Sorbitol Destroyed Intestinal Microfold Cells (M Cells) Development through Inhibition of PDE4-Mediated RANKL Expression.

Objective: Microfold cells (M cells) are specific intestinal epithelial cells for monitoring and transcytosis of antigens, microorganisms, and pathogens in the intestine. However, the mechanism for M-cell development remained elusive. Materials and Methods: Real-time polymerase chain reaction, immunofluorescence, and western blotting were performed to analyze the effect of sorbitol-regulated M-cell differentiation in vivo and in vitro, and luciferase and chromatin Immunoprecipitation were used to reveal the mechanism through which sorbitol-modulated M-cell differentiation. Results: Herein, in comparison to the mannitol group (control group), we found that intestinal M-cell development was inhibited in response to sorbitol treatment as evidenced by impaired enteroids accompanying with decreased early differentiation marker Annexin 5, Marcksl1, Spib, sox8, and mature M-cell marker glycoprotein 2 expression, which was attributed to downregulation of receptor activator of nuclear factor kappa-В ligand (RANKL) expression in vivo and in vitro. Mechanically, in the M-cell model, sorbitol stimulation caused a significant upregulation of phosphodiesterase 4 (PDE4) phosphorylation, leading to decreased protein kinase A (PKA)/cAMP-response element binding protein (CREB) activation, which further resulted in CREB retention in cytosolic and attenuated CREB binds to RANKL promoter to inhibit RANKL expression. Interestingly, endogenous PKA interacted with CREB, and this interaction was destroyed by sorbitol stimulation. Most importantly, inhibition of PDE4 by dipyridamole could rescue the inhibitory effect of sorbitol on intestinal enteroids and M-cell differentiation and mature in vivo and in vitro. Conclusion: These findings suggested that sorbitol suppressed intestinal enteroids and M-cell differentiation and matured through PDE4-mediated RANKL expression; targeting to inhibit PDE4 was sufficient to induce M-cell development.

New findings in prognostic factor assessment for adenocarcinoma of transverse colon: a comparison study between competing-risk and COX regression analysis.

Purpose: Competing-risk analysis was used to accurately assess prognostic factors for cancer-specific death in patients with adenocarcinoma of transverse colon (ATC), and the results were compared with those from a conventional Cox regression analysis. Materials and Methods: Patients diagnosed with ATC between 2000 and 2019 were selected from the Surveillance, Epidemiology, and End Results database. The crude mortality rates of patients with ATC were calculated and their differences were tested using the Gray's test, respectively. In performing multivariate analysis, the Cox regression model and the subdistribution hazard function (SD) in competing risk analysis were utilized, respectively. Results: This study included 21,477 eligible patients. The SD model indicated that age, etc. are actual independent prognostic factors. In contrast to previous recognition, the results of the Cox regression showed false-positives for sex and Carcinoembryonic antigen, and underestimated point-estimates in the stage and American Joint Committee on Cancer stage due to competing events. A detailed comparison of treatment revealed that the larger surgical scopes were prognostic risk factors compared with the smaller scope of local tumor excision, partial colectomy, or segmental resection. Patients treated with external proton beam radiotherapy had an increased risk compared with those with no radiotherapy and internal radiotherapy. Conclusions: After comparing the results of the two methods and mitigating the significant bias introduced by Cox regression, we found independent factors that really affect the prognosis of ATC. On the other hand, in terms of ATC, a larger surgical scope and external proton beam radiotherapy may not improve the long-term survival of patients. Therefore, when faced with ATC patients, these differences should be noted and treated differently from common colorectal cancer patients. Thus, clinicians are able to give more targeted treatment plans and prognostic assessments.

The novel mechanism of human norovirus induced diarrhea: Activation of PKD2 caused by HuNoVs destroyed AQP3 expression through AP2γ in intestinal epithelial cells.

