LATS1/2 control TGFB-directed epithelial-to-mesenchymal transition in the murine dorsal cranial neuroepithelium through YAP regulation.

Hippo signaling, an evolutionarily conserved kinase cascade involved in organ size control, plays key roles in various tissue developmental processes, but its role in craniofacial development remains poorly understood. Using the transgenic Wnt1-Cre2 driver, we inactivated the Hippo signaling components Lats1 and Lats2 in the cranial neuroepithelium of mouse embryos and found that the double conditional knockout (DCKO) of Lats1/2 resulted in neural tube and craniofacial defects. Lats1/2 DCKO mutant embryos had microcephaly with delayed and defective neural tube closure. Furthermore, neuroepithelial cell shape and architecture were disrupted within the cranial neural tube in Lats1/2 DCKO mutants. RNA sequencing of embryonic neural tubes revealed increased TGFB signaling in Lats1/2 DCKO mutants. Moreover, markers of epithelial-to-mesenchymal transition (EMT) were upregulated in the cranial neural tube. Inactivation of Hippo signaling downstream effectors, Yap and Taz, suppressed neuroepithelial defects, aberrant EMT and TGFB upregulation in Lats1/2 DCKO embryos, indicating that LATS1/2 function via YAP and TAZ. Our findings reveal important roles for Hippo signaling in modulating TGFB signaling during neural crest EMT.

Effect of LncRNA LOC106505926 on myogenesis and Lipogenesis of porcine primary cells.

BACKGROUND: Skeletal muscle development and fat deposition have important effects on meat quality. The study of regulating skeletal muscle development and fat deposition is of great significance in improving the quality of carcass and meat. In the present study, whole transcriptome sequencing (including RNA-Seq and miRNA-Seq) was performed on the longissimus dorsi muscle (LDM) of Jinfen White pigs at 1, 90, and 180 days of age. RESULTS: The results showed that a total of 245 differentially expressed miRNAs were screened in any two comparisons, which may be involved in the regulation of myogenesis. Among them, compared with 1-day-old group, miR-22-5p was significantly up-regulated in 90-day-old group and 180-day-old group. Functional studies demonstrated that miR-22-5p inhibited the proliferation and differentiation of porcine skeletal muscle satellite cells (PSCs). Pearson correlation coefficient analysis showed that long non-coding RNA (lncRNA) LOC106505926 and CXXC5 gene had strong negative correlations with miR-22-5p. The LOC106505926 and CXXC5 were proven to promote the proliferation and differentiation of PSCs, as opposed to miR-22-5p. In terms of mechanism, LOC106505926 functions as a molecular sponge of miR-22-5p to modulate the expression of CXXC5, thereby inhibits the differentiation of PSCs. In addition, LOC106505926 regulates the differentiation of porcine preadipocytes through direct binding with FASN. CONCLUSIONS: Collectively, our results highlight the multifaceted regulatory role of LOC106505926 in controlling skeletal muscle and adipose tissue development in pigs and provide new targets for improving the quality of livestock products by regulating skeletal muscle development and fat deposition.

Individualized network analysis reveals a link between the gut microbiome, diet intervention and Gestational Diabetes Mellitus.

Gestational Diabetes Mellitus (GDM), a serious complication during pregnancy which is defined by abnormal glucose regulation, is commonly treated by diabetic diet and lifestyle changes. While recent findings place the microbiome as a natural mediator between diet interventions and diverse disease states, its role in GDM is still unknown. Here, based on observation data from healthy pregnant control group and GDM patients, we developed a new network approach using patterns of co-abundance of microorganism to construct microbial networks that represent human-specific information about gut microbiota in different groups. By calculating network similarity in different groups, we analyze the gut microbiome from 27 GDM subjects collected before and after two weeks of diet therapy compared with 30 control subjects to identify the health condition of microbial community balance in GDM subjects. Although the microbial communities remain similar after the diet phase, we find that the structure of their inter-species co-abundance network is significantly altered, which is reflected in that the ecological balance of GDM patients was not "healthier" after the diet intervention. In addition, we devised a method for individualized network analysis of the microbiome, thereby a pattern is found that GDM individuals whose microbial networks are with large deviations from the GDM group are usually accompanied by their abnormal glucose regulation. This approach may help the development of individualized diagnosis strategies and microbiome-based therapies in the future.

