To investigate the therapeutic effect of peroxiredoxin-6(PRDX6)on ultraviolet-induced corneal injury in rats and explore the mechanism.The rat model of corneal injury was established by exposing to ultravio-let.Male wister rats were randomly divided into control groups,dexamethasone (DXM)groups and PRDX6 groups,the rats were administered four times a day and for 12 days.The corneal opacity was observed with a slit-lamp microscope.Histopathologic changes were observed with light microscopy.The content of corneal malonalde-hyde(MDA)was determined by thiobarbituric acid test and the total antioxidative capacity(TAOC)was detected by chemical colorimetric test.P38 MAPK signal pathway was detected with the method of Western blot and the gene expression of cytokines were measured by RT-PCR method.Compared with the control group,PRDX6 treat-ment significantly reduced corneal opacity,improved corneal pathology injury,decreased the MDA content and in-creased the TAOC.In the PRDX6 group the level of phosphorylated p38 protein was significantly lower than that in the control group.The gene expression of cytokine were different between control and PRDX6 groups(P <0.05).PRDX6 showed therapeutic effect in the rat model of ultraviolet-induced corneal injury.This maybe be concerned with that it could alleviated the oxidative damage,suppressed p38 MAPK phosphorylation and regulate the gene expression of cytokine.

Objective:To evaluate the role of nuclear factor E2-related factor 2 (Nrf2)/heme oxidase-1 (HO-1) in reduction of renal ischemia-reperfusion (I/R) injury by the human umbilical cord mesenchymal stem cells (hucMSCs)-derived exosomes (hucMSCs-exo) in mice.Methods:The hucMSCs were cultured, and exosomes were extracted and identified by transmission electron microscopy, nanoparticle tracking analysis and Western blot. Thirty-six male SPF-grade C57BL/6 mice, weighing 20-25 g, were used. Thirty mice were selected and divided into 5 groups ( n=6 each) by a random number table method: sham operation group (Sham group), sham operation + Nrf2 inhibitor ML385 group (Sham + ML385 group), renal I/R group (I/R group), renal I/R + exosome group (I/R+ EXO group), and renal I/R + exosome + Nrf2 inhibitor ML385 group (I/R+ EXO+ ML385 group). A model of renal I/R injury was prepared by clamping the bilateral renal pedicles for 45 min followed by perfusion in anesthetized animals. ML385 30 mg/kg was intraperitoneally injected at 45 min before preparing the model in Sham+ ML385 group and I/R+ EXO+ ML385 group, and hucMSCs-exo 100 μg was injected via the tail vein at 15 min before reperfusion in I/R+ EXO group and I/R+ EXO+ ML385 group. Serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations were detected at 24 h of reperfusion. The renal tissues were obtained for examination of the pathological changes and for determination of contents of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and malondialdehyde (MDA), superoxide dismutase (SOD) activity and reactive oxygen species (ROS) levels and expression of Nrf2 and HO-1 protein and mRNA (by Western blot and quantitative real-time polymerase chain reaction). The left 6 mice were allocated to sham operation group (Sham-IM group, n=3) and renal I/R group (I/R-IM group, n=3) by a random number table method for VISQUE in living imaging observation. Results:The exosomes showed a typical cup-shaped morphology with a transmission electron microscope, the nanoparticles tracked and analyzed the average diameter of the exosome, with an average diameter of 96.7 nm, and the positive expression of surface markers CD9, CD63 and TSG101 was detected using Western blot. The renal fluorescence intensity value was significantly increased in I/R-IM group as compared with Sham-IM group ( P<0.05). Compared with Sham group, the serum BUN and Cr concentrations were significantly increased, the contents of IL-6, TNF-α and MDA and ROS levels were increased, the activity of SOD was decreased, the expression of Nrf2 and HO-1 protein and mRNA was down-regulated ( P<0.05), and the pathological changes of renal tissues were aggravated in I/R group, and no significant change was found in serum BUN and Cr concentrations in Sham+ ML385 group ( P>0.05). Compared with I/R group, the serum BUN and Cr concentrations were significantly decreased, the contents of IL-6, TNF-α and MDA and ROS levels were decreased, the activity of SOD was increased, the expression of Nrf2 and HO-1 protein and mRNA was up-regulated ( P<0.05), and the pathological changes of renal tissues were significantly attenuated in I/R+ EXO group. Compared with I/R+ EXO group, the serum BUN and Cr concentrations were significantly increased, the contents of IL-6, TNF-α and MDA and ROS levels were increased, the activity of SOD was decreased, the expression of Nrf2 and HO-1 protein and mRNA was down-regulated ( P<0.05), and the pathological changes of renal tissues were aggravated in I/R+ EXO+ ML385 group. Conclusions:The mechanism by which hucMSCs-exo reduces renal I/R injury may be related to activation of the Nrf2/HO-1 signaling pathway in mice.

