Hypermethylation Loci of ZNF671, IRF8, and OTX1 as Potential Urine-Based Predictive Biomarkers for Bladder Cancer.

Bladder cancer (BCa) is a significant health issue and poses a healthcare burden on patients, highlighting the importance of an effective detection method. Here, we developed a urine DNA methylation diagnostic panel for distinguishing between BCa and non-BCa. In the discovery stage, an analysis of the TCGA database was conducted to identify BCa-specific DNA hypermethylation markers. In the validation phase, DNA methylation levels of urine samples were measured with real-time quantitative methylation-specific PCR (qMSP). Comparative analysis of the methylation levels between BCa and non-BCa, along with the receiver operating characteristic (ROC) analyses with machine learning algorithms (logistic regression and decision tree methods) were conducted to develop practical diagnostic panels. The performance evaluation of the panel shows that the individual biomarkers of ZNF671, OTX1, and IRF8 achieved AUCs of 0.86, 0.82, and 0.81, respectively, while the combined yielded an AUC of 0.91. The diagnostic panel using the decision tree algorithm attained an accuracy, sensitivity, and specificity of 82.6%, 75.0%, and 90.9%, respectively. Our results show that the urine-based DNA methylation diagnostic panel provides a sensitive and specific method for detecting and stratifying BCa, showing promise as a standard test that could enhance the diagnosis and prognosis of BCa in clinical settings.

Applying the Technology Acceptance Model to Explore Nursing Students' Behavioral Intention to Use Nursing Information Smartphones in a Clinical Setting.

The integration of mobile devices and nursing information systems has become a trend in modern clinical practice with various information and communication technologies available. Smartphones are gradually replacing notebooks in clinical practice as a medium for nursing information systems. Clinical nursing practicums are a necessary means for nursing students to foster their professional competence. In addition to professional skills, nursing students must also learn to apply information technologies in clinical settings. This study aimed to understand nursing students' behavioral intention toward nursing information smartphones and to further identify the factors influencing nursing students' behavioral intentions based on the technology acceptance model. A cross-sectional research design was used in this study. Eighty nursing students were recruited from a regional teaching hospital in Central Taiwan. The findings demonstrated that subjects' perceived ease of use and perceived usefulness of nursing information smartphones, as well as their attitude toward using and behavioral intention to use the smartphones, were positive, and they provided constructive feedback and suggestions to improve nursing information systems in hospitals. The findings can serve as a reference for hospitals and clinical training institutions seeking to integrate nursing information systems in clinical nursing education.

Integrative Multiomics Evaluation of IIDH1 Metabolic Enzyme as a Candidate Oncogene That is Correlated with Poor Prognosis and Immune Infiltration in Prostate Adenocarcinoma.

Mutations in the isocitrate dehydrogenase gene (IDH1) are involved in the progression of tumors. Although IDH1 has a role in various tumors, its clinical relevance and its expression in response to the immune response have not been investigated in prostate adenocarcinoma (PRAD). In the present study, we investigated the utility of IDH1 as a prognostic biomarker for PRAD by analyzing IDH1 mRNA expression and its association with patient survival and immune cell infiltration. IDH1 mRNA expression was significantly higher in PRAD tissue than in normal tissue, and Kaplan-Meier survival analysis showed that IDH1 expression was significantly associated with poor prognosis in PRAD patients. To elucidate the mechanisms involved, the correlation between IDH1 expression and the level of immune cell infiltration, in particular of immunosuppressive cells such as CD8+ T-cells, CD4+ T-cells, and macrophages, was further analyzed by single-cell RNA sequencing. We also screened a pharmacogenetic database for IDH1-specific drugs that inhibited high expression in PRAD. In the present study, we used a combination of databases to identify a significant correlation between IDH1 expression and cellular infiltration and to explain the mechanism by which IDH1 confers poor prognosis in PRAD, thus demonstrating the relevance of IDH1 expression as a prognostic biomarker with clinical utility in PRAD patients.

Inhibiting casein kinase 2 overcomes paclitaxel resistance in gastric cancer.