A Ratiometric Benzimidazole-Based Fluorescent Probe for The Recognition of Phosgene in Solution and Gaseous Phases.

Considering the high toxicity and widespread application of phosgene, there is an urgent need to develop a simple and sensitive method for detecting phosgene. In this work, we designed and synthesized a novel ratiometric fluorescent probe 1 containing fluorophores of benzimidazole and benzothiazole. Probe 1 showed excellent sensitivity (< 30 s) and selectivity (LOD = 3.82 nM) for phosgene and significant ratiometric fluorescence changes. In addition, 1-loaded polystyrene membrane test strips were used to conveniently and efficiently detect phosgene gas (0.5 ppm) via the naked eye and the RGB APP of the smartphone, indicating that this probe has great potential for phosgene detection in the gaseous phase.

Sex difference in nocturnal blood pressure dipping in adolescents with varying degrees of adiposity.

BACKGROUND: For adolescents, abnormal dipping patterns in blood pressure (BP) are associated with early-onset organ damage and a higher risk of cardiovascular disorders in adulthood. Obesity is one of the most common reasons for abnormal BP dipping in young people. However, it is unknown whether the severity of obesity is associated with BP dipping status and whether this association is sex-dependent. METHODS: 499 participants between 12 and 17 years old with overweight or obesity underwent ambulatory blood pressure monitoring (ABPM) between April 2018 and January 2019 in Beijing and Baoding. Participants were grouped by body mass index (BMI) into overweight (BMI 85th-95th percentile), obese (BMI ≥ 95th percentile) and severely obese (BMI ≥ 120% of 95th percentile or ≥ 35 kg/m2) groups. Non-dipping was defined as a < 10% reduction in BP from day to night. The interaction effect between sex and obesity degree was also analyzed. RESULTS: 326 boys and 173 girls were included, of whom 130 were overweight, 189 were obese, and 180 were severely obese. Girls with severe obesity had a higher prevalence of non-dipping, but boys showed no significant differences in BP dipping status between obesity categories. In addition, as obesity severity went up, a more evident increase in night-time SBP was observed in girls than in boys. CONCLUSIONS: Severely obese is associated with a higher prevalence of non-BP dipping patterns in girls than in boys, which suggests that the relationship between the severity of obesity and BP dipping status might be sex-specific.

Toluene Tolerated Li9.88GeP1.96Sb0.04S11.88Cl0.12 Solid Electrolyte toward Ultrathin Membranes for All-Solid-State Lithium Batteries.

Sulfide solid electrolyte membranes employed in all-solid-state lithium batteries generally show high thickness and poor chemical stability, which limit the cell-level energy density and cycle life. In this work, Li9.88GeP1.96Sb0.04S11.88Cl0.12 solid electrolyte is synthesized with Sb, Cl partial substitution of P, S, possessing excellent toluene tolerance and stability to lithium. The formed SbS43- group in Li9.88GeP1.96Sb0.04S11.88Cl0.12 exhibits low adsorption energy and reactivity for toluene molecules, confirmed by first-principles density functional theory calculation. Using toluene as the solvent, ultrathin Li9.88GeP1.96Sb0.04S11.88Cl0.12 membranes with adjustable thicknesses can be well prepared by the wet coating method, and an 8 µm thick membrane exhibits an ionic conductivity of 1.9 mS cm-1 with ultrahigh ionic conductance of 1860 mS and ultralow areal resistance of 0.68 Ω cm-2 at 25 °C. The obtained LiCoO2|Li9.88GeP1.96Sb0.04S11.88Cl0.12 membrane|Li all-solid-state lithium battery shows an initial reversible capacity of 125.6 mAh g-1 with a capacity retention of 86.3% after 250 cycles at 0.1 C under 60 °C.

The key to sustainability: In-depth investigation of environmental quality in G20 countries through the lens of renewable energy, economic complexity and geopolitical risk resilience.

