Differential DNA methylation of steatosis and non-alcoholic fatty liver disease in adolescence.

BACKGROUND AND AIMS: Epigenetic modifications are associated with hepatic fat accumulation and non-alcoholic fatty liver disease (NAFLD). However, few epigenetic modifications directly implicated in such processes have been identified during adolescence, a critical developmental window where physiological changes could influence future disease trajectory. To investigate the association between DNA methylation and NAFLD in adolescence, we undertook discovery and validation of novel methylation marks, alongside replication of previously reported marks. APPROACH AND RESULTS: We performed a DNA methylation epigenome-wide association study (EWAS) on DNA from whole blood from 707 Raine Study adolescents phenotyped for steatosis score and NAFLD by ultrasound at age 17. Next, we performed pyrosequencing validation of loci within the most 100 strongly associated differentially methylated CpG sites (dmCpGs) for which ≥ 2 probes per gene remained significant across four statistical models with a nominal p value < 0.007. EWAS identified dmCpGs related to three genes (ANK1, MIR10a, PTPRN2) that met our criteria for pyrosequencing. Of the dmCpGs and surrounding loci that were pyrosequenced (ANK1 n = 6, MIR10a n = 7, PTPRN2 n = 3), three dmCpGs in ANK1 and two in MIR10a were significantly associated with NAFLD in adolescence. After adjustment for waist circumference only dmCpGs in ANK1 remained significant. These ANK1 CpGs were also associated with γ-glutamyl transferase and alanine aminotransferase concentrations. Three of twenty-two differentially methylated dmCpGs previously associated with adult NAFLD were associated with NAFLD in adolescence (all adjusted p < 2.3 × 10-3). CONCLUSIONS: We identified novel DNA methylation loci associated with NAFLD and serum liver biochemistry markers during adolescence, implicating putative dmCpG/gene regulatory pathways and providing insights for future mechanistic studies.

[Efficacy of selective bronchial occlusion in the treatment of biliary bronchial fistula].

Objective: To analyze the effect of selective bronchial occlusion (SBO) in the treatment of biliary bronchial fistula (BBF). Methods: Eight patients with BBF that without biliary obstruction admitted to the Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Naval Medical University from January 1, 2015 to December 31, 2021 were included in this study. Bronchial silicone plug (6 cases) and autologous blood+thrombin (2 cases) were used as sealing materials for SBO treatment for the first time. Among the 7 patients who underwent subsequent closure treatment, 5 cases were blocked by bronchial silicone plug, 1 case was blocked by "bronchial silicone plug+bullet-covered stent" and 1 case was blocked by "bronchial silicone plug+bronchial one-way valve". The clinical data related to SBO treatment were collected and patients were followed up, and the therapeutic effect of SBO on BBF was analyzed. Results: The age of BBF onset was (58±9) years old, including 6 males. Among the 6 patients who used bronchial silicone plug as plugging material in the first SBO treatment, 1 case was successfully plugged, 2 cases did not achieve symptoms relief after plugging, 2 cases coughed up the plugging device immediately after surgery, and 1 case developed a new fistula. Autologous blood and thrombin were used as sealing materials in 2 patients, and both failed. Among the 7 patients who received subsequent closure treatment, bronchial silicone plug+bullet-covered stent (1 case) and bronchial silicone plug+bronchial unidirectional valve (1 case) were successful. After 2-6 times of bronchial silicone plug (5 cases), fistula were successfully blocked in 3 cases, and the frequency and volume of bile-like sputum decreased by 50% or more in 2 cases. The main postoperative complications were fever and cough (expectoration) in 7 and 6 cases, respectively. During the follow-up period, 2 patients were lost to follow-up, and the remaining 6 patients were followed up for 2-31 months. During the follow-up period, the effect of closure treatment was basically stable, and there was no death case. Conclusion: SBO therapy provides a safe and feasible palliative treatment for BBF.

Comparison of the cardiometabolic profiles of adolescents conceived through ART with those of a non-ART cohort.