PURPOSE: Casein kinase (CK) 2 activation has been implicated in the proliferation of various tumor types and resistance to chemotherapy. We investigated the mechanistic basis for the association between CK2 activation and paclitaxel resistance in a gastric cancer (GC). EXPERIMENTAL DESIGN: CK2 expression was evaluated in 59 advanced GC patients treated with paclitaxel as the second-line therapy. The efficacy of a CK2 inhibitor, CX-4945, and paclitaxel was evaluated in GC cell lines and a xenograft model. RESULTS: Patients with high CK2 expression (29/59, 39%) showed lower disease control rates (47.7% vs. 72.3%, p = 0.017) and shorter progression-free survival (2.8 vs. 4.8 months, p = 0.009) than patients with low CK2 expression. CK2 protein expression was associated with sensitivity to paclitaxel in 49 GC cell lines. Combination therapy with CX-4945 and paclitaxel exerted synergistic antiproliferative effects and inhibited the downregulation of phosphatidylinositol 3-kinase/AKT signaling in SNU-1 cells. In the SNU-1 xenograft model, the combination treatment was significantly superior to either single agent, suppressing tumor growth without notable toxicities. CONCLUSIONS: These results demonstrated that CK2 activation was related to paclitaxel resistance and that CX-4945 in combination with paclitaxel could be used as a potential treatment for paclitaxel resistance in GC.

Casein Kinase 2 Inhibitor, CX-4945, as a Potential Targeted Anticancer Agent in Gastric Cancer.

BACKGROUND/AIM: Casein kinase 2 (CK2) is involved in multiple cellular processes. Furthermore, its overexpression in several human cancers has been associated with tumor progression. In this study, we evaluated the efficacy of the CK2 inhibitor, CX-4945, in gastric cancer cell lines and explored the potential predictive biomarkers for CX-4945 sensitivity. MATERIALS AND METHODS: The sensitivity to CX-4945 was screened in 49 gastric cancer cell lines by the MTT assay. The mRNA and protein expression of CK2 subunits (α and α') were determined using qRT-PCR and western blot. Furthermore, the activity of CK2α was measured by ELISA. Gene expression and mutations were analyzed via whole-exome and RNA sequencing. RESULTS: The sensitivity to CX-4945 was determined by the inhibition rate (%) at the effective dose (10 µM) which ranged from -1% to 89% in 49 gastric cancer cell lines. CK2α', but not CK2α, mRNA expression was correlated with CX-4945 sensitivity. CONCLUSION: In this study, CX-4945 showed modest antitumor efficacy in gastric cancer cell lines. CK2 might represent a potential therapeutic target for gastric cancer.

Glycyrrhizic acid induces human MDA-MB-231 breast cancer cell death and autophagy via the ROS-mitochondrial pathway.

Glycyrrhizic acid (GA), the main component of licorice root extracts, has been shown to suppress cell proliferation and induce apoptosis in various types of cancers. However, the molecular mechanism of its anticancer activity remains poorly understood and warrants further investigation. MDA-MB­231 cells were treated with various concentrations of GA and the cytotoxic effects of GA were determined using the CCK-8 assay. Apoptosis and cell cycle status were detected by flow cytometry. Reactive oxygen species (ROS) levels and mitochondrial membrane potential (∆Ψm) were assessed using DCFDA, MitoSOX and JC-1 staining. Western blot analysis was used to quantify the expression of autophagy-related proteins. In the present study, induction of autophagic cell death was observed in GA-treated MDA-MB­231 cells. Downregulation of p62- and beclin 1-associated proteins occurred after GA treatment, and, the conversion of LC3 and increased ROS without significant changes in caspase­associated proteins and intracellular responses were detected. Furthermore, loss of mitochondria and its membrane potential in cells demonstrated that mitochondria were involved in the GA-regulated MDA-MB-231 cell death. The addition of a pan-caspase inhibitor (z-VAD-fmk) did not suppress the GA-induced apoptotic effect, and GA-induced apoptosis was not accompanied by processing of procaspase-8, -9 and -3. However, GA triggered the translocation of the apoptosis-inducing factor (AIF) from the mitochondria into the nucleus. In contrast, GA-induced LC3 conversion was significantly inhibited by 3 methlyadenine (3MA), an inhibitor of PI3K­regulated autophagy. Therefore, these results suggest that enhancement of both AIF- and LC3-dependent GA-derived ROS generation plays an important role in the inhibition of the growth of MDA-MB-231 human breast cancer cells.

