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Vascular endothelial growth factor receptors (VEGFRs) have been demonstrated to play a critical role in ischemic retinal diseases, as VEGFRs mediate hypoxia-induced neovascularization. Not only hypoxia, ischemia also induces the deficiency of glucose, yet its effects on VEGFR signal and neovascularization have seldom been studied. Bioinformatics analysis predicted that VEGFRs may be regulated by O-GlcNAcylation, while glucose deficiency influences the O-GlcNAcylation.In this study, we treated human retinal microvascular endothelial cells with low glucose (LG) alone or in combination with low oxygen (oxygen and glucose deprivation, OGD). Cell viability and apoptosis rate were used to evaluate cell growth characters.LG (2.8 mmol/L) treatment induced mRNA and protein levels of VEGFR1, 2, 3 even in the presence of the protein synthesis inhibitor, cycloheximide (CHX), suggesting that the increase in VEGFR proteins is partially associated with post-translational modifications. Immunoprecipitation analysis showed that O-GlcNAc level was decreased by LG in both VEGFR1, 2, but a de-O-GlcNAc glycosylase inhibitor restored the O-GlcNAc levels. This inhibitor also abolished the LG-induced increase in VEGFR2 protein, whereas this effect was not disappeared in the presence of the proteasome inhibitor, MG132. Similar results were also observed under OGD condition. VEGFR2 knockdown more significantly retarded the growth of hRMECs and HUVECs than VEGFR1, 3 knockdown under LG and OGD conditions.A relatively low glucose suppressed O-GlcNAcylation in VEGFR2, whereby inhibiting its proteasome degradation; up-regulated VEGFR2 promoted the proliferation of vascular endothelial cells under ischemic condition.

Supplementary Figure 1. IL17RD potentiates EGF-induced EGFR signaling. A) Cellular proliferation of HT29 and HCT116 cells 96 hours after transfection with a plasmid encoding IL-17RD compared to an EV plasmid. B) Cellular proliferation of HT29 and HCT116 cells treated with EGF for 96 hrs following transfection with a plasmid encoding IL-17RD or EV plasmid. C) pEGFR, EGFR, pAKT, and pERK expression in HT29 cells 15 minutes after treatment with EGF ( ) or media without EGF (-) and transfected with Flag-tagged IL17RD, EV plasmid, or no transfection (vehicle) (representative images of two blots containing 3 samples per group). D) pAKT expression in HT29 cells 30 minutes after treatment with media with or without EGF and transfected with a Flag-tagged IL17RD plasmid or EV plasmid. E) EGFR expression in untreated cells 96 hours after transfection with IL17RD.

Patient demographics and disease specific characteristics.

Supplementary Material, Methods, and Figure Legend

Purpose: Patients with ulcerative colitis are at increased risk for colorectal cancer, although mechanisms underlying neoplastic transformation are poorly understood. We sought to evaluate the role of microRNAs in neoplasia development in this high-risk population.Experimental Design: Tissue from 12 controls, 9 ulcerative colitis patients without neoplasia, and 11 ulcerative colitis patients with neoplasia was analyzed. miRNA array analysis was performed and select miRNAs assayed by real-time PCR on the discovery cohort and a validation cohort. DNA methylation of miR-193a was assessed. Following transfection of miR-193a-3p, proliferation, IL17RD expression, and luciferase activity of the 3′UTR of IL17RD were measured. Tumor growth in xenografts as well as EGFR signaling were assessed in HCT116 cells expressing IL17RD with either a mutant 3′ untranslated region (UTR) or wild-type (WT) 3′UTR.Results: miR-31, miR-34a, miR-106b, and miR-193a-3p were significantly dysregulated in ulcerative colitis-neoplasia and adjacent tissue. Significant down-regulation of miR-193a-3p was also seen in an independent cohort of ulcerative colitis cancers. Changes in methylation of miR-193a or expression of pri-miR-193a were not observed in ulcerative colitis cancer. Transfection of miR-193a-3p resulted in decreased proliferation, and identified IL17RD as a direct target of miR-193a-3p. IL17RD expression was increased in ulcerative colitis cancers, and miR-193a-3p treatment decreased growth and EGFR signaling of HCT116 cells in xenografts expressing both IL17RD with WT 3′UTR compared with cells expressing IL17RD with mutant 3′UTR.Conclusions: miR-193a-3p is downregulated in ulcerative colitis neoplasia, and its loss promotes carcinogenesis through upregulation of IL17RD. These findings provide novel insight into inflammation-driven colorectal cancer and could suggest new therapeutic targets in this high-risk population. Clin Cancer Res; 23(17); 5281–91. ©2017 AACR.

