The underlying causes of esophageal cancer (EC) are unknown. To explore the molecular mechanisms that lead to EC, gene expression profiles of large cohorts of patients with EC were obtained from The Cancer Genome Atlas and the Gene Expression Omnibus (GEO) databases . The present study identified 83 upregulated and 22 downregulated genes between EC and normal tissue using R statistical software and the GEO2R web tool. The Database for Annotation, Visualization and Integrated Discovery was used to identify the associated pathways, and for functional annotation of the differentially expressed genes (DEGs). Protein-protein interactions of these DEGs were analyzed based on the Search Tool for the Retrieval of Interacting Genes database, and hub genes were visualized using Cytoscape software. An online Kaplan-Meier plotter survival analysis tool was utilized to evaluate the prognostic value of hub gene expression in patients with EC. Further analysis of an additional dataset from GEO (GSE21293) revealed that these genes were associated with infiltration and metastasis in EC. In addition, the Gene Expression Profiling Interactive Analysis tool was used to evaluate expression levels of hub genes in patients with EC for different pathological stages. The Ualcan analysis tool was used to evaluate the expression levels of hub genes for different histological types. Overall, ubiquitin conjugating enzyme E2 C, cyclin dependent kinase inhibitor 3, CDC28 protein kinase regulatory subunit 2, kinesin family member 20A (KIF20A) and RAD51 associated protein 1 (RAD51AP1) were upregulated in EC tissues compared with normal tissues, and upregulation of these genes was a poor prognostic factor for patients with EC, indicating that these genes may mediate EC cell infiltration and metastasis. Among the hub genes, KIF-20A had potential value for predicting the pathological stage of EC. KIF20A and RAD51AP1 were more informative biomarkers of esophageal squamous cell carcinoma. Further studies are required to explore the value of these genes in the treatment of EC. Gastrointestinal cancer is a major health problem with 4.8 million new cases and 3.4 million mortality cases reported worldwide in 2018 . TherefoThe present study aimed to identify differentially expressed genes (DEGs) between normal esophageal and EC tissues, and to subsequently classify these DEGs based on motifs and modules at a functional level according to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, which could provide a model suggesting mechanisms of how these DEGs contribute to EC. A protein-protein interaction (PPI) network was used to identify hub genes in EC. The present study identified which hub genes influenced the prognosis of EC, and explored the association among hub genes with prognostic value and invasive, metastatic, pathological and histological characteristics of EC. Finally, biomarkers that may be involved in the occurrence and development of EC were identified, and provide a basis for further research.http://tcga-data.nci.nih.gov; accessed, November 10, 2018). The ‘RTCGAToolbox’ package (version 1.0) in the R software environment was used to download quantitative gene expression data and clinical characteristics of patients with EC in the TCGA database (https://www.ncbi.nlm.nih.gov/geo/). Following reading and screening, the three gene expression datasets GSE23400  from The Cancer Genome Atlas (TCGA) database  was used to evaluate DEGs between EC and normal tissue samples in data from the GEO database. Due to the small sample size, the parameters used to indicate significant DEGs were adjusted P-value =0.05 and |log FC| ≥1.0.The GEO2R online analysis tool (http://bioinformatics.psb.ugent.be/webtools/Venn/) was used to identify the intersection of DEGs between the TCGA and GEO datasets. The present study first identified the intersection of the three GEO datasets, and then cross-identified the DEGs common to the TCGA and GEO datasets. These intersecting genes were considered to be the DEGs between EC and normal tissue samples.Finally, a Venn diagram web tool (http://david.ncifcrf.gov/) (GO  and KEGGrf.gov/) . P<0.05 http://string-db.org/)   . PPI pai.03.6.1) . In the .03.6.1) . The maxhttp://kmplot.com/analysis/) to assess the impact of differential expression of the hub genes on the prognosis of EC . This dataset contained gene expression data for invasive and non-invasive EC cells. Fragments per kilobase million values for ubiquitin conjugating enzyme E2 C (UBE2C), cyclin dependent kinase inhibitor 3 (CDKN3), CDC28 protein kinase regulatory subunit 2 (CKS2), kinesin family member 20A (KIF20A) and RAD51 associated protein 1 (RAD51AP1) genes were log2 transformed and the resulting heatmap was constructed using HemI software (version 1.0)  . Graph dhttp://gepia.cancer-pku.cn/) was used to analyze the hub gene expression data from the TCGA EC dataset, which contained clinical staging information  was used to analyze the hub gene expression data from the TCGA EC dataset, which contained histological subtype information  were used to identify DEGs. It was observed that samples from the same tissue type aggregated into a single group, and each tissue type exhibited one or more unique tissue-specific mRNA blocks. The top 50 genes were significantly upregulated in EC tissues compared with in normal tissues .Subsequently, gene expression in three microarray datasets [GSE5364 , GSE2034Based on the significance criteria of P<0.05 and |log FC| ≥2, a total of 2,000 DEGs were identified, including 1,156 upregulated genes and 844 downregulated genes in TCGA datasets . A comprTo investigate the effects of DEGs on the occurrence of EC at a functional level, the DAVID web tool was used to perform GO functional and KEGG pathway enrichment analysis of the DEGs. The enriched GO terms were divided into cellular components (CCs), biological processes (BPs) and molecular functions (MFs). The BPs associated with upregulated DEGs were ‘collagen catabolic process’, ‘cell division’, ‘extracellular matrix disassembly’, ‘mitotic nuclear division’ and ‘cell proliferation’. For CCs, the upregulated DEGs were mainly enriched in ‘midbody’, ‘spindle’, ‘proteinaceous extracellular matrix’, ‘spindle microtubule’ and ‘kinetochore’. MF analysis demonstrated that the upregulated DEGs were mainly enriched in ‘metalloendopeptidase activity’, ‘protein binding’, ‘ATP binding’, ‘serine-type endopeptidase activity’ and ‘protein kinase binding’. KEGG pathway analysis revealed that the upregulated DEGs were mainly enriched in ‘cell cycle’, ‘DNA replication’, ‘ECM-receptor interaction’, ‘oocyte meiosis’ and ‘progesterone-mediated oocyte maturation’ pathways .The downregulated DEGs were mainly enriched in BPs, including ‘muscle contraction’, ‘complement activation, alternative pathway’ and ‘regulation of smooth muscle contraction’. The downregulated DEGs were significantly enriched in the CC terms ‘focal adhesion’, ‘extracellular exosome’, ‘myosin filament’ and ‘apical plasma membrane’. Enrichment analysis of MF terms revealed that downregulated DEGs were mainly enriched in ‘calmodulin binding’ and ‘structural constituent of muscle’. There was no significant KEGG pathway enrichment for the downregulated DEGs .Cytoscape software was used to predict protein interactions among DEGs. The PPI network comprised 82 nodes and 982 edges . After aScreening of the PPI network using MCODE identified two modules. In module 1, the MCODE score was 40.7, including 41 nodes and 814 edge connection lines . To inveTo determine whether these DEGs were associated with survival of patients with EC, the DEGs were submitted to the Kaplan-Meier plotter bioinformatics analysis platform. Kaplan-Meier plotter database data is mainly derived from GEO (Affymetrix microarray only), EGA and TCGA. All possible cutoff values between the lower and upper quartiles were computed, and the best performing threshold was used as a cutoff. The log-rank test was used to analyze OS (161 EC samples) and RFS (73 EC samples). The OS prognosis of patients with EC with high expression levels of UBE2C , CDKN3 , CKS2 F, KIF20A Poor prognosis of EC is closely associated with early invasion and metastasis . To detePathological stage is a major indicator of the progression of EC . The preTo determine if the expression of the five prognostic hub genes differed in different histological subtypes of EC, the expression levels of these genes in ESCC and EAC were analyzed based on the TCGA database. The expression levels of UBE2C , CDKN3 , CKS2 F, KIF20A in vitro and in vivo, and regulation of a stem cell-like population . RAD51AP1 facilitates the repair of damaged DNA strands by binding to RAD51 ,53. InhiIn conclusion, a comprehensive bioinformatic analysis of DEGs that may be associated with the pathogenesis of EC was presented. A total of 105 DEGs and 36 hub genes were identified, and the hub genes UBE2C, CDKN3, CKS2, KIF20A and RAD51AP1 were associated with poor prognosis of patients with EC, and may be involved in invasion of EC. Differential expression of KIF20A was associated with different pathological stages of EC. The expression levels of KIF20A and RAD51AP1 in ESCC were higher than those in EAC, suggesting an EC subtype-specific expression pattern. The role of these genes in the progression of EC merits further investigation.


