Importance:Lung cancer (LC) remains the leading cause of cancer-related mortality worldwide, with tobacco smoking being the primary risk factor. However, the long-term LC risk among individuals with negative low-dose computed tomography (LDCT) findings and the role of tobacco smoking in risk stratification remain poorly understood, limiting evidence-based guidance for subsequent screening intervals. Objective:To evaluate the association of tobacco smoking with long-term LC risk after a negative baseline LDCT finding and to inform optimized screening strategies. Design, Setting, and Participants:This population-based, prospective cohort study was conducted under the Cancer Screening Program in Urban China. Individuals aged 40 to 74 years with negative baseline LDCT findings (October 1, 2013, to December 31, 2021) were included, with follow-up until December 2023. All participants were monitored for LC incidence. Exposures:Self-reported smoking status, pack-years, and time since quitting. Main Outcomes and Measures:The primary outcome was LC incidence, analyzed using Kaplan-Meier methods and multivariable Cox proportional hazards regression models. The association between smoking exposure and LC risk was assessed, with time-stratified analyses and dose-response associations. Results:Among 30 565 participants (14 761 never smokers and 15 804 smokers; mean [SD] age, 57.1 [7.7] years; 15 693 [51.3%] female), 76 LC cases occurred during 139 011.51 person-years (crude incidence rate, 54.67 of 100 000 person-years). Smokers had higher LC risk than never smokers (adjusted hazard ratio [AHR], 2.73; 95% CI, 1.49-5.01), driven by those with a smoking history of 20 pack-years or more (eg, ≥30 pack-years: AHR, 3.22; 95% CI, 1.85-5.58). There was no elevated risk at 2 years (AHR, 2.07; 95% CI, 0.91-4.69), but risk was significantly increased at 3 years (AHR, 2.54; 95% CI, 1.19-5.41) and onward. A nonlinear dose-response association was found between pack-years and LC risk, with risk surpassing clinically relevant thresholds at approximately 20 pack-years (eg, 20 to <30 pack-years: AHR, 2.48; 95% CI, 1.14-5.40). Females exhibited higher susceptibility than males at comparable exposure (≥30 pack-years: AHR, 5.78 [95% CI,1.87-17.83] for females vs 1.36 [95% CI, 0.18-10.39] for males). Significant risk was seen in those aged 50 to 54 years (≥30 pack-years) and 55 to 74 years (≥20 pack-years). Short-term cessation (<15 years) was not significantly associated with reduced LC risk. Conclusions and Relevance:In this cohort study, smokers with negative baseline LDCT findings exhibited a substantially elevated long-term LC risk, which became significant only after 2 years after screening. These findings suggest support for extending the initial screening interval and implementing personalized long-term monitoring based on smoking history.

Cardiovascular-kidney-metabolic (CKM) syndrome has been proposed, yet its utility for cancer risk stratification and behavior modification remains unclear. The study included 227,330 cancer-free participants from UK Biobank. CKM stages were defined using the American Heart Association (AHA) criteria, and lifestyle factors included smoking, alcohol consumption, physical activity, and diet. The hazard ratios (HRs) and lifetime risks of cancer were estimated using the Cox model and Fine and Gray's model, respectively. During a median follow-up of 11.46-13.65 years, 24,916 cancer cases and 7791 deaths were identified. Risks of cancer incidence and mortality increased progressively with advancing CKM stages (P trend < 0.001), with HRs (95% CIs) of 1.23 (1.15-1.30), and 1.58 (1.41-1.78) for stage 4. Compared with participants with unfavorable lifestyle, those with favorable lifestyle were associated with decreased risks across stages 0-4, with risk reductions of 14%-24% for cancer incidence, and 22%-40% for cancer mortality. Joint analyses showed that the increased cancer risks associated with CKM stages were eliminated or attenuated by adopting favorable lifestyle, particularly in stages 1-2. Participants in stage 4/unfavorable lifestyle had the highest risks of cancer incidence and mortality, with HRs (95% CI) of 1.54 (1.39-1.70) and 2.43 (1.97-2.99), and cumulative risks by age 80 of 34.77% and 11.65%, respectively. Furthermore, CKM stage 4/unfavorable lifestyle was associated with higher incidence of breast, colorectal, lung, kidney, pancreas, bladder, head and neck, esophagus, and liver cancers. These findings support the potential utility of CKM stages for cancer risk stratification and highlight the importance of lifestyle intervention.

