Epidemiology of acute hepatitis C and hepatitis C virus-related cirrhosis in reproductive-age women, 1990-2019: An analysis of the Global Burden of Disease study.

Background: The current study uniquely focuses on the global incidence and temporal trends of acute hepatitis C (AHC) and hepatitis C virus (HCV)-related cirrhosis among women of reproductive age (15-49 years) from 1990-2019. The risk of vertical transmission and adverse perinatal outcomes associated with HCV infection underscores the importance of prioritising these women in HCV prevention efforts. Methods: Leveraging the Global Burden of Disease 2019 data, we calculated age-standardised incidence rates (ASIR) and assessed temporal trends via the average annual percent change from joinpoint regression. The age-period-cohort model was employed to understand further the effects of age, period, and birth cohort. Results: Over the 30 years, global incidences of AHC and HCV-related cirrhosis in reproductive-age women increased by 46.45 and 72.74%, respectively. The ASIR of AHC was highest in low sociodemographic index regions but showed a declining trend. Conversely, the ASIR of HCV-related cirrhosis displayed unfavourable trends in low, low-middle, and high sociodemographic index regions. Special attention is necessary for sub-Saharan Africa, high-income North America, Eastern Europe, and Central Asia due to their high incidence rates or increasing trends of AHC and HCV-related cirrhosis. Notably, the age-period-cohort model suggests a recent resurgence in AHC and HCV-related cirrhosis risk. Conclusions: The current study is the first to thoroughly evaluate the trends of AHC and HCV-related cirrhosis among reproductive-age women, shedding light on previously unexplored aspects of HCV epidemiology. Our findings identify critical areas where health care systems must adapt to the changing dynamics of HCV infection. The detailed stratification by region and nation further enables the development of localised prevention and treatment strategies.

Circularizing PET-G Multimaterials: Life Cycle Assessment and Techno-Economic Analysis.

The recycling of multimaterials such as payment or access cards poses significant challenges. Building on previous experimental work demonstrating the feasibility of chemically recyclable payment cards made from glycol-modified poly(ethylene terephthalate) (PET-G), we use life cycle assessment and techno-economic analysis to investigate two chemical recycling scenarios and evaluate their potential environmental and economic benefits. Recovering all components from the depolymerized products (Scenario 1) achieves substantial environmental benefits across most categories, reducing global warming by up to 67% compared to only recovering major components (Scenario 2). However, the environmental benefits in Scenario 1 incur 69% higher total annualized costs, causing its profitability to be dependent on a minimum selling price of £13.4/kg for cyclohexanedimethanol and less than a 10% discount rate. In contrast, Scenario 2 is less sensitive to discount rate variation and thus a lower risk and more economically feasible option, albeit less environmentally sustainable.

Economical Production of Phenazine-1-carboxylic Acid from Glycerol by Pseudomonas chlororaphis Using Cost-Effective Minimal Medium.

Phenazine compounds are widely used in agricultural control and the medicine industry due to their high inhibitory activity against pathogens and antitumor activity. The green and sustainable method of synthesizing phenazine compounds through microbial fermentation often requires a complex culture medium containing tryptone and yeast extract, and its cost is relatively high, which greatly limits the large-scale industrial production of phenazine compounds by fermentation. The aim of this study was to develop a cost-effective minimal medium for the efficient synthesis of phenazine compounds by Pseudomonas chlororaphis. Through testing the minimum medium commonly used by Pseudomonas, an ME medium for P. chlororaphis with a high production of phenazine compounds was obtained. Then, the components of the ME medium and the other medium were compared and replaced to verify the beneficial promoting effect of Fe2+ and NH4+ on phenazine compounds. A cost-effective general defined medium (GDM) using glycerol as the sole carbon source was obtained by optimizing the composition of the ME medium. Using the GDM, the production of phenazine compounds by P. chlororaphis reached 1073.5 mg/L, which was 1.3 times that achieved using a complex medium, while the cost of the GDM was only 10% that of a complex medium (e.g., the KB medium). Finally, by engineering the glycerol metabolic pathway, the titer of phenazine-1-carboxylic acid reached the highest level achieved using a minimum medium so far. This work demonstrates how we systematically analyzed and optimized the composition of the medium and integrated a metabolic engineering method to obtain the most cost-effective fermentation strategy.

