ZFHX3 variants cause childhood partial epilepsy and infantile spasms with favourable outcomes.

BACKGROUND: The ZFHX3 gene plays vital roles in embryonic development, cell proliferation, neuronal differentiation and neuronal death. This study aims to explore the relationship between ZFHX3 variants and epilepsy. METHODS: Whole-exome sequencing was performed in a cohort of 378 patients with partial (focal) epilepsy. A Drosophila Zfh2 knockdown model was used to validate the association between ZFHX3 and epilepsy. RESULTS: Compound heterozygous ZFHX3 variants were identified in eight unrelated cases. The burden of ZFHX3 variants was significantly higher in the case cohort, shown by multiple/specific statistical analyses. In Zfh2 knockdown flies, the incidence and duration of seizure-like behaviour were significantly greater than those in the controls. The Zfh2 knockdown flies exhibited more firing in excitatory neurons. All patients presented partial seizures. The five patients with variants in the C-terminus/N-terminus presented mild partial epilepsy. The other three patients included one who experienced frequent non-convulsive status epilepticus and two who had early spasms. These three patients had also neurodevelopmental abnormalities and were diagnosed as developmental epileptic encephalopathy (DEE), but achieved seizure-free after antiepileptic-drug treatment without adrenocorticotropic-hormone/steroids. The analyses of temporal expression (genetic dependent stages) indicated that ZFHX3 orthologous were highly expressed in the embryonic stage and decreased dramatically after birth. CONCLUSION: ZFHX3 is a novel causative gene of childhood partial epilepsy and DEE. The patients of infantile spasms achieved seizure-free after treatment without adrenocorticotropic-hormone/steroids implies a significance of genetic diagnosis in precise treatment. The genetic dependent stage provided an insight into the underlying mechanism of the evolutional course of illness.

Identification of patients with nonischemic dilated cardiomyopathy at risk of malignant ventricular arrhythmias: insights from cardiac magnetic resonance feature tracking.

BACKGROUND: Patients with nonischemic dilated cardiomyopathy (NIDCM) are prone to arrhythmias, and the cause of mortality in these patients is either end-organ dysfunction due to pump failure or malignant arrhythmia-related death. However, the identification of patients with NIDCM at risk of malignant ventricular arrhythmias (VAs) is challenging in clinical practice. The aim of this study was to evaluate whether cardiovascular magnetic resonance feature tracking (CMR-FT) could help in the identification of patients with NIDCM at risk of malignant VAs. METHODS: A total of 263 NIDCM patients who underwent CMR, 24-hour Holter electrocardiography (ECG) and inpatient ECG were retrospectively evaluated. The patients with NIDCM were allocated to two subgroups: NIDCM with VAs and NIDCM without VAs. From CMR-FT, the global peak radial strain (GPRS), global longitudinal strain (GPLS), and global peak circumferential strain (GPCS) were calculated from the left ventricle (LV) model. We investigated the possible predictors of NIDCM combined with VAs by univariate and multivariate logistic regression analyses. RESULTS: The percent LGE (15.51 ± 3.30 vs. 9.62 ± 2.18, P < 0.001) was higher in NIDCM patients with VAs than in NIDCM patients without VAs. Furthermore, the NIDCM patients complicated with VAs had significantly lower GPCS than the NIDCM patients without VAs (- 5.38 (- 7.50, - 4.22) vs.-9.22 (- 10.73, - 8.19), P < 0.01). Subgroup analysis based on LGE negativity showed that NIDCM patients complicated with VAs had significantly lower GPRS, GPCS, and GPLS than NIDCM patients without VAs (P < 0.05 for all). Multivariate analysis showed that both GPCS and %LGE were independent predictors of NIDCM combined with VAs. CONCLUSIONS: CMR global strain can be used to identify NIDCM patients complicated with VAs early, specifically when LGE is not present. GPCS < - 13.19% and %LGE > 10.37% are independent predictors of NIDCM combined with VAs.