STUDY QUESTION: Is the cardiometabolic health of adolescents conceived through ART worse than that of their counterparts conceived without ART? SUMMARY ANSWER: The majority of cardiometabolic and vascular health parameters of adolescents conceived through ART are similar or more favourable, than those of their counterparts of similar age and conceived without ART. WHAT IS KNOWN ALREADY: It has been proposed that the cardiometabolic health of offspring conceived with ART may be unfavourable compared to that of their counterparts conceived without ART. The literature pertaining to cardiometabolic health of offspring conceived after ART is contradictory, but generally suggests unfavourable cardiometabolic health parameters, such as an increase in blood pressure (BP), vascular dysfunction and adiposity, as well as unfavourable glucose and lipid profiles. With over 8 million children and adults born through ART worldwide, it is important to investigate whether these early signs of adverse cardiometabolic differences persist into adolescence and beyond. STUDY DESIGN, SIZE, DURATION: The Growing Up Healthy Study (GUHS) is a prospective cohort study that recruited 303 adolescents and young adults conceived after ART (aged 13-21 years) and born between 1991 and 2001 in Western Australia. Their health parameters, including cardiometabolic factors, were assessed and compared with counterparts from the Raine Study Generation 2 (Gen2). The 2868 Gen2 participants were born 1989-1992 and are representative of the Western Australian adolescent population. At ∼17 years of age (2013-2017), 163 GUHS participants replicated assessments previously completed by Gen2 at a similar age. PARTICIPANTS/MATERIALS, SETTING, METHODS: Cardiometabolic parameters were compared between a total of 163 GUHS and 1457 Gen2 adolescents. Separate male (GUHS n = 81, Gen2 n = 735) and female (GUHS n = 82, Gen2 n = 722) analyses were conducted. Assessments consisted of a detailed questionnaire including health, lifestyle and demographic parameters, anthropometric assessments (height, weight, BMI, waist circumference and skinfold thickness), fasting serum biochemistry, arterial stiffness and BP (assessed using applanation tonometry). Abdominal ultrasonography was used to assess the presence and severity of hepatic steatosis, and thickness of abdominal fat compartments. Non-alcoholic fatty liver disease (NAFLD) was diagnosed if there was sonographic fatty liver in the absence of significant alcohol consumption. Chi2, Fisher's exact and Mann-Whitney U tests, performed in SPSS V25, examined cohort differences and generalized estimating equations adjusted for the following covariates: singleton vs non-singleton pregnancy, birthweight (z-score), gestational age, BMI, smoking, alcohol consumption in the past 6 months and parent cardiovascular status. Arterial stiffness measures and waist circumference were additionally adjusted for height, and female analyses were additionally adjusted for use of oral contraceptives in the preceding 6 months. MAIN RESULTS AND THE ROLE OF CHANCE: In adjusted analyses, GUHS females had a lower BMI (22.1 vs 23.3 kg/m2, P = 0.014), and thinner skinfolds (triceps, subscapular, mid-abdominal; 16.9 vs 18.7 mm, P = 0.021, 13.4 vs 15.0 mm, P = 0.027, 19.7 vs 23.2 mm, P < 0.001, respectively), whereas males were not significantly different. Waist circumference was lower in GUHS adolescents (males: 78.1 vs 81.3 cm, P = 0.008, females: 76.7 vs 83.3 cm, P = 0.007). There were no significant differences between the two groups in glucose, insulin, homeostatic model assessment for insulin resistance, low-density lipoprotein cholesterol, non-high-density lipoprotein cholesterol (non-HDL-C), total cholesterol (TC), alanine aminotransferase and high-sensitivity C-reactive protein in both sexes. In females, serum triglycerides were lower in GUHS adolescents (1.0 vs 1.2 mmol/l, P = 0.029). GUHS males had higher serum HDL-C (1.1 vs 1.0 mmol/l, P = 0.004) and a lower TC/HDL-C ratio (3.2 vs 3.6, P = 0.036). There were no significant differences in the prevalence of NAFLD or steatosis severity scores between the cohorts in males and females. GUHS females had less subcutaneous adipose tissue (9.4 vs 17.9 mm, P < 0.001), whereas GUHS males had greater visceral adipose thickness (44.7 vs 36.3 mm, P < 0.001). There was no significant difference in pre-peritoneal adipose thickness. Pulse wave velocity was lower in GUHS males (5.8 vs 6.3 m/s, P < 0.001) and heart rate corrected augmentation index was lower in GUHS females (-8.4 vs -2.7%, P = 0.048). There were no significant differences in BP or heart rate in males or females between the two groups. LIMITATIONS, REASONS FOR CAUTION: Despite the substantial study size and the unique study design of the ART cohort, we were unable to differentiate between different types of ART, due to the low number of ICSI cycles (e.g. IVF vs ICSI), draw definite conclusions, or relate the outcomes to the cause of infertility. Considering the differences in time points when both cohorts were studied, external factors could have changed, which could not be accounted for. Given the observational nature of this study, causation cannot be proven. WIDER IMPLICATIONS OF THE FINDINGS: Contrary to our hypothesis and previous findings focussing mainly on childhood, this study reports mostly similar or favourable cardiometabolic markers in adolescents conceived with ART compared to those conceived without ART. The greater visceral adipose thickness, particularly present in males, requires further investigation. While these findings are generally reassuring, future well-designed and appropriately powered studies are required to definitively address the issue of cardiometabolic health in ART adults. STUDY FUNDING/COMPETING INTEREST(S): This project was supported by NHMRC project grant number 1042269 and R.J.H. received education grant funding support from Ferring Pharmaceuticals. R.J.H. is the Medical Director of Fertility Specialists of Western Australia and a shareholder in Western IVF. He has received educational sponsorship from MSD, Merck-Serono and Ferring Pharmaceuticals. P.B. is the Scientific Director of Concept Fertility Centre, Subiaco, Western Australia. J.L.Y. is the Medical Director of PIVET Medical Centre, Perth, Western Australia. TRIAL REGISTRATION NUMBER: N/A.

Adiposity associated DNA methylation signatures in adolescents are related to leptin and perinatal factors.

Epigenetics links perinatal influences with later obesity. We identifed differentially methylated CpG (dmCpG) loci measured at 17 years associated with concurrent adiposity measures and examined whether these were associated with hsCRP, adipokines, and early life environmental factors. Genome-wide DNA methylation from 1192 Raine Study participants at 17 years, identified 29 dmCpGs (Bonferroni corrected p < 1.06E-07) associated with body mass index (BMI), 10 with waist circumference (WC) and 9 with subcutaneous fat thickness. DmCpGs within Ras Association (RalGDS/AF-6), Pleckstrin Homology Domains 1 (RAPH1), Musashi RNA-Binding Protein 2 (MSI2), and solute carrier family 25 member 10 (SLC25A10) are associated with both BMI and WC. Validation by pyrosequencing confirmed these associations and showed that MSI2 , SLC25A10 , and RAPH1 methylation was positively associated with serum leptin. These were  also associated with the early environment; MSI2 methylation (ß = 0.81, p = 0.0004) was associated with pregnancy maternal smoking, SLC25A10 (CpG2 ß = 0.12, p = 0.002) with pre- and early pregnancy BMI, and RAPH1 (ß = -1.49, p = 0.036) with gestational weight gain. Adjusting for perinatal factors, methylation of the dmCpGs within MSI2, RAPH1, and SLC25A10 independently predicted BMI, accounting for 24% of variance. MSI2 methylation was additionally associated with BMI over time (17 years old ß = 0.026, p = 0.0025; 20 years old ß = 0.027, p = 0.0029) and between generations (mother ß = 0.044, p = 7.5e-04). Overall findings suggest that DNA methylation in MSI2, RAPH1, and SLC25A10 in blood may be robust markers, mediating through early life factors.

