Novel Insights into Ethanol-Soluble Oyster Peptide-Zinc-Chelating Agents: Structural Characterization, Chelation Mechanism, and Potential Protection on MEHP-Induced Leydig Cells.

Numerous studies have reported that mono-(2-ethylhexyl) phthalate (MEHP) (bioactive metabolite of Di(2-ethylhexyl) phthalate) has inhibitory effects on Leydig cells. This study aims to prepare an oyster peptide-zinc complex (PEP-Zn) to alleviate MEHP-induced damage in Leydig cells. Zinc-binding peptides were obtained through the following processes: zinc-immobilized affinity chromatography (IMAC-Zn2+), liquid chromatography-mass spectrometry technology (LC-MS/MS) analysis, molecular docking, molecular dynamic simulation, and structural characterization. Then, the Zn-binding peptide (PEP) named Glu-His-Ala-Pro-Asn-His-Asp-Asn-Pro-Gly-Asp-Leu (EHAPNHDNPGDL) was identified. EHAPNHDNPGDL showed the highest zinc-chelating ability of 49.74 ± 1.44%, which was higher than that of the ethanol-soluble oyster peptides (27.50 ± 0.41%). In the EHAPNHDNPGDL-Zn complex, Asn-5, Asp-7, Asn-8, His-2, and Asp-11 played an important role in binding to the zinc ion. Additionally, EHAPNHDNPGDL-Zn was found to increase the cell viability, significantly increase the relative activity of antioxidant enzymes and testosterone content, and decrease malondialdehyde (MDA) content in MEHP-induced TM3 cells. The results also indicated that EHAPNHDNPGDL-Zn could alleviate MEHP-induced apoptosis by reducing the protein level of p53, p21, and Bax, and increasing the protein level of Bcl-2. These results indicate that the zinc-chelating peptides derived from oyster peptides could be used as a potential dietary zinc supplement.

High-sensitivity miniaturized spectrometers using photonic crystal slab filters.

Miniaturized spectrometers have emerged as pivotal tools in numerous scientific and industrial applications, offering advantages such as portability, cost-effectiveness, and the capability for onsite analysis. Despite these significant benefits, miniaturized spectrometers face critical challenges, particularly in sensitivity. Reduced dimensions often lead to compromises in optical path length and component quality, which can diminish detection limits and limit their applications in areas such as low-light-level measurements. Here we developed a compact spectrometer that integrates an array of photonic crystal slab filters with band-stop spectral transmission characteristics into an image sensor. Compared to traditional gratings or bandpass filter strategies, where each detector can only read light of a single wavelength component, our band-stop strategy allows each detector to read the light of all wavelengths except the band-stop wavelength. This maximizes energy extraction from incident signals, significantly improving the sensitivity of the spectrometer. Spectral reconstruction is achieved mathematically using pre-calibrated band-stop responses combined with a single coded image. Our spectrometer delivers a spectral resolution of 1.9 nm and demonstrates sensitivity more than ten times greater than that of conventional grating spectrometers during fluorescence spectroscopy of Ascaris lumbricoides. The design is fully compatible with complementary metal-oxide-semiconductor (CMOS) technology, allowing for mass production at low costs and thus promising broad deployment in sensitive applications.

Nonreciprocal Acoustic Devices with Asymmetric Peierls Phases.

Nonreciprocity in acoustics is of paramount importance in many practical applications and has been experimentally realized using nonlinear media, moving fluids, or time modulation, which regrettably suffer from large volumes and high-power consumption, difficulty in integration, and inevitable vibrations or phase noise. In modern Hamiltonian theory, the violation of system's reciprocity can be achieved via asymmetric Peierls phases, which typically involves with non-Hermiticity or time-reversal symmetry breaking. Here, we propose a framework for designing nonreciprocal acoustic devices based on the asymmetric Peierls phases that can be fully controlled via active acoustic components. The fully controlled Peierls phases enable various high-performance acoustic devices, including non-Hermitian extensions of isolators, gyrators, and circulators, which are otherwise impossible in previous approaches that are bound by Hermiticity or passivity. We reveal that the transmission phases in isolators are equivalent to the Peierls phase plus a constant. The nonreciprocal phase delay in gyrators and the unirotational transmission behavior in circulators result from the gauge-invariant Aharonov-Bohm phases determined by Peierls phases. Our work not only uncovers multiple intriguing physics related to Peierls phases but also provides a general approach to compact, integratable, nonreciprocal acoustic devices.

Effect of resveratrol on spermatogenesis in breeding boars and the proteomic analysis for testes.