Rasch analysis of the Hospital Anxiety andDepression Scale (HADS) in patients with major depression / Análisis de Rasch de la Escala Hospitalaria de Ansiedad y Depresión (Hospital Anxiety and Depression Scale, HADS) en pacientes con depresión mayor

La Hospital Anxiety and Depression Scale (HADS) [Escala Hospitalaria de Ansiedad y Depresión] es una herramienta ampliamente utilizada para detección de la depresión y la ansiedad en pacientes con enfermedades médicas. Este estudio tuvo como objetivo explorar las propiedades psicométricas de la HADS en la detección de pacientes con depresión mayor utilizando el método de la teoría de respuesta al ítem. Un total de 460 pacientes con depresión mayor completaron el HADS. Se utilizó el análisis de Rasch para examinar la unidimensionalidad, el ajuste de los ítems, la dependencia local, la confiabilidad, el orden de las categorías, el funcionamiento diferencial de los ítems (DIF) y la focalización. La HADS mostró una construcción bidimensional. Todos los ítems se ajustaban al modelo de Rasch. Tres pares de ítems mostraron una dependencia local menor pero desconsiderada. Ambas subescalas tuvieron una confiabilidad aceptable. Ninguno de los ítems mostró categorías desordenadas o DIF. Todos los ítems estaban bien dirigidos y los participantes con niveles altos y bajos de angustia fueron menos objetivo que aquellos con niveles moderados de angustia. Finalmente, se generó una tabla de conversión para transformar las puntuaciones brutas en medidas de intervalo. El HADS demostró propiedades psicométricas adecuadas para evaluar la depresión y la ansiedad en pacientes con depresión mayor. Fue más apropiado para evaluar niveles de angustia moderados que altos o bajos. La tabla de conversión se puede utilizar para una medición más precisa. Estos resultados pueden allanar el camino para métodos eficientes y sensibles para analizar la respuesta a los síntomas de depresión en la investigación y en la práctica clínica.(AU)

The Hospital Anxiety and Depression Scale (HADS) is a widely used screening tool for depression and anxiety in patients with medical ill-nesses. This study aimed to explore the psychometric properties of the HADS in screening for patients with major depression using item response theory method.A total of 460 patients with major depression completed the HADS. Rasch analyses were used to examine unidimensionality, item fit, local dependency, reliability, ordering of categories, differential item functioning (DIF)and targeting. The HADS showed a two-dimensional construct.All items fit the Rasch model.Three pairs of items showed mi-norbut inconsiderate local dependency.Both subscales had acceptable re-liability.None of the items displayed disordered categoriesor DIF.All items werewelltargeted, and participants with high and low levels of dis-tress were less targeted than those with moderate levels of distress.Finally, a conversion table to transform the raw scores into interval measures was generated. The HADS demonstrated adequate psychometric properties in assessing depression and anxiety in patients with major depression. It was more appropriate for assessing moderate than high or low levels of dis-tress.The conversion table can be used for more precise measurement.These results may pave the way for efficient and sensitive methods of ana-lyzing depression symptom response in research and in clinical practice.(AU)

The p53/ZEB1-PLD3 feedback loop regulates cell proliferation in breast cancer.

Breast cancer is the most prevalent cancer globally, endangering women's physical and mental health. Phospholipase D3 (PLD3) belongs to the phosphodiesterase family (PLD). PLD3 is related to insulin-mediated phosphorylation of the AKT pathway, suggesting that it may play a role in the occurrence and development of malignant tumors. This study may further explore the molecular mechanism of PLD3 inhibiting breast cancer cell proliferation. In this study, we demonstrated that PLD3 and miR-6796 are co-expressed in breast cancer. PLD3 can bind with CDK1 and inhibit its expression, leading to mitotic arrest and inhibiting breast cancer proliferation. Wild-type p53 regulates PLD3 and miR-6796 expression by competitively binding to the PLD3 promoter with ZEB1. DNMT3B, as the target gene of miR-6796, is recruited into the PLD3 promoter by combining with ZEB1 to regulate the DNA methylation of the PLD3 promoter and ultimately affect PLD3 and miR-6796 expression. In conclusion, we revealed the role and molecular mechanism of PLD3 and its embedded miR-6796 in breast cancer proliferation, providing clues and a theoretical foundation for future research and development of therapeutic targets and prognostic markers for breast cancer.

Correlation analysis of structural and functional changes in the carotid artery in patients with H-type hypertension using ultrasound radiofrequency.

