Age effect on the shared etiology of glycemic traits and serum lipids: evidence from a Chinese twin study.

PURPOSE: Diabetes and dyslipidemia are among the most common chronic diseases with increasing global disease burdens, and they frequently occur together. The study aimed to investigate differences in the heritability of glycemic traits and serum lipid indicators and differences in overlapping genetic and environmental influences between them across age groups. METHODS: This study included 1189 twin pairs from the Chinese National Twin Registry and divided them into three groups: aged ≤ 40, 41-50, and > 50 years old. Univariate and bivariate structural equation models (SEMs) were conducted on glycemic indicators and serum lipid indicators, including blood glucose (GLU), glycated hemoglobin A1c (HbA1c), total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C), in the total sample and three age groups. RESULTS: All phenotypes showed moderate to high heritability (0.37-0.64). The heritability of HbA1c demonstrated a downward trend with age (HbA1c: 0.50-0.79), while others remained relatively stable (GLU: 0.55-0.62, TC: 0.58-0.66, TG: 0.50-0.63, LDL-C: 0.24-0.58, HDL-C: 0.31-0.57). The bivariate SEMs demonstrated that GLU and HbA1c were correlated with each serum lipid indicator (0.10-0.17), except HDL-C. Except for HbA1c and LDL-C, as well as HbA1c and HDL-C, differences in genetic correlations underlying glycemic traits and serum lipids between age groups were observed, with the youngest group showing a significantly higher genetic correlation than the oldest group. CONCLUSION: Across the whole adulthood, genetic influences were consistently important for GLU, TC, TG, LDL-C and HDL-C, and age may affect the shared genetic influences between glycemic traits and serum lipids. Further studies are needed to elucidate the role of age in the interactions of genes related to glycemic traits and serum lipids.

[The diagnostic and evaluation value of plasma interleukin 9 in the mucosal healing in patients with inflammatory bowel disease treated with biological agents].

Objective: To investigate the diagnostic and evaluation value of plasma interleukin 9 (IL9) in the mucosal healing (MH) in patients with inflammatory bowel disease (IBD) treated with biological agents. Methods: Cohort study. IBD patients (137 cases) treated in the Affiliated Suzhou Hospital to Nanjing Medical University (Suzhou Municipal Hospital) from September 2019 to January 2022 were prospective selected. Each patient was treated with biological agents [Infliximab (IFX, 56 cases), Adalimumab (ADA, 20 cases), Ustekinumab (UST, 18 cases), Vedolizumab (VDZ, 43 cases)]. According to different therapeutic drugs, the IFX, ADA, UST, and VDZ group were divided. Clinical symptoms, inflammatory indicators and imaging examinations etc. were evaluated every 8 weeks, and the degree of MH was evaluated by endoscopy at the 54th week. The expression of plasma IL9 was detected by ELISA after initial enrollment (W 0) and 8 weeks of biological treatment (W 8). Receiver operating characteristic curve (ROC) was used to evaluate the diagnostic efficacy of IL9 for MH. Select the cut off value for the optimal ROC threshold based on the highest value of the Youden index. Spearman's rank correlation was used to analyze the correlation between IL9 and Simple Endoscopic Score for CD (SES-CD) and Mayo Endoscopic Score (MES), so as to evaluate the predictive value of IL9 for MH in IBD patients treated with biologic agents. Results: Among the 137 patients, there were 97 Crohn's disease (CD) patients, 53 males and 44 females, aged (31.6±10.3) years (18-60 years). There were 40 ulcerative colitis (UC) patients, 22 males and 18 females, aged (37.5±14.7) years (18-67 years). Among the CD patients, 42 cases (43.3%) achieved MH on endoscopy at the 54th week, and 60 patients (61.9%) achieved clinical remission. Among the UC patients, 22 cases (55.0%) achieved MH and 30 cases (75.0%) achieved clinical remission. At W 0, the relative expression of IL9 in patients in IBD patients who achieved MH after 54 weeks of biological treatment was lower than that in the non-MH patients [x¯±s, (127.42±34.43) vs (146.82±45.64) ng/L, (113.01±44.88) vs (146.12±48.66) ng/L, respectively, both P<0.05]. At W 8, the relative expression of IL9 in the MH group was lower than that in the non-MH patients (both P<0.05). The relative expression of IL9 in the MH patients after IFX treatment was lower than that in the non-MH group (P<0.05). There was no significant difference among the other groups between MH and non-MH patients (all P>0.05). IL9 at W 8 showed high value in predicting MH in IBD [CD patients: area under curve (AUC)=0.716(95%CI: 0.616-0.817, P<0.001), sensitivity and specificity were 80.77%(95%CI:67.64%-88.45%) and 48.89%(95%CI: 35.53%-64.47%), respectively; UC patients: AUC=0.821, sensitivity and specificity were 77.78% and 72.73%, respectively]. At W 8, the cut off values for CD and UC patients were IL9>80.77 ng/L and IL9>77.78 ng/L, respectively. IL9 was positively correlated with endoscopic MH score parameters [M(Q1,Q3),SES-CD: 3.0(8.5, 18.5); MES: 2.0(1.0, 3.0)] (r=0.55, 0.72, respectively, both P<0.001) at W8. Conclusion: The plasma IL-9 at the week 8 after biological agents treatment can be used to diagnose and evaluate the MH of patients with IBD.