Economical downstream processing of microbial polyunsaturated fatty acids.

Polyunsaturated fatty acids (PUFAs) are important nutrients for humans and animals. Microorganisms, such as yeast, filamentous fungi, and microalgae, have successfully been modified to produce PUFAs. Apart from strain improvement and fermentation optimization, efficient and cost-effective downstream processing will determine whether production can advance from the laboratory to the factory.

[Characteristics of climate change and its impacts on water resources in Qilian Mountains, China]. / ç¥è¿žå±±æ°”å€™å˜åŒ–ç‰¹å¾åŠå ¶å¯¹æ°´èµ„æºçš„å½±å“.

Qilian Mountains, is an important ecological function area, an important ecological security barrier, the river runoff region in Northwest China, as well as a sensitive area to global climate change and fragile area of ecological environment. The ecological environment in this area played an important role in the economic development of Northwest China. Based on the observation data of temperature and precipitation in Qilian Mountains, MOD10A2 snow products and the flow data of Shiyang River, Heihe River and Shule River, we systematically analyzed the characteristics of climate change from 1961 to 2020, and the impacts of climate change on water resources under the scenario of climate warming. The results showed that, from 1961 to 2020, the annual average temperature increased significantly, with the rate reaching 0.39 ℃·(10 a)-1. The warming rate was the highest in the western part of Qilian Mountains, followed by the middle and eastern regions. The warming trend was the strongest in winter and the lowest in spring. The average temperature changed abruptly in 1997. The annual average precipitation increased with flucturation, with a rate of 10 mm·(10 a)-1, which increased most obviously in the middle of Qilian Mountains. After 2004, it entered a rainy period, with a warm and humid trend. The precipitation in the four seasons showed an increasing trend and the increase of precipitation in summer contributed the most to the annual precipitation. Annual precipitation was dominated by interannual scale change, and the contribution rate of 2.8-year was approximately 64.3%. The snow cover of Qilian Mountains was obviously affected by temperature and snowfall, which was negatively correlated with summer temperature and positively correlated with snowfall. From 2016 to 2020, the temperature increase had slowed down in Qilian Mountains, the snowfall had increased, and the snow cover tended to increase. After 2000, the temperature and precipitation increased more obviously, the meltwater from glacier and snow increased, the mountainous runoff of Shiyang River, Heihe River and Shule River had an increasing trend. Our findings are of great significance to the construction of ecological civilization and coping with climate change in Qilian Mountains.

Integrating the impact of large-scale hydraulic engineering with a sustainable groundwater development strategy: A case study of Zhengzhou City, China.

With the rapid growth of China's economy, the increase in water demand has threatened the sustainable development of groundwater. Construction of the South-to-North Water Diversion Project alleviated this problem. Zhengzhou, with a large population and high-intensity energy consumption, is a water-receiving city of the South-to-North Water Diversion Central Line Project (CLP). A series of ecological risks caused by the excessive exploitation of groundwater have been exposed. It is urgent to strengthen the assessment and management of groundwater to ensure sustainable development. In this study, the multi-criteria decision analysis (MCDA) underpinned the assessment of the sustainable groundwater development (ASGD) framework. Eight assessment factors were established based on the resource supply function (RSF) and eco-environment stability function (ESF). The novelty of this study lies in the integration of ASGD results with the impact of the CLP on the evolution of groundwater levels in Zhengzhou. Thus, more comprehensive and scientific management suggestions for groundwater development in Zhengzhou were obtained. GIS technology was integrated with the ASGD framework to identify five visualized areas: centralized groundwater supply area (8.61%), decentralized groundwater supply area (27.91%), vulnerable eco-environment area (14.34%), recharge protection area (45.67%), and unsuitable exploitation area (3.47%). The CLP changed the groundwater evolution pattern in Zhengzhou. The results showed that the operation of the CLP effectively slowed the decline in groundwater levels, thus confirming that the CLP has a positive impact on the rational utilization of groundwater. The disuse of two groundwater sources (G1 and G9) were able to enhance sustainable groundwater development. Meanwhile, five groundwater sources in the plain area proved unsuitable. Overall, this study provides a scientific basis for groundwater management in Zhengzhou City, while generating new ideas for sustainable groundwater development in cities affected by large-scale hydraulic projects worldwide.