Plastic food containers used in food delivery may pose underrecognized health risks by releasing micro- and nanoplastics (MNPs), especially when in contact with oil-rich foods. Here, we show that cooking oil dramatically increased MNP release from polypropylene (PP) and polyethylene (PE)-coated containers within 3 min of microwave heating, reaching up to 1.52 × 1014 particles/L, approximately 125-fold higher than with water. Concurrently, the co-release of plastic additives and heavy metals increased by up to 309-fold. Soybean oil-derived PP nanoplastics were four times more cytotoxic (IC50 = 18.7 µg/mL) than water-derived particles (IC50 = 77.1 µg/mL), likely due to oil-film coatings, a positive surface charge (+7.37 mV), altered surface properties, and co-extracted additives. Fluorescence and transmission electron microscopy revealed rapid membrane disruption within 5 min of exposure at 100 µg/mL. In vivo, oral exposure of mice to oil-derived nanoplastics (50 mg/kg) for two weeks caused severe intestinal barrier damage and mucosal immune dysfunction. These adverse effects were partially alleviated by immune- and barrier-targeted interventions. Benchmark dose modeling established safety thresholds of 0.4 µg/mL at the molecular level and 15.83 mg/kg for tissue injury. Alarmingly, modeled five-year gastrointestinal burdens and reported human tissue MNP levels exceeded molecular safety thresholds by up to 1107-fold, though not tissue injury thresholds. These findings highlight the urgent need for stricter regulations on oil-contact plastics and the development of safer food packaging materials.

BACKGROUND:Sleep health is multidimensional, yet existing research has largely examined isolated sleep characteristics, specific population groups, or only one direction of the sleep-depressive symptom relationship. As a result, the bidirectional associations between multidimensional sleep patterns and depressive symptoms remain insufficiently understood. METHODS:The study recruited 75,445 adults between July 2021 and December 2024 through the smart health management digital platform for primary cancer prevention (SmartHMDP-PCP) with 34,344 participants completed the follow-up survey. Sleep was assessed across eight dimensions-sleep midpoint, duration, pre-sleep activities, insomnia, sleep problems, daytime symptoms, medication use, and subjective quality-to construct a composite sleep score (healthy vs. unhealthy). Depressive symptoms were measured using the Self-Rating Depression Scale. Multivariate logistic regression estimated odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS:Healthy sleep was associated with reduced odds of incident depressive symptoms (OR, 95%CI: 0.62, 0.58-0.67) and increased likelihood of symptom remission (1.55, 1.41-1.69). Maintaining healthy sleep conferred the strongest protection against incident depressive symptoms (0.38, 0.35-0.42) and the greatest likelihood of remission (2.72, 2.42-3.06). In bidirectional analyses, absence of baseline depressive symptoms (0.60, 0.56-0.66), particularly when sustained over time (0.41, 0.37-0.46), was also negatively associated with subsequent deteriorations in sleep patterns, with effect sizes numerically comparable to those observed in the reverse direction. CONCLUSIONS:Multidimensional sleep and depressive symptoms exhibit bidirectional associations of numerically similar magnitude. These findings underscore the importance of integrating sleep health promotion into mental health strategies and highlight the need for interventional and long-term longitudinal research to clarify causal pathways.