DNA methylation patterns within whole blood of adolescents born from assisted reproductive technology are not different from adolescents born from natural conception.

STUDY QUESTION: Do the epigenome-wide DNA methylation profiles of adolescents born from ART differ from the epigenome of naturally conceived counterparts? SUMMARY ANSWER: No significant differences in the DNA methylation profiles of adolescents born from ART [IVF or ICSI] were observed when compared to their naturally conceived, similar aged counterparts. WHAT IS KNOWN ALREADY: Short-term and longer-term studies have investigated the general health outcomes of children born from IVF treatment, albeit without common agreement as to the cause and underlying mechanisms of these adverse health findings. Growing evidence suggests that the reported adverse health outcomes in IVF-born offspring might have underlying epigenetic mechanisms. STUDY DESIGN, SIZE, DURATION: The Growing Up Healthy Study (GUHS) is a prospective study that recruited 303 adolescents and young adults, conceived through ART, to compare various long-term health outcomes and DNA methylation profiles with similar aged counterparts from Generation 2 from the Raine Study. GUHS assessments were conducted between 2013 and 2017. The effect of ART on DNA methylation levels of 231 adolescents mean age 15.96 ± 1.59 years (52.8% male) was compared to 1188 naturally conceived counterparts, 17.25 ± 0.58 years (50.9% male) from the Raine Study. PARTICIPANTS/MATERIALS, SETTING, METHODS: DNA methylation profiles from a subset of 231 adolescents (13-19.9 years) from the GUHS, generated using the Infinium Methylation Epic Bead Chip (EPIC) array were compared to 1188 profiles from the Raine Study previously measured using the Illumina 450K array. We conducted epigenome-wide association approach (EWAS) and tested for an association between the cohorts applying Firth's bias reduced logistic regression against the outcome of ART versus naturally conceived offspring. Additionally, within the GUHS cohort, we investigated differences in methylation status in fresh versus frozen embryo transfers, cause of infertility as well as IVF versus ICSI conceived offspring. Following the EWAS analysis we investigated nominally significant probes using Gene Set Enrichment Analysis (GSEA) to identify enriched biological pathways. Finally, within GUHS we compared four estimates (Horvath, Hanuum, PhenoAge [Levine], and skin Horvath) of epigenetic age and their correlation with chronological age. MAIN RESULTS AND THE ROLE OF CHANCE: Between the two cohorts, we did not identify any DNA methylation probes that reached a Bonferroni corrected P-value < 1.24E-0.7. When comparing IVF versus ICSI conceived adolescents within the GUHS cohort, after adjustment for participant age, sex, maternal smoking, multiple births, and batch effect, three methylation probes (cg15016734, cg26744878 and cg20233073) reached a Bonferroni correction of 6.31E-08. After correcting for cell count heterogeneity, two of the aforementioned probes remained significant and an additional two probes (cg 0331628 and cg 20235051) were identified. A general trend towards hypomethylation in the ICSI offspring was observed. All four measures of epigenetic age were highly correlated with chronological age and showed no evidence of accelerated epigenetic aging within their whole blood. LIMITATIONS, REASONS FOR CAUTION: The small sample size coupled with the use of whole blood, where epigenetic differences may occur in other tissue. This was corrected by the utilized statistical method that accounts for imbalanced sample size between groups and adjusting for cell count heterogeneity. Only a small portion of the methylome was analysed and rare individual differences may be missed. WIDER IMPLICATIONS OF THE FINDINGS: Our findings provide further reassurance that the effects of the ART manipulations occurring during early embryogenesis, existing in the neonatal period are indeed of a transient nature and do not persist into adolescence. However, we have not excluded that alternative epigenetic mechanisms may be at play. STUDY FUNDING/COMPETING INTEREST(S): This project was supported by NHMRC project Grant no. 1042269 and R.J.H. received funding support from Ferring Pharmaceuticals Pty Ltd. R.J.H. is the Medical Director of Fertility Specialists of Western Australia and a shareholder in Western IVF. He has received educational sponsorship from Merck Sharp & Dohme Corp.- Australia, Merck-Serono Australia Pty Ltd and Ferring Pharmaceuticals Pty Ltd. P.B. is the Scientific Director of Concept Fertility Centre, Subiaco, Western Australia. J.L.Y. is the Medical Director of PIVET Medical Centre, Perth, Western Australia. The remaining authors have no conflicts of interest.

Machine learning and clinical epigenetics: a review of challenges for diagnosis and classification.

BACKGROUND: Machine learning is a sub-field of artificial intelligence, which utilises large data sets to make predictions for future events. Although most algorithms used in machine learning were developed as far back as the 1950s, the advent of big data in combination with dramatically increased computing power has spurred renewed interest in this technology over the last two decades. MAIN BODY: Within the medical field, machine learning is promising in the development of assistive clinical tools for detection of e.g. cancers and prediction of disease. Recent advances in deep learning technologies, a sub-discipline of machine learning that requires less user input but more data and processing power, has provided even greater promise in assisting physicians to achieve accurate diagnoses. Within the fields of genetics and its sub-field epigenetics, both prime examples of complex data, machine learning methods are on the rise, as the field of personalised medicine is aiming for treatment of the individual based on their genetic and epigenetic profiles. CONCLUSION: We now have an ever-growing number of reported epigenetic alterations in disease, and this offers a chance to increase sensitivity and specificity of future diagnostics and therapies. Currently, there are limited studies using machine learning applied to epigenetics. They pertain to a wide variety of disease states and have used mostly supervised machine learning methods.