Objective To investigate the effect of over-expression of nudix hydrolase 21 (NUDT21) on the proliferation of colon cancer HCT-116 cells and its mechanism. Methods The NUDT21 over-expression plasmid was constructed by Gibson assembly. The colony formation assay and CCK-8 assay were used to detect cell proliferation. The cell cycle of HCT-116 cells was detected by flow cytometry. Western blot was performed to detect the expressions of P53, cyclin-dependent kinase 2 (CDK2), phosphorylated retinoblastoma protein at serine 780 (p-Rb-Ser780), and p-Rb-Ser608. Results The sequencing results showed that the NUDT21 over-expression plasmid was successfully constructed. After the NUDT21 over-expression plasmid was transfected into HCT-116 cells, the expressions of NUDT21 mRNA and protein in the cells were significantly increased. The over-expression of NUDT21 inhibited the proliferation of HCT-116 cells and arrested the cell cycle in G0/G1 phase. The expressions of CDK2, p-Rb-Ser608, and p-Rb-Ser780 proteins decreased while the expression of P53 protein increased. Conclusion Over-expression of NUDT21 inhibits the proliferation of HCT-116 cells by blocking P53/CDK2/Rb signal pathway.

Clostridium difficile (C. difficile) is a Gram-positive, spore-forming, toxin-producing anaerobe that can cause nosocomial antibiotic-associated intestinal disease. Autolysin is a lytic enzyme that hydrolyzes peptidoglycans of the bacterial cell wall, with a catalytic domain and cell wall-binding domains, proven to be involved in bacterial cell wall remodeling and cell division. Although autolysins in C. difficile have been reported, the autolysins have failed to yield impressive results when used as exogenous lytic agents. In this study, we expressed and characterized the binding domains (Cwp19-BD and Acd-BD) and catalytic domains (Cwp19-CD, Acd-CD, and Cwl-CD) of C. difficile autolysins, and the domains with the best binding specificity and lytic activity were selected towards C. difficile to design a novel lytic protein Cwl-CWB2. Cwl-CWB2 showed good biosafety with significantly low hemolysis and without cytotoxicity. The results of fluorescence analysis and lytic assay demonstrated that Cwl-CWB2 has higher binding specificity and stronger lytic activity with a minimum inhibitory concentration at 13.39 ± 5.80 μg/mL against living C. difficile cells, which is significantly stronger than commercial lysozyme (3333.33 ± 1443.37 μg/mL) and other reported C. difficile autolysins. Besides, Cwl-CWB2 exhibited good stability as about 75% of the lytic activity was still retained when incubated at 37 °C for 96 h, which is considered to be a potential antimicrobial agent to combat C. difficile. KEY POINTS: • Several binding domains and catalytic domains, deriving from several Clostridium difficile autolysins, were expressed, purified, and functionally characterized. • A novel C. difficile lytic protein Cwl-CWB2 was designed from C. difficile autolysins. • The binding specificity and lytic activity of Cwl-CWB2 against C. difficile showed advantages compared with other reported C. difficile autolysins. • Cwl-CWB2 exhibited significantly low hemolysis and cytotoxicity against normal-derived colon mucosa 460 cell.

Because ADAM17 promotes colonic tumorigenesis, we investigated potential miRNAs regulating ADAM17; and examined effects of diet and tumorigenesis on these miRNAs. We also examined pre-miRNA processing and tumour suppressor roles of several of these miRNAs in experimental colon cancer. Using TargetScan, miR-145, miR-148a, and miR-152 were predicted to regulate ADAM17. miR-143 was also investigated as miR-143 and miR-145 are co-transcribed and associated with decreased tumour growth. HCT116 colon cancer cells (CCC) were co-transfected with predicted ADAM17-regulating miRNAs and luciferase reporters controlled by ADAM17-3’UTR. Separately, pre-miR-143 processing by colonic cells was measured. miRNAs were quantified by RT-PCR. Tumours were induced with AOM/DSS in WT and transgenic mice (Tg) expressing pre-miR-143/miR-145 under villin promoter. HCT116 transfection with miR-145, −148a or −152, but not scrambled miRNA inhibited ADAM17 expression and luciferase activity. The latter was suppressed by mutations in ADAM17-3’UTR. Lysates from colonocytes, but not CCC, processed pre-miR-143 and mixing experiments suggested CCC lacked a competency factor. Colonic miR-143, miR-145, miR-148a, and miR-152 were downregulated in tumours and more moderately by feeding mice a Western diet. Tg mice were resistant to DSS colitis and had significantly lower cancer incidence and tumour multiplicity. Tg expression blocked up-regulation of putative targets of miR-143 and miR-145, including ADAM17, K-Ras, XPO5, and SET. miR-145, miR-148a, and miR-152 directly suppress colonocyte ADAM17 and are down-regulated in colon cancer. This is the first direct demonstration of tumour suppressor roles for miR-143 and miR-145 in an in vivo model of colonic tumorigenesis.