Assessment of groundwater sustainable development considering geo-environment stability and ecological environment: a case study in the Pearl River Delta, China.

Groundwater resources have an important impact on the geo-environment and ecological environment. The exploitation of groundwater resources may induce geo-environmental issues and has a negative impact on the ecological environment. The assessment of groundwater sustainable development can provide reasonable suggestions for the management of groundwater resources in coastal cities. In this study, an assessment method for groundwater sustainable development based on the resource supply function, geo-environment stability function, and ecological environment function was provided. Considering the groundwater quantity and quality; the vulnerability of karst collapse, land subsidence, and seawater intrusion; and the distribution of groundwater-dependent ecosystems (GDEs) and soil erosion, the groundwater in the Pearl River Delta was divided into concentrated groundwater supply area (21.97%) and decentralized groundwater supply area (48.22%), ecological protection area (20.77%), vulnerable geo-environment area (8.94%), and unsuitable to exploit groundwater area (0.10%). ROC curve and single-indicator sensitivity analysis were applied in the assessment of geo-environment vulnerability, and the results showed that the VW-AHP model effectively adjusted the weights of the indicators so that the assessment results were more in line with the actual situation in the Pearl River Delta, and the accuracy of the VW-AHP model was higher than that of the AHP model. This study provides a scientific basis for groundwater management in the Pearl River Delta and an example for the assessment of groundwater sustainable development in coastal cities.

Assessment of Non-invasive Markers for the Prediction of Esophageal Variceal Hemorrhage.

Background: Esophageal variceal (EV) hemorrhage is a life-threatening consequence of portal hypertension in cirrhotic patients. Screening upper endoscopy and endoscopic variceal ligation to identify and treat EVs have contraindications, complications, and high costs. We sought to identify non-invasive tests (NITs) as alternatives to endoscopic EV screening. Methods: In this case-control study, we retrospectively analyzed 286 cirrhotic patients treated for EVs at the Second People's Hospital of Fuyang City, China from January to December 2019. We applied ROC curve analysis to assess the accuracy of various NITs in predicting EV hemorrhage. Results: There were significant differences between the hemorrhage and non-hemorrhage groups in median serum albumin (ALB) (p < 0.001), median bilirubin (TBIL) (p < 0.046), prothrombin (PT) time (p < 0.001), Golgi protein 73 (GP73; p = 0.012) and Child-Pugh (C-P) scores (p < 0.001). For ALB (cutoff <33.2g/L), PT time (cutoff > 14.2 seconds), GP73 (cutoff > 126.4 ng/ml), and C-P scores, the areas under the ROC curves (AUCs) were 73.4% (95% CI: 67.5-79.2), 68.6% (95% CI: 62.4-74.8), 62.2% (95% CI: 52.8-71.5) and 69.8% (95%CI: 63.8-75.8), respectively, with corresponding sensitives of 71.5, 59.8, 69.8, and 92.2% and specificities of 65.6%, 70.1%, 56.5%, and 38.6%. When ALB was combined with GP73, the AUC was 74.3% (95% CI: 66.1-82.5) with a sensitivity of 65.1% and specificity of 76.5%. When ALB, PT, and C-P scores were combined, the AUC was 76.5% (95% CI: 70.9-82.1) with a sensitivity of 79.5% and specificity of 64.3%. When ALB, PT, GP73, and C-P scores were combined, the AUC was 75.2% (95% CI: 67.3-83.1) with a sensitivity of 54.0% and specificity of 86.9%. Conclusion: ALB, TBIL, GP73, and C-P scores, may be used to predict EV hemorrhage in cirrhotic patients. The combination of multiple NITs is better than a single index and can increase diagnostic performance.