BACKGROUND:Lung cancer is the leading cause of cancer-related deaths worldwide, with non-smokers in China accounting for over 40% of cases. Despite the proven efficacy of low-dose computed tomography (LDCT) in early detection and reduction of lung cancer mortality, the current paradigm of lung cancer screening, heavily focused on smoking status and age, may inadequately address the unique risk factors associated with non-smokers, particularly those with a family history of the disease. This study evaluates the cost-effectiveness of LDCT screening for non-smokers with a first-degree relative (FDR) history of lung cancer, a group at particularly high-risk. METHODS:We developed a state-transition Markov model to evaluate the incremental cost-effectiveness ratios (ICERs) of 16 screening strategies for a hypothetical cohort of 100,000 non-smoking individuals aged 50 with a FDR history of lung cancer, considering various starting ages (50, 55, 60, 65 years) and intervals (one-off, annual, biennial, triennial). The willingness-to-pay (WTP) threshold was set at three times China's 2022 per-capita GDP. Sensitivity analyses, scenario analyses and subgroup analysis by sex, were conducted. RESULTS:Compared to no screening, all strategies except one-off screening at age 50, were cost-effective for both sexes. Biennial LDCT starting at age 55 was found to be most effective, with an ICER of CNY 68,932/QALY for males, and CNY 80,056/QALY for females. This cost-effectiveness probability for this strategy was approximately 90% for both sexes. Sensitivity analyses indicated that annual screening at age 55 was optimal without discounting. For males, biennial at age 60 was optimal if the FDR-related odds ratio for lung cancer incidence was below 1.492. Triennial screening at age 55 was optimal for females at full adherence. Ignoring disutility from false-positive results, annual at age 55 was optimal for both sexes. CONCLUSIONS:LDCT screening for non-smokers with a FDR history of lung cancer is cost-effective, especially biennial screening at 55. These findings support the development of more inclusive screening guidelines, which could enhance early detection and reduce mortality rates.

BACKGROUND:Although guidelines stress the importance of early screening for individuals at high risk of lung cancer in China, there is a lack of data on risk-adapted starting ages for screening. This study aims to determine the appropriate starting age for lung cancer screening in China, considering various risk factors associated with the disease. METHODS:The data used were from the Cancer Screening Program in Urban China. A total of 413,725 eligible participants aged 40-74 years from eight cities in China were enrolled between 2013 and 2021. The outcomes of the study included lung cancer diagnosis and age at diagnosis. The risk-adapted starting age for screening was defined as the age at which individuals with varying levels of lung cancer risk reached a 10-year cumulative risk level similar to that of those aged 50 years in the general population. RESULTS:Among the 413,725 individuals who participated in the study, 1607 were diagnosed with lung cancer with a median follow-up of 4.90 (3.01, 6.84) years. The participants were categorized into different risk groups based on their lung cancer risk scores, which were determined by various risk factors, such as gender, education level, body mass index, vegetable intake, smoking pack-years, and tea consumption. In the study, the optimal starting age for lung cancer screening was determined on the basis of an individual's risk level. Using the 10-year cumulative risk of lung cancer at age 50 years in the general population as a benchmark (0.59% [95% confidence interval, 0.52-0.63%]), the study revealed that individuals with high, medium, or low risk of lung cancer should start screening at ages 46, 48, or 54 years and older, respectively. CONCLUSIONS:This study establishes the age at which lung cancer screening should begin on the basis of the principle of equal management and risk management. These findings have the potential to contribute to updates in the current screening guidelines.

The gut microbiota-cancer interaction functions through multi-level biological mechanisms, forming the basis for both diagnostic and therapeutic applications. Current technical and biological challenges drive the field toward precision medicine approaches, aiming to integrate multi-dimensional data for optimized, personalized cancer treatments.

AIM:Effective prevention of periodontitis requires addressing multiple modifiable health behaviours (MHBs). This study aimed to examine how MHBs interact and to identify the key behaviours that could serve as targets for intervention. MATERIALS AND METHODS:We analysed data from a large, population-based survey using questionnaires conducted in China during 2021-2023. Logistic regression was used to examine associations between self-reported periodontitis and multiple MHBs. Network analysis was applied to assess interrelationships among MHBs and identify central MHBs. RESULTS:Among 85,483 participants, 12.8% reported periodontitis. Ten MHBs were associated with self-reported periodontitis. Ever smoking, ever drinking, unhealthy bedtime, frequent consumption of sugar-sweetened snacks and beverages, insufficient vegetable or fruit intake and toothbrushing less than twice daily were linked to higher prevalence, while insufficient physical activity, no daily flossing and absence of annual dental checkups were linked to lower prevalence. Network analysis identified ever smoking, ever drinking, toothbrushing frequency and dental checkups as the most central behaviours, with variations observed across different subgroups. CONCLUSION:This study identified key MHBs and revealed differences in central behaviours across subgroups. Ever smoking, ever drinking, toothbrushing frequency and annual dental checkups may represent priority targets for tailored prevention strategies. Future research is warranted to inform integrated interventions.

