CEP55 correlates with a poor prognosis by regulating tumor cell proliferation, migration and invasion via the PI3K/Akt pathway. It can serve as a promising prognostic biomarker and therapeutic target of pN0 ESCC after Ivor-Lewis esophagectomy.

FOXM1: A potential indicator to predict lymphatic metastatic recurrence in stage IIA esophageal squamous cell carcinoma.

BACKGROUND: Previous studies have elucidated that FOXM1 may predict poor prognosis in patients with multiple solid malignant tumors. In this study we explored the differential expression of FOXM1 in stage IIA esophageal squamous cell carcinoma (ESCC) and investigated its prognostic value. METHODS: Immunohistochemistry (IHC) and Western blot were used to detect FOXM1 expression in ESCC. Correlations between FOXM1 expression and clinicopathological variables, and five-year lymphatic metastatic recurrence (LMR) and overall survival (OS) of patients were analyzed. RESULTS: FOXM1 was aberrantly expressed in ESCC. Statistical analysis revealed a close relationship between FOXM1 expression and tumor size (P = 0.024), depth of invasion (P = 0.048), and degree of differentiation (P = 0.043). The five-year LMR of patients in the FOXM1 overexpression group was significantly increased compared to the low expression group (P = 0.001). The five-year OS of patients in the FOXM1 overexpression group was significantly reduced compared to the low expression group (P = 0.007). Log-rank tests demonstrated that large tumor size (P = 0.044), poor differentiation degree (P = 0.005), deep invasion (P = 0.000), and FOXM1 overexpression (P = 0.007) may indicate poor prognosis in stage IIA ESCC. Cox multivariate regression analysis revealed that all of these variables were independent predictors of unfavorable outcome (P < 0.05). CONCLUSION: FOXM1 could be a predictor of lymphatic metastatic recurrence in stage IIA ESCC after Ivor Lewis esophagectomy.

STMN1, a prognostic predictor of esophageal squamous cell carcinoma, is a marker of the activation of the PI3K pathway.

The esophageal squamous cell carcinoma (ESCC) subtype with STMN1 overexpression has a high likelihood of lymphatic metastatic recurrence. However, the underlying mechanism remains to be further elucidated. We assessed the expression level of STMN1 and PTEN in 96 pN0 ESCC patient tissues using immunohistochemistry and western blot analysis. Then, the association between STMN1 overexpression and postoperative lymphatic metastatic recurrence was evaluated. In addition, the relationship between STMN1 and PTEN was also assessed. The results showed that STMN1 expression was significantly higher in tumor tissues (P=0.013). STMN1 overexpression was related to tumor length (P=0.003) and depth of invasion (P=0.019). In addition, STMN1 overexpression was significantly associated with postoperative lymphatic metastatic recurrence in pN0 ESCC patients. Patients with STMN1-overexpressing tumors had a higher 3­year lymphatic metastatic recurrence rate (P=0.024). Furthermore, in laboratory experiments, STMN1 expression was stably silenced using lentiviral vector delivery of shRNA in Eca109 and EC9706 cell lines to assess the functional effect of STMN1 in vitro. The results indicated that stable silencing of STMN1 expression significantly inhibited cell proliferation, migration and invasion. Moreover, we inactivated the PI3K pathway in ESCC cell lines with the PI3K inhibitor LY294002 and then detected STMN1 expression by western blot analysis. STMN1 levels were robustly reduced consistent with the downregulation of p­Akt (S473) by PI3K pathway inhibition. STMN1 can act as a marker to quantitatively measure the activation of the PI3K pathway and stratify patients accordingly.

Overexpression of IFITM3 predicts poor prognosis in stage IIA esophageal squamous cell carcinoma after Ivor Lewis esophagectomy.

BACKGROUND: Recent research has shown that IFITM3 plays an important role in the tumorigenesis of many malignancies. We investigated the clinicopathological variables and prognostic value of IFITM3 in stage IIA esophageal squamous cell carcinoma (ESCC) patients. METHODS: Immunohistochemistry and Western blot analysis were used to examine IFITM3 expression in tumor specimens. The relationships between IFITM3 expression and clinicopathological variables, as well as the five-year survival and recurrence status of patients, were analyzed. RESULTS: IFITM3 was aberrantly expressed in tumor tissue. Statistical analysis showed a close correlation of IFITM3 expression with T tumor status (P = 0.004). Additionally, IFITM3 overexpression, advanced T status, poor degree of differentiation, and large tumor size were not only associated with poor survival but were high lymphatic metastatic recurrence predictors in ESCC patients (P < 0.05). CONCLUSION: Our data indicated that IFITM3 overexpression may predict poor prognosis in stage IIA ESCC patients after Ivor Lewis esophagectomy.