OBJECTIVES: To perform a correlation analysis on the structural and functional changes of the carotid artery in patients with H-type hypertension. METHODS: Outpatients and inpatients with hypertension in our hospital between 2017 and 2018 were selected and divided into the H-type hypertension group (primary hypertension + plasma homocysteine ≥ 10 umol/l) (n = 30) and the simple hypertension group (primary hypertension + plasma Hcy < 10 umol/l) (n = 30) based on the plasma homocysteine (Hcy), and 30 healthy people were included in the control group. Thickness and stiffness parameters of the intima of the carotid artery (compliance coefficient [CC], stiffness index [ß], and pulse wave velocity [PWV]) were measured for all study participants using ultrasound radiofrequency signal-based quality intima-media thickness (QIMT) and quantitative arterial stiffness (QAS) for contrast analysis. RESULTS: Indexes such as QIMT, ß, and PWV of the carotid artery were significantly higher, and the CC was significantly lower in the H-type hypertension group and simple hypertension group than the control group (p < .05), and the difference was statistically significant; these indexes were significantly higher in the H-type hypertension group than in the simple hypertension group, and the CC was significantly lower than in the control group (p < .05), and the difference was statistically significant. CONCLUSIONS: Hypertension can accelerate structural and functional changes of the carotid artery intima, with these changes being more significant in H-type hypertension. The ultrasound radiofrequency technique can be used to quantitatively evaluate the structure and function of the carotid artery in patients with H-type hypertension.

Rasch analysis of the Chinese version of the clinically useful depression outcome scale in patients with major depressive disorder.

OBJECTIVE: To examine the psychometric properties of the Chinese version of the Clinically Useful Depression Outcome Scale (CUDOS) in the Chinese patients with major depressive disorder (MDD) using Rasch analysis. METHODS: The sample consisted of 283 patients with MDD (69% females). The Rasch model was applied to examine the overall fit of the Chinese version of CUDOS and the fit of the 18 items. Dimensionality, item-model fit, differential item functioning (DIF), reliability, ordering of response category and targeting were tested to examine the psychometric properties of the Chinese version of CUDOS. RESULTS: Rasch analysis demonstrated the unidimensionality of the Chinese version of CUDOS. Of the 18 items, three items (item 4, item 5, item 6) showed misfit in the model. After merging item 4 into item 3 and item 6 into item 5, the overall model fit improved. The person separation index (PSI) was 3.0 and the person reliability coefficient was 0.90. No evidence of significant DIF was found when associated with gender and age. No disordered category and threshold of the rating response were observed, which meant the response category setting was reasonable. The mean ability of person was - 0.53. CONCLUSION: The results suggested that the Chinese version of CUDOS has acceptable psychometric properties. In order to improve the quality and applicability of the Chinese version of CUDOS, the merging of item 4 into item 3 and item 6 into item 5 are suggested.

PAX5-miR-142 feedback loop promotes breast cancer proliferation by regulating DNMT1 and ZEB1.

BACKGROUND: Breast cancer is one of the most common malignancies occurred in female around the globe. Recent studies have revealed the crucial characters of miRNA and genes, as well as the essential roles of epigenetic regulation in breast cancer initiation and progression. In our previous study, miR-142-3p was identified as a tumor suppressor and led to G2/M arrest through targeting CDC25C. However, the specific mechanism is still uncertain. METHODS: We identified PAX5 as the upstream regulator of miR-142-5p/3p through ALGGEN website and verified by series of assays in vitro and in vivo. The expression of PAX5 in breast cancer was detected by qRT-PCR and western blot. Besides, bioinformatics analysis and BSP sequencing were performed to analyze the methylation of PAX5 promoter region. Finally, the binding sites of miR-142 on DNMT1 and ZEB1 were predicted by JASPAR, and proved by luciferase reporter assay, ChIP analysis and co-IP. RESULTS: PAX5 functioned as a tumor suppressor by positive regulation of miR-142-5p/3p both in vitro and in vivo. The expression of PAX5 was regulated by the methylation of its promoter region induced by DNMT1 and ZEB1. In addition, miR-142-5p/3p could regulate the expression of DNMT1 and ZEB1 through binding with their 3'UTR region, respectively. CONCLUSION: In summary, PAX5-miR-142-DNMT1/ZEB1 constructed a negative feedback loop to regulate the progression of breast cancer, which provided emerging strategies for breast cancer therapy.

CMTM7 inhibits breast cancer progression by regulating Wnt/ß-catenin signaling.