DNA 5-hydroxymethylcytosine (5hmC) modification is known to be associated with gene transcription and frequently used as a mark to investigate dynamic DNA methylation conversion during mammalian development and in human diseases. However, the lack of genome-wide 5hmC profiles in different human tissue types impedes drawing generalized conclusions about how 5hmC is implicated in transcription activity and tissue specificity. To meet this need, we describe the development of a 5hmC tissue map by characterizing the genomic distributions of 5hmC in 19 human tissues derived from ten organ systems. Subsequent sequencing results enabled the identification of genome-wide 5hmC distributions that uniquely separates samples by tissue type. Further comparison of the 5hmC profiles with transcriptomes and histone modifications revealed that 5hmC is preferentially enriched on tissue-specific gene bodies and enhancers. Taken together, the results provide an extensive 5hmC map across diverse human tissue types that suggests a potential role of 5hmC in tissue-specific development; as well as a resource to facilitate future studies of DNA demethylation in pathogenesis and the development of 5hmC as biomarkers., DNA 5-hydroxymethylcytosine (5hmC) modification is associated with gene transcription and used as a mark of mammalian development. Here the authors report a comprehensive 5hmC tissue map and analysis of 5hmC genomic distributions in 19 human tissues derived from 10 organ systems, thus providing insights into the role of 5hmC in tissue-specific development.

Carbon dioxide can change the heat balance of the atmosphere. To study the relationship between CO2 and temperature change, we use the given CO2 concentrations to implement linear regression model, gray time series forecasting model, back-propagation model, and auto-regressive and moving average model to establish the growth function of CO2 concentration. Errors are evaluated to choose the most suitable model. then we use the most suitable model in step one to further predict future land-ocean temperature. Finally, we use grey relational analysis to analyze the relationship between CO2 concentrations and land-ocean temperatures since 1959.

Background: Arsenide, a kind of pollution widely existing in the environment, and the mechanism of occurrence and development of lung cancer leading by long-term arsenic exposure still needs further research up to present. We investigated how high and low doses of arsenic induce human bronchial epithelial cell transformation and the molecular mechanisms involvedMethods: After exposing human bronchial epithelial cells(Beas-2B) to different doses of sodium arsenite for 24 h, RNA-seq analysis was performed to detect arsenic-regulated genes; for cell viability and status and for cell cycle changes and mitochondrial function; long-term exposure to 1μM sodium arsenite after 20 consecutive passages for cell cycle changes and mitochondrial function.Results: In the study, human bronchial epithelial cells were exposed to varying high-dose sodium arsenite in short term or treated with low dose in a long term, and the data showed that both short and long terms treatment promoted G1/S transition of Beas-2B cells, inducing significant increases of expression of AKAP95, cyclin D1, cyclin D2 and cyclin E1. However, silencing AKAP95 by treating cells with siAKAP95 showed a protective function that inhibited G1/S transition, suggesting that regulatory mechanism of AKAP95 on cell cycle during cell malignant transformation induced by sodium arsenite. In addition, we also noticed some mitochondrial dysfunctions occurred during sodium arsenite exposure, including the disappearance of mitochondrial double membrane structure, the formation of vacuole structure, the decrease of mitochondrial membrane potential, the change of expression levels of mitochondrial related proteins Tim22, Tim23, Tom40 and OPA1, the release of lactate dehydrogenase (LDH) and the decrease of reduced glutathione.Beas-2B cells, which exposed to low-dose sodium arsenite for a long term, were subcultured for 20 generations, and it was found that the exposure time was positively proportional to the growth and migration rate of cells. The exposed cells were used in tumor bearing transplantation experiment (mice), and results showed that the longer the exposure time, the faster the tumor volume growth rate of As-Beas-2B cells. Tumor tissues were taken out for HE staining, and it was found that the cell morphology changed and the volume increasedConclusion: high and low doses of sodium arsenite induced malignant transformation of human bronchial epithelial cells by promoting G1/S turnover through AKAP95 with associated cyclins, accompanied by the development of mitochondrial dysfunction.

Purpose: To explore the relationship between ocular position control ability and stereopsis recovery in children with intermittent exotropia, and to analyze the influencing factors of distance stereopsis recovery. Methods: In this retrospective study, 78 children with small angle intermittent exotropia received vision training for 3 months. All patients were examined for distance stereopsis with the synoptophore and for near stereopsis with the Titmus stereogram before and after the training. The patients were divided into low and high Newcastle Control Score (NCS) groups. The stereopsis of the two groups was compared. Logistic regression analysis was used to analyze the influencing factors of distance stereopsis recovery. Results: Among 78 children with intermittent exotropia, 33 had near stereopsis (42.3%) and 22 had distance stereopsis (28.2%); the difference was significant ( P < .05). After 3 months of training, there were statistically significant differences between distance and near stereopsis in the low NCS group and the high NCS group (chi-square = 7.127, P = .008; chi-square = 13.005, P < .001). The number of children with distance and near stereopsis in the low NCS group increased significantly compared with before training (chi-square = 13.471, P < .001; chi-square = 22.244, P < .001). Multivariate logistic regression analysis showed that age of onset (odds ratio [OR] = 3.768, P = .001), near point of convergence (OR = 0.347, P = .002), and NCS (OR = 0.142, P = .002) were risk factors that affected stereopsis recovery in children with small angle intermittent exotropia. Conclusions: Control ability is one of the important indicators to assess the severity of intermittent exotropia. The worse the control ability, the more difficult the recovery of stereopsis. Age of onset, near point of convergence, and NCS are risk factors that affect the recovery of distance stereopsis. [ J Pediatr Ophthalmol Strabismus . 2021;58(6):350–354.]