INTRODUCTION:Previous studies have highlighted the importance of blood lipid levels in lung cancer. However, evidence of the association between remnant cholesterol and lung cancer remains scarce. This study aimed to investigate the association of remnant cholesterol with lung cancer morbidity and mortality and to evaluate their joint effects with C-reactive protein in women. METHODS:This prospective cohort study included 198,154 women initially without cancer from the UK Biobank. Remnant cholesterol was calculated as non-high-density lipoprotein cholesterol minus the measured low-density lipoprotein cholesterol. Cox models were adopted to estimate hazard ratios and 95% CIs for the incidence of lung cancer. Data were collected between 2006 and 2022 and analyzed in 2025. RESULTS:During a median follow-up of 11.80-13.90 years, 1,552 lung cancer cases and 1,074 related deaths were identified. Remnant cholesterol was positively associated with lung cancer morbidity and mortality in a linear manner, with respective hazard ratios (95% CIs) of 1.50 (1.23, 1.82) and 1.40 (1.11, 1.77) in Quartile 4. Compared with the low remnant cholesterol/low C-reactive protein group, the risk of incident lung cancer and lung cancer mortality increased by 115% and 102%, respectively, in the high remnant cholesterol/high C-reactive protein group. The cumulative risks of lung cancer by age 80 years were higher in the high remnant cholesterol/high C-reactive protein group than in the low remnant cholesterol/low C-reactive protein group (morbidity=3.64% vs 1.56%; mortality=1.97% vs 0.82%). CONCLUSIONS:This study found linear and positive associations of remnant cholesterol with lung cancer morbidity and mortality among women. The combination of high remnant cholesterol and C-reactive protein conferred the highest relative and absolute risks. These findings highlighted the importance of considering the combination of remnant cholesterol and C-reactive protein levels for the primary prevention of lung cancer and selection of high-risk populations for lung cancer screening among women.

This review examines global human genetic resources management, focusing on genetic data policies and repositories in high- and middle-low-income countries. A comprehensive search strategy was employed across multiple databases, including official government websites and Google, to gather relevant literature on human genetic resources management policies and genetic resource databases. Documents were screened for relevance, focusing on high-income countries (United States, United Kingdom, Japan) and middle-low-income countries (China, India, Kenya). Data were extracted, coded, and analyzed to identify common themes and differences in genetic resource management practices. High-income countries benefit from robust legal frameworks and advanced technological infrastructures. The United States enforces the Health Insurance Portability and Accountability Act and the Genetic Information Nondiscrimination Act to protect privacy and facilitate data sharing, while Japan relies on the Act on the Protection of Personal Information and ethical guidelines. Additionally, high-income countries host a variety of genetic databases and biobanks that support scientific research. In contrast, middle-low-income countries like China, India, and Kenya are still developing their frameworks. China has regulations such as the Biosecurity Law and the Regulations on the Management of Human Genetic Resources, but still requires more unified standards. India’s policies focus on genetic research and data protection through the Biological Diversity Act, while Kenya seeks to improve data management through the 2019 Data Protection Act. Significant disparities exist in human genetic resources management between high-income and middle-low-income countries. High-income countries have robust systems balancing privacy protection with research facilitation, supported by comprehensive and large-scale databases for scientific research. Middle-low-income countries need to enhance legal frameworks and build population-specific databases. Promoting equitable data sharing and adopting best practices from high-income countries are essential for advancing global scientific discovery and ensuring fair management of genetic resources.

BACKGROUND:Colorectal cancer (CRC) is prevalent in China, but many features of CRC patients remain to be better characterized. We aimed to describe the characteristics of clinical epidemiology in CRC patients in China and to evaluate the changes in health-related quality of life (HRQOL) before and after treatment. METHODS:A hospital-based survey was conducted among CRC patients from 2020 to 2021, covering 14 cities in China. Data on demographic and clinical characteristics, disease knowledge, medical service utilization, medical expenditure, and HRQOL before and after treatment were surveyed. The multivariable regressions and structural equation model (SEM) were used to assess the association between characteristics and HRQOL. RESULTS:In the survey of 4589 patients, nearly 80% were diagnosed at an age older than 50, with 59.5% being men and 54.5% having rectal cancer. Approximately 35.2% were diagnosed at stage IV and 37.5% had metastases. Prior to diagnosis, only 2.6% of patients had a colonoscopy screening, with the primary barrier being unawareness. After treatment, there was a significant decline in the overall HRQOL scores (66.98 vs. 65.39, P < 0.001) and a significant association between patients' awareness of CRC treatment and higher HRQOL changes (beta: 0.98, 95% CI 0.39-1.57; P = 0.001). The SEM model showed good model fit, revealing that awareness of CRC treatment was significantly associated with changes in HRQOL (beta: 0.06, 95% CI 0.04-0.09; P = 0.009). CONCLUSION:Our study highlighted a low CRC screening rate in China, largely due to limited public awareness. After treatment, HRQOL declined, and treatment awareness was significantly associated with this change.