Purpose: Gastric cancer (GC) has substantial biological differences between Asian and non-Asian populations, which makes it difficult to have a unified predictive measure for all people. We aimed to identify novel prognostic biomarkers to help predict the prognosis of Asian GC patients. Materials and Methods: We investigated the differential gene expression between GC and normal tissues of GSE66229. Univariate, multivariate and Lasso Cox regression analyses were conducted to establish a four-gene-related prognostic model based on the risk score. The risk score was based on a linear combination of the expression levels of individual genes multiplied by their multivariate Cox regression coefficients. Validation of the prognostic model was conducted using The Cancer Genome Atlas (TCGA) database. A nomogram containing clinical characteristics and the prognostic model was established to predict the prognosis of Asian GC patients. Results: Four genes (RBPMS2, RGN, PLEKHS1, and CT83) were selected to establish the prognostic model, and it was validated in the TCGA Asian cohort. Receiver operating characteristic analysis confirmed the sensitivity and specificity of the prognostic model. Based on the prognostic model, a nomogram containing clinical characteristics and the prognostic model was established, and Harrell’s concordance index of the nomogram for evaluating the overall survival significantly higher than the model only focuses on the pathologic stage (0.74 vs. 0.64, p < 0.001). Conclusion The four-gene-related prognostic model and the nomogram based on it are reliable tools for predicting the overall survival of Asian GC patients.

Objective: To investigate the incidence and clinical characteristics of left atrial appendage (LAA) thrombus in patients with hypertrophic cardiomyopathy (HCM) and non-valvular atrial fibrillation (AF) . Methods: Data from 10 440 patients with AF who had undergone transesophageal echocardiography (TEE) before cardioversion or catheter ablation at Beijing Anzhen Hospital from April 2006 to December 2018 were retrospectively screened. Two hundred and five HCM patients were included, 820 AF patients with the same CHA₂DS₂-VASc score over the same period were selected as the control group. HCM patients were divided into two subgroups based on presence or absence of LAA thrombus/sludge. The baseline of clinical information, transthoracic echocardiographic and TEE measures were compared among all the groups. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of left atrial diameter (LAD) for LAA thrombus/sludge. Multivariate logistic regression analysis was applied to analyze the correlative factors of LAA thrombus/sludge in HCM patients. Results: The incidences of LAA thrombus or sludge were higher in HCM group than in control group (10.7% (22/205) vs. 0.7% (6/820); 8.8% (18/205) vs.7.0% (57/820), P<0.001) . In HCM patients, LAD was significantly larger in LAA thrombus/sludge subjects than in those without thrombus/sludge ((48.9±5.1)mm vs. (45.2±6.1) mm, P<0.001). CHA₂DS₂-VASc score was similar between the two subgroups ((2.0±1.4) vs. (1.8±1.4), P>0.05). There was no difference in the rate of patients with a CHA₂DS₂-VASc scores ≥2 between the subgroups(62.5% (25/40) vs. 57.0% (94/165), P=0.525). The incidences of LAA thrombus in HCM and AF patients with CHA₂DS₂-VASc scores of 0, 1 and 2 were 8.8% (3/34) , 9.6% (5/52) , 11.8% (11/119) , respectively; and the rate of LAA sludge were 8.8% (3/52) , 7.7% (4/52) , 9.2% (11/119) , respectively. The cut off value of LAD for the diagnosis of LAA thrombus/sludge was 44.5 mm. Multivariate logistic regression analysis showed that LAD≥44.5 mm (OR=5.134, 95%CI 1.862-14.156, P=0.002) , non-paroxysmal AF (OR=2.782, 95%CI 1.238-6.252, P=0.013) , previous thromboembolism or stroke (OR=1.820, 95%CI 0.774-4.227, P=0.017) were independent determinants of LAA thrombus/sludge. Conclusions The incidence of LAA thrombus/sludge is higher in patients with HCM and AF than in AF patients without HCM. The CHA₂DS₂-VASc score is similar between HCM and AF patients with LAA thrombus/sludge and those without thrombus/sludge. Patients with CHA₂DS₂-VASc score 0-1 are also likely to suffer LAA thrombus/sludge. Left atrial enlargement is associated with LAA thrombus/sludge.