Gut epithelial morphogenesis is maintained by intestinal stem cells. Here, we report that depletion of N

Arsenic is a widely existing pollutant in the environment, but the mechanism of occurrence and development of lung cancer by long-term arsenic exposure needs to be elucidated further. How the high and low doses of arsenic induce human bronchial epithelial cell transformation is yet to be elucidated. In the present study, human bronchial epithelial cells were exposed to varying high-dose sodium arsenite (NaAsO

Aimed at the input parallel and output parallel (IPOP) dual-active bridge (DAB) dc-dc converters, this paper proposes an unbalanced-power-distribution scheme (UPDS) that considers the practical parameter deviation between DAB converters for minimizing the total backflow power. Firstly, combined with the characteristics of extended-phase-shift (EPS) control, the popular balanced-power-distribution scheme (BPDS) cannot achieve minimum backflow power due to the parameter deviation. In this regard, UPDS is established by detailed mathematical derivation, dividing the system operation into three sections. Simulation model and two-parallel DAB converters protype were built, and the results substantiated the validity of the theoretical analysis and the effectiveness of the proposed method.

Electrical vehicles (EV) may breaks down on the road when they don’t have enough energy driving to nearby charging pots. In this paper, an isolated DC/DC converter using dual-active-bridge (DAB) topology is designed for a car to be a portable battery, which can drive where the EV needs and charge it in emergency. Firstly, the actual switching status using optimized Triple-phase-shift (TPS) control is analyzed considering dead-time. The improved TPS control which can realize all- ZVS (Zero Voltage Switch) operation is presented. Finally, a 10- kW prototype is built to verify the enhancement in efficiency.

To minimize the power loss of dual active bridge (DAB) converters, the issue of current stress minimization based on bidirectional inner dual phase shift (BIDPS) control for DAB is analyzed and an optimal control strategy is proposed in this paper. Firstly, the operating principle of BIDPS control is introduced. Then based on this control method, the minimum current stress under different operating conditions is derived and an optimal control algorithm is presented. Finally, experiments are implemented to validate the correctness of the theoretical analysis and the effectiveness of the proposed control strategy.

Purpose: Cx43 (connexin 43) has been found to inhibit cell cycle of tumor cells. No nuclear localization sequence of Cx43 was found but the protein has been found in nucleus. The process remains unclear. We are aiming at providing the model that AKAP95 (A-kinase anchoring protein) binds to and transports Cx43 into nucleus of lung cancer cells.Methods: Lung cancer cells were arrested at preliminary stage (P), middle and late stage (M) of G1 phase and restriction point (R). Immunocoprecipitation (Co-IP) and Western blot (WB) were used to analyze expression and binding of AKAP95, Cx43, cyclin D1 and cyclin E1 at different stages of lung cancer cells during G1 phase. Confocal laser scanning microscopy (CLSM) was used to detect the location of proteins in arrested cells. Transmission electron microscope (TEM) with higher resolution was used to detect binding/aggregating and location of the above four proteins at subcellular level.Results: 1) AKAP95 transports Cx43 into the nucleus through the nuclear pore mainly at R stage. Some AKAP95 and Cx43 form protein aggregates during the process. 2) Cyclin E1 was found to be bind/aggregate with AKAP95-Cx43 complex and involved in the nuclear transport of Cx43 mainly at R stage. 3)Cyclin D1 was found to bind with AKAP95 and Cx43 respectively but not to be involved in their aggregating and nuclear transport of Cx43.Conclusions: 1) AKAP95 and Cx43 bind/aggregate to each other during G1 phase, forming AKAP95-Cx43 complexes/aggregates. After their binding/aggregating, AKAP95 targets nuclear pore and matrix, transporting Cx43 into nucleus. Formed AKAP95-Cx43 complexes/ aggregates do not dissociate in nucleus. The process can be summarized as ‘forming AKAP95-Cx43 complexes/aggregates; transporting Cx43 into nucleus; functioning remaining binding/aggregating’. 2) Cyclin D1 was only found to bind to AKAP95 and Cx43 respectively and function in cytoplasm while cyclin E1 was involved in the binding/aggregation of AKAP95 and Cx43, as well as the nuclear transport of Cx43.