[Application of meticulous anatomy skills with straight bipolar electric coagulation forceps in thyroid surgery].

Objective: To compare the efficacies of the two techniques of "micro-hemostasis and micro-cutting" with straight bipolar electrocoagulation forceps and traditional clamp-ligation for hemostasia in thyroid surgery. Methods: A total of 228 patients who underwent surgical treatment for thyroid neoplasms in our hospital between January 2015 and December 2018 were retrospectively analyzed, including 50 males and 178 females, aged 23-68 years old. Of those, 150 cases as electric knife group received traditional thyroid surgery between January 2015 and December 2018 and 78 cases as bipolar electrocoagulation group received thyroid surgery by using the technique of bipolar electrocoagulation with meticulous anatomy between January 2018 and December 2018. The total operation time, single operation time, intraoperative hemorrhage, postoperative drainage volume on the first day, postoperative hoarseness and hypocalcemia were compared between the two groups. SPSS 16.0 was used to analyze the data. Results: The total operation time and intraoperative hemorrhage in the bipolar electrocoagulation group were significantly lower than those in the electric knife group ((59.33±18.29)min vs (77.21±25.39)min, (14.83±9.22)ml vs (36.86±11.80)ml, all P<0.01). The single operation time of the bipolar electrocoagulation group was shorter than that of the electric knife group((10.25±6.16) min vs (20.34±7.24)min, (16.25±7.15)min vs (35.68±8.25)min, (12.12±5.25)min vs (20.68±7.26)min, t value was 3.948,16.262,8.238, all P<0.01).There was no significant difference between the two groups in postoperative drainage volume on the first day (P>0.05) and the incidence of postoperative hoarseness (P>0.05), while the incidence of hypocalcemia in the bipolar electrocoagulation group(10.26%) was lower than that in the electric knife group(21.33%,χ(2)=4.353, P<0.05). Conclusions: The fine dissection for thyroid operation can be achieved by using straight bipolar electrocoagulation tweezers. The use of "micro-hemostasis" and "micro-cutting" technique with bipolar electrocoagulation tweezers can greatly reduce intraoperative bleeding, operation time and postoperative complication.

Comparison of cardiac MRI with PET for assessment of myocardial viability in patients with coronary chronic total occlusion.

AIM: To compare cardiac magnetic resonance imaging (MRI) and positron-emission tomography (PET) assessment of myocardial viability in patients with coronary chronic total occlusion (CTO). MATERIALS AND METHODS: Eighty patients with coronary CTO underwent cardiac MRI and PET. Cardiac MRI images were analysed using a 17-segment model, and late gadolinium enhancement (LGE) and wall motion were scored. PET was used to classify myocardial viability via myocardial perfusion and 18F-fluorodeoxyglucose, digital superscript uptake. RESULTS: With PET as the reference standard, the sensitivity of cardiac MRI in detecting myocardial viability was 95.3%, specificity was 87.5%, positive predictive value was 96.8%, negative predictive value was 84.2%, and accuracy was 93.8% on a per patient basis. The receiver operator characteristic curve was used to analyse the performance of cardiac MRI in the detection of myocardial viability on a per-patient basis and the area under the curve was 0.910 (95% confidence interval 0.805 to 1). Cardiac MRI had the highest sensitivity and specificity for differentiating viable and non-viable myocardium as defined by PET when the cut-off value of LGE was 50%. The motion consistency and correlation of cardiac MRI and PET were analysed and kappa was 0.788 (r=0.825; p<0.001). CONCLUSION: Compared with PET, cardiac MRI assessment of myocardial viability in patients with coronary CTO has high sensitivity, specificity, and accuracy. Therefore, cardiac MRI can be used as an important method for evaluating myocardial viability in coronary CTO patients.

Impact of maternal body mass index and gestational weight gain on pregnancy complications: an individual participant data meta-analysis of European, North American and Australian cohorts.

OBJECTIVE: To assess the separate and combined associations of maternal pre-pregnancy body mass index (BMI) and gestational weight gain with the risks of pregnancy complications and their population impact. DESIGN: Individual participant data meta-analysis of 39 cohorts. SETTING: Europe, North America, and Oceania. POPULATION: 265 270 births. METHODS: Information on maternal pre-pregnancy BMI, gestational weight gain, and pregnancy complications was obtained. Multilevel binary logistic regression models were used. MAIN OUTCOME MEASURES: Gestational hypertension, pre-eclampsia, gestational diabetes, preterm birth, small and large for gestational age at birth. RESULTS: Higher maternal pre-pregnancy BMI and gestational weight gain were, across their full ranges, associated with higher risks of gestational hypertensive disorders, gestational diabetes, and large for gestational age at birth. Preterm birth risk was higher at lower and higher BMI and weight gain. Compared with normal weight mothers with medium gestational weight gain, obese mothers with high gestational weight gain had the highest risk of any pregnancy complication (odds ratio 2.51, 95% CI 2.31- 2.74). We estimated that 23.9% of any pregnancy complication was attributable to maternal overweight/obesity and 31.6% of large for gestational age infants was attributable to excessive gestational weight gain. CONCLUSIONS: Maternal pre-pregnancy BMI and gestational weight gain are, across their full ranges, associated with risks of pregnancy complications. Obese mothers with high gestational weight gain are at the highest risk of pregnancy complications. Promoting a healthy pre-pregnancy BMI and gestational weight gain may reduce the burden of pregnancy complications and ultimately the risk of maternal and neonatal morbidity. TWEETABLE ABSTRACT: Promoting a healthy body mass index and gestational weight gain might reduce the population burden of pregnancy complications.