The prevalence rate of deviations in body constitutions and related factors in follow-up stage breast cancer patients-A nationwide study.

OBJECTIVES: The aim of this study was to apply a rigorous traditional Chinese medicine (TCM) body constitution questionnaire (BCQ) to survey the prevalence rate of deviations in body constitution and to explore the health-related lifestyle behavior factors of deviations in body constitution. DESIGN: A cross-sectional study was administered through postal mail. Subjects were recruited from a national organization for breast cancer patients (Taiwan Breast Cancer Alliance). SETTING/MAIN OUTCOME MEASURES: Data were obtained from 311 breast cancer patients by questionnaires including a demographic record sheet, lifestyle behavior scales and the BCQ (Yang-Xu, Yin-Xu, and Stasis). Differences concerning the presence of body constitutions were analyzed by Chi-square tests and analyses of variance, and potential predictive factors were analyzed using multivariate logistic regression. RESULTS: In total, 55.3% of the subjects had a Yang-Xu constitution, 61.0% had a Yin-Xu constitution, and 47.6% had a Stasis constitution. A total of 42.8% of the patients displayed a combination of the three types of body constitutions. Feeling stressed, physical exercise, and favoring fried food were predictors of the combined Yang-Xu, Yin-Xu and Stasis constitutions (p<0.05). Staying up late was significantly associated with Yin-Xu (p=0.017), and favoring salty food was significantly associated with Stasis (p=0.019). CONCLUSIONS: A high prevalence of deviations in body constitutions was observed in the follow-up stage of breast cancer patients. Increasing the adherence to healthy lifestyle behaviors might strengthen and balance body constitution, which could improve supportive care in breast cancer survivors.

CX-5461 is a DNA G-quadruplex stabilizer with selective lethality in BRCA1/2 deficient tumours.

G-quadruplex DNAs form four-stranded helical structures and are proposed to play key roles in different cellular processes. Targeting G-quadruplex DNAs for cancer treatment is a very promising prospect. Here, we show that CX-5461 is a G-quadruplex stabilizer, with specific toxicity against BRCA deficiencies in cancer cells and polyclonal patient-derived xenograft models, including tumours resistant to PARP inhibition. Exposure to CX-5461, and its related drug CX-3543, blocks replication forks and induces ssDNA gaps or breaks. The BRCA and NHEJ pathways are required for the repair of CX-5461 and CX-3543-induced DNA damage and failure to do so leads to lethality. These data strengthen the concept of G4 targeting as a therapeutic approach, specifically for targeting HR and NHEJ deficient cancers and other tumours deficient for DNA damage repair. CX-5461 is now in advanced phase I clinical trial for patients with BRCA1/2 deficient tumours (Canadian trial, NCT02719977, opened May 2016).

Hydrogen peroxide inducible clone-5 mediates reactive oxygen species signaling for hepatocellular carcinoma progression.