Our previous work has demonstrated protein kinase D2 (PKD2) played a critical influence in experimental colitis in animal. However, the role of PKD2 in human norovirus (HuNoVs)-induced diarrhea remained unknown. Aquaporin 3 (AQP3) expression, a critical protein mediating diarrhea, was assessed by western blot, qRT-PCR in intestinal epithelial cells (IECs). Luciferase, IF, IP and ChIP assay were used to explore the mechanism through which HuNoVs regulated AQP3. Herein, we found that AQP3 expression was drastically decreased in IECs in response to VP1 transfection, the major capsid protein of HuNoVs, or HuNoVs infection. Mechanistically, HuNoVs triggered phosphorylation of PKD2 through TLR2/MyD88/IRAK4, which further inhibited AP2γ activation and nuclear translocation, leading to suppress AQP3 transactivation in IECs. Most importantly, PKD2 interacted with MyD88/IRAK4, and VP1 overexpression enhanced this complex form, which, in turn, to increase PKD2 phosphorylation. In addition, endogenous PKD2 interacted with AP2γ, and this interaction was enhanced in response to HuNoVs treatment, and subsequently resulting in AP2γ phosphorylation inhibition. Moreover, inhibition of PKD2 activation could reverse the inhibitory effect of HuNoVs on AQP3 expression. In summary, we established a novel mechanism that HuNoV inhibited AQP3 expression through TLR2/MyD88/IRAK4/PKD2 signaling pathway, targeting PKD2 activity could be a promising strategy for prevention of HuNoVs-induced gastroenteritis.

Competing-risks analysis for evaluating the prognosis of patients with microinvasive cutaneous squamous cell carcinoma based on the SEER database.

BACKGROUND: Utilizing the traditional Cox regression model to identify the factors affecting the risk of mortality due to microinvasive cutaneous squamous cell carcinoma (micSCC) may produce skewed results. Since cause-specific mortality can guide clinical decision-making, this study employed the Fine-Gray model based on the Surveillance, Epidemiology, and End Results (SEER) database to identify significant predictive variables for the risk of micSCC-related mortality. METHODS: This study used the information of patients with micSCC who were listed in the SEER database during 2000-2015. Cox regression and Fine-Gray models were utilized for the multivariable analysis, and Gray's test and the cumulative incidence function were used for the univariable analyses. RESULTS: There were 100 patients who died from other reasons and 38 who died from micSCC among the 1259 qualified patients with micSCC. Most were female, white, married, had localized metastasis, etc. According to the univariable Gray's test (P < 0.05), the cumulative incidence rate for events of interest was strongly associated with age, sex, marital status, American Joint Committee on Cancer staging, radiation status, summary stage, chemotherapy status, surgery status, and tumor size. Multivariable Cox regression analysis and multivariable competing-risks analysis indicated that age, tumor size, and income were independent risk variables for the prognosis of patients with micSCC. In both age and tumor size variables, the competing-risks model showed a slight decrease in the hazard ratio and a slight narrowing of the 95% confidence interval compared with the Cox regression model. However, this pattern is not evident in the income variable. CONCLUSIONS: This study established a Fine-Gray model for identifying the independent risk factors that influence the risk of mortality among patients with micSCC. This study uncovers that, in the context of competing risks, age, tumor size, and income serve as independent risk factors influencing the risk of mortality due to micSCC among patients. Our findings have the potential to provide more accurate risk assessments for patient outcomes and contribute to the development of individualized treatment plans.

Prediction of death probability in adenocarcinoma of the transverse colon: competing-risk nomograms based on 21,469 patients.