BACKGROUND: Breast cancer is the major cause of death in females globally. Chemokine-like factor like MARVEL transmembrane domain containing 7 (CMTM7) is reported as a tumor suppressor and is involved in epidermal growth factor receptor degradation and PI3K/AKT signaling in previous studies. However, other molecular mechanisms of CMTM7 remain unclear. METHODS: The expression level of CMTM7 in breast cancer cells and tissues was detected by qRT-PCR and western blot, and the methylation of CMTM7 promoter was detected by BSP sequencing. The effect of CMTM7 was verified both in vitro and in vivo, including MTT, colony formation, EdU assay, transwell assay and wound healing assay. The interaction between CMTM7 and CTNNA1 was investigated by co-IP assay. The regulation of miR-182-5p on CMTM7 and TCF3 on miR-182-5p was detected by luciferase reporter assay and ChIP analysis. RESULTS: This study detected the hypermethylation levels of the CMTM7 promoter region in breast cancer tissues and cell lines. CMTM7 was performed as a tumor suppressor both in vitro and in vivo. Furthermore, CMTM7 was a direct miR-182-5p target. Besides, we found that CMTM7 could interact with Catenin Alpha 1 (CTNNA1) and regulate Wnt/ß-catenin signaling. Finally, transcription factor 3 (TCF3) can regulate miR-182-5p. We identified a feedback loop with the composition of miR-182-5p, CMTM7, CTNNA1, CTNNB1 (ß-catenin), and TCF3, which play essential roles in breast cancer progression. CONCLUSION: These findings reveal the emerging character of CMTM7 in Wnt/ß-catenin signaling and bring new sights of gene interaction. CMTM7 and other elements in the feedback loop may serve as emerging targets for breast cancer therapy.

miR-92a-3p promotes breast cancer proliferation by regulating the KLF2/BIRC5 axis.

BACKGROUND: Breast cancer remains the most common malignancy in females around the world. Recently, a growing number of studies have focused on gene dysregulation. In our previous study, Krüppel-like factors (KLFs) were found to play essential roles in breast cancer development, among which KLF2 could function as a tumor suppressor. Nevertheless, the underlying molecular mechanism remains unclear. METHODS: miR-92a-3p was identified as the upstream regulator of KLF2 by starBase v.3.0. The regulation of KLF2 by miR-92a-3p was verified by a series of in vitro and in vivo assays. Further exploration revealed that Baculoviral IAP Repeat Containing 5 (BIRC5) was the target of KLF2. ChIP assay, dual-luciferase reporter analysis, quantitative real-time PCR, and western blot were performed for verification. RESULTS: miR-92a-3p functioned as a tumor promoter by inhibiting KLF2 by binding to its 3'-untranslated region (3'-UTR). In addition, KLF2 could transcriptionally suppress the expression of BIRC5. CONCLUSION: Collectively, our results uncovered the miR-92a-3p/KLF2/BIRC5 axis in breast cancer and provided a potential mechanism for breast cancer development, which may serve as promising strategies for breast cancer therapy.

Equating the PHQ-9 and GAD-7 to the HADS depression and anxiety subscales in patients with major depressive disorder.

OBJECTIVE: The present study aimed to equate the 9-item Patient Health Questionnaire (PHQ-9) and 7-item Generalized Anxiety Disorder Scale (GAD-7) to the Hospital Anxiety and Depression Scale (HADS) depression and anxiety subscales (HADS-D,HADS-A) respectively in patients with major depressive disorder (MDD) and generate crosswalks of raw scores. METHODS: As it is a single group design that adopts common-person equating method, a total of 460 patients with MDD completed the PHQ-9, GAD-7 and HADS at the same time. Rasch analysis was used to filter out invalid participants, investigate the psychometric properties of test items and participants, link the PHQ-9 and HADS-D as well as GAD-7 and HADS-A, and produce conversion tables respectively. The differences between original scores and converted scores were analyzed to validate the crosswalks. RESULT: 401 samples of depression part and 396 samples of anxiety part were left for final samples. Both the PHQ-9 / HADS-D and GAD-7 / HADS-A combined analysis adequately fit the unidimensional Rasch model, demonstrated acceptable reliability and item-person targeting and showed no disordering category. Slight differential item functioning across gender was found in item PHQ9 and item GAD6. The crosswalks were generated and verified to be validity. LIMITATIONS: The results might be restricted to patients with MDD recruited in a single mental health center. CONCLUSION: The PHQ-9, GAD-7 and HADS depression and anxiety subscales were successfully linked, producing conversion tables that could be used for directly converting raw score from one instrument to the other.