A Cost-Effective and Non-Invasive pfeRNA-Based Test Differentiates Benign and Suspicious Pulmonary Nodules from Malignant Ones.

The ability to differentiate between benign, suspicious, and malignant pulmonary nodules is imperative for definitive intervention in patients with early stage lung cancers. Here, we report that plasma protein functional effector sncRNAs (pfeRNAs) serve as non-invasive biomarkers for determining both the existence and the nature of pulmonary nodules in a three-stage study that included the healthy group, patients with benign pulmonary nodules, patients with suspicious nodules, and patients with malignant nodules. Following the standards required for a clinical laboratory improvement amendments (CLIA)-compliant laboratory-developed test (LDT), we identified a pfeRNA classifier containing 8 pfeRNAs in 108 biospecimens from 60 patients by sncRNA deep sequencing, deduced prediction rules using a separate training cohort of 198 plasma specimens, and then applied the prediction rules to another 230 plasma specimens in an independent validation cohort. The pfeRNA classifier could (1) differentiate patients with or without pulmonary nodules with an average sensitivity and specificity of 96.2% and 97.35% and (2) differentiate malignant versus benign pulmonary nodules with an average sensitivity and specificity of 77.1% and 74.25%. Our biomarkers are cost-effective, non-invasive, sensitive, and specific, and the qPCR-based method provides the possibility for automatic testing of robotic applications.

Microplastics in seawater and two sides of the Taiwan Strait: Reflection of the social-economic development.

Microplastic abundance, distribution and source characteristics were investigated for the surface seawaters from the Taiwan Strait as well as those of sediments along its west and east coasts. The microplastic abundances were in the range of 28-208 (mean 90) and 10-246 (mean 69) items/kg (d.w.) along the west and east coasts respectively. The higher microplastic abundance on the west coast might be related to the different local economic development, population, land-use and other human activities. Combined with microplastic pollution and socio-economic development, regression analysis results showed that urbanization level is negatively correlated with foams while positively correlated with fibers. This study, as the first report of microplastics in the Taiwan Strait, suggested further research on microplastics cross-strait transportation and the relationship with economic developments.

Antidelaminating, Thermally Stable, and Cost-Effective Flexible Kapton Platforms for Nitrate Sensors, Mercury Aptasensors, Protein Sensors, and p-Type Organic Thin-Film Transistors.

The inkjet printing of metal electrodes on polymer films is a desirable manufacturing process due to its simplicity but is limited by the lack of thermal stability and serious delaminating flaws in various aqueous and organic solutions. Kapton, adopted worldwide due to its superior thermal durability, allows the high-temperature sintering of nanoparticle-based metal inks. By carefully selecting inks (Ag and Au) and Kapton substrates (Kapton HN films with a thickness of 135 µm and a thermal resistance of up to 400 °C) with optimal printing parameters and simplified post-treatments (sintering), outstanding film integrity, thermal stability, and antidelaminating features were obtained in both aqueous and organic solutions without any pretreatment strategy (surface modification). These films were applied in four novel devices: a solid-state ion-selective (IS) nitrate (NO3-) sensor, a single-stranded DNA (ssDNA)-based mercury (Hg2+) aptasensor, a low-cost protein printed circuit board (PCB) sensor, and a long-lasting organic thin-film transistor (OTFT). The IS NO3- sensor displayed a linear sensitivity range between 10-4.5 and 10-1 M (r2 = 0.9912), with a limit of detection of 2 ppm for NO3-. The Hg2+ sensor exhibited a linear correlation (r2 = 0.8806) between the change in the transfer resistance (RCT) and the increasing concentration of Hg2+. The protein PCB sensor provided a label-free method for protein detection. Finally, the OTFT demonstrated stable performance, with mobility values in the linear (µlin) and saturation (µsat) regimes of 0.0083 ± 0.0026 and 0.0237 ± 0.0079 cm2 V-1 S-1, respectively, and a threshold voltage (Vth) of -6.75 ± 3.89 V.