PURPOSE: Tumors in parts of the colon other than the transverse colon have been well studied, but little is known about adenocarcinoma of the transverse colon (ATC).The aim of this study was to construct nomograms using competing-risk model for accurately predicting the probability of cancer-specific and non-cancer-specific death in patients with ATC. METHODS: Data on eligible patients recorded during 2000-2019 in the Surveillance, Epidemiology, and End Results database were extracted and screened. Factors that influencing prognosis were screened for death from ATC (DATC) and death from other causes (DOC) using competing-risk analysis, including univariate and multivariate analyses based on Gray's test and the Fine-Gray model, respectively. Independent prognostic factors were identified and nomograms were constructed. For comparison, we also constructed a Cox model and an AJCC stage-only competing-risk model (AJCC model) for patients with DATC. Performance evaluations of the nomograms and comparison between the models were performed using calibration plots, Harrell's concordance index (C-index), receiver operating characteristic (ROC) curves, and the areas under the ROC curve (AUCs). The nomograms and models were validated using a validation cohort. The net reclassification index, integrated discrimination improvement, decision curves, and risk stratification were not assessed because no accepted methods were suited for competing-risk model. RESULTS: This study included 21,469 patients with ATC, and 17 and 9 independent influencing factors were identified for the construction of DATC nomograms (DATCN) and DOC nomograms (DOCN), respectively. In both the training and validation cohorts, the calibration curves indicated good agreement between the nomogram-based predictions and the actual observations in the two nomograms, respectively. The C-index of the DATCN was higher than 80% (80.3-83.3%) at 1, 3 and 5 years in both the training and validation cohorts, significantly outperforming the AJCC (76.7-78%) and Cox (75.4-79.5%) model. The C-index of the DOCN was also higher than 69% (69.0-73.6%). In terms of ROC curves at each time point, those of the DATCN were very close to the upper-left corner of the coordinate axis in both the training and validation cohorts, and their AUCs were larger than 84% (84.2-85.4%).The AUCs of the AJCC (78.4-81.1%) and Cox (79.4-81.5%) models were significantly lower (p < 0.05), and the curves were closer to the diagonal. The ROC curves of the DOCN was similar to those of the DATCN, and the AUCs were 68.5-74%. The DATCN and DOCN therefore had good consistency, accuracy, and stability, respectively. CONCLUSION: This study was the first to construct competing-risk nomograms for ATC. These nomograms have proved useful for accurately assessing patient prognoses and allowing more-individualized follow-up strategies, thereby reducing the mortality.

Prognosis of the Keratinizing Squamous Cell Carcinoma of the Tongue Based on Surveillance, Epidemiology, and End Results Database.

Background: The objective of this study is to determine the prognostic factors of keratinizing squamous cell carcinoma of the tongue (KTSCC) and to establish a prognostic nomogram of KTSCC to assist clinical diagnosis and treatment. Methods: This study identified 3874 patients with KTSCC from the Surveillance, Epidemiology, and End Results (SEER) database, and these patients were randomly divided into the training (70%, (n = 2711) and validation (30%, n = 1163) cohorts. Cox regression was then used to filter variables. Nomograms were then constructed based on meaningful variables. Finally, the concordance index (C-index), net reclassification index (NRI), integrated discrimination improvement (IDI), calibration charts, and decision-curve analysis (DCA), were used to evaluate the discrimination, accuracy and effectiveness of the model. Results: A nomogram model was established for predicting the 3-, 5-, and 8-year overall survival (OS) probabilities of patients with KTSCC. The model indicated that age, radiotherapy sequence, SEER stage, marital status, tumor size, American Joint Committee on Cancer (AJCC) stage, radiotherapy status, race, lymph node dissection status, and sex were factors influencing the OS of patients with KTSCC. Verified by C-index, NRI, IDI, calibration curve, and DCA curve, our model has better discrimination, calibration, accuracy and net benefit compared to the AJCC system. Conclusions: This study identified the factors that affect the survival of KTSCC patients and established a prognostic nomogram that can help clinicians predict the 3-, 5-, and 8-year survival rates of KTSCC patients.

Retraction: Down-regulation of HOXB5 inhibits TGF-ß-induced migration and invasion in hepatocellular carcinoma cells via inactivation of the PI3K/Akt pathway.

[This retracts the article DOI: 10.1039/C8RA06860G.].