Supplementary Data from Risk of Liver Cirrhosis in HBV/HCV-Infected Individuals with First-Degree Relatives Who Have Liver Cancer: Development and Validation of a Simple Model

Low dose computed tomography (LDCT) screening has been proven to be effective in reducing lung cancer mortality, but the ensuing high false-positive and overdiagnosis rates shackle the effectiveness of lung cancer screening (LCS) in China. Nodule malignancy prediction models may be an applicable solution. We conducted a prospective cohort study to develop and internally validate the model using data from the ongoing Henan province Cancer Screening Program in Urban China (CanSPUC). From 2013 to 2021, 23,031 heavy smokers underwent baseline screening with LDCT; 2553 participants were diagnosed with pulmonary nodules. Detailed questionnaire, physical assessment and follow-up were completed for all participants. Multivariable Cox proportional risk regression models were used to identify and integrate key prognostic factors for the development of a nomogram model. Data from the National Lung Screening Trial (NLST) were utilized for external validation. A total of 111 lung cancer cases with a median follow-up duration of 3.7 years occurred in the Henan CanSPUC. Age, gender, physical activity, consumption of pickled food, history of silicosis or pneumoconiosis, nodule type, size, calcification, and pleural retraction sign were included into the model. The AUC was 0.855, 0.844, and 0.863 for the 1-, 3- and 5-year lung cancer risk in the training set, respectively. Compared with Mayo model, VA model, PKU model, and Brock model, the Henan CanSPUC model yield statistically better discriminatory performance (all P values < 0.05). The model calibrated well across the deciles of predicted risk in both the overall population and all subgroups. The model demonstrated good calibration and discrimination in the internal validation cohort, while the external validation cohort showed lower predictive performance, indicating that further external validation is needed. The model developed and validated in this study may be used to estimate the probability of lung cancer in nodules detected at baseline LDCT, allowing more efficient risk-adapted follow-up in population-based LCS programs. However, further external validation in broader and more diverse populations is warranted.

Previous research has demonstrated that in pregnant mice deficient in l-methionine (Met), the mixture of the dipeptide l-methionyl-l-methionine (Met-Met) with Met was more effective than Met alone in promoting mammogenesis and lactogenesis. This study aimed to investigate the role of a novel long noncoding RNA (lncRNA), named mammary gland proliferation-associated lncRNA (MGPNCR), in these processes. Transcriptomic analysis of mammary tissues from Met-deficient mice, supplemented either with a Met-Met/Met mixture or with Met alone, revealed significantly higher MGPNCR expression in the Met group compared to the mixture group, a finding recapitulated in a mammary epithelial cell model. Our findings suggested that MGPNCR hindered mammogenesis and milk protein synthesis by binding to eukaryotic initiation factor 4B (eIF4B). This interaction promoted the dephosphorylation of eIF4B at serine-422 by enhancing its association with protein phosphatase 2A (PP2A). Our study sheds light on the regulatory mechanisms of lncRNA-mediated dipeptide effects on mammary cell proliferation and milk protein synthesis. These insights underscore the potential benefits of utilizing dipeptides to improve milk protein in animals and potentially in humans.