The world is currently facing urgent climate and environmental issues, such as global warming, ecosystem collapse, and energy shortages. In this context, this study selected data from 2000 to 2021 and employed the Method of Moment Quantile Regression (MMQR) to thoroughly investigate the impact of renewable energy consumption, economic complexity, and geopolitical risks on the ecological footprint of the Group of Twenty (G20) countries. The results indicate that in countries with lower quantiles, renewable energy consumption significantly reduces the ecological footprint, whereas its effect is not prominent in countries with higher quantiles. Economic complexity has a negative impact on the ecological footprint, and this impact becomes stronger as the quantile of the ecological footprint rises. Additionally, economic complexity moderates the effect of renewable energy on the ecological footprint. Geopolitical risks facilitate the growth of the ecological footprint. Likewise, robustness tests such as DOLS, FMOLS, and quantile regression confirm these estimates in the same framework. This study has conducted a profound analysis of global environmental issues, offering innovative perspectives and recommendations for achieving goals related to sustainable energy utilization, mitigating climate change, and improving the ecological environment. The findings of this research will guide policymakers in G20 countries to adopt more effective environmental protection measures, thereby contributing to the construction of a sustainable future.

Hippo-Yap Signaling Maintains Sinoatrial Node Homeostasis.

BACKGROUND: The sinoatrial node (SAN) functions as the pacemaker of the heart, initiating rhythmic heartbeats. Despite its importance, the SAN is one of the most poorly understood cardiac entities because of its small size and complex composition and function. The Hippo signaling pathway is a molecular signaling pathway fundamental to heart development and regeneration. Although abnormalities of the Hippo pathway are associated with cardiac arrhythmias in human patients, the role of this pathway in the SAN is unknown. METHODS: We investigated key regulators of the Hippo pathway in SAN pacemaker cells by conditionally inactivating the Hippo signaling kinases Lats1 and Lats2 using the tamoxifen-inducible, cardiac conduction system-specific Cre driver Hcn4CreERT2 with Lats1 and Lats2 conditional knockout alleles. In addition, the Hippo-signaling effectors Yap and Taz were conditionally inactivated in the SAN. To determine the function of Hippo signaling in the SAN and other cardiac conduction system components, we conducted a series of physiological and molecular experiments, including telemetry ECG recording, echocardiography, Masson Trichrome staining, calcium imaging, immunostaining, RNAscope, cleavage under targets and tagmentation sequencing using antibodies against Yap1 or H3K4me3, quantitative real-time polymerase chain reaction, and Western blotting. We also performed comprehensive bioinformatics analyses of various datasets. RESULTS: We found that Lats1/2 inactivation caused severe sinus node dysfunction. Compared with the controls, Lats1/2 conditional knockout mutants exhibited dysregulated calcium handling and increased fibrosis in the SAN, indicating that Lats1/2 function through both cell-autonomous and non-cell-autonomous mechanisms. It is notable that the Lats1/2 conditional knockout phenotype was rescued by genetic deletion of Yap and Taz in the cardiac conduction system. These rescued mice had normal sinus rhythm and reduced fibrosis of the SAN, indicating that Lats1/2 function through Yap and Taz. Cleavage Under Targets and Tagmentation sequencing data showed that Yap potentially regulates genes critical for calcium homeostasis such as Ryr2 and genes encoding paracrine factors important in intercellular communication and fibrosis induction such as Tgfb1 and Tgfb3. Consistent with this, Lats1/2 conditional knockout mutants had decreased Ryr2 expression and increased Tgfb1 and Tgfb3 expression compared with control mice. CONCLUSIONS: We reveal, for the first time to our knowledge, that the canonical Hippo-Yap pathway plays a pivotal role in maintaining SAN homeostasis.

Unidirectional hybrid diode laser through the integration of a hook-shaped traveling-wave semiconductor optical amplifier and Taiji ring resonator.

We demonstrate a unidirectional ring diode laser based on hybrid integration of a hook-shaped traveling-wave semiconductor optical amplifier (SOA) and a Taiji ring resonator. The additional crossover bending waveguide inside the silicon nitride Taiji ring introduces a non-reciprocal loss in the laser cavity while the gain is provided by a multiple depth etched hook-shaped SOA. We present the detailed design flow for both active and passive components of the unidirectional hybrid diode laser. This work paves the way to use a hook-shaped SOA-based hybrid platform for various applications including optical sensing, all-optical switching, photonic memories, and topological optics.

Yap Promotes Noncanonical Wnt Signals From Cardiomyocytes for Heart Regeneration.

[Figure: see text].

Association between length of daily siesta and brachial-ankle pulse wave velocity (baPWV): a community-based cross sectional study in North China.