Objective: To investigate the effect of non-vitamin K antagonist oral anticoagulants (NOAC) on left atrial or atrial appendage (LA/LAA) thrombi in patients with nonvalvular atrial fibrillation (NVAF). Method: Data from 3 042 patients with atrial fibrillation(AF), who underwent transesophageal echocardiography (TEE) examination before cardioversion or catheter ablation for the detection of LA/LAA thrombus in our department from March 2016 to January 2018 were prospectively analyzed. Among these patients, LA/LAA thrombus was detected by TEE in 57 patients. A total of 19 patients who received dabigatran or rivaroxaban for ≥3 weeks and underwent repeated TEE were included, 38 patients were excluded (7 patients with rheumatic heart disease, 1 patient treated with pericardial decortication, 1 patient treated with surgical repair of endocardial cushion defect, 1 patient with LA thrombus associated with the atrial septal occluder device, 14 patients received warfarin therapy, 14 patients did not receive repeated TEE). Results: First repeated TEE results showed that LA/LAA thrombus was not completely resolved in 4 out of 4 patients treated with dabigatran (110 mg bid) for a median time of 119 (47, 258) days, whereas LA/LAA thrombus was completely resolved in 5 out of 11 patients treated with dabigatran (150 mg bid) for a median time of 80 (58, 147) days. Thrombus was completely resolved in 2 out of 2 patients treated with rivaroxaban (15 mg qd) for 110 days and 95 days respectively, and in 1 out of 2 patients treated with rivaroxaban (20 mg qd) for 91 days. Second repeated TEE was performed in 8 patients. Thrombus was resolved completely in 2 out of 3 patients with undissolved thrombus treated by dabigatran (110 mg bid) after increasing the dabigatran dosage (150 mg bid). Thrombus was resolved in 3 (1 patient prolonged treatment with dabigatran 150 mg bid and 2 patients switched to rivaroxaban 20 mg qd) out of 4 patients with undissolved thrombus under the dabigatran 150 mg bid regimen, whereas the thrombus remained unresolved in 1 patient switched to rivaroxaban (15 mg qd). After receiving rivaroxaban 15 mg bid treatment, the thrombus was finally resolved in 1 patient with undissolved thrombus treated by rivaroxaban 20 mg qd. There was no clinical thromboembolism or major bleeding events during the median follow up time of 462 (305, 558) days. Conclusions Our data show that NOAC is an effective therapeutic option for the treatment of LA/LAA thrombi. When eligible, a higher NOAC dosage may be preferred due to the higher efficacy on thrombus resolvement.