One of the signaling components involved in hepatocellular carcinoma (HCC) progression is the focal adhesion adaptor paxillin. Hydrogen peroxide inducible clone-5 (Hic-5), one of the paralogs of paxillin, exhibits many biological functions distinct from paxillin, but may cooperate with paxillin to trigger tumor progression. Screening of Hic-5 in 145 surgical HCCs demonstrated overexpression of Hic-5 correlated well with intra- and extra-hepatic metastasis. Hic-5 highly expressed in the patient derived HCCs with high motility such as HCC329 and HCC353 but not in the HCCs with low motility such as HCC340. Blockade of Hic-5 expression prevented constitutive migration of HCC329 and HCC353 and HGF-induced cell migration of HCC340. HCC329Hic-5(-), HCC353Hic-5(-), HCC372Hic-5(-), the HCCs stably depleted of Hic-5, exhibited reduced motility compared with each HCC expressing Scramble shRNA. Moreover, intra/extrahepatic metastasis of HCC329Hic-5(-) in SCID mice greatly decreased compared with HCC329Scramble. On the other hand, ectopic Hic-5 expression in HCC340 promoted its progression. Constitutive and HGF-induced Hic-5 expression in HCCs were suppressed by the reactive oxygen species (ROS) scavengers catalase and dithiotheritol and c-Jun N-terminal kinase (JNK) inhibitor SP600125. On the contrary, depletion of Hic-5 blocked constitutive and HGF-induced ROS generation and JNK phosphorylation in HCCs. Also, ectopic expression of Hic-5 enhanced ROS generation and JNK phosphorylation. These highlighted that Hic-5 plays a central role in the positive feedback ROS-JNK signal cascade. Finally, the Chinese herbal derived anti-HCC peptide LZ-8 suppressed constitutive Hic-5 expression and JNK phosphorylation. In conclusion, Hic-5 mediates ROS-JNK signaling and may serve as a therapeutic target for prevention of HCC progression.

Increased yin-deficient symptoms and aggravated autonomic nervous system function in patients with metastatic cancer.

OBJECTIVES: The objectives of this study were to investigate the differences in severity of yin-deficiency syndrome (YDS) and function of the autonomic nervous system (ANS) between patients with cancer with metastasis and those without metastasis. SETTING: The setting was an outpatient clinic in a teaching hospital in central Taiwan. SUBJECTS: The subjects were a total of 124 patients who had been diagnosed with cancer on the basis of pathologic and clinical findings. Among them, 61 had distant metastasis, and the other 63 had no evidence of metastasis. The two groups were similar in terms of age and gender. INTERVENTIONS: The severity of YDS in each subject was evaluated using a questionnaire containing 12 items about symptoms and signs related to YDS. The severity of each symptom or sign was rated on a 4-point scale. OUTCOME MEASURES: The total score on the questionnaire represented the severity of YDS. ANS function in each subject was evaluated by measuring heart rate variability (HRV), including time and frequency domains. The questionnaire data were coded, and statistical analysis was performed using SPSS version 12.0. Data were analyzed using the Student's t test or the χ(2) test. RESULTS: The patients with metastasis had significantly higher average total YDS score and heart rate compared with the patients without metastasis. In contrast, they had significantly lower HRV, including standard deviation of the 5-minute average R-R interval, total power, very-low-frequency power, and low frequency (LF) power, but not high-frequency (HF) power and LF/HF ratio. CONCLUSIONS: The results of this study indicate that patients with metastatic cancer have more severe YDS and impaired ANS function than those without metastasis.

Severity of Yin deficiency syndrome and autonomic nervous system function in cancer patients.

OBJECTIVE: To evaluate the distribution of symptoms related to Yin deficiency syndrome (YDS), and to analyze the relationship between the severity of YDS and the function of the autonomic nervous system (ANS) in cancer patients. SETTING: Outpatient clinic in a teaching hospital in central Taiwan. SUBJECTS: Eighty (80) patients had been diagnosed with cancer by pathologic and clinical findings. METHOD: The severity of YDS in each subject was evaluated by a questionnaire consisting of 12 items concerning symptoms and signs related to YDS, scored from 1 to 4 points. OUTCOME MEASURES: The total score for all 12 items represented the severity of YDS. ANS function in each subject was evaluated by measuring heart rate variability (HRV), including time-frequency analysis. We coded the collected questionnaire material and performed statistical analysis (description analysis, ANOVA, and Pearson's correlation coefficients) using SPSS v.12.0 software. RESULTS: The highest total YDS score was 36 points and the lowest was 10 points. The 3 most common YDS signs were dry mouth (58.8%), sleeplessness with annoyance (56.3%), and flush over face in the afternoon (22.5%). The total YDS scores had a significantly positive correlation with heart rate (HR), but had significantly negative correlation with the standard deviation of the 5-minute mean R-R intervals (SDANN), total HRV power, power in the very low frequency band, and in the low frequency band. CONCLUSIONS: The above results suggest that the severity of YDS in cancer patients was associated with increased HR and decreased ANS activity. There is a possibility that the disturbance of ANS function may contribute to the occurrence of YDS in cancer patients.