[The mechanism by which hsa_circRNA_103124 highly expressed in peripheral blood of patients with active Crohn's disease regulates macrophage differentiation, pyroptosis and inflammation].

Objective: To investigate the role and related mechanism of the highly expressed circular RNA molecule 103124 (hsa_circRNA_103124) in macrophage differentiation, pyroptosis and inflammation in peripheral blood mononuclear cells (PBMC) of patients with active Crohn's disease (CD). Methods: Patients with active CD (CD group) admitted to the Affiliated Suzhou Hospital of Nanjing Medical University from April to September 2018 and healthy people (control group) from the physical examination center of the hospital from July to October 2018 were retrospectively selected. The levels of hsa_circRNA_103124 and Toll-like receptor 4 (TLR4) in PBMC of the two groups were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Tohoku hospital pediatrics-1 (THP1) cell line was used as a model for the study of hsa_circRNA_103124 regulating macrophage differentiation. Lentivirus infection was used to construct hsa_circRNA_103124 overexpressed or down-regulated THP1 cells to induce macrophage-like differentiation. According to the expression level of hsa_circRNA_103124, THP1 cell lines were divided into the following four groups: pLC5-ciR was overexpression control group; hsa_circRNA_103124 OE was the overexpression group; ShRNActrl was down-regulated expression control group; hsa_circRNA_103124 ShRNA was the down-regulated expression group. Flow cytometry was used to detect levels cluster of differentiation (CD) 68, CD80, interleukin (IL)-6, tumor necrosis factor α (TNF-α) and reactive oxygen species (ROS). The expression levels of IL-6, TNF-α, IL-1ß, TLR4 and myeloid differentiation factor 88 (MyD88) were detected by RT-qPCR. The levels of gasdermin D (GSDMD), IL-18 and NOD-like receptor thermal protein domain associated protein 3 (NLRP3) were determined by immunofluorescence and RT-qPCR. Pearson correlation analysis was used to analyze the correlation between the abundance of hsa_circRNA_103124 and TLR4 expression level or Crohn's disease activity index (CDAI). Results: A total of 50 patients were included in the CD group, including 36 males and 14 females, aged (35±10) (19-64) years. A total of 30 subjects were included in the control group, including 22 males and 8 females, aged (38±9) (24-64) years. hsa_circRNA_103124 [(0.009±0.016) vs (0.003±0.002), P=0.042] and TLR4 [(0.005±0.003) vs (0.001±0.001), P<0.001] were all upregulated in the PBMC of patients in the CD group, compared with the control group. And hsa_circRNA_103124 was positively correlated with TLR4 (r=0.40, P=0.004). hsa_circRNA_103124 level was positively correlated with CDAI (r=0.32, P=0.024). The expression of CD68 (P=0.002) and CD80 (P<0.001) were enhanced. hsa_circRNA_103124 promoted production of ROS and the expression of IL-6, TNF-α, IL-1ß, TLR4, MyD88, GSDMD, IL-18 and NLRP3 in macrophage-like M1 differentiated THP1 cells (all P<0.05). Conclusion: High expresion of hsa_circRNA_103124 in PBMC of patients with active CD may promote macrophage M1 differentiation, pyroptosis and inflammation through enhancing the expression of TLR4, MyD88, NLRP3 and GSDMD.

[Heritability and genetic correlation of body mass index and coronary heart disease in Chinese adult twins].