Effect of resveratrol (RSV) on spermatogenesis and the mechanism of resveratrol in promoting spermatogenesis of breeding boars was explored by feeding sexually mature Duroc boars with normal diet and 20 mg/kg resveratrol diet for 14 days to the control group and experimental group, respectively. Semen volume, sperm density, motility, viability and abnormality rate were analyzed on day 0, 7, and 14. Blood samples were collected, and levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) in serum were analyzed. On day 14, the testis tissue was collected for antioxidant and proteomics analysis etc. The semen volume, sperm density, motility, and viability of the experimental group and the contents of serum FSH, LH, T and plasma SOD activity were significantly higher than those in the control group. However, the serum IL-6, TNF-α and plasma MDA were remarkably lower in experimental group. The above results showed that resveratrol can simulate spermatogenesis in breeding boars. Proteomic results demonstrated that three differentially expressed proteins (DEPs) were up-regulated and 12 DEPs were down-regulated; ODF1, calmodulin, Cabs1, and Hp were involved in spermatogenesis; and the main enriched metabolic pathway is steroid hormone synthesis pathway. Therefore, the improvement in sperm quality by resveratrol may be achieved by regulating the changes in outer dense fiber 1, calmodulin, spermatid specific 1, and haptoglobin expression and steroid synthesis pathway.

Immune escape of avian oncogenic Marek's disease herpesvirus and antagonistic host immune responses.

Marek's disease virus (MDV) is a highly pathogenic and oncogenic alpha herpesvirus that causes Marek's disease (MD), which is one of the most important immunosuppressive and rapid-onset neoplastic diseases in poultry. The onset of MD lymphomas and other clinical diseases can be efficiently prevented by vaccination; these vaccines are heralded as the first demonstration of a successful vaccination strategy against a cancer. However, the persistent evolution of epidemic MDV strains towards greater virulence has recently resulted in frequent outbreaks of MD in vaccinated chicken flocks worldwide. Herein, we provide an overall review focusing on the discovery and identification of the strategies by which MDV evades host immunity and attacks the immune system. We have also highlighted the decrease in the immune efficacy of current MD vaccines. The prospects, strategies and new techniques for the development of efficient MD vaccines, together with the possibilities of antiviral therapy in MD, are also discussed.

Compound probiotics regulate the NRF2 antioxidant pathway to inhibit aflatoxin B1-induced autophagy in mouse Sertoli TM4 cells.

This study investigated the effects of compound probiotics (CP) on AFB1-induced cytotoxicity in Sertoli TM4 cells. The L9 (3 × 3) orthogonal test was conducted to determine the optimal CP required for high AFB1 degradation in the artificial gastrointestinal fluid in vitro. The maximal AFB1 degradation rate was 40.55 % (P < 0.05) when the final viable count was 1.0 × 105 CFU/mL for Bacillus subtilis, Lactobacillus casein, and Saccharomyces cerevisiae. The effects of CP and the CP supernatant (CPS) on TM4 cell viability were evaluated to achieve the optimal protective conditions. When CPS4 (corresponding to CP viable counts of 1.0 × 104 CFU/mL) was added to the TM4 cells for 24 h, the cell viability reached 108.86 % (P < 0.05). AFB1 reduced TM4 cell viability in a concentration- and time-dependent manner at an AFB1 concentration ranging from 0 to 1.5 µM after 48-h AFB1 exposure. The optimal AFB1 concentration/times for low- and high damage models were 0.5 and 1.25 µM both for 24 h, which decreased viability to 76.04 % and 65.35 %, respectively. however, CPS4 added to low- and high-damage models increased the cell viability to 97.43 % and 75.12 %, respectively (P < 0.05). Transcriptome sequencing was performed based on the following designed groups: the control, 0.5 µM AFB1, 1.25 µM AFB1, CPS4, and CPS4+0.5 µM AFB1. The Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was further performed to identify significantly enriched signaling pathways, which were subsequently verified. It was shown that AFB1 induced apoptosis by blocking the PI3K-AKT-mTOR pathway and upregulating autophagy proteins such as LC3B, Beclin1, and ATG5 while inhibiting autophagic flux. CPS4 promoted AFB1 degradation, activated the p62-NRF2 antioxidant, and inhibited ROS/TRPML1 pathways, thereby reducing ROS production and inflammation and ultimately alleviating AFB1-induced autophagy and apoptosis. These findings supports the potential of probiotics to protect the male reproductive system from toxin damage.

Multiple Brillouin Zone Winding of Topological Chiral Edge States for Slow Light Applications.

Photonic Chern insulators are known for their topological chiral edge states (CESs), whose absolute existence is determined by the bulk band topology, but concrete dispersion can be engineered to exhibit various properties. For example, the previous theory suggested that the edge dispersion can wind many times around the Brillouin zone to slow down light, which can potentially overcome fundamental limitations in conventional slow-light devices: narrow bandwidth and keen sensitivity to fabrication imperfection. Here, we report the first experimental demonstration of this idea, achieved by coupling CESs with resonance-induced nearly flat bands. We show that the backscattering-immune hybridized CESs are significantly slowed down over a relatively broad bandwidth. Our work thus paves an avenue to broadband topological slow-light devices.

Real higher-order Weyl photonic crystal.