INTRODUCTION: To date, many studies have shown a link between siesta and cardiovascular events. Little is known regarding the connection between siesta and brachial-ankle pulse wave velocity (baPWV) levels, even though baPWV can determine the degree of atherosclerosis and vascular stiffness. Thus, we examined the relationship between siesta time and baPWV in a cross-sectional study. METHODS: Interviews, physical examinations, lab testing, and electron beam computed tomography were all part of the baseline evaluation for participants aged older than 35. Baseline data were compared for 3 different siesta habits: irregular or no siestas, daily short siestas (1 h or less), and daily long siestas (> 1 h). Utilizing logistic regression models and multivariate linear regression, the link between siesta time and baPWV was determined. RESULTS: Among all 6566 participants, the different siesta groups had a significant difference of the degrees of AS (P < 0.001). The same outcome was true for both males (P < 0.001) and females (P < 0.001). Numerous cardiovascular risk variables and markers of subclinical atherosclerosis were positively correlated with daily extended siestas. Results from the fully adjusted model showed that long siestas (> 60 min, OR = 1.18, 95%CI: 1.06-1.31, P = 0.002) were linked to a more severe level of the baPWV. For age or gender stratification, we found significant differences between non-siesta and > 60 min siesta groups. Multiple linear regression analysis revealed a positive connection between siesta duration and baPWV (ß = 0.197, P = 0.038). CONCLUSIONS: An elevated risk of atherosclerosis was shown to accompany prolonged siestas. These results need to be followed up on with prospective studies and additional lab work.

A Dielectric Polymer/Metal Oxide Nanowire Composite for Self-Adaptive Charge Release.

Electrostatic discharge is a fatal threat to a variety of electronic products. Here we report on a polymer nanocomposite composed of a dielectric polymer embedded with aligned core-shell structured nanowires for highly efficient distributed electrostatic discharge protection. The dielectric nanocomposite is capable of self-adaptive charge release, stemming from the nonlinear interface built in the Bi/Co oxide coated ZnO nanowires that leads to a "hand-in-hand" double-Schottky barrier. The ultralow filler concentration (e.g., 0.5 vol %) endows the nanocomposite with low permittivity (close to the pristine polymer matrix) and high optical transmittance (75%) in the visible light wavelength range, which is desirable in packaging materials and display coatings for portable electronics. The effectiveness for electrostatic discharge protection was validated with an organic light-emitting diode screen protected by the nanocomposite that survived multiple rounds of electrostatic discharge.

Watt-level beam combined diode laser systems in a chip-scale hybrid photonic platform.

Scaling up the power of on-chip diode lasers is of great importance for many emerging applications, such as integrated nonlinear optics, remote sensing, free space communication, infrared countermeasure, and light detection and ranging (LIDAR). In this manuscript, we introduce and demonstrate photonic integrated circuits (PIC) based beam combining methods to create power scalable, integrated direct diode laser systems. Traditional laser beam combining, including coherent beam combining (CBC) and wavelength beam combining (WBC), usually requires free space or fiber optical components, leading to bulky and complex systems. Instead, PIC based beam combining methods can greatly reduce the cost, size, weight, and power consumption (CSWaP) of next generation direct diode laser systems. We experimentally demonstrate four channel chip-scale CBC and WBC with watt-level on-chip power by using III/V-Si3N4 hybrid integration. Our results show that PIC based beam combining is very suitable for power scaling in a chip-scale platform.

Tricyano-Methylene-Pyridine Based High-Performance Aggregation-Induced Emission Photosensitizer for Imaging and Photodynamic Therapy.

Photosensitizers equipped with high reactive oxygen species (ROS) generation capability and bright emission are essential for accurate tumor imaging and precise photodynamic therapy (PDT). However, traditional aggregation-caused quenching (ACQ) photosensitizers cannot simultaneously produce desirable ROS and bright fluorescence, resulting in poor image-guided therapy effect. Herein, we report an aggregation-induced emission (AIE) photosensitizer TCM-Ph with a strong donor-π-acceptor (D-π-A) structure, which greatly separates the HOMO-LUMO distribution and reduces the ΔEST, thereby increasing the number of triplet excitons and producing more ROS. The AIE photosensitizer TCM-Ph has bright near-infrared emission, as well as a higher ROS generation capacity than the commercial photosensitizers Bengal Rose (RB) and Chlorine e6 (Ce6), and can effectively eliminate cancer cells under image guidance. Therefore, the AIE photosensitizer TCM-Ph has great potential to replace the commercial photosensitizers.