BackgroundAKAP95(A-kinase anchoring protein) and Cx43 (connexin 43) express abnormally in lung cancer cells. As potential tumor therapeutic targets, specific process of bindings and dynamic changes of Cx43-AKAP95 complex in lung cancer cells may guide further treatments and detections of lung cancer. However, the process remains unclear now. We are aiming at investigating the dynamic changes of expression, localization, and binding of AKAP95 and Cx43, as well as their interaction with cyclin D1 and cyclin E1 in lung cancer cells during G1 phase.MethodsA549 and Beas-2B cells were arrested at preliminary stage(P), middle stage(M) and restriction point(R) of G1 phase and Western blot(WB), Confocal laser scanning microscopy (CLSM) and Transmission electron microscope (TEM) were used in our study to detect proteins to provide further evidence of the correlation of these proteins.Results: 1) AKAP95 carries Cx43 into the nucleus through the nuclear pore by binding to it during G1 phase. Some AKAP95 and Cx43 can aggregate and form larger protein aggregates. The process happens mainly during R stage. 2) AKAP95 and Cx43 mainly bind to cyclin D1 during P and M stage while bind to cyclin E1 during R stage respectively. Complexes of AKAP95-cyclin D1 and Cx43-cyclin D1 cannot enter nucleus while AKAP95-cyclin E1 and Cx43-cyclin E1 can.Conclusions1) Binding process of AKAP95-Cx43 complexes/ aggregates can be summarize as ‘bind and aggregate -target and enter nucleus- keep binding/aggregating in nucleus’. 2) Cyclin E1 was involved in the binding/aggregation and nuclear entry of AKAP95 and Cx43 while cyclin D1 only binds to them respectively in the cytoplasm.

Background This study aimed to overexpress or silence connexin 43 (Cx43) and A‐kinase anchoring protein 95 (AKAP95) in human A549 cells to explore their effects on cyclins and on G1/S conversion when the interrelationship of Cx43, AKAP95, and cyclin E1/E2 changes. Methods The study mainly used Western blot analysis and Co‐immuno precipitation to detect the target protein in Cx43/AKAP95 over expressed human A549 cells, and the relationship of proteins Cx43, AKAP95 and Cyclin E during G1‐S phase was explored with qualitative and quantitative analysis. Results The overexpression of Cx43 inhibited the expression of cyclin D1 and E1 by accelerating their degradation and reduced the Cdk2 activity that blocked the DNA transcription activity. However, the overexpression of AKAP95 increased the expression of cyclin D1 and E1 and inhibited their degradation, and enhanced the Cdk2 activity that promoted the DNA transcription activity. Cx43 and AKAP95 competitively bound to cyclin E1/E2, and the competitive binding affected the Cdk2 activity, Rb phosphorylation, DNA transcription activity, and G1/S conversion. Conclusions This study showed that the expression of ERK1/2, PKA, and PKB increased when BEAS‐2B cells were treated with PDGF‐BB, suggesting that ERK1/2, PKA, and PKB might be involved in the binding of AKAP95 with cyclin E, or the separation of AKAP95 from Cx43 from cyclin E1/E2. The specific mechanism underlying this process still needs further exploration.

In the present study, we developed a nano-integrated diagnostic and therapeutic platform with oxidation–reduction reactions in tumor microenvironments (TMEs). The proposed platform resolved the contradiction of particle size between the enhanced permeability and retention (EPR) effect and tumor interstitial penetration, as well as poor circulation and low drug-loading efficiency. Flower-like MnO2 NPs were used as the core and modified with hyaluronate (HA) and H2PtCl6 to obtain MnO2–HA@H2PtCl6 (MHP). The maximum drug-loading efficiency rate of H2PtCl6 reached 35% due to its chelation with HA. MHP showed satisfactory integrity and stability during circulation and can also be used as a magnetic resonance imaging (MRI) contrast agent. In addition, MHP as a radiosensitizer achieved an excellent tumor inhibition effect in combination with radiotherapy. Importantly, MHP released ultra-small nanoparticles, USNPs, (∼20 nm) through the supramolecular self-assembly abilities of Mn2+, HA, and H2PtCl6 in TMEs, leading to the increase of penetration into multicellular spheres and solid tumors (Scheme), as well as prolonging its retention in tumors.

Carbon fiber reinforced polymer (CFRP) plates are widely used to retrofit or reinforce steel structures, and the debonding damage between the steel structure and the CFRP plate is a typical failure in strengthening steel structures. This paper proposes a new approach to detecting debonding between a steel beam and a reinforcing CFRP plate by using removable lead zirconate titanate (PZT)-based transducers and active sensing. The removable PZT-based transducers are used to implement the active sensing approach, in which one transducer, as an actuator, is used to generate stress wave, and another transducer, as a sensor, is used to detect the stress wave that propagates across the bonding between the steel beam and the reinforcing CFRP plate. The bonding condition significantly influences the received sensor signal, and a wavelet-packet-based energy index (WPEI) is used to quantify the energy of the received signal to evaluate the severity of debonding between the steel beam and the reinforcing CFRP plate. To validate the proposed approach, experimental studies were performed, and two removable PZT-based transducers were designed and fabricated to detect the debonding between a steel beam and the reinforcing CRFP plate. The experimental results demonstrate the feasibility of the proposed method in detecting the debonding between a steel beam and the reinforcing CFRP plate using removable PZT-based transducers.