Mage = 41.70, 50.9% female). More than 50% of parents reported being stressed by social distancing and the closure of schools and childcare facilities. Parental stress increased significantly during the pandemic (d = 0.21). Subgroups of parents also reported very high levels of depressive symptoms (12.3%) and anxiety (9.7%). Up to one-third of the sample reported ACEs in the child’s lifetime. In this group, 29.1% reported an increase in children witnessing domestic violence during the pandemic, and 42.2% an increase verbal emotional abuse. These families were characterized by higher parental stress, job losses, and younger parent and child age. Positive aspects of the pandemic related primarily to personal or family life . While some parents coped well, a particularly negative pattern was observed in a subgroup of families that experienced an increase in ACEs. Parental stress emerged as important target point for interventions addressing the negative sequelae of the pandemic.Parenting during the COVID-19 pandemic is highly challenging, with parents having to meet various demands simultaneously. An increase in adverse childhood experiences (ACEs) has been widely predicted, but empirical evidence is still scarce. This study aimed to (1) generate representative data on pandemic-related stress, parental stress, general stress, parental subjective and mental health, and the occurrence of ACEs; (2) identify risk factors for an increase in ACEs, and (3) provide qualitative data on parents’ experiences. A representative survey was conducted in Germany in August 2020 with 1024 parents of underage children (The online version contains supplementary material available at 10.1007/s00787-021-01739-0. The SARS-CoV-2 coronavirus was declared a “public health emergency of international concern” by the World Health Organisation on January 30, 2020, and upgraded to a pandemic on March 11, 2020. In Germany, the first peak of infections occurred in spring 2020 and comprehensive, nationwide restrictions were implemented to slow down infection rates.The restrictions are especially challenging for families, with home schooling, social distancing measures and lockdown situations having a profound and complex impact in the family context –3. TheseWhile evidence on the psychosocial sequelae of the COVID-19 pandemic for general population samples  and for The Canadian Perspectives Survey Series examined family stress and perceived risk for domestic violence due to the COVID-19 pandemic in 4627 adults . The resBy surveying a representative sample of parents with underage children, this study aims to (1) generate representative data on pandemic-related stress, parental stress, parental subjective and mental health and the occurrence of ACEs; (2) describe risk factors for an increase in ACEs and (3) provide qualitative data on parents’ negative and positive experiences during the pandemic.n = 402) with a computer-assisted web survey . For the recruitment of the CATI subsample, a dual-frame-design was used to include households with landline and parents with mobile phone numbers. Dual-frame design weighting accounted for different selection chances of parents available by landline vs. mobile phones and for the specific characteristics of landline users and mobile phone users. The telephone surveys were conducted by trained and supervised staff. Participants in the web survey were recruited from an active online-access panel. Participants received incentives from the panel provider to compensate their participation in the survey according to a fixed scheme. Respondents with an unrealistically short completion time (n = 66) were excluded from the CAWI data set. To provide representative data, the total sample was recruited according to current micro-census quota for the German population for age, sex, household size, educational level and residency.The study population were German speaking households with underage children. Data were collected by the Berlin-based market research company INFO Marktforschungsinstitut using the survey software keyingress (Ingress GmbH). The survey was run between August 3rd and 11th 2020. We used a mixed mode design that combined computer-assisted telephone interviews . In this paper, sample characteristics were presented both for the unweighted data and weighted data; statistical analyses were conducted with the weighted data set. The results and conclusion do not change when using the unweighted data.Data were collected on parent age and sex, parental status , marital status, number of children in the household, and children’s age and sex. Family socioeconomic status (SES) was classified as low, medium, or high on the Winkler Index according to German population-based reference data .In addition, parent-related risk factors for an increase in ACEs were assessed: the parent’s risk of alcohol abuse during the pandemic was assessed by the alcohol abuse module of the German version of the Patient Health Questionnaire (PHQ-D) , 17; theThe Pandemic Stress Scale  was deveThe Parental Stress Scale  is an 18The stress module of the PHQ-D  was usedThe PHQ-4 , a four-We used a well-established single-item measure  to eliciTo provide a comprehensive assessment of ACEs, we collected data on child abuse, neglect and household dysfunction , 24. We In addition to the questionnaires, we asked two open questions on perceived highest burden and positive aspects of the pandemic: “Overall, what caused you the most stress during the pandemic?” and “What has changed for the better during the pandemic?”. Parents responded by typing their answers into a box (online survey) or by telling the interviewer (telephone survey).M = 40.89, SD = 8.17, range 18–73). Mean child age was 9.41 years . Table The sample comprised 1024 parents with a mean age of 41.7 years .First, we analyzed data on pandemic-related stress and the other outcome measures. The level of pandemic-related stress varied across domains see Fig. . ParentsM = 36.93, SD = 10.45, range 18–71 vs. pre-COVID-19 M = 34.72, SD = 10.63, range = 18–70; t(1023) = 12.474, p < 0.001); the effect size was small (d = 0.21). However, parental stress at the time of the subjectively highest burden was not higher than in two reference samples  and was significantly lower than in clinical reference samples [n = 51, M = 43.2, SD = 9.1; parents in inpatient psychiatric treatment: n = 83, M = 41.9, SD = 9.4).Parental stress at the time of the subjectively highest burden was significantly higher than pre-COVID-19 levels  and thus in the low range [The mean score for parents’ general stress at the time of the subjectively highest burden was ow range .M = 6.80, SD = 2.21 vs. pre-COVID-19 M = 7.34, SD = 2.04; t(1023) = 10.33, p < 0.001). The effect size was small (d = 0.31).Parents rated their overall health status at the time of the subjectively highest burden as significantly worse than pre-COVID-19 levels . 12.3% of the sample scored above the 95th percentile of the PHQ-4 [Regarding parent mental health, our sample reported significantly higher symptoms of anxiety and depression than the German normative data  see Fig; the effd = 0.32, p < 0.001), anxiety , and depression  and showed poorer subjective health . No sex differences emerged for pandemic-related stress  and parental stress .As women usually report higher levels of subjective distress than men, we checked for gender differences. Mothers scored higher than fathers on general stress . Coefficients of significant correlations were in the small to medium range (r = − 0.077 to r = 0.393).To gain a more detailed picture of the associations between pandemic-related stress and parental outcomes, we conducted an item-wise analysis of the Pandemic Stress Scale . Results showed a homogenous pattern: all outcomes were significantly related to the specific restrictions, with two exceptions  of parents reported on their children’s lifetime occurrence of severe stressful life experiences including violence, abuse, or neglect. Of these, 34.8% reported an increase in occurrence during the pandemic . Concerning the specific subtypes of ACE, rates of lifetime occurrence were mainly higher. The highest lifetime occurrence was reported for children witnessing domestic violence  and for verbal emotional abuse against the children . Figure n = 14) and physical neglect (n = 11). These subtypes were therefore excluded from further analyses, as conclusions would be limited. Note that across the subtypes, 27.1–46.2% of cases reported no change in occurrence and that in 11.6 − 34.3% of cases, a decrease was reported.6.5%  reported that their child(ren) had been exposed to mental illness in the household in their lifetime; of these, 10.9% reported that domestic problems related to the mental illness had significantly increased during the pandemic . 37 parents (3.7%) reported alcohol or substance abuse in the household over the child(ren)’s lifetime; in 5.1% of these cases, problems significantly increased during the pandemic .We examined the relationships between the parental outcomes and an increase in the two ACEs with the highest lifetime occurrence: witnessing domestic violence and verbal emotional abuse. Results are shown in Figs. p = 0.013), and 62.1% reported an increase in VEA . In families reporting significant financial losses during the pandemic, 53.0% reported an increase in VEA ; the differences with respect to WDV were not statistically significant.With respect to sociodemographic characteristics, analyses showed that the parents and children in the subgroups reporting a pandemic-related increase in witnessing domestic violence (WDV) or verbal emotional abuse (VEA) were significantly younger . Concerning pandemic-specific factors, job losses and financial losses were related to an increase in WDV and VEA: 48.4% of the families with job losses during the pandemic reported an increase in WDV . 61.5% of parents reporting the experience of physical or sexual violence in adulthood indicated an increase in VEA . The other risk factors were not significantly related to an increase in the ACEs .Concerning parent-related risk factors, 37.8% of parents with a history of child physical or sexual abuse reported an increase in WDV  provided answers to the open question asking what had caused them most stress during the pandemic  answered the question on the positive aspects of the pandemic  was “nothing”, indicating that participants had difficulty seeing any kind of silver lining. Most responses related to positive outcomes for the family and the parents themselves, e.g., slower pace of life , gratitude, and an increase in family time.A total of 932 participants . Including the score for pre-pandemic parental stress as covariate in the analyses did not change the results. Other studies conducted during the COVID-19 pandemic have also highlighted the relevance of parental stress for child-related outcomes, such as child behavior problems [Severe ACEs were reported by 6.5% of the sample, which is in line with current prevalence rates . For theproblems  and pareproblems . ParentaParental stress was not the only factor related to an increase in the two most prevalent ACEs: witnessing domestic violence and verbal emotional abuse. Families with younger children were especially at risk, which may reflect the impact of childcare and school closures . Contrarth percentile of the PHQ-4. A recent meta-analysis of 17 studies conducted mostly in Asia found a much more pronounced increase in rates of anxiety and depression, of up to 18.7–50.9% [Parental stress and general stress were not much higher than usual in our sample, in line with other recent findings , 36. How.7–50.9% . Further.7–50.9% , which mThe qualitative data on the aspects that caused parents most stress validated and added to the quantitative data from the Pandemic Stress Scale. Participants reported being particularly stressed by social distancing, restrictions and childcare at home. About one-third of participants could not identify any positive aspects of the pandemic; however, parents perceived more time for themselves and for the family and an improvement in family cohesion as positive aspects of the pandemic , 39.In sum, parental stress emerged as important target point for interventions. The qualitative data also identified promising areas for resource-oriented interventions to mitigate the detrimental effects of the pandemic . ProspecOne strength of this study is the mixed mode approach to data collection, reducing the bias associated with collecting data online only. Unlike other parent samples, in which mothers tend to be heavily overrepresented, our sample included equal proportions of mothers and fathers. Mothers reported higher general stress, anxiety and depression, in line with both the literature in general  and dataConcerning the assessment of ACEs, the parent-report on lifetime occurrence of ACEs provided plausible data, as with 6.5% for severe experiences up to 32% for the less severe forms, the data correspond to prevalence estimations in the general population , 28. TheFor reasons of parsimony, we did not assess pre-COVID-19 data on all measures, which prohibited analyses of change in general stress and mental health. Furthermore, pre-COVID-19 data were assessed retrospectively and we only have parent’s self-reported data and child-related outcome measures were not included in the study. Given the cross-sectional design of the survey, the results are solely correlational in nature and causal inferences cannot be drawn. Prospective studies are needed to replicate the predictive role of the risk factors for an increase in witnessing domestic violence and verbal emotional abuse, as well as the interplay between the predictors.The results of this study confirm that the pandemic has had a profound impact on families. As school closures and social distancing from family and friends were among the most burdening restrictions for parents, great caution is necessary in applying these measures. Furthermore, additional efforts need to be put into helping families deal with the restrictions and “the new normal” . StructuSupplementary file1 (PDF 174 KB)Supplementary file2 (PDF 107 KB)Supplementary file3 (PDF 102 KB)Supplementary file4 (PDF 125 KB)Supplementary file5 (PDF 121 KB)Below is the link to the electronic supplementary material.