CEP55 overexpression predicts poor prognosis in patients with locally advanced esophageal squamous cell carcinoma.

Development of esophageal squamous cell carcinoma (ESCC) involves alterations in multiple genes with corresponding proteins. Recent studies have demonstrated that centrosomal protein 55 (CEP55) shares certain features with oncogenes, and CEP55 overexpression is associated with the development and progression of malignant tumors. The present study aimed to analyze, for the first time, whether CEP55 expression is related to clinicopothalogic features in the esophageal squamous cell carcinoma (ESCC), as well as patient survival. A total of 110 patients with mid-thoracic ESCC who suffered from Ivor-Lewis were enrolled. The CEP55 expression profile of these patients in tumour tissues and corresponding healthy esophageal mucosa (CHEM) was detected by immunohistochemistry and semi-quantitative reverse transcription-polymerase chain reaction analyses. Correlations between CEP55 expression and clinicopathological factors were analyzed using χ2 test. The log-rank test was employed to calculate survival rate. A Cox regression multivariate analysis was performed to determine independent prognostic factors. The results demonstrated that CEP55 expression in ESCC was significantly higher than that of CHEM (P<0.001). Overexpression of CEP55 was significantly associated with differentiation degree (P=0.022), T stage (P=0.019), lymph node metastasis (P=0.033), clinicopathological staging (P=0.002) and tumor recurrence (P=0.021) in locally advanced ESCC patients. In addition, CEP55 overexpression was significantly associated with reduced overall survival of patients after surgery (P=0.012). The 5-year survival rate of patients without CEP55 overexpression was significantly higher than that of patients with CEP55 overexpression (P=0.012). Therefore, these findings suggest that CEP55 overexpression correlates with poor prognosis in locally advanced ESCC patients.

Prognostic significance of centrosomal protein 55 in stage I pulmonary adenocarcinoma after radical resection.

BACKGROUND: Pulmonary adenocarcinoma is a predominant pathologic non-small cell lung cancer (NSCLC) with a high morbidity in China. Even at histological stage I, many patients still experience recurrence after radical surgery; therefore, it is critical to determine useful indicators to stratify patients according to recurrent risk. Centrosomal protein 55 (CEP55) shares certain characteristics with oncogenes and aberrant expression of CEP55 can lead to tumorigenesis. Therefore, we aimed to clarify the clinicopathological significance and prognostic value of CEP55 in stage I pulmonary adenocarcinoma. METHODS: We enrolled 106 patients with stage I pulmonary adenocarcinoma who had received complete resection in our study. CEP55 expression levels in the pulmonary tissues of all patients were validated by Western blot analyses and immunohistochemistry. SPSS 17.0 software was employed to analyze the correlation between CEP55 expression and clinicopathological characteristics of patients, as well as prognosis. RESULTS: CEP55 overexpression was detected in 67 patients (63.2%). Overexpression is associated with tumor differentiation (P = 0.036), T stage (P = 0.000) and visceral pleural invasion (P = 0.009). Patients with CEP55 overexpression had worse survival compared with those with low expression (P = 0.043). Univariate analysis revealed that T stage (P = 0.000), differentiation degree (P = 0.002), visceral pleural invasion (P = 0.000), and tumor size (P = 0.013) were also significant prognostic factors. CONCLUSION: CEP55 is a useful predicator to improve stratification of patients with stage I pulmonary adenocarcinoma.

Overexpression of IFITM3 predicts the high risk of lymphatic metastatic recurrence in pN0 esophageal squamous cell carcinoma after Ivor-Lewis esophagectomy.