Features of the metabolic syndrome in late adolescence are associated with impaired testicular function at 20 years of age.

STUDY QUESTION: Are early signs of metabolic disorder in late adolescence associated with features of impaired testicular function many years before the majority seek parenthood? SUMMARY ANSWER: Adolescents with features of metabolic disorder at 17 years, or insulin resistance (IR) at 20 years of age, show impaired testicular function and altered hormone levels compared to those without metabolic disorder. WHAT IS KNOWN ALREADY: Controversial evidence suggests a recent decline in sperm production potentially linked to environmental influences, but its cause remains unclear. Concomitant increases in obesity and diabetes suggest that lifestyle factors may contribute to this decline in testicular function. Although obesity has been associated with adverse testicular function in some studies, it remains unclear whether poor testicular function merely reflects, or causes, poor metabolic health. If metabolic disorder were present in adolescence, prior to the onset of obesity, this may suggest that metabolic disorder maybe a precursor of impaired testicular function. STUDY DESIGN, SIZE, DURATION: The Western Australian Pregnancy Cohort (Raine) Study is a longitudinal study of children born in 1989-1991 who have undergone detailed physical assessments since birth (1454 male infants born). At 17 years of age, 490 boys underwent a hepatic ultrasound examination, serum cytokine assessment (n = 520) and a metabolic assessment (n = 544). A further metabolic assessment was performed at 20 years (n = 608). Testicular assessment was performed at 20 years; 609 had reproductive hormones measured, 404 underwent a testicular ultrasound and 365 produced a semen sample. PARTICIPANTS/MATERIALS, SETTING, METHODS: Testicular volume was estimated by ultrasonography, and semen analysis was performed according to World Health Organization guidelines. Concentrations of LH, FSH and inhibin B (inhB) in serum were measured by immunoassay and total testosterone by liquid chromatography-mass spectrometry.At 17 years of age, a liver ultrasound examination was performed to determine the presence of non-alcoholic fatty liver disease (NAFLD), and serum analysed for the cytokines interleukin-18 and soluble tumour necrosis factor receptor 1 and 2 (sTNFR1, sTNFR2).At 17 and 20 years of age, fasting blood samples were analysed for serum liver enzymes, insulin, glucose, triglycerides (TG), total cholesterol, high density lipoprotein and low density lipoprotein cholesterol, high sensitivity C-reactive protein and uric acid. The homoeostatic model assessment (HOMA) was calculated and approximated IR was defined by a HOMA >4. Anthropometric data was collected and dual energy X-ray absorptiometry measurement performed for lean and total fat mass. As at this young age the prevalence of metabolic syndrome was expected to be low, a two-step cluster analysis was used using waist circumference, TGs, insulin, and systolic blood pressure to derive a distinct high-risk group with features consistent with the metabolic syndrome and increased cardiometabolic risk. MAIN RESULTS AND THE ROLE OF CHANCE: Men at age 17 years with increased cardiometabolic risk had lower concentrations of serum testosterone (medians: 4.0 versus 4.9 ng/mL) and inhB (193.2 versus 221.9 pg/mL) (P < 0.001 for both) compared to those within the low risk metabolic cluster. Men with ultrasound evidence of NAFLD (n = 45, 9.8%) had reduced total sperm output (medians: 68.0 versus 126.00 million, P = 0.044), testosterone (4.0 versus 4.7 ng/mL, P = 0.005) and inhB (209.1 versus 218.4 pg/mL, P = 0.032) compared to men without NAFLD.Men with higher concentrations of sTNFR1 at 17 years of age had a lower sperm output and serum concentration of inhB, with an increase in LH and FSH (all P < 0.05 after adjustment for age, BMI, abstinence and a history of cryptorchidism, varicocele, cigarette smoking, alcohol and drug use), compared to those without an elevated sTNFR1. Multivariable regression analysis, adjusting for confounders, demonstrated that men in the high-risk metabolic cluster at 20 years had a lower serum testosterone and inhB (P = 0.003 and P = 0.001, respectively). A HOMA-IR > 4 was associated with a lower serum testosterone (P = <0.001) and inhB (P = 0.010) and an increase in serum FSH (P = 0.015). LIMITATIONS, REASONS FOR CAUTION: This study is limited by the sample size and multiple comparisons, and causality cannot be proven from an observational study. Due to a 3-year interval between some metabolic assessments and assessment of testicular function, we cannot exclude the introduction of a bias into the study, as some of the participants and their testicular function will not have been fully mature at the 17-year assessment. WIDER IMPLICATIONS OF THE FINDINGS: Irrespective of a proven causation, our study findings are important in that a significant minority of the men, prior to seeking parenthood, presented co-existent features of metabolic disorder and signs of testicular impairment. Of particular note is that the presence of NAFLD at 17 years of age, although only present in a minority of men, was associated with an almost 50% reduction in sperm output at 20 years of age, and that the presence of IR at 20 years was associated with a 20% reduction in testicular volume. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by Australian NHMRC (Grant Numbers 634457, 35351417 and 403981) and received support from the Raine Medical Research Foundation, The Telethon Kids Institute, University of Western Australia, Women and Infants Research Foundation, Curtin University and Edith Cowan University. D.A.D., J.E.D., N.M., L.A.A., R.-C.H., T.A.M., J.K.O., L.J.B. have nothing to declare. R.J.H. is Medical Director of Fertility Specialists of Western Australia, has equity interests in Western IVF, and has received grant support from MSD, Merck-Serono and Ferring Pharmaceuticals. RMcL has equity interests in the Monash IVF Group. R.J.N. has equity interests in FertilitySA, and has received grant support from Merck Serono and Ferring Pharmaceuticals. D.J.H. has received institutional grant funding (but no personal income) for investigator-initiated testosterone pharmacology studies from Lawley and Besins Healthcare and has provided expert testimony to anti-doping tribunals and for testosterone litigation.This abstract was awarded the Fertility Society of Australia clinical exchange award for the oral presentation at ESHRE, Barcelona, in 2018.