A Chinese adaptation of six items, self-report Hamilton Depression Scale: Factor structure and psychometric properties.

OBJECTIVES: The objective of this research was to verify the psychometric characteristics of the Chinese Adaptation of self-report HAMD-6. METHODS: Outpatients and inpatients who met the DSM-5 criterion for major depressive disorder (MDD) were evaluated by the Chinese self-report HAMD-6, seventeen items of Hamilton Depression Rating Scale (HAMD-17), Patient Health Questionnaire Depression Scale (PHQ-9) and Improved Clinical Global Impression Scale (iCGI-S). The internal consistency reliability, retest reliability, criterion validity and construct validity of the Chinese self-report HAMD-6 were tested. Pearson correlation coefficient was used to assess the correlativity between the total score and the item scores. By drawing the Receiver Operating Characteristics (ROC) curve, the best cut-off value, sensitivity and specificity of Chinese Adaptation self-report HAMD-6 were obtained. RESULTS: Cronbach's alpha coefficient of the Chinese self-report HAMD-6 was 0.91, and the intra-group correlation coefficient (ICC) of retest reliability was 0.81(P < 0.01). The Spearman correlation coefficients of the Chinese self-report HAMD-6, Chinese clinician version of HAMD-6, PHQ-9 and HAMD-17 were 0.86, 0.81 and 0.86, respectively (all P < 0.01). Results of the confirmatory factor analysis (CFA) supported a unidimensional construct. In addition, HAMD-17 ≤ 7 and iCGI-S= 1 were taken as the remission criteria for depression disorder, and the ROC curves of the Chinese self-report HAMD-6 were plotted with a cut-off value of 3/4, the specificity and sensitivity were 0.85/0.92 and 0.96/0.93 respectively. CONCLUSION: These results suggested that the abbreviated Chinese self-report HAMD-6 has good reliability and validity among the Chinese population. This study suggested that the remission cut-off value of the scale is 3/4.

Perceived uncertainty stress and its predictors among residents in China during the COVID-19 pandemic.

The prevalence of and risk factors for uncertainty stress among residents during the COVID-19 pandemic remain unclear. An online cross-sectional survey was conducted to explore and identify the risk factors for high perceived uncertainty stress among the general public in China during the COVID-19 outbreak. Information about the respondents' socioeconomic characteristics, knowledge of and attitudes towards COVID-19, perceived uncertainty stress, social capital, anxiety, and depressive symptoms was collected and analysed. Among the 1205 respondents, 45.3% (546) reported a high level of uncertainty stress. Multiple linear regression analysis indicated that anxiety (ß=3.871,P<0.001) and depression symptoms (ß=2.458, P<0.001), family residence (in towns or rural areas) (ß=0.947, P<0.001), lack of support for local epidemic control strategies (ß=1.253, P<0.001), worry about the pandemic (ß=1.191, P<0.001), and symptoms of weakness among family members (ß=1.525, P=0.002) were positively associated with perceived uncertainty stress. Cognitive social capital (ß=-0.883, P<0.001) and social networks (ß=-0.726, P<0.001) were negatively, but social participation (ß=0.714, P<0.001) was positively associated with perceived uncertainty stress. Our findings identify factors associated with a higher level of uncertainty stress and should be helpful in the consideration of effective policies and interventions for uncertainty stress during the initial phases of public health emergencies.

SIRT7-Induced PHF5A Decrotonylation Regulates Aging Progress Through Alternative Splicing-Mediated Downregulation of CDK2.