Spatial analysis, coupling coordination, and efficiency evaluation of green innovation: A case study of the Yangtze River Economic Belt.

Green innovation is an important driving force to promote the sustainable development of urban society and economy. This paper constructs an evaluation index system containing social undesirable outputs, and uses the Super-SBM model and the Malmquist-Luenberger index to evaluate green innovation efficiency in 42 cities along the Yangtze River Economic Belt from 2013 to 2017. Additionally, spatial autocorrelation analysis is used to study the spatial correlation of green innovation efficiency. Finally, the coupling coordination degree model is used to study the coupling coordination degree between green innovation efficiency and high-tech industries. The following results were obtained. (1) The average value of green innovation efficiency increased from 1.0446 to 1.0987, and the annual average growth rate of total factor productivity of green innovation was 1.1%. (2) Green innovation efficiency of the Yangtze River Economic Belt had a significant spatial positive correlation, but the types of agglomeration among cities in different sections of the Yangtze River were quite different. (3) The coupling coordination degree between green innovation efficiency and the development level of high-tech industries in the cities of the Yangtze River Economic Belt was in the basic coordination stage. Based on the research results, we suggest that cities in this belt further enhance the interactive relationship between green innovation and economic development and promote the coordinated development of economy and society.

Spatial heterogeneity can lead to substantial local variations in COVID-19 timing and severity.

Standard epidemiological models for COVID-19 employ variants of compartment (SIR or susceptible-infectious-recovered) models at local scales, implicitly assuming spatially uniform local mixing. Here, we examine the effect of employing more geographically detailed diffusion models based on known spatial features of interpersonal networks, most particularly the presence of a long-tailed but monotone decline in the probability of interaction with distance, on disease diffusion. Based on simulations of unrestricted COVID-19 diffusion in 19 US cities, we conclude that heterogeneity in population distribution can have large impacts on local pandemic timing and severity, even when aggregate behavior at larger scales mirrors a classic SIR-like pattern. Impacts observed include severe local outbreaks with long lag time relative to the aggregate infection curve, and the presence of numerous areas whose disease trajectories correlate poorly with those of neighboring areas. A simple catchment model for hospital demand illustrates potential implications for health care utilization, with substantial disparities in the timing and extremity of impacts even without distancing interventions. Likewise, analysis of social exposure to others who are morbid or deceased shows considerable variation in how the epidemic can appear to individuals on the ground, potentially affecting risk assessment and compliance with mitigation measures. These results demonstrate the potential for spatial network structure to generate highly nonuniform diffusion behavior even at the scale of cities, and suggest the importance of incorporating such structure when designing models to inform health care planning, predict community outcomes, or identify potential disparities.

Predictors of caregiver burden in patients with neurologic Wilson disease.

OBJECTIVES: Caregiver burden in neurologic Wilson disease (NWD) has received little attention. We investigated predictors of caregiver burden in Chinese NWD patients. METHODS: Participants in this retrospective study were NWD patients admitted to The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from 1 August to 31 December 2019. Sociodemographic information was recorded for caregivers and NWD patients. Caregiver burden was evaluated using the Caregiver Burden Inventory (CBI). Cognitive impairment, functional problems, depression and anxiety were evaluated by professional interviewers. Path analysis was used to evaluate predictors of CBI scores. RESULTS: Sixty NWD patients were enrolled (mean age: 21.35 ± 4.89 years; mean NWD duration: 7.85 ± 3.11 years). The mean CBI score was 52.00 ± 17.16. Care duration had a significant direct effect on CBI score after controlling for confounders (r = 0.493). Cognitive impairment (r = -0.426), functional problems (r = 0.581), depression (r = 0.349) and anxiety (r = 0.317) had significant indirect effects on CBI score. CONCLUSION: Caregivers of NWD patients may experience a medium level of caregiver burden. NWD duration, cognitive impairment, functional problems, depression and anxiety in NWD patients may be useful predictors of caregiver burden.

Occurrence and Risk Assessment of PAHs in Surface Sediments from Western Arctic and Subarctic Oceans.