To examine the relation between long working hours and change in body mass index (BMI).n = 122,078), with a mean of 4.4-year follow-up. Working hours were measured at baseline and categorised as part time (<35 h/week), standard weekly hours , 41–48 h, 49–54 h and ≥55 h/week (long working hours). There were four outcomes at follow-up: (1) overweight/obesity (BMI ≥ 25 kg/m2) or (2) overweight (BMI 25–29.9 kg/m2) among participants without overweight/obesity at baseline; (3) obesity (BMI ≥ 30 kg/m2) among participants with overweight at baseline, and (4) weight loss among participants with obesity at baseline.We performed random effects meta-analyses using individual-participant data from 19 cohort studies from Europe, US and Australia (P for trend <0.0001). The findings were similar after multivariable adjustment and in subgroup analyses. Long working hours were associated with an excess risk of shift from normal weight to overweight rather than from overweight to obesity. Long working hours were not associated with weight loss among participants with obesity.Of the 61,143 participants without overweight/obesity at baseline, 20.2% had overweight/obesity at follow-up. Compared with standard weekly working hours, the age-, sex- and socioeconomic status-adjusted relative risk (RR) of overweight/obesity was 0.95 (95% CI 0.90–1.00) for part-time work, 1.07 (1.02–1.12) for 41–48 weekly working hours, 1.09 (1.03–1.16) for 49–54 h and 1.17 (1.08–1.27) for long working hours (This analysis of large individual-participant data suggests a small excess risk of overweight among the healthy-weight people who work long hours. Obesity is a modifiable risk factor for an array of health problems, including cardiovascular disease, type 2 diabetes, certain cancers and dementia that collectively contribute to a substantial proportion of disease burden and death worldwide , 2. ResuPsychosocial work environment may have a role in weight control although little research has been published beyond perceived stress at work. Current evidence suggests a modest association between work stress and overweight or obesity, with limited support from longitudinal studies , 6. AnotPrevious studies suggest associations of long working hours with increased incidence of cardiovascular disease and type 2 diabetes –9, for wAccordingly, with a large collaborative study of 19 cohorts from Europe, the US and Australia, we examined the association between working hours and subsequent change in body mass index (BMI), including an analysis of long working hours in relation to each stage of weight gain: from non-overweight to overweight or obesity, from overweight to obesity. Long working hours may also be associated with lower probability of weight loss among those with obesity. For the first time to our knowledge, the size of this dataset allows to examine also this issue.http://www.icpsr.umich.edu/icpsrweb/ICPSR/) or the UK Data Service (http://ukdataservice.ac.uk/). The studies were chosen according to the availability of working hours and BMI data in a prospective cohort design with the follow-up not exceeding 10 years. All studies have their own approval from their relevant local or national ethics committee, and all participants had given informed consent.We used data from 19 cohort studies (Supplementary Table Six cohort studies were from the UK: British Birth Cohort 1970 (BCS1970) , NationaIn total, there were 122,078 participants for prospective analyses who were at work and provided data on working hours, sociodemographic characteristics and repeat measurements of BMI.In all cohorts, information on working hours at baseline was obtained from self-report. We used the previously used categorisation of weekly working hours: <35 (part time), 35–40 (reference group), 41–48, 49–54 and ≥55 h (long working hours) , 39. In 2) was based on self-reported height and weight in all cohort studies except Whitehall II and Belstress, in which height and weight were measured directly during a standardised clinical examination. The following categories at baseline and at follow-up were calculated: not overweight (BMI < 25.0); overweight (25.0–29.9); obese (30.0 or more). For those with obesity, we also determined whether they lost weight to non-obesity or had weight loss of >5% [At baseline and follow-up, BMI , and psychological distress or depressive symptoms. Smoking (current vs non-smoker)  and leisWe used a two-stage approach to the present meta-analysis . First, In the second stage, we used random effects meta-analysis to obtain a pooled estimate from the first-stage study-specific estimates. We conducted our analyses in seven parts. Part 1: forest plots including cohort-specific estimates from main analysis comparing different working hours categories in relation to weight gain—that is, transition from non-overweight to overweight/obesity), adjusted for age, sex and socioeconomic status. Significance of trend was assessed by meta-regression.n = 1343; adjusted for age, sex and socioeconomic status). Part 6: subgroup analyses by sex, age groups (<50 years and ≥50 years), socioeconomic status group, and by follow-up time .Further parts were conducted comparing ≥55 weekly working hours to 35–40 h. Part 2: weight gain outcome as in analysis 1 plus adjustment for chronic somatic disease, psychological distress/depressive symptoms, smoking and physical activity. Part 3: sensitivity analysis excluding smokers . Part 4: sensitivity analysis excluding those with chronic disease at baseline . Part 5: sensitivity analysis excluding those with underweight  were without overweight or obesity at baseline; 42,965 (35.2%) had overweight and 17,970 (14.7%) had obesity. Of those without overweight/obesity, 31,703 (51.9%) worked standard 35–40 h a week, 10,568 (17.3%) worked 41–48 h, 3897 (6.4%) worked 49–54 h and 3947 (6.5%) worked 55 h or more. The number of part-time workers was 11,028 (18.0%). Of the cohort, 12,349 (20.2%) gained weight and were overweight or obese at follow-up. The duration of follow-up across studies was 1–9 years (mean 4.4 years), corresponding to 267,491 person-years at risk for the participants without overweight/obesity  or moderate (c) heterogeneity between study-specific estimates. τ-values in Panels a–d were 0, 0, 0.10 and 0.03, respectively.Figure Multivariable adjustment for chronic somatic disease, mental health and lifestyle attenuated the association between long working hours and onset of overweight/obesity, from 1.17 (95% CI 1.08–1.27) to 1.12 (95% CI 1.01–1.25) Fig. . SensitiFigure In this individual-participant data meta-analysis of 19 cohort studies with 122,078 participants, we examined the prospective associations between working hours and weight change in a greater detail than has previously been possible. We found a modest association but a suggestive dose-response relation between longer working hours at baseline and the risk of weight gain from non-overweight to overweight or obesity at follow-up. The association with weekly working hours of 55 or more was further examined and the analyses suggest that the finding is robust to adjustments and replicable across subgroups. However, our detailed analysis of the development of BMI showed that long working hours may increase the risk of a shift from normal to overweight but not from overweight to the obese category. Working hours were not associated with weight loss among those who had obesity.Our results are in line with previous prospective studies suggesting a positive association between long working hours and BMI –15 but iThe longest (≥55) weekly working hour category was associated with a 1.17-fold risk of new-onset overweight or obesity in the model adjusted for age, sex and socioeconomic status. The estimates from other working hour categories suggested a dose-response relationship although the overall association was modest. The association remained after adjustment for other lifestyle factors, chronic diseases and psychological distress, exclusion of smokers and those with chronic somatic disease or underweight, and in subgroup analyses by sex, age, socioeconomic status and length of follow-up. Thus, although weak, the association between long working hours and weight gain appeared to be robust. Our sensitivity analyses suggested that we can be most confident with the finding regarding a shift from normal to overweight category, with a RR of 1.16 with no significant heterogeneity in study-specific estimates.Possible mechanism explaining the association between long working hours and weight gain may be extended periods of sitting, because today, many occupations are sedentary. Other mechanisms may be linked to too little exercise and unhealthy diet, because people with long working hours may not have resources, time or energy to engage in a healthy lifestyle . Part-tiLong working hours were not associated with weight loss among participants with obesity. However, we could not distinguish between intentional and unintentional weight loss. One could assume that psychosocial factors, such as work-related stress, affect success in intentional attempts to lose weight among overweight individuals whereas other factors, such as the onset of physical diseases may contribute to unintentional weight loss. However, the relationship between stress and weight change has been suggested to be complex as for some people stress seem to induce weight gain whereas for others it is associated with weight loss and for still others stress has no significant effect on weight .Important strengths of our study include the first large-scale longitudinal individual participant data analysis of the association between working hours and weight change. A large sample size with harmonised variables reduces the likelihood of random error. We included cohort data from several countries in Europe, US and Australia, which increases generalisability of our findings, however, to those countries only. Notable limitations include an observational study design, which does not allow us to determine causality, and self-reported exposure to working hours and BMI in almost all cohorts. Working hours was measured only once although for some employees, long reported working hours may have had been a temporary ‘peak’ which was resolved shortly after the baseline survey whereas for others, excessive hours may have had been persistent. Self-report may also involve recall bias if the participants’ recall hours worked inaccurately or if they overestimate their height and underestimate their weight . HoweverThe PAF was 2.8% which suggests that overweight/obesity could be reduced by 2.8% if all full-time workers worked standard 35 to 40 h a week and the observed associations were causal. Given the multifactorial aetiology of obesity, this PAF is not trivial for population-wide prevention although the clinical significance at the individual level is modest. This size of PAF is comparable with those obtained for the relation of work stress (3.4%)  and deprIn conclusion, this individual participant meta-analysis of 19 cohort studies found a small excess risk of overweight among individuals who work longer hours. Detailed analyses were consistent with the hypothesis that the impact of long working hours involves the progression from normal to overweight rather than from overweight to obesity and that working long hours is not associated with weight loss among individuals with obesity. Future longitudinal studies assessing causality and underlying mechanisms, such as diet and accelerometer-measured physical activity would increase our understanding of the effects of long working hours on changes in body weight.Supplementary Table 1