Analysis and prediction of relative survival trends in patients with non-Hodgkin lymphoma in the United States using a model-based period analysis method.

Background: Survival rates are usually used to evaluate the effect of cancer treatment and prevention. This study aims to analyze the 5-year relative survival of non-Hodgkin lymphoma (NHL) in United States using population-based cancer registry data. Methods: A period analysis was used to evaluate the improvement in long-term prognosis of patients with NHL from 2004 to 2018, and a generalized linear model was developed to predict the 5-year relative survival rates of patients during 2019-2023 based on data from the SEER database stratified by age, sex, race and subtype. Results: In this study, relative survival improved for all NHL, although the extent of improvement varied by sex, age group and lymphoma subtype. Survival improvement was also noted for NHL subtypes, although the extent varied, with marginal-zone lymphoma having the highest 5-year relative survival rate (92.5%) followed by follicular lymphoma (91.6%) and chronic lymphocytic leukemia/small lymphocytic lymphoma (87.3%). Across all subtypes, survival rates were slightly higher in females than in males. Survival rates are lower in the elderly than in the young. Furthermore, the study demonstrated that black patients had lower NHL survival rates than white patients. Survival rates for NHL were higher in rural areas than in urban areas. Patients with extra-nodal NHL had a higher survival rate than patients with nodal NHL. Conclusion: Overall, patient survival rates for NHL gradually improved during 2004-2018. The trend continues with a survival rate of 75.2% for the period 2019-2023. Analysis by NHL subtype and subgroups indicating that etiology and risk factors may differ by subtype. Identification of population-specific prevention strategies and treatments for each subtype can be aided by understanding these variations.

The Incidence Trend of Papillary Thyroid Carcinoma in the United States During 2003-2017.

BACKGROUND: The rapid increase in the detection rate of thyroid cancer over the past few decades has caused some unexpected economic burdens. However, that of papillary thyroid carcinoma (PTC) seems to have had the opposite trend, which is worthy of further comprehensive exploration. METHODS: The Surveillance, Epidemiology, and End Results 18 database was used to identify patients with PTC diagnosed during 2003-2017. The incidence trends were analyzed using joinpoint analysis and an age-period-cohort model. RESULTS: The overall PTC incidence rate increased from 9.9 to 16.1 per 100 000 between 2003 and 2017. The joinpoint analysis indicated that the incidence growth rate began to slow down in 2009 (annual percentage change [APC] = 3.1%, 95% confidence interval [CI] = 1.9%-4.4%). After reaching its peak in 2015, it began to decrease by 2.8% (95% CI = -4.6% to -1.0%) per year. The stratified analysis indicated that the incidence patterns of different sexes, age groups, races, and tumor stages and sizes had similar downward trends, including for the localized (APC = -4.5%, 95% CI = -7% to -1.9%) and distant (APC = -1.3%, 95% CI = -2.7% to -.1%) stages, and larger tumors (APC = -4%, 95% CI = -12% to 4.7%). The age-period-cohort model indicated a significant period effect on PTC, which gradually weakened after 2008-2012. The cohort effect indicates that the risk of late birth cohorts is gradually stabilizing and lower than that of early birth cohorts. CONCLUSION: The analysis results of the recent downward trend and period effect for the incidence of each subgroup further support the important role of correcting overdiagnosis in reducing the prevalence of PTC. Future research needs to analyze more-recent data to verify these downward trends.

Handheld pH meter-assisted immunoassay for C-reactive protein using glucose oxidase-conjugated dendrimer loaded with platinum nanozymes.