Objective: To examine the heritability of body mass index (BMI) and coronary heart disease (CHD), and to explore whether genetic factors can explain their correlation. Methods: Participants were from 11 provinces/municipalities reqistered in the Chinese National Twin Registry (CNTR) from 2010 to 2018. Participants data were collected from face-to-face questionnaire survey. Bivariate structure equation model was used to estimate the heritability and the genetic correlation of BMI and CHD. Results: A total of 20 340 pairs of same-sex twins aged ≥25 years were included in this study. After adjusting for age and gender, the heritability of BMI and CHD was 0.52 (95%CI: 0.49-0.55) and 0.76 (95%CI: 0.69-0.81), respectively. Further, a genetic correlation was identified between BMI and CHD (rA=0.10, 95%CI:0.02-0.17). Conclusion: In Chinese adult twin population, BMI and CHD are affected by genetic factors, and their correlation can be attributed to the common genetic basis.

[Diagnostic efficiency and incremental value of myocardial blood flow quantification by CZT SPECT for patients with coronary artery disease].

Objective: To investigate the diagnostic efficiency and incremental value of quantitative myocardial blood flow measurements by Cadmium-Zine-Telluride (CZT) single photon emission computed tomography (SPECT) dynamic myocardial perfusion imaging (MPI) in patients with coronary artery disease (CAD) compared with traditional semi-quantitative measurements by MPI. Methods: This is a retrospective, cross-sectional study. We retrospectively analyzed clinical data of patients with suspected or known CAD, who underwent the dynamic MPI quantitative blood flow measurement of CZT SPECT in TEDA International Cardiovascular Hospital from October 2018 to December 2020. Clinical data, semi-quantitative parameters (stress score (SS), rest score (RS) and different score (DS)) and myocardial quantitative blood flow parameters (rest myocardial blood flow (rMBF), stress myocardial blood flow (sMBF) and myocardial flow reserve (MFR)) were analyzed. According to the results of coronary angiography, patients were divided into the stenosis group and the control group with coronary artery stenosis ≥50% or ≥75% as the diagnosis criteria. The differences of quantitative and semi-quantitative parameters between the two groups were compared, and the diagnostic efficacy was compared by receiver operating characteristic(ROC) curve. Results: A total of 98 patients with a mean age of (62.1±8.7) years were included in the study, including 66 males (67%). At the patient level, with the positive standard of coronary artery stenosis≥50%, the left ventricle (LV) stress MBF (LV-sMBF) ((1.36±0.45) ml·min-1·g-1) and LV-MFR (1.45±0.43) of the stenosis group were lower than the LV-sMBF ((2.09±0.64) ml·min-1·g-1) and LV-MFR (2.17±0.54) of control group; summed SS and summed DS were higher than control group (all P<0.05). With the positive standard of coronary artery stenosis ≥75%, the LV-sMBF ((1.19±0.34) ml·min-1·g-1) and LV-MFR (1.34±0.35) of stenosis group were lower than the LV-sMBF ((1.94±0.63) ml·min-1·g-1) and MFR (2.00±0.58) of control group; all semi-quantitative parameters were higher than control group (all P<0.05). At the vascular level, with coronary artery stenosis ≥50% as the diagnosis criteria, the sMBF ((1.26±0.49) ml·min-1·g-1) and MFR (1.35±0.46) of stenosis group were lower than the sMBF ((1.95±0.70) ml·min-1·g-1) and MFR (2.05±0.65) of control group; SS and DS were higher than control group (all P<0.05). With coronary artery stenosis≥75% as the diagnosis criteria, the sMBF ((1.12±0.41) ml·min-1·g-1) and MFR (1.25±0.38) of stenosis group were lower than the sMBF ((1.84±0.70) ml·min-1·g-1) and MFR (1.93±0.66) of control group; all semi-quantitative parameters were higher than control group (all P<0.05). With coronary artery stenosis≥50% as the diagnosis criteria and CAG as the reference standard, the AUC and 95%CI of myocardial quantitative blood flow parameters indicated by ROC curve for diagnosis of CAD were 0.830 (0.783-0.877). The sensitivity (86.1% vs. 61.5%), specificity (82.6% vs. 73.8%), positive predictive value (77.8% vs. 62.5%), negative predictive value (89.3% vs. 73.0%) and accuracy (84.0% vs. 68.7%) were all higher than the semi-quantitative parameters (all P<0.05). With coronary artery stenosis≥75% as the diagnosis criteria, the AUC and 95%CI of myocardial quantitative blood flow parameters indicated by ROC curve for diagnosis of CAD were 0.832(0.785-0.879). The sensitivity (89.2% vs. 67.6%), negative predictive value (95.5% vs. 86.2%) and accuracy (80.6% vs. 68.0%) were all higher than semi-quantitative parameters (all P<0.05). Conclusion: Compared with traditional SPECT MPI derived semi-quantitative parameters, diagnostic efficacy for CAD is higher using CZT SPECT quantitative myocardial blood flow parameters, this strategy thus has additional diagnostic benefits and incremental value on the diagnosis of CAD.