Background. Recent studies have shown that the aberrant expression of IFITM3 is implicated in the lymph node metastasis of many malignancies. Our research aimed to investigate the expression of IFITM3 in pathological N0 (pN0) esophageal squamous cell carcinoma (ESCC) and its relationship with lymph node metastatic recurrence. Methods. Immunohistochemistry (IHC) was used to examine the expression profile of IFITM3 in 104 pairs of samples. Each pair consisted of ESCC tissue and its adjacent normal mucosa (ANM). This aberrant expression was verified by reverse transcription-polymerase chain reaction (RT-PCR) with 20 tumor specimens with strong immunostaining and their mucosal tissues. In addition, 20 samples of low expression tissues and their ANMs were evaluated. Moreover, the correlations between the IFITM3 expression level and the clinicopathological variables, recurrence risk and overall survival (OS) of patients were analyzed. Results. Both IHC and RT-PCR demonstrated that the IFITM3 expression level was significantly higher in tumor tissue than in ANM. Statistical analysis showed a significant correlation of IFITM3 expression with the T status of esophageal cancer (p = 0.015). In addition, IFITM3 overexpression was demonstrated to be not only an important risk factor of lymphatic metastatic recurrence but a significant prognostic factor in pN0 ESCC (p < 0.005). Conclusions. Even pN0 ESCC patients will still experience lymphatic metastatic recurrence. The IFITM3 gene could be a predictor of lymphatic metastatic recurrence in pN0 ESCC after Ivor-Lewis esophagectomy.

The relationship between KRAS gene mutations and HLA class I antigen downregulation in the metastasis of non-small cell lung cancer.

OBJECTIVE: To investigate the association between v-Ki-ras2 Kirsten rat sarcoma viral oncogene homologue (KRAS) gene mutations and levels of human leucocyte antigen (HLA) class I antigen in primary lung tumours and metastatic lymph nodes of patients with non-small cell lung cancer (NSCLC). METHODS: Patients with NSCLC undergoing tumour resection were enrolled. KRAS codon 12 mutations were analysed in normal lung and lymph node tissue, primary lung tumours and metastatic lymph nodes using polymerase chain reaction-restriction fragment length polymorphism analysis. HLA class I antigen immunostaining was examined using flow cytometry. RESULTS: A total of 65 patients participated in the study. All normal lung tissues had positive HLA class I antigen immunostaining. The majority of primary lung tumours (56/65) and all of the metastatic lymph nodes (31/31) had downregulated HLA class I antigen immunostaining. There was a positive correlation between downregulated HLA class I antigen in primary tumours and metastatic lymph nodes. There was a negative correlation between KRAS codon 12 mutations and the level of HLA class I antigen in primary and metastatic tumours. CONCLUSIONS: KRAS codon 12 mutations appear to be important in the downregulation of HLA class I antigen in NSCLC. Abnormal activation of the oncogenic KRAS pathway might provide a new treatment target for NSCLC.

CCR7 and VEGF-C: molecular indicator of lymphatic metastatic recurrence in pN0 esophageal squamous cell carcinoma after Ivor-Lewis esophagectomy?

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a malignant tumor with a high incidence of lymph node metastasis. This study was undertaken to investigate the expression of CCR7 and VEGF-C in pN0 ESCC and its relationship with lymphatic metastatic recurrence. METHODS: The expression of CCR7 and VEGF-C was examined by RT-PCR and immunohistochemistry. The recurrence rates were calculated by the Kaplan-Meier method and their difference was determined by log rank analysis. Cox regression analysis was performed to determine the independent risk factors. RESULTS: In 99 patients, CCR7 mRNA expression was observed in 42 patients with a 3 year recurrence rate of 57.1%; VEGF-C mRNA expression was observed in 52 patients with a 3 year recurrence rate of 53.8%; and coexpression of CCR7 mRNA and VEGF-C mRNA was observed in 22 patients with a 3 year recurrence of 63.6%. Neither CCR7 mRNA nor VEGF-C mRNA expression was observed in 27 patients with a 3 year recurrence rate of 22.2%. The recurrence rates of patients with positive expression of CCR7 mRNA and/or VEGF-C mRNA were significantly higher than in patients without expression of both CCR7 mRNA and VEGF-C mRNA. We achieved better concordance between RT-PCR and immunohistochemistry detection of both markers. The Cox regression analysis showed tumor T classification, positive expression of CCR7/VEGF-C mRNA, and positive expression of CCR7/VEGF-C protein in tumor tissues to be independent risk factors for 3 year recurrence. CONCLUSIONS: Patients with positive expression of CCR7 and/or VEGF-C have a higher recurrence rate than patients without expression of both CCR7 and VEGF-C. CCR7 and VEGF-C may become molecular indicators of disease in patients vulnerable to lymphatic metastatic recurrence.