A simple and feasible pH meter-based immunoassay is reported for detection of C-reactive protein (CRP) using glucose oxidase (GOD)-conjugated dendrimer loaded with platinum nanozyme. Initially, platinum nanozymes were loaded into the dendrimers through an in situ synthetic method. Then, GOD and monoclonal anti-CRP antibody with a high molar ratio were covalently conjugated onto carboxylated dendrimers via typical carbodiimide coupling. The immunoreaction was carried out with a competitive mode in a CRP-coated microplate. Along with formation of immunocomplex, the added glucose was oxidized into gluconic acid and hydrogen peroxide by GOD, and the latter was further decomposed by platinum nanozyme, thus accelerating chemical reaction in the positive direction. The produced gluconic acid changed the pH of detection solution, which was determined using a handheld pH meter. Under optimum conditions, the pH meter-based immunoassay gave a good signal toward target CRP from 0.01 to 100 ng mL-1. The limit of detection was 5.9 pg mL-1. An intermediate precision ≤ 11.2% was acquired with batch-to-batch identification. No nonspecific adsorption was observed during a series of procedures to detect target CRP, and the cross-reaction against other biomarkers was very low. Importantly, our system gave well-matched results for analysis of human serum samples relative to a referenced ELISA kit.Graphical abstract.

MT1-MMP activatable fluorogenic probes with enhanced specificity via high-affinity peptide conjugation for tumor imaging.

Overlapping substrate specificities within the family of matrix metalloproteinases (MMPs), usually caused by their highly conserved structural topology, increase the potential for a substrate to be cleaved by multiple enzymes within this family, which leads to the decrease in the selectivity of MMP substrate-based probes. To resolve this issue, MT1-MMP activatable fluorogenic probes for tumor detection with enhanced specificity were developed by combining a fluorescence resonance energy transfer (FRET) peptide substrate and its specific binding peptide with different lengths of linkers. The specificity of the probes increased profiting from the high affinity of the MT1-MMP specific binding peptide while keeping the ability to amplify the output imaging signals in response to MMP activity with the FRET substrate. Enzyme kinetics analysis clearly demonstrated that the conjugation of P-1 and MT1-AF7p enhanced both the specificity and selectivity of the fluorogenic probes for MT1-MMP, and introducing a linker composed of 12 PEG subunits into these two fragments led to optimized specificity and selectivity of the fluorogenic probe for MT1-MMP. Both in vitro and in vivo results revealed that the imaging probe with the linker composed of 12 PEG subunits based on our designed strategy could be effectively applied for MT1-MMP positive tumor imaging. Since this strategy for enhancing the specificity of protease sensing probes can be applied to other proteases and is not just limited to MT1-MMP, it is an appealing platform to achieve selective tumor imaging.

Experimental analysis of a wavefront coding system with a phase plate in different surfaces.

The application of an integrated phase plate can greatly simplify an optical system structure due to the defocus insensitivity and aberration inhibition of the wavefront coding imaging technology. In addition, the development of free-form surface technology promotes the application of integrated phase plates. In order to view the imaging characteristics of the integrated wavefront coding system, we built two kinds of wavefront coding systems with the phase plate integrated in different surfaces of the optical system. Simulation results show that phase plates with the same parameters but in different surfaces can realize consistent results. For further observation, two separated phase plates with the same parameters are processed, and, correspondingly, experiments of microscopic objective imaging are carried out. Experimental results confirm that the phase plate can be successfully applied to increase the design flexibility of wavefront coding, leading to a potential mass application solution.

LFB: A Novel Antimicrobial Brevinin-Like Peptide from the Skin Secretion of the Fujian Large Headed Frog, Limnonectes fujianensi.