Identification of biological markers predicting the onset of neoplasia in patients with long-standing ulcerative colitis (UC) could allow for risk stratification in this population. In this study, we retrospectively identified subjects with chronic UC who developed colon neoplasia (n = 16) matched to UC patients who never developed neoplasia. RNA was extracted from archived colonic biopsies obtained at an interval of 1–2 years prior and 3–5 years prior to the onset of neoplasia. miRNA expression was assessed using Nanostring arrays in 12 subjects, and significantly up-regulated miRNAs were evaluated by real time pcr in the entire cohort of patients. Expression of miR-215 was also assessed in UC-associated colon cancers and compared to p53 expression. By array analysis, there were 17 significantly down-regulated and 7 significantly up-regulated miRNAs in subjects who later developed neoplasia. miR-215 was significantly up-regulated both 1–2 years prior to the onset of neoplasia (3.5-fold, p < 0.001) and 3–5 years prior to the onset of neoplasia (5.4-fold, p = 0.007). miR-215 expression was also increased in UC-associated colon cancers (5.3-fold, p = 0.03) and adjacent non-dysplastic UC tissue (6.2-fold, p = 0.02). p53 was expressed in 20% of patients prior to the onset of neoplasia and in 67% of UC-associated colon cancers, although was not correlated with miR-215 expression. Our data demonstrates that expression of miR-215 can discriminate patients who progressed to neoplasia from non-progressors as early as 5 years prior to the diagnosis of neoplasia, supporting that this and perhaps other miRNAs could serve as predictive biomarkers to risk stratify patients with chronic UC.

AIM This study aimed to investigate the correlation between the expression of A-kinase anchor protein95 (AKAP95), p-retinoblastoma (phosphorylated Rb, p-Rb), cyclin D2, cyclin D3 and cyclin E2 in esophageal cancer tissues and clinicopathological indexes. METHOD The protein expression levels of AKAP95, p-Rb, cyclin D2/3 and cyclin E2 in 40 esophageal cancer tissues were detected using immunohistochemistry, and the correlation between them was analyzed. RESULT The percentage of p-Rb (Ser780)-, cyclin D2-, cyclin D3- and cyclin E2-positive samples was 62.50%, 70.00%, 67.50% and 60.00%, respectively. Also, the positive expression did not correlate with the histological type, histological differentiation or lymph node metastasis. The expression of AKAP95 and p-Rb (Ser780), p-Rb (Ser780) and cyclin D2 and p-Rb (Ser780) and cyclin D3 in esophageal cancer tissues was found to be correlated (P

Background and Aims As ulcerative colitis [UC]-associated colorectal cancer [CRC] and sporadic CRC differ in presentation and molecular features, we sought to evaluate differences in the impact of DNA methylation on gene expression. Methods DNA methylation was assessed in 11 UC-CRCs and adjacent tissue and 11 sporadic CRCs and adjacent tissue, using Illumina arrays. RNA sequencing was performed on 10 UC-CRCs and adjacent tissue and eight sporadic CRCs and adjacent tissues. Differences in DNA methylation and transcript expression, as well as their correlation in the same tissues, were assessed. Immunohistochemistry was performed for three proteins, ANPEP, FAM92A1, and STK31, all of which exhibited an inverse correlation between DNA methylation and transcript expression in UC. Results Thirty three loci demonstrated differences in DNA methylation between UC-CRC and adjacent tissue. In contrast, there were 4204 differentially methylated loci between sporadic colon cancer and adjacent tissue. Eight hundred eighty six genes as well as 10 long non-coding RNAs [lncRNA] were differentially expressed between UC-CRC and adjacent tissues. Although there were no differentially methylated loci between UC and sporadic CRC, 997 genes and 38 lncRNAs were differentially expressed between UC-CRC and sporadic CRC. In UC, 18 genes demonstrated a negative correlation between DNA methylation and transcript expression. Evaluation of protein expression related to three genes, ANPEP, FAM92A1, and STK31, confirmed down-regulation of ANPEP and up-regulation of STK31 in UC-CRC. Conclusions Regulation of transcript expression by DNA methylation involves genes key to colon carcinogenesis and may account for differences in presentation and outcomes between inflammatory bowel disease and sporadic colon cancer.