Dysregulation of protein posttranslational modification (PTM) can lead to a variety of pathological processes, such as abnormal sperm development, malignant tumorigenesis, depression, and aging process. SIRT7 is a NAD+-dependent protein deacetylase. Besides known deacetylation, SIRT7 may also have the capacity to remove other acylation. However, the roles of SIRT7-induced other deacylation in aging are still largely unknown. Here, we found that the expression of SIRT7 was significantly increased in senescent fibroblasts and aged tissues. Knockdown or overexpression of SIRT7 can inhibit or promote fibroblast senescence. Knockdown of SIRT7 led to increased pan-lysine crotonylation (Kcr) levels in senescent fibroblasts. Using modern mass spectrometry (MS) technology, we identified 5,149 Kcr sites across 1,541 proteins in senescent fibroblasts, and providing the largest crotonylome dataset to date in senescent cells. Specifically, among the identified proteins, we found SIRT7 decrotonylated PHF5A, an alternative splicing (AS) factor, at K25. Decrotonylation of PHF5A K25 contributed to decreased CDK2 expression by retained intron (RI)-induced abnormal AS, thereby accelerating fibroblast senescence, and supporting a key role of PHF5A K25 decrotonylation in aging. Collectively, our data revealed the molecular mechanism of SIRT7-induced k25 decrotonylation of PHF5A regulating aging and provide new ideas and molecular targets for drug intervention in cellular aging and the treatment of aging-related diseases, and indicating that protein crotonylation has important implications in the regulation of aging progress.

Psychometric Properties of the Chinese Version of the Clinically Useful Depression Outcome Scale for Patients with Major Depressive Disorder.

OBJECTIVE: This study aimed to evaluate the psychometric properties of the Chinese version of the Clinically Useful Depression Outcome Scale (CUDOS). METHODS: One hundred ninety patients with major depressive disorder (MDD) according to Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) criteria were recruited to the study. The English version of the CUDOS was translated into Chinese using a forward and backward translation method, which was according to the guidelines of adaptation and validation of instruments in cross-cultural health care research. The Chinese version of the CUDOS, the 17-item Hamilton Rating Scale for Depression (HRSD) and the improved Clinical Global Impression-Severity Scale (iCGI-S) were used to evaluate depressive symptoms in one hundred ninety patients with MDD. One week after the first evaluation, sixteen patients were selected randomly for a second assessment. Reliability and validity tests and receiver operating characteristic curves were performed. RESULTS: The internal consistency of the CUDOS was 0.95, and the split-half reliability coefficient of the CUDOS was 0.92. The correlation coefficient of the retest in sixteen patients was 0.77 (P < 0.01). There was a significant difference in the total score of the Chinese version of the CUDOS between the different levels of depression severity groups (P < 0.01). The ability of the CUDOS to identify patients in remission was high (area under ROC curve= 0.97). A cut-off score of 14/15 yielded 90.20% sensitivity and 93.60% specificity when iCGI-S=1. CONCLUSION: The Chinese version of the CUDOS is valuable as a brief and reliable instrument to assess depressive symptoms and clinical outcome. The findings suggest that the optimal cut-off score to identify patients in remission was 14/15.

Relationship between alcohol use, blood pressure and hypertension: an association study and a Mendelian randomisation study.

BACKGROUND: Past studies have found a strong relationship between alcohol drinking and human health. METHODS: In this study, we first tested the association of rs671 with alcohol use in 2349 participants in southeast China. We then evaluated the causal impact between alcohol use and cardiovascular traits through a Mendelian randomisation (MR) analysis. RESULTS: We found strong evidence for the association of rs671 in the ALDH2 gene with alcohol drinking (p=6.08×10-47; ORadj G=4.50, 95% CI 3.67 to 5.52). We found that female G carriers of rs671 had a higher proportion of non-drinkers than male G carriers (88.01% vs 38.70%). In non-drinkers, the female G allele frequency was higher than the male G allele frequency (71.1% vs 55.2%). MR analysis suggested that alcohol use had a causal effect on blood pressure (increasing 9.46 mm Hg for systolic blood pressure (p=9.67×10-4) and 7.50 mm Hg for diastolic blood pressure (p=9.62×10-5)), and on hypertension in men (p=0.011; OR =1.19, 95% CI 1.04 to 1.36) and in pooled samples (p=0.013; OR =1.20, 95% CI 1.04 to 1.39), but not in women. We did not observe a causal effect of alcohol use on body mass index and lipid levels; further studies are needed to clarify the non-causal relationship. CONCLUSIONS: Compared to never-drinkers, current and previous alcohol use had a causal effect on blood pressure and hypertension in pooled samples and in men. These results reflect Chinese culture which does not encourage women to drink.

Roles of reactive oxygen species (ROS) in the photocatalytic degradation of pentachlorophenol and its main toxic intermediates by TiO2/UV.