Genome-wide association studies of leaf angle in maize.

Compact plant-type with small leaf angle has increased canopy light interception, which is conducive to the photosynthesis of the population and higher population yield at high density planting in maize. In this study, a panel of 285 diverse maize inbred lines genotyped with 56,000 SNPs was used to investigate the genetic basis of leaf angle across 3 consecutive years using a genome-wide association study (GWAS). The leaf angle showed broad phenotypic variation and high heritability across different years. Population structure analysis subdivided the panel into four subgroups that correspond to the four major empirical germplasm origins in China, i.e., Tangsipingtou, Reid, Lancaster and P. When tested with the optimal GWAS model, we found that the Q + K model was the best in reducing false positive. In total, 96 SNPs accounting for 5.54-10.44% of phenotypic variation were significantly (P < 0.0001) associated with leaf angle across three years. According to the linkage disequilibrium decay distance, 96 SNPs were binned into 43 QTLs for leaf angle. Seven major QTLs with R2 > 8% stably detected in at least 2 years, and BLUP values were clustered in four genomic regions (bins 2.01, 2.07, 5.06, and 10.04). Seven important candidate genes, Zm00001d001961, Zm00001d006348, Zm00001d006463, Zm00001d017618, Zm00001d024919, Zm00001d025018, and Zm00001d025033 were predicted for the seven stable major QTLs, respectively. The markers identified in this study can be used for molecular breeding for leaf angle, and the candidate genes would contribute to further understanding of the genetic basis of leaf angle. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-021-01241-0.

Development of hydrophilic magnetic molecularly imprinted polymers for the dispersive solid-phase extraction of sulfonamides from animal-derived samples before HPLC detection.

Highly hydrophilic magnetic molecularly imprinted polymers were prepared through a surface imprinting technique for dispersive solid-phase extraction coupled with high-performance liquid chromatography to detect trace levels of ten sulfonamides in animal-derived samples. The obtained imprinted polymers were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, thermogravimetric analysis, and adsorption experiments, which exhibited excellent specific adsorption for template sulfamethazine in aqueous solution. Moreover, the adsorption process could be completed within 25 min. Under the optimum conditions, the method exhibits good linear performance in the range of 5-to 10 mg/L, limits of detection ranging from 0.57 to 1.50 µg/L, and good recoveries of 85.09-110.93% in the spiked samples (chicken, cow milk, and goat milk). The proposed detection method not only avoids the use of organic solvents but also simplifies the pretreatment procedure via excellent magnetic properties. Furthermore, the method shows great potential for the rapid detection of drug residues.

Development and Validity Testing of a Type 1 Diabetes Resource for 10-19-Years Old Adolescents in China.

PURPOSE: This phase 1 study aimed to develop a Type 1 diabetes (T1DM) educational resource for adolescents (10-19 years) in China, and to test its validity for improving diabetes knowledge, self-efficacy and adherence. DESIGN AND METHODS: This phase 1 study was carried out in four phases: preliminary development of the Type 1 diabetes educational resource in English based on the health belief model (HBM) and the International Society for Pediatric and Adolescent Diabetes (ISPAD) Consensus Guidelines 2014; translation of the resource into Chinese; readability testing of the resource; and face and content validity testing for scientific and clinical accuracy. RESULTS: Type 1 diabetes educational resource with 17 "topics" in four sections was developed for 10-19-years old adolescents in China. Cartoon story, images, stories, questions & answers, and quizzes were used to attract adolescents' interest. Furthermore, the educational resource was confirmed to be understandable among adolescents, clinically accurate, and consistent with its purpose. CONCLUSIONS: This resource with good readability and content validity is a potential facilitator for diabetes education and self-management in adolescents with T1DM in China. PRACTICAL IMPLICATIONS: This type 1 diabetes resource can be used to educate adolescents with T1DM as a validated resource after further testing in an effectiveness study.

Effectiveness of Internet and Phone-Based Interventions on Diabetes Management of Children and Adolescents With Type 1 Diabetes: A Systematic Review.