Amphibians are a natural source of abundant antimicrobial peptides and thus have been widely investigated for isolation of such biomolecules. Many new antimicrobial peptide families have been discovered from amphibians. In this study, a novel antimicrobial peptide named Limnonectes fujianensis Brevinvin (LFB) has been identified in the skin secretion from the Fujian large headed frog, Limnonectes fujianensis. The cDNA sequence was cloned from a skin secretion library and the predicted mature peptide was identified through MS/MS fragmentation sequencing of reverse phase HPLC fractions on the same sample. LFB was predicted to be an amphipathic, hydrophobic, alpha helical, and beta turn peptide that inserts into a lipid bilayer in order to kill the cells. In antimicrobial assays, a synthetic replicate of this novel antimicrobial peptide demonstrated significant activity against the Gram-positive bacterium Staphylococcus aureus, the Gram-negative bacterium Escherichia coli and the yeast, Candida albicans. This novel peptide was highly potent (MIC 4.88 uM) against Gram-negative bacterium, and also has the ability to inhibit the growth of human cancer cell lines with IC50 values ranging from 18.9 µM down to 2.0 µM. These findings help to enrich our understanding of Brevinin-like peptides. Moreover, the data presented here validate frog secretion as a source of potential novel antimicrobial peptides, that also exhibit anti-tumor properties, that could be useful for the treatment of cancer.

Design and Fabrication of Wafer-Level Microlens Array with Moth-Eye Antireflective Nanostructures.

Wafer-level packaging (WLP) based camera module production has attracted widespread industrial interest because it offers high production efficiency and compact modules. However, suppressing the surface Fresnel reflection losses is challenging for wafer-level microlens arrays. Traditional dielectric antireflection (AR) coatings can cause wafer warpage and coating fractures during wafer lens coating and reflow. In this paper, we present the fabrication of a multiscale functional structure-based wafer-level lens array incorporating moth-eye nanostructures for AR effects, hundred-micrometer-level aspherical lenses for camera imaging, and a wafer-level substrate for wafer assembly. The proposed fabrication process includes manufacturing a wafer lens array metal mold using ultraprecise machining, chemically generating a nanopore array layer, and replicating the multiscale wafer lens array using ultraviolet nanoimprint lithography. A 50-mm-diameter wafer lens array is fabricated containing 437 accurate aspherical microlenses with diameters of 1.0 mm; each lens surface possesses nanostructures with an average period of ~120 nm. The microlens quality is sufficient for imaging in terms of profile accuracy and roughness. Compared to lenses without AR nanostructures, the transmittance of the fabricated multiscale lens is increased by ~3% under wavelengths of 400-750 nm. This research provides a foundation for the high-throughput and low-cost industrial application of wafer-level arrays with AR nanostructures.

MiR-6991-3p is identified as a novel suppressor in the expansion and activation of myeloid-derived suppressor cells in hepatoma-bearing mice.

OBJECTIVE: Myeloid-derived suppressor cells (MDSCs) are a heterogeneous group of cells derived from bone marrow, which has a significant ability in inhibition of immune cell response. In this study, the role of miR-6991-3p in regulating function of MDSCs was investigated. METHODS: MDSCs were isolated from different tissues of the control and hepatoma-bearing mice, and then expression of miR-6991-3p was detected with qPCR. Then, the miR-6991-3p mimic and inhibitor were respectively transfected into MDSCs, and behaviors of MDSCs were evaluated, including expansion, apoptosis, and production of inflammatory factors. Furthermore, we explored the underlying mechanism from which miR-6991-3p regulated MDSC functions. RESULTS: Expression miR-6991-3p was markedly decreased in the MDSCs derived from spleen and further decreased in the MDSCs derived from the tumor tissue. MiR-6991-3p mimic transfection suppressed expansion and promoted apoptosis of MDSCs, accompanied by a significant decrease in the production of IL-6 and GM-CSF that are identified as stimulators in MDSC expansion. In contrast, miR-6991-3p inhibitor transfection displayed the opposite effect. miR-6991-3p bound with and negatively regulated expression of LGALS9, a newly identified immune checkpoint gene and activator of STAT3, suppressing production of multiple factors that were customarily used to characterize the activation of MDSCs. MiR-6991-3p-accommodated MDSCs displayed less suppression on T cells, while miR-6991-3p inhibitor enhanced the suppression of MDSCs on T cells. CONCLUSION: MiR-6991-3p is identified as a novel suppressor in the expansion and activation of myeloid-derived suppressor cells, which may be regarded as a promising target for modulating the function of MDSCs.