The disposable paper cups (DPCs) release millions of microplastics (MPs) when used for hot beverages. However, the tissue-specific deposition and toxic effects of MPs and associated toxins remain largely unexplored, especially at daily consumption levels. We administered MPs and associated toxins extracted from leading brand DPCs to pregnant mice, revealing dose-responsive harmful effects on fetal development and maternal physiology. MPs were detected in all 13 examined tissues, with preferred depositions in the fetus, placenta, kidney, spleen, lung, and heart, contributing to impaired phenotypes. Brain tissues had the smallest MPs (90.35 % < 10 µm). A dose-responsive shift in the cecal microbiome from Firmicutes to Bacteroidetes was observed, coupled with enhanced biosynthesis of microbial fatty acids. A moderate consumption of 3.3 cups daily was sufficient to alter the cecal microbiome, global metabolic functions, and immune health, as reflected by tissue-specific transcriptomic analyses in maternal blood, placenta, and mammary glands, leading to neurodegenerative and miscarriage risks. Gene-based benchmark dose framework analysis suggested a safe exposure limit of 2 to 4 cups/day in pregnant mice. Our results highlight tissue-specific accumulation and metabolic and reproductive toxicities in mice at DPC consumption levels presumed non-hazardous, with potential health implications for pregnant women and fetuses.

The health implications of human exposure to microplastics (MPs) have raised significant concerns. While evidence indicates MPs can accumulate in closed human organs like the heart, placenta, and blood, there is no available data on MP exposure specifically within the human bone marrow. To fill the research gap, this study detected the concentration of microplastics (MPs) in bone marrow samples by pyrolysis gas chromatography-mass spectrometry (Py-GC/MS) and assessed the size range and morphological characteristics of MPs by Laser Direct Infrared Spectroscopy (LD-IR) and scanning electron microscopy (SEM). Our study shows that MPs were present in all 16 bone marrow samples, with an average concentration of 51.29 µg/g ranging from 15.37 µg/g to 92.05 µg/g. Five polymer types-polyethylene (PE), polystyrene (PS), polyvinyl chloride (PVC), polyadiohexylenediamine 66 (PA66), and polypropylene (PP), were identified. PE was the most frequent polymer detected in the bone marrow, with an average concentration of 30.02 µg/g ranging from 14.77 µg/g to 52.57 µg/g, with a detection rate of 93.75 %. PS had the highest detection rate at 100 % of bone marrow samples, while PVC and PA66 were found in 75 % of samples each. LD-IR analysis revealed the identification of 25 polymer types, with an average abundance of 19.72 particles/g. Of these, 89.82 % of the MPs were smaller than 100 µm. In summary, this study has, for the first time, demonstrated the presence of MPs are deeply embedded within human bone marrow, providing a basis for future investigations into their potential toxicological effects and underlying mechanisms affecting the hematopoietic system.

Objective: To analyze the detection of colorectal advanced neoplasms in the population who underwent colonoscopy screening in Henan Province as part of the Urban China Cancer Screening Program and its influencing factors. Methods: A cross-sectional study design was employed. Based on the Cancer Screening Program conducted in Henan Province, the study enrolled 7 454 urban residents who manifested no symptoms and were recruited from eight cities in the province, including Zhengzhou, Zhumadian, Anyang, Luoyang, Nanyang, Jiaozuo, Xinxiang, and Puyang from October 2013 to October 2019, and participated in colonoscopy screening. The χ2 test was used to compare the detection rates of colorectal advanced neoplasms among participants with different characteristics, and a multivariate logistic stepwise regression model was used to analyze the factors affecting the detection rates. Results: A total of 7 454 subjects underwent colonoscopy screening, and 112 cases of colorectal advanced neoplasms were detected. Multivariate logistic regression analysis suggested that older age, smoking, higher meat intake, history of diabetes, and family history of colorectal cancer in a first-degree relative were risk factors for colorectal advanced neoplasms. The detection rate was significantly higher in people aged 60-74 years compared with those aged 40-49 years, with an odds ratio (OR) of 2.04 (95% CI: 1.23-3.38).The rates were higher in people who smoked than those who did not smoke, with an OR of 2.21 (95% CI: 1.48-3.31), and in people who consumed more meat than those who consumed less, with an OR of 1.53 (95% CI: 1.04-2.26). Those with diabetes had a higher detection rate compared with those without, with an OR of 1.69 (95% CI: 1.07-2.69), and those with a first-degree family history of colorectal cancer had a higher detection rate than those without, with an OR of 1.64 (95% CI: 1.09-2.46). Conclusion: The detection rate of colorectal advanced neoplasms through colonoscopy screening in Henan Province covered by the Urban China Cancer Screening Program is 1.50%. Older age, smoking, higher meat intake, history of diabetes, and family history of colorectal cancer in a first-degree relative are identified as risk factors for colorectal advanced neoplasms.