195 Background: Noninvasive tests for peritoneal metastasis (PM) detection lack sensitivity. Genome-wide mapping of 5-hydroxymethylcytosine (5hmC) on nanogram quantities of peripheral plasma circulating cell-free DNA (PcfDNA) was previously shown to differentiate non-metastatic colorectal cancer from healthy subjects. We aimed to examine if patients with colorectal cancer (CRC), high grade appendiceal cancer (HGA) or low grade appendiceal cancer (LGA) with PM have distinct signatures of 5hmC in PcfDNA compared to each other and to patients matched for tumors without PM. Methods: We analyzed plasma samples from a prospectively collected tissue bank. To correlate 5hmC signatures with intraoperative findings, only patients who underwent abdominal surgery in proximity to plasma collection were selected. Key steps of PcfDNA processing included extraction from plasma, nano-hmC-Seal chemical labeling and enrichment of 5hmC-modified fragments, next-generation sequencing, and mapping to the reference human genome. DESeq2 R package was finally used to compare relative 5hmC enrichment and detect distinct 5hmC signatures according to disease histology and PM presence. Results: Plasma samples were collected between 11/2016 – 3/2019 from 46 patients with CRC (n = 21), HGA (n = 17) and LGA (n = 8). Of those, 32 (70%) had PM based on intraoperative findings (median peritoneal cancer index score = 15, range 2-39) and 14 did not have PM. Most samples (91%) were collected on the same day as surgery. An average of 24 million paired-end reads were sequenced for each sample. Four samples (8.7%) were excluded from the analysis due to low sequencing coverage or high duplication level. Unique 5hmC enrichment patterns were found to discriminate with p < 0.05 between CRC PM and HGA PM (n = 616 differentially hydroxymethylated genes (DHMGs)), CRC/HGA PM and LGA PM (n = 1074 DHMGs), and CRC/HGA PM and CRC/HGA patients without PM (n = 1576 DHMGs). Conclusions: Distinct signatures of 5hmC in PcfDNA could differentiate patients with CRC/HGA/LGA PM from each other and from patients with similar tumor histologies without PM. Thus 5hmC signatures in PcfDNA might potentially serve as a sensitive marker of occult PM.

To investigate the prevalence of dry eye disease (DED) and distribution of associated risk factors among a hospital-based population.In this cross-sectional study, we collected detailed information of clinically defined moderate-to-severe patients with dry eye among a consecutive hospital-based population, including age trend, gender structure, frequency of symptoms, and distribution of associated environmental/occupational risk factors.Of 6,657 consecutive outpatients aged older than 20 years, symptomatic dry eye presented in 635 subjects (9.54%). Five hundred thirty-two of these 635 subjects (7.99%) were clinically diagnosed as defined DED that combined with positive signs. Women (10.41%) were significantly higher than men (5.21%) (P0.001). Overexposure to visual display terminal was a major risk factor for DED among young men and women (56.2%). Our study also found occupational conditions with the risk of exposure to adverse environment made up over half of all 532 patients with dry eye. The use of contact lenses was closely associated with DED in young women, and history of ocular surgeries might be another factor associated with DED in old people. One hundred sixty-three of 371 female patients with dry eye (43.9%) were associated with hormonal changes. The incidence of meibomian gland dysfunction-related DED increased gradually with age. There were only 10 patients with dry eye (1.9%) associated with Sjögren syndrome, and all of them were women.Environmental and occupational factors were strongly associated with DED and constituted the major proportion in a hospital-based population. A classification of DED based on the distribution of risk factors was recommended for clinical use.

Purpose: Patients with ulcerative colitis are at increased risk for colorectal cancer, although mechanisms underlying neoplastic transformation are poorly understood. We sought to evaluate the role of microRNAs in neoplasia development in this high-risk population. Experimental Design: Tissue from 12 controls, 9 ulcerative colitis patients without neoplasia, and 11 ulcerative colitis patients with neoplasia was analyzed. miRNA array analysis was performed and select miRNAs assayed by real-time PCR on the discovery cohort and a validation cohort. DNA methylation of miR-193a was assessed. Following transfection of miR-193a-3p, proliferation, IL17RD expression, and luciferase activity of the 3′UTR of IL17RD were measured. Tumor growth in xenografts as well as EGFR signaling were assessed in HCT116 cells expressing IL17RD with either a mutant 3′ untranslated region (UTR) or wild-type (WT) 3′UTR. Results: miR-31, miR-34a, miR-106b, and miR-193a-3p were significantly dysregulated in ulcerative colitis-neoplasia and adjacent tissue. Significant down-regulation of miR-193a-3p was also seen in an independent cohort of ulcerative colitis cancers. Changes in methylation of miR-193a or expression of pri-miR-193a were not observed in ulcerative colitis cancer. Transfection of miR-193a-3p resulted in decreased proliferation, and identified IL17RD as a direct target of miR-193a-3p. IL17RD expression was increased in ulcerative colitis cancers, and miR-193a-3p treatment decreased growth and EGFR signaling of HCT116 cells in xenografts expressing both IL17RD with WT 3′UTR compared with cells expressing IL17RD with mutant 3′UTR. Conclusions: miR-193a-3p is downregulated in ulcerative colitis neoplasia, and its loss promotes carcinogenesis through upregulation of IL17RD. These findings provide novel insight into inflammation-driven colorectal cancer and could suggest new therapeutic targets in this high-risk population. Clin Cancer Res; 23(17); 5281–91. ©2017 AACR.