Predictive Value of Nutritional Risk Screening 2002 and Prognostic Nutritional Index for Esophageal Cancer Patients Undergoing Definitive Radiochemotherapy.

OBJECTIVES: The present study identified the prognostic nutritional factors and their relationships with survival outcome in patients with esophageal cancer treated with chemoradiotherapy (CRT). METHODS: A total of 97 esophageal cancer patients previously treated with CRT were enrolled in the study. The nutritional status was assessed by Nutrition Risk Screening 2002 (NRS-2002). Weight, total serum protein, albumin, prealbumin level, red blood cell, total lymphocyte count, and hemoglobin were also recorded. The prognostic nutritional index (PNI) was calculated. RESULTS: The proportion of patients at nutritional risk from baseline until the sixth week of radiotherapy was increased. In univariate analysis, the NRS-2002 cutoff score ≤3 at baseline was associated with improved 2-year overall survival (OS) than that ≥4. The maximum NRS-2002 cutoff score ≤2 during treatment was associated with an improved 2-year OS that ≥3. The baseline PNI or PNI at the end of CRT ≥45 was associated with improved 2-year OS than that <45. Cox regression analyses revealed that the TNM stage, NRS-2002 score at baseline, and PNI at the third week of CRT were independent risk factors for prognosis. CONCLUSIONS: The NRS2002 scores and PNI are simple and useful markers for predicting the long-term outcome in patients with esophageal cancer after CRT.

MMP-2-responsive fluorescent nanoprobes for enhanced selectivity of tumor cell uptake and imaging.

It is difficult to develop highly selective substrate-based fluorescent nanoprobes for specific matrix metalloproteinases (MMPs) due to overlapping substrate specificities among the family of MMP enzymes. To resolve this issue, we have developed novel fluorescent nanoprobes that are highly selective for soluble MMP-2. Herein, MMP-2-responsive nanoprobes were prepared by immobilizing fluorescent fusion proteins on nickel ferrite nanoparticles via the His-tag nickel chelation mechanism. The fusion protein consisted of a fluorescent mCherry protein with a cell penetrating peptide (CPP) moiety. An MMP-2 cleavage site was also introduced within the fusion protein, which was directly linked to the nickel ferrite nanoparticles. The selectivity of nanoprobes was modulated by hiding the cleavage site of MMP-2 substrates deeply inside the system, which could result in strong steric hindrance between the nanoprobes and MMPs, especially for membrane-tethered MMPs such as MMP-14. A cell-based assay demonstrated that the nanoprobes could only be activated by tumor cells secreting soluble MMP-2, but not membrane-tethered MMP-14. To further evaluate the contribution of the steric hindrance effect on the nanoprobes, a truncated recombinant MMP-14 was employed to confer their cleavage activity as compared to native membrane-tethered MMP-14. Furthermore, a designed probe with a diminished steric hindrance effect was proved to be activated by membrane-tethered type MMP-14. The results indicated that the design of fluorescent nanoprobes employing the steric hindrance effect can greatly enhance the selectivity of MMP-responsive nanoprobes realizing the specific detection of soluble MMP-2 in a tumor microenvironment. We believe that highly selective MMP-2-responsive fluorescent nanoprobes have broad impacts on biomedical applications including molecular imaging and labeling for tumor detection.

Long non-coding RNA ZFAS1 sponges miR-484 to promote cell proliferation and invasion in colorectal cancer.