Yin-tonic herbal medicines have been shown to possess properties that make skin healthy by nourishing within various organs of the body. However, the antiphotoaging effect of these medicines on the skin has not been fully studied. Photoaging occurs with prolonged sun exposure and causes skin damage and aging, with depletion of the dermal extracellular matrix and chronic alterations in skin structure, such as wrinkles. In this study, we assessed the antiphotoaging effects of eight yin-tonic herbal medicines on human skin cells and skin equivalents. The levels of type I procollagen and matrix metalloproteinase-1 (MMP-1) in ultraviolet B- (UVB-) irradiated CCD-986sk fibroblasts were measured, and then three medicines were chosen based on screening results. Using UVB-irradiated human skin equivalents, we evaluated the effect of three yin-tonic herbal medicines on histological changes of skin, epidermal and dermal thickness, and MMP-1 production. Furthermore, we observed collagen fiber content and protein expression of filaggrin in UVB-irradiated human skin equivalents. Yin-tonic herbal medicines increased type I procollagen levels and decreased the production of MMP-1 in UVB-irradiated CCD-986sk fibroblasts. The three selected yin-tonic herbal medicines recovered the collagen content and filaggrin expression via MMP-1 downregulation in UVB-irradiated human skin equivalents. Our results show that yin-tonic herbal medicines can prevent skin photoaging by reduction of MMP-1 levels and increasing the expression of moisturizing factors. Based on these results, we suggest that yin-tonic herbal medicines have the potential to be used as helpful agent for skin photoaging. Skin aging is a complex biological process that progresses as a person ages or is exposed to sun. Skin aging can be divided into two types, intrinsic or chronological aging and extrinsic or photoaging, based on the physiological and environmental factors, respectively . The intUVA 320–400 nm) is considered to have a minor effect on skin, although studies have shown that these rays can penetrate deep into the skin. UVC radiation (100–290 nm) is almost completely absorbed by the ozone layer and does not affect the skin. UVB (290–320 nm) has high energy to cross the epidermis and reach the upper dermis where it interacts with cellular chromophores, leading to increased oxidative stress and DNA damage . In othe00 nm is In the extracellular matrix (ECM), type I collagen is the predominant form of structural fibrous proteins. Procollagen, collagen precursor molecules, are synthesized by dermal fibroblasts and secreted into extracellular spaces, where it is enzymatically processed to mature collagen. Mature collagen pack together to form fibrils, which are largely responsible for the tensile strength and mechanical resilience of connective tissues in skin. The loss of collagen in the ECM can be attributed to the reduced synthesis and abnormal breakdown of dermal collagen . Under tin vitro and human skin equivalent in vivo model.The concept of yin and yang is a fundamental principle of traditional medicine and the harmony of body can be understood through the philosophy of yin and yang . AccordiIscove's modified Dulbecco's medium (IMDM), Dulbecco's Modified Eagles medium (DMEM), Dulbecco's phosphate-buffered saline (DPBS), fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Life Technologies Inc. . 3--2,5-diphenyltetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) were purchased from Junsei Chemical Co., Ltd. . Procollagen Type I C-peptide (PIP) EIA kit was purchased from Takara Bio Inc. . Human Total MMP-1 DuoSet ELISA kit was obtained from R&D Systems .in vitro experiment, the extract powder that resulted from the drying process was dissolved in distilled water.The eight yin-tonic herbal medicines used in this study are listed in μg/mL) in a humidified atmosphere with 5% CO2. HaCaT keratinocytes were provided by Professor Jae-Young Um  and were incubated at 37°C in DMEM supplemented with 10% FBS, penicillin (100 U/mL), and streptomycin (100 μg/mL) in a humidified atmosphere with 5% CO2.CCD-986sk fibroblasts were purchased from Korea Cell Line Bank . The cells were incubated at 37°C in IMDM supplemented with 10% FBS, penicillin (100 U/mL) and streptomycin  for 4 h at 37°C. After discarding the supernatant, the insoluble formazan product was dissolved in DMSO. Cell viability was measured at 540 nm using a microplate reader .2 was used according to the previous reports [As a UVB source, a UVB lamp , having an emission spectrum of 280–320 nm and the spectral peak of UVB, was set at 312 nm in this study. The programmed microprocessor controlled the constant UVB light emission with precise irradiation in energy or time. After CCD-986sk, fibroblasts were seeded and precultured with serum-free medium for 24 h and then treated with samples in serum-free medium for 24 h. Before UV irradiation, the cells were washed twice with DPBS and submerged in a minimum volume of DBPS, followed by exposure to UVB light using the UVB lamp. The UVB dose of 20 mJ/cm reports –18. AfteHarvested cell supernatants and culture media from the human skin equivalent model were used for the experiments. Type I procollagen and MMP-1 production measurement was performed using an ELISA kit in accordance with the manufacturer's protocol.®-ED which is a human skin equivalent model was purchased from TEGO Science . Briefly, human dermal fibroblasts were cultured in the collagen matrix for 1 day before keratinocytes were seeded on top of the collagen matrix and cocultured for 4 days. The keratinocytes were then lifted, and the human dermal fibroblasts block was exposed to the air. Samples were treated for 1 h, and the cells were irradiated with 50 mJ/cm2 solar UVB twice daily for 8 days. The skin equivalent was incubated at 37°C in an atmosphere of 5% CO2. The MTT assay was performed in accordance with the manufacturer's protocol. Briefly, human skin equivalent was pretreated with samples and incubated for 24 h, followed by incubation with MTT solution (0.3 mg/mL) for 3 h at 37°C. Separated skin equivalent samples were taken using 8 mm biopsy punch and were decolorized by stirring in 0.04 N HCl-isopropanol solution for 4 h. Cell viability was measured at 570 nm using a microplate reader.Neoderm2 UVB twice daily for 8 days. The skin equivalent was incubated at 37 °C in an atmosphere of 5% CO2.UVB irradiation and treatment with the samples were performed according to a method previously described –22 with μm thick, and hematoxylin and eosin (H&E) and Masson's trichrome staining were performed. For immunohistochemical staining, a portion of the skin samples from the back of mice in each group was fixed in 10% formalin. After paraffin embedding, sections were cut and the slides were deparaffinized by xylene, rehydrated in ethanol, and rehydrated in water. Endogenous peroxidase activity was blocked using 0.6% H2O2 in 50% methanol, and the slides were then treated with 0.3 triton in DPBS for permeabilization and problocked with 10% normal goat serum (NGS) for 1 h, followed by overnight incubation with a specific antibody at 4°C. The sections were then washed and incubated with horseradish peroxidase-conjugated secondary antibodies for 1 h at room temperature. The activity was visualized with 3,3′-diaminobenzidinechromogen and counterstained with H&E. Pathological changes of all stained skin sections were observed using a DM IL LED microscope  and photographed using a DFC295 . Digital images were taken from each slide (2 per group) and measured using Leica Application Suite .Human skin equivalent samples were fixed in 10% buffered formalin, embedded in paraffin, sectioned into 4 p values <0.05 were considered statistically significant. Statistical analysis was performed using SPSS statistical analysis software .Data are expressed as the mean ± SD for triplicate experiments. Statistically significant values were compared using ANOVA and Dunnett's post hoc test, and μg/mL doses of the eight yin-tonic herbal medicines did not affect the viability of both of the cell lines, rather skin constituting cells were proliferated by the treatment. Thus, the extracts concentrations 125, 250, and 500 μg/mL were used for subsequent studies.To test whether the eight yin-tonic herbal medicines could affect cell viability on skin cells, an MTT assay was used. The cell cytotoxicity of the eight yin-tonic herbal medicines against human keratinocytes (HaCaT cells) and human skin fibroblasts (CCD-986sk cells) is shown in Tables As a key indicator of photoaging, a decreased collagen production is observed in dermal fibroblasts. UV irradiation causes alternations of dermal collagen through stimulation of collagen breakdown, resulting in a fragmented and disorganized collagen fibers . Therefo2) for 30, 60, 90, or 120 seconds, respectively, twice a day for 8 days. UVB irradiation decreased the cell viability, showing a rapid decline from UVB irradiation for 60 seconds in the human skin equivalent (2) for 40 seconds considered the effect of UVB on the human skin equivalent and then conducted subsequent studies. The schematic diagram of UVB irradiation and treatment with yin-tonic herbal medicines using the human skin equivalent model is shown in To examine the effect of DB, MA, and EP on 3-dimensional skin, human skin equivalent was prepared. The MTT assay was used to determine the permissible concentration of DB, MA, and EP to be used on the human skin equivalent. The treatment with DB, MA, and EP at concentrations of 62.5, 125, 250, and 500 µg/ml increased cell viability, compared to nontreated cells . To deteuivalent . We alsoin vitro results, collagen content was restored in the UVB-irradiated human skin equivalent  substances that maintain skin hydration and skin barrier function . In the Dioscorea batata Decaisne is neutral in nature, is sweet in flavor, and is utilized to cure yin deficiency in metabolic disorders, such as diabetes and hyperthyroidism, by tonifying and replenishing organs [Morus alba L. is commonly used as a tonic, sedative, and laxative herbal medicine [Eclipta prostrata L. is also added to medicinal tonics that are usually used for treating dizziness, hemoptysis, hematuria, tinnitus, and uterine bleeding [in vitro study. Human keratinocytes are located in the outermost skin layer and therefore continuously exposed to UVR. It has been reported that the deleterious effects of UVB radiation on keratinocytes induce inflammation, cell death, senescence, and epithelial tumors [D. batata [M. alba [E. prostrata [D. batata, anthocyanin from M. alba, and wedelolactone from E. prostrata, exert the antioxidant and anti-inflammatory effects [The root of g organs . The frumedicine . Ecliptableeding . Considel tumors . Considel tumors  and vital tumors  have beel tumors , 36 as wl tumors , implyinl tumors . In addil tumors . Since al tumors . Recentll tumors , 41. Ind. batata –44, M. a[M. alba , 46, androstrata , 48 have effects –51, suppin vitro and showed a significant antiphotoaging effect including the restoration of collagen and filaggrin expression on human skin equivalent in vivo. Overall, the results obtained in this study suggested the promising complementary materials for improving skin and contributed to elucidating the mechanisms of antiphotoaging properties of yin-tonic herbal medicines. We believe that our findings could be used to help better understand the antiaging effect of yin-tonic herbal medicine and identify the scientific basis of herbal medicine background knowledge.In conclusion, the yin-tonic herbal medicines provided potential antiphotoaging and antiwrinkle activities


We have previously shown in 41 patients with PsA (Caspar criteria) and obesity  and to study the effects on cardiovascular risk factors.The patients were assessed with 66/68 joints count, Leeds enthesitis index (LEI), body surface area, blood pressure, BMI, questionnaires and fasting blood samples at the 12- and 24-month visits.p = 0.008) and 45.7%  at the 12- and 24-month visits.In total, 39 and 35 PsA patients attended the 12- and the 24-month visits, respectively. Median weight loss since baseline was 16.0% (IQR 10.5–22.4) and 7.4% (IQR 5.1–14.0) at the 12- and 24-months follow-up. The 66/68 swollen/tender joints score, LEI, CRP and HAQ score were still significantly reduced at the 12- and 24-month visits compared to baseline. The number of patients with Minimal Disease Activity increased from 28.2% (11/39) at baseline, to 38.5% (15/39; The weight loss was also associated with improved levels of serum lipids, glucose and urate and the antihypertensive treatment was reduced or stopped in five patients during the follow-up.Weight loss treatment, with VLED included in the program, was associated with long-term improvement of measures of disease activity, self-reported function and markers of the metabolic syndrome after 24-months follow-up.NCT02917434, Registered September 28, 2016- Retrospectively registered.ClinicalTrials.gov identifier:  Psoriatic arthritis (PsA) is an inflammatory disease characterized by psoriasis, arthritis, enthesitis and dactylitis . Obesityp = 0.002) [We have previously in an open interventional study shown that weight loss treatment with Very Low Energy Diet (VLED) in 41 patients with PsA and obesity resulted in a substantial weight loss of median 18.7 kg (interquartile range: IQR 14.6–26.5) after six months, which was associated with positive effects on the disease activity in joints, entheses and skin . Signifi= 0.002) .Short term weight loss can however easily be regained. Moreover, the anti-inflammatory effects seen in our previous study could have been caused by, both metabolic effects due to energy restriction during the initial phase of rapid weight loss, as well as loss of adipose tissue and lessened production of pro-inflammatory cytokines and adipokines therein.The primary aim of this follow-up was to study the association between weight loss and disease activity in longer term; 12 and 24 months in patients