Antifungal mechanism of Streptomyces ma. FS-4 on fusarium wilt of banana.

AIM: Research on prevention and cure of banana wilt is important to ensure the healthy development of the banana industry. In this study, antifungal mechanism of Streptomyces ma. FS-4 on fusarium wilt of banana was investigated. METHODS AND RESULTS: The physiological strain of banana fusarium pathogen Fusarium oxysporum f. sp. cubense Race 4 (FOC.4) was used as the target fungus, and the antifungal mechanism of the crude extract of Streptomyces ma. FS-4 was investigated. Eighteen different compounds identified by gas chromatography-mass spectrometry were composed of aldehydes, methyl, hydrocarbons, amides, esters and acids. FS-4 significantly inhibited the spore germination of the target fungi, with an EC50 of 22·78 µg ml-1 . After treatment with 100 µg ml-1 FS-4 crude extract, the N-acetylglucosamine content in the mycelium increased 1·95-fold. However, the extract had no significant effect on ß-1,3-glucanase. At the FS-4 crude extract dose of 100 µg ml-1 , the total sugar and protein contents decreased by 28·6 and 29·1% respectively, and the fat content was 41·3%. FS-4 significantly inhibited the activity of the mitochondrial complex III of Foc4, which was reduced by 52·45%. Moreover FS-4 reduced the activity of succinate dehydrogenase, a key enzyme in the Krebs cycle, by 60·2%. However, FS-4 had no significant effect on malate dehydrogenase. The membrane potential on the mitochondrial inner membrane was significantly reduced at the test concentration of 100 µg ml-1 . ROS gradually accumulated in the Foc4 hypha, and the burst was 3·97 times higher than the control. CONCLUSIONS: This study demonstrated that the antifungal mechanism of Streptomyces ma. FS-4 against Foc4 includes the destruction of the plasma membrane and mitochondrial dysfunction and finally induction of cell apoptosis. SIGNIFICANCE AND IMPACT OF THE STUDY: These results may indicate the prevention and control of banana wilt, which is of great significance to the healthy development of banana industry system.

Integrated microRNA-mRNA analysis reveals the roles of microRNAs in the muscle fat metabolism of Yanbian cattle.

Fat deposition is an important economic trait in farm animals. However, it is difficult to genetically improve intramuscular fat deposition via trait-based cattle breeding. The main objectives of this study were to analyze the factors about beef flavor, and to detect functional microRNA (miRNA, miR) associated with intramuscular fat deposition in Yanbian cattle. Longissimus dorsi samples from six steers were separated into high- and low-fat groups (n = 3 each) based on the marbling score, and transcriptomic analysis was performed using miRNA sequencing. A total of 33 miRNAs and 38 genes were found to be differentially expressed in the high- and low-fat groups. Quantitative real-time polymerase chain reaction was performed to validate the sequencing results. Integrated miRNA-mRNA analysis revealed that miRNA-associated target genes were primarily associated with skeletal muscle development. However, some of the miRNAs (miR-424 etc.) and genes (ATF3 etc.) were also associated with fat metabolism. A targeted relationship between miR-22-3p and the WFIKKN2 gene and its involvement in adipocyte differentiation were confirmed experimentally. The study findings may provide potential candidate molecular targets for the selection of cattle with improved meat quality.

[Correlation between fasting plasma glucose, HbA1c and DNA methylation in adult twins].