Identification and characterization of a novel fibril forming peptide in fungal starch binding domain.

Scanty information is available regarding the chemical basis for structural alterations of the carbohydrate-binding modules (CBMs). The N-terminal starch binding domain (SBD) of Rhizopus oryzae glucoamylase (GA) forms fibrils under thermal stress, presenting an unusual conformational change from immunoglobulin-like to beta-sheet-rich structure. Site-directed mutagenesis revealed that the C-terminal Lys of SBD played a crucial role in the fibril formation. The synthetic peptide (DNNNSANYQVSTSK) representing the C-terminal 14 amino acid residues of SBD was further demonstrated to act as a fibril-forming segment, in which terminal charges and an internal NNNxxNYQ motif were key fibril-forming determinants. The formation of fibril structure in a fungal SBD, caused by its chemical and biophysical requirements, was demonstrated for the first time.

The distribution of Yin-Deficient symptoms and their relationship on survival rate in cancer patients with Yin-Deficiency.

Yin-Deficiency (YD), representing a status of the human body under lack of nutrition and fluid in traditional Chinese medicine, is commonly seen in late stage of cancer patients. It is not known whether the severity of YD related symptoms/signs can predict the survival rate of cancer patients. This study evaluated the distribution of Yin-deficiency symptoms/signs (YDS) in cancer patients with YD, and investigated whether the severity of YDS can predict the survival rate of cancer patients with YD. From 5 January 2007 to 5 May 2007, we selected 43 cancer patients with diagnosis of YD from hospitalized patients and outpatients. The severity of YD was evaluated by a questionnaire. We further estimated the cumulative probabilities of the survival rates over 4 months since the start of study by the Kaplan-Meier product-limit method, and compared the differences among groups with various severities in each symptom/sign with the use of the log-rank test. The results revealed that, the 3 most common YDS were sleeplessness with annoyance, less or non-coated tongue with or without redness and dry mouth. In the survival rate analysis, only 2 parameters, rapidly small pulse (p = 0.002) and less-or non-coated tongue with paleness (p = 0.017), were found to be related to the decrease of cancer patients with YD. This suggests that, both rapidly small pulse and less-or non-coated tongue without redness may be used as predictors for the estimation of survival rate in cancer patients with YD.

Characterization of molecular interactions between eosinophil cationic protein and heparin.

Eosinophil cationic protein (ECP) is currently used as a biomarker for airway inflammation. It is a heparin-binding ribonuclease released by activated eosinophils. Its cytotoxicity toward cancer cell lines is blocked by heparin. The objective of this study was to locate the heparin binding site of ECP by site-directed mutagenesis and construction of a synthetic peptide derived from this region. Synthetic heparin with > or =5 monosaccharide units showed strong inhibition of ECP binding to the cell surface. Analysis of ECP mt1 (R34A/W35A/R36A/K38A) showed that these charged and aromatic residues were involved in ECP binding to heparin and the cell surface. A potential binding motif is located in the loop L3 region between helix alpha2 and strand beta1, outside the RNA binding domain. The synthetic peptide derived from the loop L3 region displayed strong pentasaccharide binding affinity and blocked ECP binding to cells. In addition, ECP mt1 showed reduced cytotoxicity. Thus, the tight interaction between ECP and heparin acts as the primary step for ECP endocytosis. These results provide new insights into the structure and function of ECP for anti-asthma therapy.

[A nursing experience with a patient with esophagus cancer combined with tumor ulcer].

The purpose of this article is to describe a nursing experience with a patient with esophagus cancer combined with tumor ulcer. The author assessed the patient's health condition by observation, interview, medical history and the Gorden functional health assessment guide. The client had three nursing problems: impaired tissue integrity, diarrhea and body image disturbance. The author used the nursing process and empathy to assist the patient to face disease and treatment. Nursing intervention was also performed to enhance the patient's comfort and provide psychological support.