AIMS: To synthesize evidence about the impact of Internet and phone-based diabetes education and management on metabolic control, self-management behavior changes, and psychological effects among children and adolescents with type 1 diabetes mellitus (T1DM). BACKGROUND: Internet and mobile technologies were commonly used to improve diabetes management among children and adolescents with type 1 diabetes mellitus. The effectiveness of new technology-based diabetes education and management has previously not been synthesized. METHODS: PubMed, EBSCO, Cochrane Library, Web of Science, Joanna Briggs Institute Library, and the Chinese databases CNKI and Wanfang were searched from 1989 to March 2020. Two reviewers independently selected randomized controlled trials (RCTs), in English and Chinese, which compared an intervention group of new technology-based diabetes education and management with a control group of usual care. The primary outcomes were metabolic control, such as glycated hemoglobin (HbA1c), and secondary outcomes consisted of behavior changes and psychological effects, such as self-efficacy and quality of life. RESULTS: A total of 23 RCTs with 1,824 participants met the inclusion criteria. The meta-analysis showed that phone calls could significantly reduce HbA1c (MD = -.17; 95% CI [-.33, -.01]; I2  = 0%) in children and adolescents with T1DM. New technology-based diabetes education and management could significantly improve self-efficacy (SMD = 0.37; 95% CI [.07, .67]; I2  = 0%). No benefits on behavior changes and quality of life were identified. LINKING EVIDENCE TO ACTION: New technology-based diabetes education has potential benefits for children and adolescents with T1DM, such as improving metabolic control through phone calls and increasing their self-efficacy of diabetes management. Well-designed RCTs with larger sample sizes and longer intervention duration should be conducted, especially in developing countries.

Study of the Expression Transition of Cardiac Myosin Using Polarization-Dependent SHG Microscopy.

Detection of the transition between the two myosin isoforms α- and ß-myosin in living cardiomyocytes is essential for understanding cardiac physiology and pathology. In this study, the differences in symmetry of polarization spectra obtained from α- and ß-myosin in various mammalian ventricles and propylthiouracil-treated rats are explored through polarization-dependent second harmonic generation microscopy. Here, we report for the, to our knowledge, first time that α- and ß-myosin, as protein crystals, possess different symmetries: the former has C6 symmetry, and the latter has C3v. A single-sarcomere line scan further demonstrated that the differences in polarization-spectrum symmetry between α- and ß-myosin came from their head regions: the head and neck domains of α- and ß-myosin account for the differences in symmetry. In addition, the dynamic transition of the polarization spectrum from C6 to C3v line profile was observed in a cell culture in which norepinephrine induced an α- to ß-myosin transition.

Imprinting aberrations of SNRPN, ZAC1 and INPP5F genes involved in the pathogenesis of congenital heart disease with extracardiac malformations.

Congenital heart disease (CHD) with extracardiac malformations (EM) is the most common multiple malformation, resulting from the interaction between genetic abnormalities and environmental factors. Most studies have attributed the causes of CHD with EM to chromosomal abnormalities. However, multi-system dysplasia is usually caused by both genetic mutations and epigenetic dysregulation. The epigenetic mechanisms underlying the pathogenesis of CHD with EM remain unclear. In this study, we investigated the mechanisms of imprinting alterations, including those of the Small nuclear ribonucleoprotein polypeptide N (SNRPN), PLAG1 like zinc finger 1 (ZAC1) and inositol polyphosphate-5-phosphatase F (INPP5F) genes, in the pathogenesis of CHD with EM. The methylation levels of SNRPN, ZAC1, and INPP5F genes were analysed by the MassARRAY platform in 24 children with CHD with EM and 20 healthy controls. The expression levels of these genes were detected by real-time polymerase chain reaction (PCR). The correlation between methylation regulation and gene expression was confirmed using 5-azacytidine (5-Aza) treated cells. The methylation levels of SNRPN and ZAC1 genes were significantly increased in CHD with EM, while that of INPP5F was decreased. The methylation alterations of these genes were negatively correlated with expression. Risk analysis showed that abnormal hypermethylation of SNRPN and ZAC1 resulted in 5.545 and 7.438 times higher risks of CHD with EM, respectively, and the abnormal hypomethylation of INPP5F was 8.38 times higher than that of the control group. We concluded that abnormally high methylation levels of SNRPN and ZAC1 and decreased levels of INPP5F imply an increased risk of CHD with EM by altering their gene functions. This study provides evidence of imprinted regulation in the pathogenesis of multiple malformations.