OBJECTIVE: To explore the cytidine-phosphate-guanosine (CPG) sites associated with fas-ting plasma glucose (FPG) and glycated haemoglobin (HbA1c) in twins. METHODS: In the study, 169 pairs of monozygotic twins were recruited in Qingdao, Zhejiang, Jiangsu, Sichuan and Heilongjiang in June to December of 2013 and June 2017 to October 2018. The methylation was detected by Illumina Infinium HumanMethylation450 BeadChip and Illumina Infinium MethylationEPIC BeadChip. According to the Linear Mixed Effect model (LME model), fasting plasma glucose and HbA1c were taken as the main effects, the methylation level (ß value) was taken as the dependent variable, continuous variables, such as age, body mass index (BMI), blood pressure, components of blood cells, surrogate variables generated by SVA, and categorical variables, such as gender, smoking and drinking status, hypoglycemic drugs taking, were included in the fixed effect model as covariates, and the identity numbers (ID) of the twins was included in the random effect model. The intercept was set as a random. Regression analysis was carried out to find out the CpG sites related to fasting blood glucose or HbA1c, respectively. RESULTS: In this study, 338 monozygotic twins (169 pairs) were included, with 412 459 CpG loci. Among them, 114 pairs were male, and 55 pairs were female, with an average age of (48.2±11.9) years. After adjustment of age, gender, BMI, blood pressure, smoking, drinking, blood cell composition, and other covariates, and multiple comparison test, 7 CpG sites (cg19693031, cg01538969, cg08501915, cg04816311, ch.8.1820050F, cg06721411, cg26608667) were found related to fasting blood glucose, 3 of which (cg08501915, ch.8.1820050f, cg26608667) were the newly found sites in this study; whereas 10 CpG sites (cg19693031, cg04816311, cg01538969, cg01339781, cg01676795, cg24667115, cg09029192, cg20697417, ch.4.1528651F, cg16097041) were found related to HbA1c, and 4 of which(cg01339781, cg24667115, cg20697417, and ch.4.1528651f) were new. We found that cg19693031 in TXNIP gene was the lowest P-value site in the association analysis between DNA methylation and fas-ting plasma glucose and HbA1c (PFPG=2.42×10-19, FDRFPG<0.001; PHbA1c=1.72×10-19, FDRHbA1c<0.001). CONCLUSION: In this twin study, we found new CpG sites related to fasting blood glucose and HbA1c, and provided some clues that partly revealed the potential mechanism of blood glucose metabolism in terms of DNA methylation, but it needed further verification in external larger samples.

The Chinese National Twin Registry: a 'gold mine' for scientific research.

The Chinese National Twin Registry (CNTR) currently includes data from 61 566 twin pair from 11 provinces or cities in China. Of these, 31 705, 15 060 and 13 531 pairs are monozygotic, same-sex dizygotic and opposite-sex dizygotic pairs, respectively, determined by opposite sex or intrapair similarity. Since its establishment in 2001, the CNTR has provided an important resource for analysing genetic and environmental influences on chronic diseases especially cardiovascular diseases. Recently, the CNTR has focused on collecting biologic specimens from disease-concordant or disease-discordant twin pairs or from twin pairs reared apart. More than 8000 pairs of these twins have been registered, and blood samples have been collected from more than 1500 pairs. In this review, we summarize the main findings from univariate and multivariate genetic effects analyses, gene-environment interaction studies, omics studies exploring DNA methylation and metabolomic markers associated with phenotypes. There remains further scope for CNTR research and data mining. The plan for future development of the CNTR is described. The CNTR welcomes worldwide collaboration.

The PNKD gene is associated with Tourette Disorder or Tic disorder in a multiplex family.

Tourette Disorder (TD) is a childhood-onset neuropsychiatric and neurodevelopmental disorder characterized by the presence of both motor and vocal tics. The genetic architecture of TD is believed to be complex and heterogeneous. Nevertheless, DNA sequence variants co-segregating with TD phenotypes within multiplex families have been identified. This report examines whole exomes of affected and unaffected individuals in a multiplex TD family to discover genes involved in the TD etiology. We performed whole exome sequencing on six out of nine members in a three-generation TD multiplex family. Putative deleterious sequence variants co-segregating with TD patients were identified by our in-house bioinformatics pipeline. Induced pluripotent stem cells (iPSCs) were generated from one unaffected and two TD affected individuals. Neurons were derived from the iPSCs and biochemical assays were conducted to evaluate possible molecular differences between affected and unaffected. A rare heterozygous nonsense mutation in PNKD was co-segregated with TD in this multiplex family. Transcript and protein levels of the PNKD long isoform were reduced in neurons derived from the individuals with TD due to the nonsense mutation, indicating nonsense-mediated mRNA decay. We demonstrated that the PNKD long isoform monomer oligomerizes with itself as well as interacts with the synaptic active zone protein RIMS1α. We concluded that reduced PNKD long isoform levels are detected in all affected individuals and we provide evidence for a mechanism whereby this might contribute to the TD phenotype.

The CODATwins Project: The Current Status and Recent Findings of COllaborative Project of Development of Anthropometrical Measures in Twins.