with PsA and obesity. A secondary aim was to study the effects of weight loss on aspects of the MetS in patients with PsA and matched controls.2 or more, and age 25–75 years [The patients were recruited from the departments of Rheumatology at Sahlgrenska University Hospital and the hospitals of Borås and Alingsås. Inclusion criteria were having PsA fulfilling the Classification for Psoriatic Arthritis (CASPAR) criteria, a body mass index (BMI) of 33 kg/m75 years . If treaThe controls were patients with obesity planned for treatment with VLED and matched for sex, age and body weight to the PsA patients, and they were recruited from the Regional Obesity Centre of the Region Western Sweden at Sahlgrenska University Hospital. Exclusion criteria for controls were having psoriasis, PsA or any other inflammatory rheumatic disease in addition to the above stated exclusion criteria for the PsA patients. The controls were used as comparators for vital signs, biochemical measures and effects on cardiovascular risk factors of the VLED treatment.The partcipants´ treatments for hypertension, diabetes and hyperlipidemia were monitored and could be changed throughout the duration of the study.All the participants in the study gave their written informed consent. The study was approved by the Regional Ethics Committee in Gothenburg and carried out in accordance with the Helsinki declaration.2, the strict VLED treatment was maintained during 12 or 16 weeks. Non-energy-containing beverages were allowed ad libitum. After the strict VLED period, food was gradually reintroduced during a period of 12 weeks and each participant was given a personal dietary advise based on individual energy requirements for weight stability with a reduction of 30% to achieve further weight loss. The macronutrient distribution in the recommended diet was approximately 20 energy percent (E%) protein, 30–35 E% fat and 45–50 E% carbohydrates. Each patient was given a written meal plan including three main meals and a snack meal. The dietary advice focus on healthy food selection including vegetables, fruits, whole-grain cereals, low-fat dairy products and vegetable fats and oils, fish, poultry and lean meat and limited amount of energy-dense and nutrient-sparse foods such as chocolate, sweets, bakery foods, sugar-sweetened beverages. The food selection was modified based on individual food preferences for each patient. The overall aim of the dietary advice was to restrict total energy intake, saturated fats and added sugar and increase dietary fiber, unsaturated fats as well as essential nutrients. After 12 months the patients were no longer followed at the Regional Obesity Centre, but at the departments of Rheumatology. In addition, all patients were seen by a physiotherapist at baseline and after six, 12 and 24 months and were instructed to engage in health-enhancing physical activity at least 150 min per week.Weight loss treatment with VLED was provided at the Regional Obesity Centre of the Region Western Sweden at Sahlgrenska University Hospital, within a framework of medical follow-up, dietary advice and support during 12 months. The VLED provides a mixture of carbohydrates, proteins and fats, with added recommended doses of vitamins, minerals, trace elements and essential fatty acids, in powder formulas that require mixing with hot or cold water before consumption as shakes or soups. The VLED treatment in our study consisted of four doses of VLED per day, providing a daily intake of 640 kcal. . Depending on baseline BMI, < 40 or ≥ 40 kg/mThe patients with PsA were assessed at baseline and after three, six, 12 and 24 months. Body height was measured at baseline, and weight was measured at baseline and at every follow-up and BMI was calculated. Waist circumference was measured in standing position with a tape measure midway between the lower rib and iliac crest. Joints were examined with 66/68 swollen/tender joints count and entheses with Leeds enthesitis index . The extAt the 24 month visit the questionnaire Patient Global Impression of Change (PGIC) was used, where patients are asked to rate their overall status from the start of the study using the following scale: (1) very much improved, (2) much improved (3) minimally improved, (4) no change, (5) minimally worse, (6) much worse, (7) very much worse .The controls were assessed at baseline and at three, six and 12 months, but not at 24 months, with measurements of height, weight, waist circumference and blood pressure. Information about medical history and current medication was obtained via questionnaires.Blood samples were drawn from the participants in the morning after ≥8 h of fasting and analyzed for hemoglobin (Hb), white blood cell count (WBC), platelet count (PLT), C-reactive protein (CRP), alanine transaminase (ALT), creatinine, urate, glucose, glycosylated hemoglobin (HbA1c), total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol and triglycerides (TG) using standard laboratory techniques at Sahlgrenska University Hospital.1elevated waist circumference (≥80 cm for women and ≥ 94 cm for men), 2elevated TG (TG ≥1.7 mmol/L or drug treatment for elevated TG), 3reduced HDL (HDL < 1.3 mmol/L for women and < 1.0 mmol/L for men or drug treatment for reduced HDL), 4elevated blood pressure (BP) (systolic BP ≥130 mmHg or diastolic BP ≥85 mmHg or antihypertensive drug treatment), 5elevated fasting glucose .Occurrence of MetS was defined as exhibiting three or more of the following five criteria : 1elevatS). All tests were two-tailed and p ≤ 0.05 was considered statistically significant. Only the 39 PsA patients and 39 controls who attended the 12- month visit and the 35 PsA patients who attended the 24-month visit, were included in the statistical analyses.Statistical analyses were made using SPSS Statistics version 25 . Descriptive statistics are presented as median and IQR. Wilcoxon Signed Rank Test was used to compare continuous related samples and McNemar’s test to compare categorical related samples. Correlations were calculated using Spearman’s correlation . The demographics and the medication of the completers are shown in Table In total, 46 patients with PsA were included and started VLED treatment, whereof 41, 39 and 35 patients attended the six, 12- and 24-month visits respectively. The 11 patients (8 women and 3 men) lost to follow-up during the whole study were significantly younger than the patients who continued in the study . In median, the patients had lost 16.1 (10.5–22.8) % of their baseline weight. In total, 77% (N = 30/39) of the patients had a weight loss of 10% or more compared to baseline, and 17.9% (N = 7/39) a weight loss of 5–10%. The majority of patients (34/39) had however regained some weight between the six- and 12-month visits, in median 3.9 (IQR 1.5–6.6) kg.There were 39 patients with PsA who attended the 12-month visit. At baseline their median weight was 106.0 (IQR 93.5–112.5) kg and at 12 months 87.5 (80.6–95.5) kg (p = 0.008). However, no significant difference was found in VAS pain and DLQI at 12 months follow-up, compared to baseline.  at baseline and 38.5% (15/39) at the 12-months follow-up (9 at basep = 0.041), and the men a greater reduction in swollen joints (∆swollen/66) than the women (p = 0.047), but there was no significant difference in change in BMI, DAPSA, DA28CRP, BSA, HAQ between the sexes. PsA patients on a bDMARD had a greater reduction of tender joints  than those without, but otherwise the change in disease activity was similar between the groups at 12 months. The weight loss and change in CRP was also comparable in the PsA patients and controls at the six and 12-month visits  compared to the men p = 0.01, and thN = 14/35) of the patients still had a weight loss of 10% or more compared to baseline, and 37.1% (N = 13/35) had a weight loss of 5–10%. All patients had however regained some weight since the 12-month visit, in median 6.3 (IQR 3.4–9.3) kg.In total, 35 patients were examined at the 24-month visit. The median weight was 92.7 (IQR 85.7–100.7) kg and the median weight loss 7.4 (IQR 5.1–14.0) % of the baseline weight. Forty percent . There were however no significant differences in BSA, DLQI, BASFI and VAS global health, pain and fatigue between baseline and 24 months.At 24 months the swollen/66 and tender/68 joints count, Leeds enthesitis index, CRP, HAQ, DAS28-CRP and DAPSA scores were still significantly lower compared to baseline. (Table p = 0.007) and DAS28CRP (∆DAS28CRP) (p = 0.011) than the men, but there was no significant difference in the change in BMI, CRP, swollen/tender joints, DAPSA, BSA, HAQ between the sexes. Patients treatment with a bDMARD had a greater reduction in tender joints  and DAPSA  compared to patients without a bDMARD at 24 months. No comparisons between PsA patients and controls could be made at the 24-month visit.Subgroup analyses showed that the women demonstrated a greater reduction in VAS Global health , ∆DAPSA , ∆CRP , ∆BASFI  and ∆VAS for global health  in the PsA patients.  and ∆VAS for fatigue .  at 24 months.At 12-months follow-up the change in BMI compared to baseline (∆BMI) was significantly correlated with ∆DAS28CRP  and ∆DAS28CRP  at the 12 months visit. In addition, ∆LDL was correlated with ∆DAPSA  at the 24-month visit.The change in serum levels of TG (∆TG) was correlated with ∆DAPSA  and controls (p = 0.001). The decrease in serum urate (Δ-urate) was however not significantly correlated with Δ-BMI at any timepoint in the female and male patients and controls except in the PsA women at the 24-month visit .Serum urate was also significantly reduced during the study in both PsA patients and controls. Table . BaselinIn this prospective, open intervention study, we analyzed the long-term effects of weight loss on disease activity and self-reported physical function in patients with PsA and obesity. After 12 months of structured weight loss treatment, a median weight reduction of 16% was associated with improvement of a majority of disease activity measures compared to baseline, including swollen/tender joints count, enthesitis, extent of psoriasis, CRP, HAQ, BASFI and VAS global health and fatigue. After 24 months, all patients had regained some weight, but a significant improvement could still be shown in swollen/tender joints count, enthesitis, CRP and HAQ, compared to baseline. The results of the study support the hypothesis of obesity as a promotor of disease activity in PsA, showing what could be attained by adding a weight loss program to routine medical care in patients with PsA and obesity.A secondary aim was to study the effects on variables associated with the MetS in PsA patients and matched controls. The magnitude of the weight loss was similar in the PsA patients and controls and associated with comparable reductions in waist circumference and serum levels of CRP, HBA1c, glucose, TC, LDL, TG and urate and increase in HDL, showing that the beneficial effects of weight loss treatment on inflammation and cardiovascular risk factors is general.In the present study the weight loss treatment with VLED was judged as easy to implement by most PsA patients, but the transition from VLED to normal food was perceived as harder . A substTotally 11 PsA patients and 13 controls were lost to follow-up during the study. It is possible that the participants who did not come to follow-up had a poorer weight maintenance than the participants who continued in the study.Studies on the effects of weight loss on disease activity in PsA are scarce, but there are a few studies on psoriasis. Effect on disease severity in psoriasis has been shown after weight loss by bariatric surgery , 39 and Obesity is a pro-inflammatory state, where the adipose tissue is invaded by activated M1-type macrophages and B- and T-lymphocytes, with an over-production of cytokines, such as tumor necrosis factor-α (TNFα) and interleukin (IL)1, IL6, IL17 and IL23, which are all involved in the pathogenesis of psoriasis and PsA , 50. MorIn the present study, weight loss was associated with a reduction of other cardiovascular risk factors such as hypertension and dyslipidemia, and additionally lowered levels of blood glucose and HbA1c. MetS is defined as a set of cardiovascular risk factors that cluster together; enlarged waist circumference, high blood glucose levels, elevated blood pressure and TG, and reduced HDL . InsulinPatients with psoriasis and PsA are at increased risk of developing gout . The parStrengths of the study are the prospective design with a long follow-up and the powerful intervention which resulted in a considerable weight loss.There are also limitations of the present study to be acknowledged. Firstly, this is not a randomized controlled study and the absence of an untreated control group with PsA is a major short-coming. A control group consisting of patients with