Objective To investigate the incidence and clinical characteristics of left atrial appendage (LAA) thrombus in patients with hypertrophic cardiomyopathy (HCM) and non?valvular atrial fibrillation (AF). Methods Data from 10 440 patients with AF who had undergone transesophageal echocardiography (TEE) before cardioversion or catheter ablation at Beijing Anzhen Hospital from April 2006 to December 2018 were retrospectively screened. Two hundred and five HCM patients were included, 820 AF patients with the same CHA2DS2?VASc score over the same period were selected as the control group. HCM patients were divided into two subgroups based on presence or absence of LAA thrombus/sludge. The baseline of clinical information, transthoracic echocardiographic and TEE measures were compared among all the groups. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of left atrial diameter (LAD) for LAA thrombus/sludge. Multivariate logistic regression analysis was applied to analyze the correlative factors of LAA thrombus/sludge in HCM patients. Results The incidences of LAA thrombus or sludge were higher in HCM group than in control group (10.7% (22/205) vs. 0.7% (6/820); 8.8% (18/205) vs.7.0% (57/820), P<0.001). In HCM patients, LAD was significantly larger in LAA thrombus/sludge subjects than in those without thrombus/sludge ((48.9±5.1)mm vs. (45.2±6.1) mm, P<0.001). CHA2DS2?VASc score was similar between the two subgroups ((2.0 ± 1.4) vs. (1.8 ± 1.4), P>0.05). There was no difference in the rate of patients with a CHA2DS2?VASc scores ≥2 between the subgroups (62.5% (25/40) vs. 57.0% (94/165), P=0.525). The incidences of LAA thrombus in HCM and AF patients with CHA2DS2?VASc scores of 0, 1 and 2 were 8.8% (3/34), 9.6% (5/52), 11.8% (11/119), respectively; and the rate of LAA sludge were 8.8% (3/52), 7.7% (4/52), 9.2% (11/119), respectively. The cut off value of LAD for the diagnosis of LAA thrombus/sludge was 44.5 mm. Multivariate logistic regression analysis showed that LAD≥44.5 mm ( OR=5.134, 95%CI 1.862-14.156, P=0.002), non?paroxysmal AF ( OR＝2.782, 95%CI 1.238-6.252, P=0.013), previous thromboembolism or stroke ( OR＝1.820, 95%CI 0.774-4.227, P=0.017) were independent determinants of LAA thrombus/sludge. Conclusions The incidence of LAA thrombus/sludge is higher in patients with HCM and AF than in AF patients without HCM. The CHA2DS2?VASc score is similar between HCM and AF patients with LAA thrombus/sludge and those without thrombus/sludge. Patients with CHA2DS2?VASc score 0-1 are also likely to suffer LAA thrombus/sludge. Left atrial enlargement is associated with LAA thrombus/sludge.

Objective:To investigate the clinical efficacy of middle-preserving pancreatectomy (MPP).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of two patients who underwent MPP in Fujian Provincial Hospital from July 2019 to October 2019 were collected. Case 1 was a 52-year-old woman with multiple serous cystic neoplasms of the pancreatic head and tail. Case 2 was a 20-year-old man with chronic pancreatitis and multiple pancreatic duct stones. After comprehensive evaluation, two patients were performed laparoscopic pancreatic head tumor resection+ pancreatic tail resection+ splenectomy+ pancreaticojejunostomy and duodenum-preserving pancreatic head resection+ pancreatic tail resection+ lithotomy by longitudinal pancreatic duct incision+ side-to-side pancreaticojejunostomy, respectively. Observation indicators: surgery, operation time, volume of intraoperative blood loss, blood transfusion, postoperative fasting blood glucose, postoperative complications, duration of postoperative hospital stay, postoperative pathological examination, and follow-up. Follow-up was performed using outpatient examination and telephone interview to detect new-onset diabetes mellitus, pancreatic enzyme replacement therapy and disease recurrence up to March 2020.Results:Two patients successfully underwent MPP. The operation time of case 1 and case 2 were 470 minutes and 400 minutes, the volume of intraoperative blood loss were 200 mL and 100 mL, respectively. No blood transfusion was performed in either patient. The postoperative fasting blood glucose fluctuated between 5.4 and 11.8 mmol/L in case 1, and fluctuated between 5.9 and 11.3 mmol/L in case 2. Case 1 developed abdominal infection after operation, and was discharged after anti-infective treatment. Case 2 had good recovery with no complication. No pancreatic fistula or perioperative death occurred in two patients. The duration of postoperative hospital stay were 12 days and 8 days of case 1 and case 2, respectively. The pathological examination of case 1 showed serous cystadenoma of the pancreatic head and pancreatic tail. The pathological examination of case 2 showed that slightly dilated pancreatic ducts of the pancreatic head and tail with multiple stones and chronic inflammatory cells infiltration around the pancreatic ducts. The length of pancreas preserved was 8.5 cm and 8.3 cm of case 1 and case 2 on postoperative computed tomography (CT) reexamination. Two patients were followed up for 5 months and 7 months, respectively. During the follow-up, both patients had no new-onset diabetes and they didn′t require pancreatic enzyme replacement therapy. Both patients underwent upper abdominal CT examination at postoperative 5 months, which showed good blood supply in middle pancreas and no signs of recurrence of cystadenoma or stones.Conclusions:MPP is a safe and feasible procedure for the treatment of multifocal pancreatic lesions. The procedure can eradicate the lesions and ensure good control of blood glucose in patients.