The COllaborative project of Development of Anthropometrical measures in Twins (CODATwins) project is a large international collaborative effort to analyze individual-level phenotype data from twins in multiple cohorts from different environments. The main objective is to study factors that modify genetic and environmental variation of height, body mass index (BMI, kg/m2) and size at birth, and additionally to address other research questions such as long-term consequences of birth size. The project started in 2013 and is open to all twin projects in the world having height and weight measures on twins with information on zygosity. Thus far, 54 twin projects from 24 countries have provided individual-level data. The CODATwins database includes 489,981 twin individuals (228,635 complete twin pairs). Since many twin cohorts have collected longitudinal data, there is a total of 1,049,785 height and weight observations. For many cohorts, we also have information on birth weight and length, own smoking behavior and own or parental education. We found that the heritability estimates of height and BMI systematically changed from infancy to old age. Remarkably, only minor differences in the heritability estimates were found across cultural-geographic regions, measurement time and birth cohort for height and BMI. In addition to genetic epidemiological studies, we looked at associations of height and BMI with education, birth weight and smoking status. Within-family analyses examined differences within same-sex and opposite-sex dizygotic twins in birth size and later development. The CODATwins project demonstrates the feasibility and value of international collaboration to address gene-by-exposure interactions that require large sample sizes and address the effects of different exposures across time, geographical regions and socioeconomic status.

[Correlation between blood pressure and DNA methylation in adult twins].

OBJECTIVE: To explore the DNA methylation sites correlated with blood pressure (systolic blood pressure, diastolic blood pressure, mean arterial pressure, pulse pressure) in adult twin population. METHODS: A total of 476 twins from the Chinese National Twin Registry were selected as the research population. Questionnaires were used to collect demographic characteristics, lifestyle, disease status and other information, and blood pressure, height, weight and other anthropometric indicators were measured. The genome-wide DNA methylation of whole blood samples was detected by using Infinium HumanMethylation450 BeadChip. The DNA methylation sites correlated with blood pressure were analyzed by constructing mixed effect model with adjusting potential confounding factors, and the significant level was false discovery rate <0.05. RESULTS: After data quality control, 465 twins (122 pairs of monozygotic twins, 104 pairs of dizygotic twins, 13 individuals from 13 pairs of twins) aged (44.8±13.2) years were finally enrolled. There were more males and more monozygotic twins, and the current smokers and current regular drinkers both accounted for more than 30%. No significant CpG site was found after multiple testing in the correlation study between genome-wide DNA methylation and blood pressure by using the collected twins. However, the cg07761116 located on chromosome 10 had low P value in the correlation analysis of 3 blood pressure indices (systolic blood pressure, diastolic blood pressure, mean arterial pressure), suggesting that this site might be correlated with blood pressure. The other 7 sites had low P value in the correlation analysis of the two blood pressure indices, respectively, which pointed to genes involved in neurological development, protein homeostasis, inflammatory reaction and other pathways. CONCLUSION: There is no sufficient evidence to support any DNA methylation site correlated with blood pressure, which may be caused by insufficient sample size and other reasons. This study could provide a reference for subsequent similar twin studies, and subsequent studies can focus on the cg07761116 located on chromosome 10 and other sites with low P values.

Exposure to famine in early life and the risk of obesity in adulthood in Qingdao: Evidence from the 1959-1961 Chinese famine.

BACKGROUND AND AIMS: We aimed to evaluate the association between famine exposure during early life and obesity and obesitymax (obese at the highest weight) in adulthood. METHODS AND RESULTS: Data were from two population-based cross-sectional surveys conducted in 2006 and 2009 in Qingdao, China. A total of 8185 subjects born between 1/1/1941 and 12/31/1971 were categorized into unexposed (born between 01/01/1962 and 12/31/1971), fetal/infant exposed (born between 01/01/1959 and 12/31/1961), childhood exposed (born between 01/01/1949 and 12/31/1958) and adolescence exposed (born between 01/01/1941 and 12/31/1948) according to their age when exposed to the Chinese famine from 1959 to 1961. Obesity was defined as BMI (body mass index) ≥28.0 and obesitymax was defined as BMImax (BMI at the highest weight) ≥28.0. We compared fetal/infant exposed, childhood exposed and adolescence exposed to the unexposed using logistic regression models to assess the effect of famine exposure on later obesity and obesitymax. Fetal/infant exposed (OR = 1.59, P < 0.001), childhood exposed (OR = 1.42, P < 0.01) and adolescence exposed (OR = 1.86, P < 0.01) all had higher risks of obesity than the unexposed. Exposure groups were more likely to be obese at their highest weight than the unexposed, and ORs (95%CIs) for obesitymax in the fetal/infant exposed, childhood exposed and adolescence exposed were 1.49(1.20-1.86), 1.24(1.02-1.49) and 1.64 (1.40-1.93), respectively. Similar results were found in both men and women. CONCLUSION: Exposure to famine in early life was associated with increased risks of obesity and obesitymax in adulthood. Preventing undernutrition in early life appears beneficial to reduce the prevalence of later obesity.