obesity, but otherwise healthy, matched for sex, age and bodyweight, undergoing the same VLED treatment was however included in the study, to enable comparisons of vital signs and biochemical measures. Secondly, a threshold for disease activity was not among the inclusion criteria of the study. The study included patient from every day practice, whereof some fulfilled criteria for MDA already at baseline and some were near fulfillment of MDA. The data concerning MDA should therefor be interpreted with caution, since it is possible that even a minor improvement can have turned some of the non-MDA patients into MDA. In addition, the number of included patients was small. Nevertheless, the study was able to show significant association between weight loss and reduced disease activity.Thirdly, treatment with cs/bDMARDs was held constant from three month prior to baseline until the six-month visit, but change in the cs/bDMARD treatment was allowed after the six-month visit, due to ethical reasons. We cannot exclude that change in treatment may have affected the results regarding disease activity at the 12- and 24-month visits. In the vast majority of patients, the medication with cs/bDMARDs was however held unchanged during the whole study period. Fourthly, pharmacological treatment against hypertension, hyperlipidemia and diabetes could not be held constant throughout the study, which may have affected levels of blood pressure, blood lipids and glucose metabolism during the follow-up. During the initial period with intense weight loss, some patients experienced hypotension and antihypertensive treatment had to be tapered. Fifthly, there were no treatment or follow-up visits during the second year that also may have affected the weight regain among most PsA patients. The results indicate that prolonged treatment periods for weight maintenance or further weight loss may be needed.Weight loss treatment with VLED in patients with PsA and obesity was associated with sustained lowered disease activity after 12 months and 24 months of follow-up, with concurrent improvements in cardiovascular risk factors and lowered serum urate. The results of the study provide support to the hypothesis of obesity as a promotor of disease activity in PsA and shows what can be achieved by 5–15% weight loss in patients with PsA and obesity.Additional file 1: Suppl Table 1. Parameters associated with the metabolic syndrome at baseline (BL) and after weight loss at 6 months (M6) and 12 months (M12) in controls with obesity. Values are median and inter-quartile range (IQR).


DNA damage accumulates over the course of cancer development. The often-substantial amount of somatic mutations in cancer poses a challenge to traditional methods to characterize tumors based on driver mutations. However, advances in machine learning technology can take advantage of this substantial amount of data.https://github.com/jessada/pyCancerSig.We developed a command line interface python package, pyCancerSig, to perform sample profiling by integrating single nucleotide variation (SNV), structural variation (SV) and microsatellite instability (MSI) profiles into a unified profile. It also provides a command to decipher underlying cancer processes, employing an unsupervised learning technique, Non-negative Matrix Factorization, and a command to visualize the results. The package accepts common standard file formats . The program was evaluated using a cohort of breast- and colorectal cancer from The Cancer Genome Atlas project (TCGA). The result showed that by integrating multiple mutations modes, the tool can correctly identify cases with known clear mutational signatures and can strengthen signatures in cases with unclear signal from an SNV-only profile. The software package is available at pyCancerSig has demonstrated its capability in identifying known and unknown cancer processes, and at the same time, illuminates the association within and between the mutation modes. Cancer is a genomic disorder, involving different kinds of DNA damage. DNA damage and imperfect repair occurs frequently in human cells. Changes accumulate over time, starting with our first cell, the fertilized egg, and progressively over the course of cell division . CellulaBRCA1/BRCA2 null mutation  G in the feature output file.SV profiling scans the VCF record for the INFO field SVTYPE to classify the mutation into structural duplication, structural deletion, inversion, and translocation. Then it checks another INFO field, END, to calculate the length of the event, in order to subclassify the event based on its approximated size in log10 scale . In total, there were 32 mutation types (4 types of variation × 8 length groups). For example, if a VCF record has an INFO field SVTYPE set to “DUP” and a length of 15,000 bp, this would be counted as one mutation for structural duplication of length between 10 K–100 K.MSI profiling quantifies all possible repeat patterns of a repeat unit with size between 1 and 3 nucleotides . For repeat unit sizes of 4 and 5 base pairs, it performs quantification without checking repeat patterns. The profile merger merges all sample profiles into one ready to be used by the deciphering- and visualization processes.The purpose of this step is to use an unsupervised learning model to find mutational signature components in the tumors. The process was developed based on a well-established framework  and usesThe purpose of this step is to decompose the tumor mutational signature components and visualize them together with mutation profiles of the tumor. The visualized profiles are generated in pdf format files consisting of 4 pages Fig. , adaptedA cohort of 92 breast cancer cases, TCGA-BRCA, and 38 colorectal cancer cases, TCGA-COAD and TCGA-READ, from The Cancer Genome Atlas project (TCGA) was used to evaluate the tool. These 130 cases were all of TCGA breast cancer and colorectal cancer cases where we could obtain both whole genome and whole exome sequencing data from dbGAP. The data consists of tumor-normal pair whole genome sequence in BAM format and SNV exome sequencing data called by MuTect2  in VCF fFor SNVs, the downloaded somatic VCF files called by MuTect2 were in a format ready to be used for SNV profiling in the next stage. For SVs, a singularity container implementation of FindSV was used with downloaded aligned bam files. Briefly, balanced structural rearrangements were called by TIDDIT . Copy nuClinical information on the cases was provided by TCGA. Annotation of somatic SNVs in genes of interest , and SV-only. The SNV-only signatures used in the package verification were from COSMIC Mutational Signatures (v2 - March 2015) . pyCancePOLE as proposed etiology. All of these cases were found to have somatic mutation in POLE and POLD1  truncating variants in BRCA1, p.Val1734Ter, p.Glu23ValfsTer17 (both germline) and p.Glu720Ter, c.4739-1G > A (both somatic). One case has a somatic truncating variant in BRCA2, c.7805 + 1G > A. One breast cancer case had bi-allelic truncating variants in BARD1. The rest had truncating variants in BLM, ATM and ATRX. In addition, 30 other cases (27 breast cancer and 3 CRC) had in-frame or missense variants in 22 DSB genes  repair by homologous recombination. Using SNV profiling by pyCancerSig, 78 breast cancer– and 10 colorectal cancer cases were found to be associated with signature SBS3. Among these, eleven breast cancer cases and one colorectal case had pathogenic truncating variants in DSB-pathway genes. Four cases have truncating variants in Visual comparison with a leading tool specialized on profiling, deconstruction and visualisation (deconstructSigs) showed effectively identical results in the five cases with signature SBS10.E and I were only associated with colorectal cancer cases, while signatures C, D, G and H were only associated with breast cancer cases and signatures. A, B and F were found to be associated with both cancer types  as well as a high TMB (881/Mbps).Signature I was the only signature with a pattern of MSI identified by msisensor [CG > TG backgrounds. The majority of SV events were structural deletion of 1 K–10 K. Signature I was associated with four colorectal cancer cases, two of which were the two cases associated with signature SBS6.Signature sisensor . It exhiI. TCGA-AZ-6601 had both a germline truncating variant in MSH6 (p.Lys537Ilefs*33) and a somatic in frame change in MSH6 suggesting a mismatch repair defect, while no pathogenic variants were found in TCGA-AD-6964.  and one was not associated with any signatures. The median SV burden in these eleven cases (1004 events) did not differ significantly from the median of the entire cohort . , signature a for structural deletion 1 K–10 K, signature c for structural inversion of size 1 K–10 K, signature d for structural deletion of size 10 K–100 K and signature e for structural duplication of size 10 K–100 K. The rest were not associated with any signatures . SV profiler can process 1755 variants in 0.1 s . MSI profiler can process 65,535 loci in 0.3 s . Deciphering a combined profile for 130 samples took 33 min. Visualizer took 9 s to generate the pdf file for one sample.In the present study, a command line tool was developed and tested on two different cancer types. We report a working tool, and study the use of a combined SV, SNV and MSI profile, contrasting to SNV or SV only profiles.POLE and POLD1: signature E, and loss-of-function in MMR genes: signature I. Moreover, beside backward compatibility to SNV-only signature SBS6, signature I generated by the combined profile has identified two additional cases, TCGA-AD-6964 and TCGA-AZ-6601, with limited association to signature SBS6 (23 and 18%) which were also MSI positive according to msisensor. In principle, it could be possible to reduce the required level of association to signature SBS6 in order to increase the sensitivity of the SNV-only profile. However, this also reduces the specificity. In total, 10 colorectal cancer and 20 breast cancer cases displayed a contribution from signature SBS6, ranging from weight 6 to 42% of the total mutational profile. Four of them had weights in the range 20–30%, including one MSI and three MSS cases. Integrating SNV, SV and MSI into one single profile significantly increased the strength of the signal (signature I) due to a specific SV pattern  and MSI.This study has shown that combining SNV, SV and MSI into one single profile can correctly identify mutational signature processes caused by altered activity of the DNA polymerases BRCA1/2-mutated samples displayed a nonspecific SV pattern and did not contain a high number of SV events. This is in contrast to previous work where BRCA1/2 deficient structural variation-only profiles were detected as clustered events [COSMIC signature SBS3 is generally described as associated with deficiency in homologous recombination repair of double stranded breaks . SNV-onld events , 28, 29.Whether to use a combined or SV-only signature depends on the problem at hand. Combining an informative profile with a non-informative one may weaken the signal, depending on the amount of training data available. On the other hand, combining two informative profiles of two different mutation modes will increase accuracy, as in the cases with MMR gene mutation.When analyzing SVB, a clear difference between cases with high SVB and low SVB . It is useful for systematic interpretation for multiple tumors and can be worked on with basic data processing operations, like “sort”, “not zero”, “more than”, etc.The aim of this python package was to have a complete integrated set of tools written in Python to work with and answer questions related to mutational signatures of multiple variant types. Even though the package was benchmarked with the recommended variant callers, other bioinformaticians may use different variant callers which may produce output in different formats, which may not be compatible with our tools. So, we have designed our workflow Fig.  to be flThis pattern-based study has shown that applying an unsupervised learning method on joint information from multiple variant types can identify the cases suggested by the SNV-only profile and demonstrated that integrating SV and MSI into the profile can strengthen previously unclear signals. This illuminated the association within and between the mutation modes. The tool presented, pyCancerSig, accepts common file formats, and is easily incorporated into a genome center pipeline.Project name: pyCancerSigProject home page:https://github.com/jessada/pyCancerSigOperating system(s): Platform independentProgramming language: PythonOther requirements: Python 3.0 or higherLicense: GNU v.3.0.Any restrictions to use by non-academics: None.Additional file 1.Additional file 2.Additional file 3.Additional file 4.Additional file 5.Additional file 6.Additional file 7.