Pentachlorophenol (PCP) caused water quality problems owe to its past widespread application and stability, harmful to human health. Photocatalysis, which was mainly involved in the reactive oxygen species (ROS) reaction, has large potential as water treatment process. However, the roles of ROS on the degradation process of PCP are not yet clearly defined. The main objectives of this work were to investigate the roles of ROS involved in the whole degradation of PCP and main toxic intermediates and elucidate the degradation mechanisms. Tetrachloro-1,4-benzo/hydroquinone (TCBQ/TCHQ), trichlorohydroxy-1,4-benzoquinone (OH-TrCBQ) and 2,5-dichloro-3,6-dihydroxy-1,4-benzoquinone (OH-DCBQ) were identified as main intermediates. The roles of generated ROS including OH, O2- and H2O2 were systematically explored for the degradation of PCP and its main intermediates using radical quenchers. The results showed that, OH played the dominant role for the degradation of PCP, O2- played more contributing roles for the degradation of TCBQ, H2O2 exhibited major contribution for the degradation of OH-TrCBQ and OH-DCBQ. These results offered us an insight into the degradation mechanism of PCP involved with ROS. It can also serve as the basis for controlling and blocking the generation of highly toxic substances through regulating the ROS generation during the PCP degradation.

Identification of amentoflavone as a potent highly selective PARP-1 inhibitor and its potentiation on carboplatin in human non-small cell lung cancer.

BACKGROUND: Nuclear protein poly (ADP-ribose) polymerase-1 (PARP-1) is a key enzyme in the repair of DNA and is a promising target in the development of chemosensitizers. This study first investigated the inhibitory effects of amentoflavone (AMF) and its derivatives on PARP-1 and the potentiation of AMF on carboplatin (CBP) in non-small cell lung cancer (NSCLC). PURPOSE: This study aims to evaluate the inhibitory effect of AMF against PARP-1 and its potentiation on CBP in lung cancer both in vitro and in vivo. STUDY DESIGN: The inhibitory effect of AMF on PARP-1 was investigated using molecular docking and cell-free model of PARP-1 assay. Its potentiation on CBP in lung cancer was also evaluated. METHODS: Fluorescence resonance energy transfer assay was used to detect the inhibitory effects of AMF and its analogues on PARP-1. Molecular docking was employed to predict the binding mode of AMF and PARP-1. MTT assay, isobologram analysis, Hoechst staining, and Annexin V-PI double staining were used to confirm the potentiation of AMF on CBP in vitro. siRNA (PARP-1)-A549 cells were used to reveal the action target of AMF. Western blot analysis, immunohistochemistry, and Tunnel assay were employed to evaluate the potentiation of AMF on CBP in A549 xenograft mice. RESULTS: AMF and its analogues exerted excellent inhibitory effects on PARP-1 with IC50 values ranging from 0.198  µM to 0.409  µM. Docking experiment showed that AMF can stably bind to PARP-1 with a comparable binding energy to olaparib. AMF can decrease the expression of PAR induced by H2O2in vitro. AMF synergistically increased the CBP anti-proliferative effect in A549. However, its potentiation nearly disappeared when the cells were transfected with siRNAs against PARP-1. Oral administration of AMF (100  mg/kg), combined with CBP, remarkably inhibited A549 tumor growth and ki67 expression, and increased apoptosis compared with CBP-alone group. CONCLUSION: All results suggest that AMF can be a potential PARP-1 inhibitor and a candidate adjuvant agent to boost the anticancer effect of CBP in NSCLC.

Dynamic Tracking of Highly Toxic Intermediates in Photocatalytic Degradation of Pentachlorophenol by Continuous Flow Chemiluminescence.