In the fourth Chinese National Arctic Research Expedition (from July to September, 2010), 14 surface sediment samples were collected from the Bering Sea, Chukchi Sea, and Canadian Basin to examine the spatial distributions, potential sources, as well as ecological and health risk assessment of polycyclic aromatic hydrocarbons (PAHs). The ∑PAH (refers to the sum of 16 priority PAHs) concentration range from 27.66 ng/g to 167.48 ng/g (dry weight, d.w.). Additionally, the concentrations of ∑PAH were highest in the margin edges of the Canadian Basin, which may originate from coal combustion with an accumulation of Canadian point sources and river runoff due to the surface ocean currents. The lowest levels occurred in the northern of Canadian Basin, and the levels of ∑PAH in the Chukchi Sea were slightly higher than those in the Being Sea. Three isomer ratios of PAHs (Phenanthrene/Anthracene, BaA/(BaA+Chy), and LMW/HMW) were used to investigate the potential sources of PAHs, which showed the main source of combustion combined with weaker petroleum contribution. Compared with four sediment quality guidelines, the concentrations of PAH are much lower, indicating a low potential ecological risk. All TEQPAH also showed a low risk to human health. Our study revealed the important role of the ocean current on the redistribution of PAHs in the Arctic.

Geochemical Assessment and Spatial Analysis of Heavy Metals in the Surface Sediments in the Eastern Beibu Gulf: A Reflection on the Industrial Development of the South China Coast.

The Beibu Gulf (also named the Gulf of Tonkin), located in the northwest of the South China Sea, is representative of a bay suffering from turbulence and contamination associated with rapid industrialization and urbanization. In this study, we aim to provide the novel baseline levels of heavy metals for the research area. Concentrations of five heavy metals (i.e., Cu, Pb, Zn, Cd and Cr) were determined in surface sediments from 35 sites in the eastern Beibu Gulf. The heavy metal content varied from 6.72 to 25.95 mg/kg for Cu, 16.99 to 57.98 mg/kg for Pb, 73.15 to 112.25 mg/kg for Zn, 0.03 to 0.12 mg/kg for Cd, and 20.69 to 56.47 mg/kg for Cr, respectively. With respect to the Chinese sediment quality criteria, sediments in the eastern Beibu Gulf have not been significantly affected by coastal metal pollutions. The results deduced from the geoaccumulation index (Igeo) showed that the study area has been slightly polluted by Pb, which might be caused by non-point sources. Relatively high concentrations of Cu, Pb and Cd were found around the coastal areas of Guangxi province, the Leizhou Peninsula and the northwest coast of Hainan Island, whereas the highest concentrations of Zn and Cr were found on the northwest coast of Hainan Island. Spatial distribution patterns of the heavy metals showed that bioavailable fractions of Pb were higher than in the residual fractions, while Cu and Cd concentrations in exchangeable and carbonate fractions were relatively higher than those in the bioavailable fractions. Hierarchical clustering analysis suggested that the sampling stations could be separated into three groups with different geographical distributions. Accompanying their similar spatial distribution in the study area, significant correlation coefficients among Cu, Cd and Pb were also found, indicating that these three metals might have had similar sources. Overall, the results indicated that the distribution of these heavy metals in the surface sediments collected from the Beibu Gulf was complex.

Assessment of iodine uptake by pancreatic cancer following chemotherapy using dual-energy CT.

Pancreatic cancer remains a major health problem, and only less than 20% of patients have resectable disease at the time of initial diagnosis. Systemic chemotherapy is often used in the patients with borderline resectable, locally advanced unresectable disease and metastatic disease. CT is often used to assess for therapeutic response; however, conventional imaging including CT may not correctly reflect treatment response after chemotherapy. Dual-energy (DE) CT can acquire datasets at two different photon spectra in a single CT acquisition, and permits separating materials and extract iodine by applying a material decomposition algorithm. Quantitative iodine mapping may have an added value over conventional CT imaging for monitoring the treatment effects in patients with pancreatic cancer and potentially serve as a unique biomarker for treatment response. In this pictorial essay, we will review the technique for iodine quantification of pancreatic cancer by DECT and discuss our observations of iodine quantification at baseline and after systemic chemotherapy with conventional cytotoxic agents, and illustrate example cases.