Cyrotrachelus buqueti (Coleoptera: Curculionidae) is a destructive forest pest and distributed widely in Southeast Asia. The 15,035 bp complete mitochondrial genome of the species consists of 13 protein-coding genes (PCGs), two ribosomal RNA genes (rRNAs), 21 transfer RNA genes (tRNAs) and a control region (GenBank accession no. MG674390). The trnl gene was not found in the C. buqueti mitogenome. The gene order and the orientation of C. buqueti were similar to those found in other Coleoptera species. The nucleotide composition was significantly biased  with A + T contents of 73.74%. ATG, ATA, ATT, AAT, and TTG were initiation codons and TAA, TAG, and T were termination codons. All the 21 tRNAs displayed a typical cloverleaf secondary structure, except for 1trnS which lacked the dihydrouridine arm. Phylogenetic analysis was performed using 13 PCGs with 14 other beetles showed that C. buqueti is closely related to Eucryptorhynchus brandti, which agree with the traditional classification.The bamboo snout beetle  Cyrtotrachelus buqueti Guerin-Meneville (Coleoptera: Curculionidae), is a highly destructive forest pest and distributed widely in Southeast Asia . Samples have been deposited in the insect specimen room of Guiyang University with an accession number GYU-Col-20170001).The bamboo snout beetle, C. buqueti was 15,035 bp in length (GenBank accession no. MG674390) and included sets of genes, including 13 protein-coding genes (PCGs), two ribosomal RNA genes (rrnL and rrnS), 21 transfer RNA genes (tRNAs), and a large non-coding region (putative control region). The trnl was not found in the C. buqueti mitogenome, as observed in Sympiezomias velatus  lacking a stable dihydrouridine arm, which were consistent with those reported in other animal mitogenomes (Wolstenholme trnC and trnE) to 71 bp (trnK), A + T content ranged from 58.46% (trnN) to 86.15% (trnD). The rrnL was located between 1trnL and trnV, and rrnS resided between trnV and the control region. The rrnL was 1298 bp in length with A + T content of 78.43%, and the rrnS was 788 bp in length with A + T content of 75.13%.All the 21 tRNAs were predicted to have typical cloverleaf secondary structures, except the gene C. buqueti were the canonical putative start codons ATN . However, cox1 and nad6 used AAT and TTG as start codon, respectively. Nine PCGs were terminated with TAA or TAG, and the remaining PCGs including cox1, cox3, nad4, and nad5 use a single T as stop codon. We analyzed the amino acid sequences of 13 PCGs with neighbour-joining method to construct the phylogenetic relationship of C. buqueti with 14 other representative bettles. The result showed that C. buqueti is closely related to Eucryptorhynchus brandti (The initial codons for 11 PCGs of  brandti , which a


Quercus spp. The volatile compounds detected when the barrels were toasted were analyzed. Five types of sorbents—carbon wide range/polydimethylsiloxane, divinylbenzene/carbon wide range/polydimethylsiloxane, divinylbenzene/polydimethylsiloxane, polydimethylsiloxane, and polyacrylate—were used for this investigation. Fifty-four volatile compounds were detected in Soju using gas chromatography/mass spectrometry. A high extraction efficiency was obtained using carbon wide range/polydimethylsiloxane. Nineteen samples were analyzed using barrels made of six species of carbonated oak  and control groups in three ways: noncharring, medium charring, and heavy charring. Ethanol, 1-propanol, isoamyl acetate, and isoamyl alcohol can be used as indicator volatile components for Soju and other such traditional Korean distilled liquors. We believe our study results can be used to design better analysis methods for Soju and other distilled liquors.The SPME Arrow technology—a novel solid phase micro-extraction technique—was used to analyze Soju, a traditional Korean distilled liquor, in barrels made of  Profiling of volatile compounds is an important step in determining the flavor of distilled spirits. Volatile compounds form during fermentation, distillation, and storage of distilled spirits, and are affected by various factors such as the type of raw material and the manufacturing and aging processes ,2. Soju,Quercus spp., a dominant broad-leafed species, is gradually spreading in the country [Q. aliena, GAL), gulchamnamu , tteoggalnamu , sangsurinamu , singalnamu , and jolchamnamu . These oaks also are part of broad-leafed Korean forests and were selected as representative oak species of Korea.Wines are typically stored in oak barrels. However, it causes their oxidation owing to the continuous flow of a small amount of air through a bunghole in the barrels; as a result, new volatile compounds are produced in the wine during aging ,6. It ha country . To prodThe conventional extraction methods for analyzing volatile food substances are vacuum simultaneous steam distillation solvent extraction (V-SED) ,12, direRecently, a new technology—SPME Arrow—has been developed that combines the benefits of both SPME and HS-SBSE. SPME Arrow protects the fiber with an outer metal tube, making the fiber more stable and extending its lifetime. As a result, the limitations of mechanical failure associated with SPME can be overcome . In addiThe present study aimed to find a suitable sorbent for Soju by analyzing the volatile substances present in different Korean distilled spirits using SPME Arrow. Five SPME Arrow fibers were analyzed for optimization. Finding a suitable adsorbent and using the results obtained, the study aims to analyze changes in the volatile profiles of Soju when aged in oak barrels made of six species of carbonated oak. Traditional Korean liquors such as Soju have not been sufficiently studied, in contrast to the distilled liquors of other countries that have been investigated extensively. This study will help improve the analysis of such Korean liquors.®/polydimethylsiloxane  adsorbent . In addition, 20-mL headspace vials and caps with PTFE/silicone septa were purchased from Thermo Fisher Scientific .Linalool, used as an internal standard (ISTD), and C7–C40 saturated alkane standard  were purchased from Sigma-Aldrich . The SPME Arrow fibers were coated with carbon wide range/polydimethylsiloxane , divinylbenzene/carbon wide range/polydimethylsiloxane , polydimethylsiloxane , divinylbenzene/polydimethylsiloxane , and polyacrylate , which were supplied by CTC Analytics AG . The SPME fiber was coated with the carboxenAspergillus luchuensis, Chungmujongguk). Saccharomyces cerevisiae 88-4 strain of Korea Food Research Institute was used as a fermentation starter. The fermentation was carried out in two stages: the first stage was performed for 5 days and the second stage was carried out for 15 days at 25–30 °C. The fermentation yielded 4180 L of liquor with 19.2% (v/v) alcohol content. The fermented liquor was then distilled under reduced pressure (100 mmHg) and 1548 L of distilled spirit with 49.3% (v/v) alcohol was obtained.Here, 1800 kg of white rice (Yeoju rice) was used as a raw material. In addition, 600 kg of rice ipguk was prepared using Baekguk  in triplicate. To extract the volatile compounds, 3 mL of the distilled liquor and 75 μg/mL of linalool (200 μg/mL) as ISTD were added into the headspace screw-top vial (20 mL) to achieve the final ISTD concentration of 4.878 μg/mL. The samples were heated and stirred at 40 °C for 10 min at 500 rpm to reach equilibrium. The SPME Arrow fiber was exposed to the headspace for 30 min at 1000 rpm. Then, it was inserted into the GC injector port operating in split mode (5:1) and desorbed at 220 °C for 5 min. After desorption, the Arrow fiber was subjected to a postconditioning treatment for 5 min to eliminate any possible contamination before performing the next analytical run.m/z and the scan rate was 0.4 scan per second in full scan mode. Electron ionization was carried out using ionization energy of 70 eV. The volatile compounds were identified by comparing the mass spectrum ratio of the sample with the data available in Wiley Registry 11th edition/NIST 2017 Mass Spectral Library. The experimental retention index was calculated using alkane standards (C7–C40) and compared with the literature retention index on DB-Wax. KI for the unknown compounds was calculated using the following equation [The analysis was performed using a TRACE 1310 gas chromatograph equipped with a TSQ 9000 triplus quadrupole mass spectrometer . All samples were analyzed using the DB-WAX capillary column . Helium gas was used as the carrier gas at a constant flow of 1.0 mL/min. The GC oven programs were held at 45 °C for 10 min, and then first increased to 105 °C at 2 °C/min and then to 220 °C at 3 °C/min and held for 5 min. The ion source and transfer line temperatures were maintained at 230 °C and 280 °C, respectively. The mass range was 35–550 equation :KIA= 100KI = Kováts index of compound An = number of carbon atoms in the smaller n-alkane eluting before compound AN = number of carbon atoms in the larger n-alkane eluting after compound ATr(A) = retention time of compound ATr(n) = retention time of the smaller n-alkane eluting before compound ATr(N) = retention time of the larger n-alkane eluting after compound A.The volatile compounds were quantified as relative concentrations calculated by comparing the peak area of each compound with that of the internal standard . The results were expressed as the average of triplicate runs for each of the 18 samples.p < 0.01) using SPSS Statistics ver. 23 . Heatmaps obtained using hierarchical clustering techniques were analyzed and visualized using the MetaboAnalyst version 4.0 software .The mean±standard deviation from triplicate measurements was presented. We assessed the results by analyzing the variance and Duncan’s multiple range test to identify significant differences  were analyzed. Samples that were not aged were used as controls. These samples were carbonized in three ways: noncharring (1), medium charring (3), and heavy charring (5) . Acids wVolatile components of food have been widely analyzed by the headspace extraction method. In this study, volatile compounds of Soju, a traditional distilled liquor of Korea, were extracted with the SPME Arrow fiber, which shows better robustness and sensitivity than the SPME fiber. The extraction efficiencies of volatile compounds were compared using five types of SPME Arrows. Fifty-four volatile compounds were identified: acetals (2), acids (1), alcohols (11), aldehydes (2), esters (20), furans (5), hydrocarbons (9), lactones (3), and phenols (1). The results showed that volatile compounds are extracted differently based on the properties of the sorbent used with the SPME Arrow fiber. The use of CAR/PDMS afforded high extraction efficiency. Using the results obtained, the volatile components of Soju stored in barrels made of different species of carbonized Korean oak were profiled. Ethanol, 1-propanol, isoamyl acetate, and isoamyl alcohol can be used as indicator volatile components. Our findings support the use of SPME Arrow for analyzing volatile compounds present in food. The results also show that future research into the volatiles of traditional Korean distilled liquors that affect their flavor and taste can be furthered by using different processing methods.