Effects of postnatal omega-3 fatty acid supplementation on offspring pro-resolving mediators of inflammation at 6 months and 5 years of age: A double blind, randomized controlled clinical trial.

BACKGROUND: Resolution of inflammation is an active process involving specialised pro-resolving mediators (SPMs) generated from the omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Omega-3 fatty acid supplementation during infancy may provide an intervention strategy to modify SPMs and reduce oxidative stress. This study evaluates the effect of omega-3 fatty acid supplementation in infancy on SPMs and F2-isoprostanes from 6 months to 5 years of age. METHODS: In a double-blind, placebo-controlled, parallel-group study design, 420 infants were randomized to a daily supplement of omega-3 fatty acids (280mg DHA and 110mg EPA) or olive oil (control), from birth to age 6 months. Blood was collected at birth (cord blood), 6 months, 12 months and 5 years. Plasma SPMs included 18-HEPE, E-series resolvins, 17-HDHA, D-series resolvins, 14-HDHA, 10S,17S-DiHDoHE, MaR1 and PD1. F2-isoprostanes were measured in plasma and urine, as markers of oxidative stress in vivo. RESULTS: The change in the concentration of 18-HEPE from birth to 6 months was greater in the omega-3 fatty acid group (Ptimepoint*group=0.04) with levels at 6 months significantly higher than controls (P=0.02). Other SPMs were not different between the groups at any time point. Plasma 18-HEPE concentration were associated with erythrocyte EPA concentrations after age and group adjustments (P<0.001), but not with allergic outcomes at 12 months. There were no between-group differences in plasma and urinary F2-isoprostanes at any time point. CONCLUSION: Omega-3 fatty acid supplementation from birth to 6 months of age increased SPM at 6 months but the effects were not sustained after supplementation ceased. Given that 18-HEPE is a biologically active metabolite, future studies should examine how the increase in 18-HEPE relates to potential health benefits of omega-3 fatty acid supplementation in infancy.

[Value of evaluating the coronary collateral circulation by transluminal attenuation gradient in patients with chronic total occlusion and related influencing factors].

Objective: To observe the value of evaluating the coronary collateral circulation of chronic total occlusion (CTO) by transluminal attenuation gradient (TAG) and Rentrop grading, and analyze the influencing factors for coronary collateral circulation. Methods: A total of 179 CTO patients admitted to Beijing Anzhen hospital during June 2013 to August 2016 were included in this study.All patients received coronary computed tomographic angiography (CCTA) examination before coronary angiography.Finally, 75 patients (79 vessels) were enrolled.Patients were divided into two groups on the basis of Rentrop classification.The Rentrop 3 was defined as a well-developed coronary collateral circulation group, including 50 CTO vessels, Rentrop 2 or below was defined as poorly-developed collateral circulation group, including 29 vessels.TAG values in patients with various Rentrop grades were analyzed.Univariate and multivariate analysis were used to determine the predictors of collateral circulation. Results: TAG increased consistently in proportion to the angiographic extent of collateral flow (TAG was (-33.6±24.4), (-16.5±15.7) and (-12.8±15.8) HU/10 mm in patients with Rentrop grade 0 or 1, 2 and 3, respectively, P=0.007). Number of good collateral circulation vessels predicted by TAG≥15.6 HU/10 mm was 45(57.0%, 45/79), and 50 (63.3%, 50/79) by Rentrop grade (P=0.383). Prevalence of diabetes was significantly lower (20.8%(10/48) vs. 48.1%(13/27), P=0.01), while history of previous myocardial infarction (MI) was significantly higher (35.4%(17/48) vs. 11.1%(3/27), P=0.02) in patients with good collateral circulation than in patients with poor collateral circulation.The TAG of the good collateral circulation group was significantly higher than in poorly collateral circulation group ((-12.8±15.8) HU/10 mm vs. (-21.2±19.6) HU/10mm, P=0.041). Multiple logistic regression analysis revealed that history of MI (OR=0.196, 95%CI 0.041-0.936, P=0.041) and diabetes(OR=6.604, 95%CI 1.726-25.274, P=0.006) were independent predictors of coronary collateral circulation. Conclusions: TAG could be used to evaluate status of coronary collateral circulation.Presence of MI history and absence of diabetes history are the two independent predictors of well-developed coronary collateral circulation in CTO patients.

[Association between collateral circulation and myocardial viability evaluated by cardiac magnetic resonance imaging in patients with coronary artery chronic total occlusion].