Photocatalytic degradation is a powerful technique for the decomposition of pollutants. However, toxic intermediates might be generated which have become a great concern recently. In the present work, a continuous flow chemiluminescence (CFCL) method was developed for dynamic monitoring of toxic intermediates generated in the photocatalytic degradation of pentachlorophenol (PCP). Among the main intermediates, tetrachloro-1,4-benzoquinone (TCBQ) and trichlorohydroxy-1,4-benzoquinone (OH-TrCBQ) showed higher or similar toxicity to PCP. As both TCBQ and OH-TrCBQ can produce chemiluminescence (CL) in the presence of H2O2, a CFCL system was established for the dynamic tracking of the two toxic intermediates. A PCP/TiO2 suspension was irradiated in a photoreactor, pumped continuously into a detection cell, and mixed with H2O2 to produce CL. The time-dependent CL response displayed two distinctive peaks at pH 7, which were attributed to the generation of OH-TrCBQ and TCBQ, respectively, by comparing with their changes measured by high-performance liquid chromatography (HPLC). Furthermore, the CL response curve of PCP/TiO2 suspension showed a pattern very similar to their bacteria inhibition. Therefore, the CFCL could be used as a simple and low-cost method for online monitoring of TCBQ and OH-TrCBQ to ensure complete removal of not only PCP but also highly toxic degradation intermediates.

HMGB1 regulates P-glycoprotein expression in status epilepticus rat brains via the RAGE/NF-κB signaling pathway.

Overexpression of P-glycoprotein (P-gp) in the brain is an important mechanism involved in drug­resistant epilepsy (DRE). High-mobility group box 1 (HMGB1), an inflammatory cytokine, significantly increases following seizures and may be involved in upregulation of P­gp. However, the underlying mechanisms remain elusive. The aim of the present study was to evaluate the role of HMGB1 and its downstream signaling components, receptor for advanced glycation end­product (RAGE) and nuclear factor­κB (NF­κB), on P­gp expression in rat brains during status epilepticus (SE). Small interfering RNA (siRNA) was administered to rats prior to induction of SE by pilocarpine, to block transcription of the genes encoding HMGB1 and RAGE, respectively. An inhibitor of NF­κB, pyrrolidinedithiocarbamic acid (PDTC), was utilized to inhibit activation of NF­κB. The expression levels of HMGB1, RAGE, phosphorylated­NF­κB p65 (p­p65) and P­gp were detected by western blotting. The relative mRNA expression levels of the genes encoding these proteins were measured using reverse transcription­quantitative polymerase chain reaction and the cellular localization of the proteins was determined by immunofluorescence. Pre­treatment with HMGB1 siRNA reduced the expression levels of RAGE, p­p65 and P­gp. PDTC reduced the expression levels of P­gp. These findings suggested that overexpression of P­gp during seizures may be regulated by HMGB1 via the RAGE/NF­κB signaling pathway, and may be a novel target for treating DRE.

Pregnane X Receptor Not Nuclear Factor-kappa B Up-regulates P-glycoprotein Expression in the Brain of Chronic Epileptic Rats Induced by Kainic Acid.

Drug-resistance epilepsy (DRE) is attributed to the brain P-glycoprotein (P-gp) overexpression. We previously reported that nuclear factor-kappa B (NF-κB) played a critical role in regulating P-gp expression at the brain of the acute seizure rats. This study was extended further to investigate the interaction effect of NF-κB and pregnane X receptor (PXR) on P-gp expression at the brain of chronic epileptic rats treated with carbamazepine (CBZ). The chronic epileptic models were induced by the micro-injection of kainic acid (KA) into rats' hippocampus. Subsequently, the successful models were treated with different intervention agents of CBZ; PMA(a non-specific PXR activity inhibitor) or PDTC(a specific NF-κB activity inhibitor) respectively. The expression levels of P-gp and its encoded gene mdr1a/b were significantly up-regulated on the brain of KA-induced chronic epilepsy rats or the epilepsy rats treated with CBZ for 1 week, meanwhile with a high expression of PXR. The treatment of PMA dramatically reduced both PXR and P-gp expressions at the protein and mRNA levels in the chronic epilepsy brain. By compared to the epilepsy model group, the P-gp expression was not markedly attenuated by the inhibition of NF-κB activity with PDTC treatment, nevertheless with a decrease of NF-κB expression in this intervention group. Higher levels of proinflammatory cytokines(IL-1ß, IL-6, TNF-α) were found both in the brain tissue and the serum in the epilepsy rats of each group. There was a declined trend of the pro-inflammatory cytokines expression of the PDTC treatment group but with no statistical significance. This study demonstrates for the first time that P-gp up-regulation is due to increase PXR expression in the chronic phase of epilepsy, differently from that NF-κB signaling may induce the P-gp expression in the acute seizure phase. Our results offer insights into the mechanism underlying the development of DRE using or not using CBZ treatment.