Distribution, sources and contamination assessment of heavy metals in surface sediments of the South Yellow Sea and northern part of the East China Sea.

Surface sediment samples collected from the South Yellow Sea and northern part of the East China Sea during spring and autumn, respectively, were analyzed for grain size, aluminum, and heavy metals (Cr, Ni, Cu, Zn, and Pb) to evaluate heavy metal levels and the contamination status. The results showed that all of the heavy metal concentrations met the standard criteria of the Chinese National Standard Criteria for Marine Sediment Quality. Both the EFs and a multivariate analysis (PCA) indicated that Cr, Ni, Cu, and Zn were mainly from natural contributions, while Pb was influenced by anthropogenic inputs, especially during autumn. The geoaccumulation index of Pb near the mouth of the Yangtze River suggested that the pollution degree in autumn was heavier than that in spring, which might be caused by the greater river discharge in summer and more heavy metal adsorption with finer grain sizes.

Gene Methylation and Cytological Atypia in Random Fine-Needle Aspirates for Assessment of Breast Cancer Risk.

Methods to determine individualized breast cancer risk lack sufficient sensitivity to select women most likely to benefit from preventive strategies. Alterations in DNA methylation occur early in breast cancer. We hypothesized that cancer-specific methylation markers could enhance breast cancer risk assessment. We evaluated 380 women without a history of breast cancer. We determined their menopausal status or menstrual cycle phase, risk of developing breast cancer (Gail model), and breast density and obtained random fine-needle aspiration (rFNA) samples for assessment of cytopathology and cumulative methylation index (CMI). Eight methylated gene markers were identified through whole-genome methylation analysis and included novel and previously established breast cancer detection genes. We performed correlative and multivariate linear regression analyses to evaluate DNA methylation of a gene panel as a function of clinical factors associated with breast cancer risk. CMI and individual gene methylation were independent of age, menopausal status or menstrual phase, lifetime Gail risk score, and breast density. CMI and individual gene methylation for the eight genes increased significantly (P < 0.001) with increasing cytological atypia. The findings were verified with multivariate analyses correcting for age, log (Gail), log (percent density), rFNA cell number, and body mass index. Our results demonstrate a significant association between cytological atypia and high CMI, which does not vary with menstrual phase or menopause and is independent of Gail risk and mammographic density. Thus, CMI is an excellent candidate breast cancer risk biomarker, warranting larger prospective studies to establish its utility for cancer risk assessment. Cancer Prev Res; 9(8); 673-82. ©2016 AACR.

[Construction of human Bcl-6 3'UTR reporter vector and expression vector and their functional assessment].

OBJECTIVE: To observe the direct regulation of miR-127 on Bcl-6 and the effect of Bcl-6 in rescuing miR-127-induced cell cycle and cell growth inhibition. METHODS: The 3'UTR and coding region of human bcl-6 gene were amplified by PCR and cloned into pcDNA3.0-Luc and pcDNA3.0-Flag vectors, respectively. Mutations were introduced into the seed sequences of the predicted miR-127 target sites within the Bcl-6 3'UTR using recombinant PCR. Luciferase assay was used to verify the direct targeted regulation of miR-127 on Bcl-6. In HepG2 cell models with overexpression or knockdown of miR-12, the changes of cell cycle and cell growth were investigated after transfection with the constructed vectors. RESULTS: The recombinant plasmids were successfully obtained as confirmed by double digestion and sequence identification. Luciferase assay showed that in 293T and HepG2 cells, miR-127 inhibited the activation of wild-type Bcl-6 3'UTR reporter vector but not mutated Bcl-6 3'UTR vector. Overexpression of miR-127 induced cell cycle arrest at G(2)/M phase and suppressed the growth of HepG2 cells, and these effects were reversed by Bcl-6 overexpression. CONCLUSION: We successfully cloned wild-type and mutated 3'UTR reporter vectors and expression vector of bcl-6 gene and confirmed their biological functions.