Objective: Late gadolinium enhancement(LGE) cardiac magnetic resonance imaging(CMR) was used to evaluate the myocardial viability of chronic total occlusion(CTO) in patients with coronary heart disease and to observe the relationship between collateral circulation and myocardium viability in these patients. Methods: This retrospective study included 40 patients with CTO diagnosed by invasive coronary angiography (CAG) from September 2015 to June 2016 in our department, all patients performed CMR examination within one week after CAG.The collateral circulation of CTO was graded with Rentrop classification as follows: poor or no collateral circulation group, moderate collateral circulation group and good collateral circulation group.According to CMR images, the delayed enhancement transmural extent of myocardial segments were scored, the ventricular wall motion of the myocardial segment were graded, and the wall motion score index (WMSI) was calculated.Spearman correlation analysis was used to analyze the relationship between the delayed enhancement transmural extent of myocardial segments and WMSI. Results: In the no or poor collateral group of 6 myocardial regions, 1 myocardial region had viable myocardium and 3 myocardial regions had no viable myocardium; in the moderate collateral group of 16 myocardial regions, 11 myocardial regions had viable myocardium and 5 myocardial regions had no viable myocardium; in the good collateral group of 24 myocardial regions, 21 myocardial regions had viable myocardium and 3 myocardial regions had no viable myocardium, there was significant difference between the groups (P=0.002). The WMSI of poor or no collateral circulation group, moderate collateral circulation group and good collateral circulation group were 1.54±0.50, 1.21±0.34 and 1.26±0.40, respectively, there was no significant difference between the groups (P=0.063). Spearman correlation analysis showed that the extent of delayed enhancement transmural extent of myocardial segment was significantly associated with WMSI (r=0.638, P<0.01). Conclusion: Collateral circulation in patients with chronic total occlusion can predict myocardial viability.Increase of Rentrop grade is linked with higher possibility of the presence of viable myocardium.

[Impact of different antithrombotic therapy strategy on prognosis in coronary heart disease patients combining with atrial fibrillation: a meta analysis].

Objective: To evaluate the impact of various anticoagulation antiplatelet therapy strategies on the prognosis of patients with coronary heart disease combining with atrial fibrillation. Methods: Present meta analysis was performed according to search results on English EMBASE database by computer retrieval, Pubmed, the Cochrane Central Register of Controlled Trials, Medline, Chinese CBM database, CNKI database, Wan Fang database, China science and technology papers online electronic databases, manual retrieval for important international conference proceedings up to April 30 2016. Trials published in English and Chinese language, which met the Cochrane system evaluation requirements were included and the inclusion and exclusion criteria were made based on Cochrane system evaluation requirements. The end point is the incident of major adverse cardiac events (MACE), ischemic stroke and major bleeding events. The patients were randomly assigned into triple antithrombotic therapy (aspirin+ clopidogrel+ warfarin) group and dual antiplatelet therapy (aspirin+ clopidogrel) group.The collected full-text literatures underwent further quality assessment of the risks of bias using RevMan 5.3 software. Impact of various antithrombotic therapeutic strategies on the outcome of coronary heart disease patients combining with atrial fibrillation were evaluated. Results: In this meta analysis, 12 randomized controlled trials with 11 353 patients were included. Among these patients, 3 486 patients received triple antithrombotic therapy and 7 867 patients received dual anti-platelet therapy. There was no significant difference in incidence rate of MACE (OR=0.93, 95%CI 0.74-1.18, P>0.05) and the incidence rate of ischemic stroke (OR=0.88, 95%CI 0.70-1.10, P=0.27) between the two patients groups. However, the incidence rate of major bleeding events in triple antithrombotic therapy group was twice higher than that in dual anti-platelet therapy (OR=1.94, 95%CI 1.33-2.82, P=0.000 6). Conclusion: Compared with dual anti-platelet therapy strategy, coronary heart disease patients combining with atrial fibrillation who were treated by triple antithrombotic therapy strategy have the similar outcome on risk of ischemic stroke, but higher risk of major bleeding events.

[The clinical efficacy analysis of the surgery of the posterior approach parotid gland region resection preserving retinal ganglion and parotid fascia].

Objective:To explore the clinical efficacy analysis the surgery of the posterior approach parotid gland resection preserving retinal ganglion and parotid fascia for the treatment of benign parotid tumour.Method:One hundred and twelve cases were randomly divided into control group and treatment group. The control group was treated by anterior approach parotid gland region resection preserving retinal ganglion and parotid fascia. The treatment group was treated by posterior approach parotid gland region resection preserving retinal ganglion and parotid fascia. The data of operation time, postoperative local numbness, Frey syndrome, facial paralysis and postoperative recurrence case were analyzed. Result:There was no postoperative recurrence case in both two groups, and there was no significant difference between two the groups in operation time and the percent of facial paralysis（all P>0.05）.However, the percent of postoperative local numbness and Frey syndrome in control group were significantly higher than those in treatment group（all P<0.05）. Conclusion:The treatment group had similar clinical efficacy with the traditional surgical operation. In addition, the operation of treatment group contributed to decreased postoperative complications.

[The relationship between myocardial bridge in mural coronary artery segment and coronary atherosclerosis].