BACKGROUND As mechanisms of neoplasia in patients with ulcerative colitis (UC) remain poorly understood, we sought to identify pathways of carcinogenesis in this high-risk population. METHODS MicroRNA (miRNA) and mRNA expression was examined in nondysplastic rectosigmoid mucosa from UC patients with (n = 19) or without remote colon neoplasia (n = 23). We developed a method to identify miRNA-regulated pathways based on differentially expressed miRNAs and their putative mRNAs targets in the same samples. One key pathway identified in the analysis, miR-4728-3p regulation of focal adhesion signaling was further evaluated in vitro and through examination of expression in UC-cancers. RESULTS There were 101 significantly up-regulated and 98 down-regulated miRNAs (adjusted P < 0.05) in the rectal mucosa of UC patients harboring proximal neoplasia. Bioinformatic analysis identified miR-4728-3p as a regulator of 3 proteins involved in focal adhesion signaling, CAV1, THBS2, and COL1A2. Real-time PCR validated down-regulation of miR-4728-3p in nondysplastic tissue remote from UC-neoplasia and in UC-associated colon cancers. miR-4728-3p transfection into colon cancer cells down-regulated expression levels and decreased luciferase activities in cells expressing a wild type 3' untranslated region compared with a mutant 3' untranslated region for all 3 genes. Exogenous transfected miR-4728-3p also delayed wound healing and decreased formation of focal adhesion complexes. CONCLUSIONS Patients with long-standing UC who harbor neoplasia can be identified based on miRNA and mRNA profiles in nondysplastic tissue. Using a method to analyze miRNA and mRNA expression from the same tissues, we identified that miR-4728-3p is likely an important tumor suppressor in UC-associated colon carcinogenesis.

With a high spatial resolution and wide coverage, satellite-based precipitation products have compensated for the shortcomings of traditional measuring methods based on rain gauge stations, such as the sparse and uneven distribution of rain gauge stations. However, the accuracy of satellite precipitation products is not high enough in some areas, and the causes of their errors are complicated. In order to better calibrate and apply the product&rsquo, s data, relevant research on this kind of product is required. Accordingly, this study investigated the spatial error distribution and spatial influence factors of the Tropical Rainfall Measuring Mission (TRMM) Multi-Satellite Precipitation Analysis (TMPA) post-process 3B42V7 (hereafter abbreviated as 3B42V7) data over mainland China. This study calculated accuracy indicators based on the 3B42V7 data and daily precipitation data from 797 rain gauge stations across mainland China over the time range of 1998&ndash, 2012. Then, a clustering analysis was conducted based on the accuracy indicators. Moreover, the geographical detector (GD) was used to perform the error cause analysis of the 3B42V7. The main findings of this study are the following. (1) Within mainland China, the 3B42V7 data accuracy decreased gradually from the southeast coast to the northwest inland, and shows a similar distribution for precipitation. High values of systematic error (&gt, 1.0) is mainly concentrated in the southwest Tibetan Plateau, while high values of random error (&gt, 1.0) are mainly concentrated around the Tarim Basin. (2) Mainland China can be divided into three areas by the spectral clustering method. It is recommended that the 3B42V7 can be effectively used in Area I, while in Area III the product should be calibrated before use, and the product in Area II can be used after an applicability study. (3) The GD result shows that precipitation is the most important spatial factor among the seven factors influencing the spatial error distribution of the 3B42V7 data. The relationships between spatial factors are synergistic rather than individual when influencing the product&rsquo, s accuracy.

In this work, we develop a ratiometric two-photon fluorescent probe, ATD@QD-E2Zn2SOD (ATD = amino triphenylamine dendron, QD = CdSe/ZnSe quantum dot, E2Zn2SOD = Cu-free derivative of bovine liver copper-zinc superoxide dismutase), for imaging and sensing the changes of intracellular Cu(2+) level with clear red-to-yellow color change based on specific biomolecular recognition of E2Zn2SOD for Cu(2+) ion. The inorganic-organic nanohybrided fluorescent probe features two independent emission peaks located at 515 nm for ATD and 650 nm for QDs, respectively, under two-photon excitation at 800 nm. Upon addition of Cu(2+) ions, the red fluorescence of QDs drastically quenches, while the green emission from ATD stays constant and serves as a reference signal, thus resulting in the ratiometric detection of Cu(2+) with high accuracy by two-photon microscopy (TPM). The present probe shows high sensivity, broad linear range (10(-7)-10(-3) M), low detection limit down to ∼10 nM, and excellent selectivity over other metal ions, amino acids, and other biological species. Meanwhile, a QD-based inorganic-organic probe demonstrates long-term photostability, good cell-permeability, and low cytotoxicity. As a result, the present probe can visualize Cu(2+) changes in live cells by TPM. To the best of our knowledge, this is the first report for the development of a QD-based two-photon ratiometric fluorescence probe suitable for detection of Cu(2+) in live cells.