Objective To examine the associations between season of birth and factors as age at menarche,age at menopause and reproductive span.Methods A total of 285 186 female from the China Kadoorie Biobank,with complete data on critical variables and had menarche at 9-18 years old,were included.A total of 132 373 female with natural menopause were included for the analysis on age at menopause and reproductive span.Multiple linear regression models were used to assess the associations of birth season and the age at menarche,menopause,and reproductive span.Subgroup analyses were performed on birth cohorts and urban/rural residence.Results Compared with the Spring-born (March,April,and May),participants who were born in Summer (June,July,and August),Autumn (September,October,and November),and Winter (December,January,and February)appeared late on both age at menarche and menopause.Multivariable-adjusted coefficients (95％ CI)appeared as 0.14 (95％CI:0.13-0.16),0.26(95％CI:0.24-0.27),0.10 (95％CI:0.08-0.12) for age at menarche respectively and 0.14 (95％CI:0.08-0.20),0.18 (95％CI:0.12-0.24),0.09 (95％CI:0.03-0.16) for age at menopause respectively.No statistically significant association was found between the season of birth and reproductive span.The association was consistent between urban and rural residents and across the birth cohorts.Conclusions female born in spring showed both earlier age on both menarche and menopause,compared to the ones born in other seasons.Our findings suggested that exposures in early life with some degree of seasonal variation might influence the development of female reproductive system.

Objective To investigate the association between metabolic associated fatty liver disease (MAFLD) and sleep disorders. Methods A total of 222 patients with MALFD who were admitted to Panjin Central Hospital from February 2020 to February 2021 and 270 healthy individuals were enrolled as subjects. According to age, the patients with MALFD were divided into youth group with 93 patients, middle-aged group with 76 patients, and elderly group with 53 patients; according to controlled attenuation parameter (CAP) of liver fat, the patients were divided into non-steatosis group with 23 patients, mild steatosis group with 85 patients, moderate steatosis group with 76 patients, and severe steatosis group with 38 patients; according to liver stiffness measurement (LSM), the patients were divided into non-progressive fibrosis group with 124 patients and progressive fibrosis group with 98 patients. Related data were collected, including general information such as age and sex and laboratory markers such as routine blood test results and biochemistry, and after informed consent was obtained, three sleep scales, i.e., Pittsburgh Sleep Quality Index (PSQI) scale, Epworth Sleepiness Scale (ESS), and Morningness-Eveningness Questionnaire (MEQ), were completed independently. The MAFLD group and the healthy control group were compared in terms of general information and laboratory markers to investigate the association between MAFLD and sleep disorders under different grouping criteria. The chi-square test was used for comparison of categorical data between two groups; the t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups; the Kruskal-Wallis H test was used for comparison between the groups of MAFLD patients with different ages and degrees of hepatic steatosis and pairwise comparison within each group. A logistic regression analysis was used to investigate the independent risk factors for MAFLD, and a Spearman correlation analysis was used to investigate the correlation of hepatic steatosis degree and fibrosis degree with sleep quality, somnolence, and circadian rhythm. Results There were significant differences in age, sex, hypertension, diabetes, and smoking between the MAFLD group and the healthy control group (all P < 0.05). Compared with the healthy control group, the MAFLD group had significantly higher hemoglobin, white blood cell count, lymphocyte percentage, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transpeptidase, serum uric acid, low-density lipoprotein, total bilirubin, total cholesterol, triglyceride, and body mass index and significantly lower albumin and high-density lipoprotein (all P < 0.05). Compared with the healthy control group, the MAFLD group had significantly higher PSQI score ( t =35.529, P < 0.001) and ESS score ( t =24.647, P < 0.001) and significantly lower MEQ score ( t =-22.416, P < 0.001) and sleep time ( t =-8.660, P < 0.001). With the increase in age in the MAFLD group, hepatic steatosis degree, liver fibrosis degree, and PSQI score showed an increasing trend, and pairwise comparison of each scoring factor between groups showed statistical significance (all P < 0.05). Compared with the healthy control group, the MAFLD group had a significantly higher proportion of patients with definitely evening type, moderately evening type, or intermediate type and a significantly lower proportion of patients with moderately morning type or definitely morning type (all P < 0.05), and MEQ score tended to decrease with the increase in hepatic steatosis degree and liver fibrosis degree, with significant differences between two groups (all P < 0.05). The multivariate logistic regression analysis showed that overweight/obesity (odds ratio [ OR ]=3.166, P =0.027), diabetes ( OR =6.811, P =0.045), increase in white blood cell count ( OR =2.301, P < 0.001), increase in lymphocyte percentage ( OR =1.316, P =0.002), poor sleep quality ( OR =8.493, P < 0.001), a high degree of somnolence ( OR =5.420, P < 0.001), and circadian rhythm disturbance ( OR =3.805, P < 0.001) were risk factors for MAFLD. The Spearman correlation analysis showed that in the MAFLD group, hepatic steatosis degree was positively correlated with PSQI score ( r =0.444, P < 0.001) and ESS score ( r =0.339, P < 0.001) and was negatively correlated with MEQ score ( r =-0.195, P =0.004), and liver fibrosis degree was positively correlated with PSQI score ( r =0.518, P < 0.001) and ESS score ( r =0.373, P < 0.001) and was negatively correlated with MEQ score ( r =-0.250, P =0.004). Conclusion Compared with healthy individuals, the patients with MAFLD often have sleep disorders, and the severity of sleep disorders increases with age, hepatic steatosis degree, and liver fibrosis degree. Obesity, diabetes, and sleep disorders are risk factors for the onset of MAFLD.

Objective:To explore the association of lifestyle and cardiometabolic risk factors with five epigenetic age acceleration (AA) indices.Methods:This study included 980 participants of China Kadoorie Biobank, for whom genome-wide DNA methylation of peripheral blood cells had been detected in baseline survey. Five indices of DNA methylation age (DNAm age) were calculated, i.e. Horvath clock, Hannum clock, DNAm PhenoAge, GrimAge and Li clock. Epigenetic AA was defined as the residual of regressing DNAm age on chronological age. Lifestyle factors studied included smoking status, alcohol consumption, eating habits, physical activity level and body shape defined by a combination of BMI and waist circumference. Cardiometabolic risk factors included blood pressure, blood glucose level and total cholesterol level. Linear regression model was used to analyze the association of lifestyle and cardiometabolic risk factors with AA ( β). Results:GrimAge_AA was associated with smoking status, alcohol consumption, physical activity level and BMI. Compared with non-smokers, non-drinkers, or participants with BMI of 18.5- 23.9 kg/m 2, the smokers who smoked 1-14 cigarettes/day ( β=0.71, 95% CI: 0.57-0.86), 15-24 cigarettes/day ( β=0.88, 95% CI: 0.73-1.03), and ≥25 cigarettes/day ( β=0.99, 95% CI: 0.81-1.18), respectively, heavy drinkers with daily pure alcohol consumption ≥60 g ( β=0.33, 95% CI: 0.11-0.55) and participants with BMI<18.5 kg/m 2 ( β=0.23, 95% CI: 0.03-0.43) showed accelerated aging. Compared with those in the lowest quintile of physical activity level, participants in the top three quintile of physical activity level showed decelerated aging ( β=-0.13, 95% CI: -0.26-0.01, β=-0.12, 95% CI: -0.26-0.02, and β=-0.14, 95% CI: -0.27- -0.00, respectively). GrimAge_AA decreased with the increase of the number of healthy lifestyle factors ( P<0.001). Compared with the participants with 0 to 1 healthy lifestyle factor, the β of those with 2, 3, or 4 to 5 healthy lifestyle factors were -0.30 (95% CI: -0.47- -0.12), -0.47 (95% CI: -0.65- -0.30) and -0.72 (95% CI: -0.90- -0.53), respectively. The other four indices were not statistically significantly associated with most lifestyle factors. None of the five indices of AA was associated with blood pressure, blood glucose level or total cholesterol level. Conclusion:People with unhealthy lifestyle showed accelerated epigenetic aging, that is, the predicted DNAm age is older than their own chronological age.