The incidence and mortality rate of colorectal cancer (CRC) have been significantly increasing. However, mechanisms involved in CRC progression are still unclear. LncRNA ZFAS1 has been verified as oncogenic molecular in a series of tumors, including CRC. However, the underlying mechanism of ZFAS1 in CRC carcinogenesis remains unclear. In the present study, our data showed that ZFAS1 expression was significantly upregulated in CRC tissues and cell lines. Correlation analysis showed that high ZFAS1 expression was significantly associated with Helicobacter pylori infection, lymph nodes metastasis, advanced TNM stage and poor overall survival of CRC patients. Loss-of-function experiments revealed that ZFAS1 inhibition could markedly suppress CRC cells proliferation and invasion both in vitro and in vivo. Bioinformatics analysis and luciferase reporter assay revealed that ZFAS1 directly interacted with miR-484. Rescue experiments showed that miR-484 inhibitor reversed the tumor suppressing roles of ZFAS1 knockdown on CRC cells. Therefore, our study suggested that ZFAS1 could act as an oncogene in CRC tumorigenesis, and discovered the functional regulatory pathway of ZFAS1 sponging miR-484.

Down-regulation of HOXB5 inhibits TGF-ß-induced migration and invasion in hepatocellular carcinoma cells via inactivation of the PI3K/Akt pathway.

HOXB5, a member of the HOX gene family, is a developmental gene which encodes homeoproteins and is known to be a crucial player in development of enteric nervous systems. Recently, HOXB5 was reported to be associated with cancer progression. However, the specific effect of HOXB5 in hepatocellular carcinoma (HCC) remains unclear. In this study, we demonstrated the important role of HOXB5 in HCC. We showed that HOXB5 was up-regulated in HCC tissues and cell lines. Furthermore, down-regulation of HOXB5 inhibited TGF-ß-induced HCC cell migration and invasion in vitro and suppressed tumor metastasis in vivo. We also found that the PI3K/Akt pathway partly accounted for the mechanisms underlying the inhibitory effect of HOXB5 down-regulation on TGF-ß-induced HCC progression. Taken together, these findings demonstrated that down-regulation of HOXB5 inhibits TGF-ß-induced migration and invasion in HCC cells via inactivation of the PI3K/Akt pathway. Thus, HOXB5 may be a novel therapeutic target for HCC treatment.

Cell-Permeable, MMP-2 Activatable, Nickel Ferrite and His-Tagged Fusion Protein Self-Assembled Fluorescent Nanoprobe for Tumor Magnetic-Targeting and Imaging.

Matrix metalloproteinases (MMPs) activatable imaging probe has been explored for tumor detection. However, activation of the probe is mainly done in the extracellular space without intracellular uptake of the probe for more sensitivity. Although cell-penetrating peptides (CPPs) have been demonstrated to enable intracellular delivery of the imaging probe, they nevertheless encounter off-target delivery of the cargos to normal tissues. Herein, we have developed a dual MMP-2-activatable and tumor cell-permeable magnetic nanoprobe to simultaneously achieve selective and intracellular tumor imaging. This novel imaging probe was constructed by self-assembling a hexahistidine-tagged (His-tagged) fluorescent fusion protein chimera and nickel ferrite nanoparticles via a chelation mechanism. The His-tagged fluorescent protein chimera consisted of a red fluorescent protein mCherry that acted as the fluorophore, the low-molecular-weight protamine peptide as the CPP, and the MMP-2 cleavage sequence fused with the hexahistidine tag, whereas the nickel ferrite nanoparticles functioned as the His-tagged protein binder and also the fluorescent quencher. Both in vitro and in vivo results revealed that this imaging probe would not only remain nonpermeable to normal tissues, thereby offsetting the nonselective cellular uptake, but was also suppressed of fluorescent signals during magnetic tumor-targeting in the circulation, primarily because of the masking of the CPP activity and quenching of the fluorophore by the associated NiFe2O4 nanoparticles. However, these properties were recovered or "turned on" by the action of tumor-associated MMP-2 stimuli, leading to cell penetration of the nanoprobes as well as fluorescence restoration and visualization within the tumor cells. In this regard, the presented tumor-activatable and cell-permeable system deems to be an appealing platform to achieve selective tumor imaging and intracellular protein delivery. Its impact is therefore significant, far-reaching, and wide-spread.