Objective: The myocardial bridging (MB) prevalence, anatomic characteristics of MB, and the relationship between characteristics of MB in mural coronary artery segment and coronary atherosclerosis were analyzed. Methods: In this perspective nonrandomized controlled study, a total of 1 132 patients who admitted to our hospital for suspected or known coronary artery disease from January 2012 to June 2013 were enrolled. All patients underwent dual-source 64-slice spiral CT coronary angiography. The general patient characteristics including gender, age, history of hypertension, diabetes, hyperlipidemia and smoking, serum level of total cholesterol (TC) and LDL-C were recorded. The length, depth and the degree of compression of myocardial bridge in systolic or diastolic phase were also analyzed in patients with MB. The relationship between MB and coronary atherosclerosis, the characteristics of MB and coronary atherosclerosis were analyzed by Spearman correlation analysis, univariate logistic regression analysis, variate logistic regression analysis and linear regression analysis. Results: Myocardial bridging was detected in 330 out of 1 132 patients, and MB was mostly located in the mural coronary artery (329/330) and at the mid-distal segment of the left anterior descending artery (LAD). Average MB length was 20.1 mm (3.3-95.5 mm) and the average depth was 2.13 mm (0.24-12.40 mm). There were 140 patients with intramyocardial MB (42.6%) and 189 patients with superficial MB (57.4%). Myocardial bridging was an independent protective factor of coronary atherosclerosis (OR=0.361, P=0.000) and the proximal segment of MB was more susceptible to atherosclerosis compared to the distal segment of MB (P=0.000). Multivariate analysis revealed that age, hypertension and the degree of compression of myocardial bridge in diastolic phase were independent factors related to the atherosclerosis (odds ratio: 1.064, 2.186 and 1.049 respectively, P value: 0.000, 0.002 and 0.000). The depth of MB was significantly correlated with systolic or diastolic narrowing(OR: 4.227, 3.398 and P value: 0.000, 0.001). Conclusions: The prevalence of myocardial bridging is 29% in this patient cohort. The proximal segment of myocardial bridging in mural coronary artery is more susceptible to atherosclerosis. In addition, the depth of myocardial bridging and the degree of compression of myocardial bridge in diastolic phase are the independent factors related to atherosclerosis.

Prenatal omega-3 fatty acid supplementation does not affect offspring telomere length and F2-isoprostanes at 12 years: A double blind, randomized controlled trial.

BACKGROUND: Oxidative stress and nutritional deficiency may influence the excessive shortening of the telomeric ends of chromosomes. It is known that stress exposure in intrauterine life can produce variations in telomere length (TL), thereby potentially setting up a long-term trajectory for disease susceptibility. OBJECTIVE: To assess the effect of omega-3 long chain polyunsaturated fatty acid (n-3 LCPUFA) supplementation during pregnancy on telomere length and oxidative stress in offspring at birth and 12 years of age (12y). DESIGN: In a double-blind, placebo-controlled, parallel-group study, 98 pregnant atopic women were randomised to 4g/day of n-3 LCPUFA or control (olive oil [OO]), from 20 weeks gestation until delivery. Telomere length as a marker of cell senescence and plasma and urinary F2-isoprostanes as a marker of oxidative stress were measured in the offspring at birth and 12y. RESULTS: Maternal n-3 LCPUFA supplementation did not influence offspring telomere length at birth or at 12y with no changes over time. Telomere length was not associated with F2-isoprostanes or erythrocyte total n-3 fatty acids. Supplementation significantly reduced cord plasma F2-isoprostanes (P<0.001), with a difference in the change over time between groups (P=0.05). However, the differences were no longer apparent at 12y. Between-group differences for urinary F2-isoprostanes at birth and at 12y were non-significant with no changes over time. CONCLUSIONS: This study does not support the hypothesis that n-3 LCPUFA during pregnancy provides sustained effects on postnatal oxidative stress and telomere length as observed in the offspring.

[Impact of adherence to statins on cardiovascular adverse events in patients with coronary artery disease: a meta-analysis].

OBJECTIVE: To evaluate the impact of different adherence mode to statins on cardiovascular adverse events in patients with coronary artery disease (CAD). METHODS: Electronic searches, including PubMed, Scopus, Ovid MEDLINE, Ovid EMBASE, Ovid EBM Reviews CENTRAL, CINAHL, The Cochrane Library, Ovid PsycInfo, Wanfang data, CNKI and Science & Technology Magazine Online, were performed and all related literatures of all languages were retrieval till to March 1, 2015. The full text was obtained through manual retrieval, inter-library loan and document delivery service, or by contacting the author directly. According to inclusion and exclusion criteria, data was extracted dependently by two raters. The high adherence to statin was use defined by the ratio of statins cover time and the total time (proportion of days covered, PDC≥80%). Data were analyzed quantitatively using RevMan 5.1. Then implement subgroup analysis was made according to different statin adherence and classification of clinical outcomes. The impact of adherence to statin on cardiovascular events (all-cause mortality, non-fatal myocardial infarction, hospitalization due to unstable angina pectoris, heart insufficiency attack) in CAD patients was evaluated. RESULTS: Present analysis enrolled eight relevant retrospective and observational studies. Because there were only few literatures describing the impact of statin adherence on clinical outcomes, we also included literatures with low adherence group (4 studies in PDC<80%, 2 studies in PDC<40% and 2 studies in PDC<20%). High adherence group includes 189 556 cases; low adherence group includes 11 384 cases. Compare with low adherence group, cardiovascular events rate reduced by 32% in high adherence group (OR=0.68, 95%CI 0.58-0.80, P<0.001). Subgroup analysis with 4 literatures with PDC≥80% or<80% showed that the cardiovascular events prominently decreased in high adherence group compared to low adherence group (OR=0.63, 95%CI 0.53-0.76, P<0.001). According to 5 literatures with all-cause mortality parameter, we also found a borderline decrease in all-cause mortality in high adherence group compared to low adherence group ( OR=0.67, 95%CI 0.44-1.02, P=0.06), while non-fatal cardiovascular events were significantly reduced by 18% in high adherence group (OR=0.82, 95%CI 0.77-0.87, P<0.001). CONCLUSIONS: High adherence to statins is related to significantly lower cardiovascular events in CAD patients.