[Significance of serum calprotectin in assessing inflammatory bowel disease activity].

Objective: To investigate the application value of serum calprotectin in assessing the activity of IBD Methods: Eighty-five Crohn's disease(CD) and eighty-five Ulcerative colitis(UC) patients at Suzhou Municipal Hospital(North Branch) from August 2015 to January 2017 were enrolled in this study, and eighty-five healthy subjects were selected as controls. Serum level of calprotectin was determined by ELISA. Correlation of serum calprotectin with clinical features of IBD was analyzed. The diagnostic efficacy for CD and UC were assessed by ROC curve. Results: The level of serum calprotectin was significantly higher in IBD patients than in healthy subjects. Positive correlations were found between serum calprotectin level and CRP, ESR, fecal calprotectin level of IBD(ρ＝0.341, P<0.001；ρ＝0.438, P<0.001；ρ＝0.542, P<0.001，respectively). Besides, positive correlations were found between serum calprotectin level with disease activity index(CDAI or Mayo score) of IBD(ρ＝0.309, P＝0.004；ρ＝0.227, P＝0.036，respectively)．Nevertheless, no correlation was found between serum calprotectin level with location of IBD and disease subtypes(P>0.05)．The area under curve(AUC) of serum calprotectin for diagnosis of CD and UC were 0.946 and 0.906, respectively. Conclusions: Serum level of calprotectin is associated with the activity of IBD. As a result，serum level of calprotectin may has the potential to be served as a clinical index of IBD activity.

[Expression of microRNA-155 in inflammatory bowel disease and its clinical significance].

Objective: To explore the expression of microRNA-155 in colonic mucosa and peripheral blood in patients with inflammatory bowel disease(IBD), and to examine the clinical value and significance of microRNA-155 in the diagnosis of IBD. Methods: Quatitative reverse-transcription PCR was performed to detect the expression of microRNA-155 in 20 patients with Crohn disease(CD), 21 patients with ulcerative colitis (UC), 18 patients with IBD type unclassified(IBDU), 25 healthy people(control group), 12 patients with infection colitis and 19 patients with ischemia colitis.Receiver operating characteristic (ROC) curve was performed to analyze the clincal value of microRNA-155 in diagnosis of IBD. Results: The expression of microRNA-155 in colonic mucosa in CD, UC and IBDU group was significantly higher than that in control group(P<0.05). MicroRNA-155 expression was also significantly higher in UC group in comparison to CD group (35.4±3.0 vs 18.6±5.9, P<0.01), IBDU group in comparison to CD group (23.0±3.7 vs 18.6±5.9, P<0.05) and UC group in comparison to IBDU group (35.4±3.0 vs 23.0±3.7, P<0.01). The plasma level of microRNA-155 in UC group (55.6±2.5) and IBDU group (48.1±6.2) was significantly higher than that in control group(P<0.05), while no significant difference in CD group was observed when compared with control group(P>0.05). ROC curve shows an AUC of 0.83 and 95%CI of 0.679-0.986 of microRNA-155 expression in colonic mucosa.The sensitivity and specificity of microRNA-155 expression in colonic mucosa in diagnosis of IBD was 68.4% and 78.6%, respectively. Conclusions: MicroRNA-155 showed high expression in colonic mucosa and peripheral blood in patients with IBD.MicroRNA-155 shows promise as a biomarker in diagnosis of IBD.Furthermore, the aberrant expression indicates that microRNA-155 may be involved in pathogenesis and progression of IBD.

[Mendelian randomization analysis of the relationship between obesity and DNA methylation].

Objective: To explore the association between DNA methylation and body mass index (BMI) using Mendelian randomization analysis. Methods: A total of 469 participants were selected from the Chinese National Twin Registry in 2013, who were living in Shandong, Jiangsu, Zhejiang, and Sichuan provinces, and at least 18 years of age. A questionnaire survey and physical examination were conducted to collect demographic, clinical, and behavioral information. Peripheral blood cells were collected to detect genotype and methylation status. Association analyses between DNA methylation and BMI and between CpGs and cis-SNP were conducted. With rs748212 as the instrumental variable, the association between cg15053022 and BMI was explored using the Mendelian randomization method. Results: A total of 469 participants were selected. The mean age of participants was (44.8±13.2) years and the BMI was (25.0±3.8) kg/m(2). Nine BMI-related DNA methylation sites were found and DNA methylation site cg15053022 in the ATP4A gene was negatively associated with cis-SNP rs748212 (ß=-0.020); the mean methylation level of AA, AC, and CC were 0.212±0.025, 0.242±0.024, and 0.264±0.028, respectively. rs748212 was associated with BMI (ß=0.04, P=0.007) and closely related to cg15053022 (F=237.66, P=0.143). Mendelian randomization analysis showed lower methylation levels at cg15053022 were associated with higher BMI (ß=-1.97, P<0.001). Conclusion: This study supported the impact of cg15053022 methylation in the ATP4A gene on BMI using Mendelian randomization analysis and provided the basis for using Mendelian randomization analysis in methylation studies.

Maternal anti-HBVs suppress the immune response of infants to hepatitis B vaccine.

It is still controversial whether maternal anti-HBV antibodies (anti-HBVs) affect the infants' immune response to hepatitis B virus (HBV) vaccination. This multicentre study aims to address this question. First, we determined whether the transplacental transfer of maternal anti-HBVs occurs by measuring the titres of 90 anti-HBVs-positive pregnant women and their newborns. The anti-HBVs-positive rates of newborns ranged from 89.7% to 100.0%, depending on the maternal anti-HBVs titres. Secondly, we investigated the effects of maternal anti-HBVs on the immune response of infants to HBV vaccination. A total of 1063 mother-and-infant pairs were enrolled and divided into three groups with maternal anti-HBVs titres of <10 IU/L (negative - 37.9%), 10-499 and ≥500 IU/L. The infants' anti-HBVs-positive rate and titres were negatively correlated with maternal anti-HBVs titres: the anti-HBVs-positive rate of infants were 88.9% (360/405), 84.5% (381/451) and 77.3% (160/207) in mothers with low, intermediate and high antibody titres, respectively, P<.0001. Median titres of anti-HBVs (IU/L) among infants were 169.1, 141.0 and 79.4, respectively, P=.020. One hundred and sixty-two infants were negative for anti-HBVs after the standard vaccination, and 120 of 131 of these infants (91.6%) reached anti-HBVs positivity after the first "booster" dose. The maternal anti-HBVs titres did not significantly affect infant response to this booster. In summary, transplacental transfer of anti-HBVs occurs and high titres of maternal anti-HBVs may suppress the immune response of infants to the standard HBV vaccination. The current schedule of the 0, 1 and 6 month may not be the optimal choice of infants with anti-HBVs-positive mothers.

AIM2 inflammasome is dispensable for the host defense against Pseudomonas aeruginosa infection.

Respiratory tract infection with Pseudomonas aeruginosa is a major cause of hospital—acquired pneumonia in immune—compromised individuals. Lung infection with P. aeruginosa is often associated with production of various inflammatory cytokines including IL—1β. Production of IL—1β requires proteolytic cleavage by a multiprotein complex termed inflammasome. AIM2 inflammasome recognizes foreign cytosolic double stranded DNA. A role of AIM2 in P. aeruginosa infection has not been reported previously. In this study, we found that P. aeruginosa infection induced degradation of AIM2 protein in macrophages and induction of AIM2 mRNA expression in macrophages and in the lung of mice. Interestingly, P. aeruginosa infection induced a similar level of IL—1β, IL—6 and TNF production in wild—type and AIM2—deficient mice. Similarly, no significant differences in bacterial clearance, neutrophil infiltration and NF—κB activation were observed between wild—type and AIM2—deficient mice following P. aeruginosa lung infection. Our data suggest that AIM2 inflammasome is dispensable for the host defense against P. aeruginosa infection.

Cardiovascular risk profiles in relation to newly diagnosed type 2 diabetes diagnosed by either glucose or HbA1c criteria in Chinese adults in Qingdao, China.

AIMS: To study the cardiovascular disease risk profiles in newly diagnosed diabetes diagnosed by either glucose or/and HbA(1c) criteria in Chinese adults. METHODS: Two population-based cross-sectional studies were conducted in 2006 and 2009, respectively. Data from 1987 men and 2815 women aged 35-74 years were analysed. Newly diagnosed diabetes was defined according to either glucose (fasting and/or 2-h glucose), HbA(1c) or both criteria. RESULTS: Ageing, positive family history of diabetes, elevated levels of waist circumference, systolic blood pressure, total cholesterol, triglycerides and γ-glutamyl transferase were independently associated with newly diagnosed diabetes defined by glucose criterion alone, but not for diabetes defined by HbA(1c) criterion alone. Only waist circumference, total cholesterol and smoking were significantly associated with the presence of diabetes defined by HbA(1c) criterion alone. CONCLUSIONS: Cardiovascular disease risk profiles were different in patients with newly diagnosed diabetes defined by the two diagnostic criteria for diabetes. This may have certain clinical implications on diabetes management and research.