Selenium, as an antioxidant, has been implicated in the development of autoimmune thyroiditis (AIT). Many studies showed selenium supplementation could decrease thyroid autoantibodies in patients with AIT. However, the underlying mechanisms have not been well determined. Therefore, we performed a clinical study to investigate the possible mechanism of beneficial effects of selenium treatment on AIT patients. μg/day selenium supplementation, and group II received a placebo over a 3-month period. Thyroid stimulating hormone (TSH), thyroid peroxidase antibody (TPOAb), antithyroglobulin antibody (TgAb), malondialdehyde (MDA), total antioxidant capacity (TAC), and superoxide dismutase (SOD) were measured before and 3 months after treatments. Additionally, twenty healthy volunteers also served as a control group for the evaluation of such parameters in basic condition.  Forty euthyroid patients with AIT were randomized into two groups. Group I was treated with 200 n = 18; group II, n = 14) completed the clinical study and were incorporated into the statistics. MDA level was higher and SOD activity and TAC were lower in patients compared to healthy individuals. After 3 months, TPOAb titer significantly decreased within group I (P < 0.001) but did not change within group II (P=0.001). There were also no statistically significant changes in TSH and TgAb titers within the two groups . Additionally, decreased MDA level  and increased TAC  and SOD activity  were simultaneously observed after 3 months' selenium treatment. Moreover, there was a negative correlation between TAC and TgAb/TPOAb and a positive correlation between MDA and TgAb/TPOAb in AIT patients.  Totally, 32 patients (group I,  Our findings support the hypothesis that selenium treatment could decrease TPOAb titer via enforcing the defense against oxidative stress in euthyroid patients with AIT, which may be a potential underlying mechanism. Autoimmune thyroiditis (AIT) is a chronic autoimmune inflammatory disease of the thyroid gland. The T and B lymphocytes contribute to the pathogenesis of AIT by the production of autoantibodies to thyroglobulin and thyroid peroxidase and excessive reactive oxygen species (ROS) under the environmental influence or thyroid dysfunction . OxidatiSelenium, as an essential trace element, is also served as an antioxidant, and selenium deficiency has been proven to be associated with many diseases, such as cancer, diabetes, and AIT , 7–9. WuThere is growing evidence that oxidative stress is closely correlated with thyroid function. Previous studies have indicated an imbalance between oxidant stress and the defense system in hyperthyroidism –20. AddiThe present study was to evaluate the oxidant/antioxidant status in euthyroid patients with AIT and determine the correlation between oxidative stress parameters  and thyroid autoantibodies. Furthermore, we investigated whether selenium supplementation could decrease thyroid autoantibodies and reduce autoimmune responses by scavenging free radicals in newly diagnosed AIT patients.This is a prospective placebo-controlled randomized study carried out in our hospital between September 2016 and August 2017. A total of 40 patients with euthyroidism, newly diagnosed with AIT, and aged over 18 years old  were enrolled in this study. AIT was diagnosed when the serum thyroid peroxidase antibody (TPOAb) and/or thyroglobulin antibody (TgAb) was positive together with the presence of parenchymal heterogeneity in high-resolution sonography.μg/day selenium supplementation  orally for 3 months, and group II received placebo.Patients who had nonthyroidal disorders including cancer, hypertension, diabetes mellitus, coronary artery disease, chronic kidney disease, liver diseases, heart failure, cerebrovascular disease and rheumatism, smokers, use of other antioxidant agents or vitamin supplements within the past 6 months and those who were pregnant were not included in this study. All patients were randomized into two groups. Group I received 200 Moreover, age, sex, and body mass index (BMI) matched 20 healthy volunteers were served as a control group for the evaluation of oxidative stress parameters in basic condition. This study is conducted in accordance with the Declaration of Helsinki and was approved by the Ethics Research Committee of Tianjin Second People's Hospital, and informed consent was obtained from each patient and control subject.Venous blood samples were obtained after overnight fasting at pretreatment (baseline) for patients and healthy volunteers and at 3 months posttreatment for patients, and measured in plasma samples (collected in tubes containing 16 IU/ml heparin) for oxidative stress parameters and in serum samples for thyroid function and autoantibodies. After centrifugation at 4000 rpm for 10 minutes, the collected samples were stored at −80°C until assay.3, reference 3.50–6.50 pmol/L), free thyroxine  and TSH (reference 0.30–5.00 μIU/mL) levels, and TgAb (reference 0–40.00 IU/mL) and TPOAb (reference 0–35.00 IU/mL) titers were measured by a fully automated ADVIA Centaur analyzer  based on chemiluminescent reaction principle. Selenium levels were determined by absorption spectroscopy .Serum free triiodothyronine (FTn-butanol. The mixture was centrifuged at 1600 ×g for 10 minutes. The absorbance of the resultant supernatant was read at 532 nm.MDA level (nmol/ml) was estimated with the method based on the reaction of MDA with thiobarbituric acid (TBA) to form thiobarbituric acid-reactive substances (TBARS) . BrieflySuperoxide dismutase (SOD) activity (U/ml) was measured using the method described previously , which cTAC was measured using a commercial kit based on the ability of antioxidants in plasma to inhibit the oxidation of 2,2′-azino-di(3-ethylbenzthiazoline sulphonate) (ABTS) to ABTS + by metmyoglobin . The abst-test or Mann–Whitney U test was used to compare the baseline parameters between the AIT patients and healthy controls. A Chi-square test was used to compare the categorical data. Changes in the variables pre- and posttreatment were assessed using a paired samples t-test or Wilcoxon signed rank test. Correlation analyses were performed with a Pearson's correlation test. A P < 0.05 was considered statistically significant.All data were analyzed by SPSS software . The normal distribution of data was evaluated with the Kolmogorov–Smirnov test. Numerical variables were expressed as mean ± standard deviation (SD) or median (25th–75th percentile) depending on normal distribution or not. A Student's The population of this study consisted of 40 patients with AIT and 20 healthy controls. During the study, 4 patients in the selenium supplementation group (group I) and 4 in the placebo group (group II) dropped out of this research because of not insisting on taking drugs. Finally, 32 patients completed this study. No significant adverse events were reported by the participants.3, FT4, TSH, and BMI . Although selenium levels in the patient group were lower than those in the control group, they were both in the normal range.The basal characteristics of patients and healthy controls are presented in P=0.004) whereas the mean TAC and SOD activities were lower in patients than the controls (both P < 0.001).The plasma oxidative stress parameters of patients and controls are shown in n = 18) or placebo  treatment are shown in 4 were still normal after treatment. With regard to the thyroid autoantibodies, after selenium supplementation, TPOAb titer significantly decreased at 3 months compared to baseline , P < 0.001) whereas there was no obvious change in the placebo group , P=0.110) (P=0.081) and group II , P=0.363)  . MoreoveP=0.363) .t-test indicated a significant decrease in MDA level  and increase in TAC  and SOD  activity in the selenium group, whereas there were no significant changes for the three indexes in the placebo group   and positive correlations between MDA and TgAb or TPOAb  in the AIT patients, but no obvious relationships were found between them in the healthy controls. Additionally, we did not find any correlation between SOD and the two thyroid autoantibodies in the two groups.At baseline, the correlation analysis between oxidative stress parameters and thyroid autoantibodies titers is demonstrated in In this prospective study, we demonstrated that the MDA level was higher, while antioxidative defense capacity was lower in euthyroid patients with AIT as compared to healthy individuals. Selenium supplementation could not only reduce the oxidative stress, but to some extent, decrease the TPOAb titer. Additionally, we found that TPOAb and TgAb correlated positively with MDA and negatively with TAC, suggesting an interdependent relationship between thyroid autoimmunity and enhanced oxidative stress in AIT patients. We suspect that selenium treatment may decrease TPOAb titer through reducing oxidative stress, and this can be served as a potential underlying mechanism of inhibiting thyroid autoimmune response in the development of AIT.Oxidative stress occurs as a result of either overproduction of ROS or insufficiency of antioxidant defense systems . GeneralAIT is a chronic autoimmune disease of the thyroid gland. Physiological ROS is essential for the synthesis of thyroid hormone . However4-treated AIT population, while reduce TPOAb and TgAb titers only at 3 months in an untreated AIT population [Selenium, as a trace element, is essential for life. Various diseases, such as cancer, diabetes, and AIT, have been proven to be associated with selenium deficiency , 8, 9. Apulation . The prepulation . This wapulation . There aGrowing evidence showed increased oxidative stress was not only closely correlated with thyroid dysfunction, including hypothyroidism and hyperthyroidism, but had significant influences on the pathogenesis of AIT , 23. PreThere are certain limitations to our study. Our sample size was small, and we only used MDA, TAC, and SOD for the evaluation of oxidative stress because the methods were established to be easy, rapid, reliable, and inexpensive, while we did not measure oxidative stress index (OSI), 8-iso-PGF2a or 8-hydroxy-2′-deoxyguanosine levels which are widely used as biomarkers of oxidative damage.In conclusion, our study demonstrates an imbalance between oxidant stress and the defense system in patients with euthyroid AIT. The correlation between thyroid antibodies and biomarkers of oxidative stress may reflect the role of oxidative stress in the development of autoimmunity. Additionally, selenium supplementation could decrease TPOAb titer, which may play a role via enforcing the defense against oxidative stress in euthyroid patients with AIT.