Higher-order Weyl semimetals are a family of recently predicted topological phases simultaneously showcasing unconventional properties derived from Weyl points, such as chiral anomaly, and multidimensional topological phenomena originating from higher-order topology. The higher-order Weyl semimetal phases, with their higher-order topology arising from quantized dipole or quadrupole bulk polarizations, have been demonstrated in phononics and circuits. Here, we experimentally discover a class of higher-order Weyl semimetal phase in a three-dimensional photonic crystal (PhC), exhibiting the concurrence of the surface and hinge Fermi arcs from the nonzero Chern number and the nontrivial generalized real Chern number, respectively, coined a real higher-order Weyl PhC. Notably, the projected two-dimensional subsystem with kz = 0 is a real Chern insulator, belonging to the Stiefel-Whitney class with real Bloch wavefunctions, which is distinguished fundamentally from the Chern class with complex Bloch wavefunctions. Our work offers an ideal photonic platform for exploring potential applications and material properties associated with the higher-order Weyl points and the Stiefel-Whitney class of topological phases.

Physiological and transcriptional studies reveal Cr(VI) reduction mechanisms in the exoelectrogen Cellulomonas fimi Clb-11.

A facultative exoelectrogen, Cellulomonas fimi strain Clb-11, was isolated from polluted river water. This strain could generate electricity in microbial fuel cells (MFCs) with carboxymethyl cellulose (CMC) as the carbon source, and the maximum output power density was 12.17 ± 2.74 mW·m-2. In addition, Clb-11 could secrete extracellular chromate reductase or extracellular electron mediator to reduce Cr(VI) to Cr(III). When the Cr(VI) concentration was less than 0.5 mM in Luria-Bertani (LB) medium, Cr(VI) could be completely reduced by Clb-11. However, the Clb-11 cells swelled significantly in the presence of Cr(VI). We employed transcriptome sequencing analysis to identify genes involved in different Cr(VI) stress responses in Clb-11. The results indicate that 99 genes were continuously upregulated while 78 genes were continuously downregulated as the Cr(VI) concentration increased in the growth medium. These genes were mostly associated with DNA replication and repair, biosynthesis of secondary metabolites, ABC transporters, amino sugar and nucleotide sugar metabolism, and carbon metabolism. The swelling of Clb-11 cells might have been related to the upregulation of the genes atoB, INO1, dhaM, dhal, dhak, and bccA, which encode acetyl-CoA C-acetyltransferase, myo-inositol-1-phosphate synthase, phosphoenolpyruvate-glycerone phosphotransferase, and acetyl-CoA/propionyl-CoA carboxylase, respectively. Interestingly, the genes cydA and cydB related to electron transport were continuously downregulated as the Cr(VI) concentration increased. Our results provide clues to the molecular mechanism of Cr(VI) reduction by microorganisms in MFCs systems.

Antitumor activity and mechanisms of dual EGFR/DNA-targeting strategy for the treatment of lung cancer with EGFRL858R/T790M mutation.

Dual- or multi-targeted EGFR inhibitors as single drugs can overcome EGFR inhibitor resistance and circumvent many disadvantages of combination therapy. In this work, fifteen 4-anilinoquinazoline derivatives bearing nitrogen mustard or hemi mustard moieties were designed and synthesized as dual EGFR-DNA targeting anticancer agents. Structures of target molecules were confirmed by 1H NMR, 13C NMR and HR-MS, and evaluated for their in vitro anti-proliferative activities using MTT assay. Compound 6g emerged as the most potent derivative against mutant-type H1975 cells with IC50 value of 1.45 µM, which exhibited 4-fold stronger potency than Chl/Gef (equimolar combination of chlorambucil and gefitinib). Kinase inhibition studies indicated that 6g showed excellent inhibitory effect on EGFRL858R/T790M enzyme, which was 8.6 times more effective than gefitinib. Mechanistic studies indicated that 6g induced apoptosis of H1975 cells in a dose-dependent manner and caused DNA damage. Importantly, 6g could significantly inhibit the expression of p-EGFR and its downstream p-AKT and p-ERK in H1975 cells. Molecular docking was also performed to gain insights into the ligand-binding interactions of 6g inside EGFRWT and EGFRL858R/T790M binding sites. Moreover, 6g efficiently inhibited tumor growth in the H1975 xenograft model without side effects.

All-Organic PTFE Coated PVDF Composite Film Exhibiting Low Conduction Loss and High Breakdown Strength for Energy Storage Applications.

Plastic film capacitors are widely used in pulse and energy storage applications because of their high breakdown strength, high power density, long lifetime, and excellent self-healing properties. Nowadays, the energy storage density of commercial biaxially oriented polypropylene (BOPP) is limited by its low dielectric constant (~2.2). Poly(vinylidene fluoride) (PVDF) exhibits a relatively high dielectric constant and breakdown strength, making it a candidate material for electrostatic capacitors. However, PVDF presents significant losses, generating a lot of waste heat. In this paper, under the guidance of the leakage mechanism, a high-insulation polytetrafluoroethylene (PTFE) coating is sprayed on the surface of a PVDF film. The potential barrier at the electrode-dielectric interface is raised by simply spraying PTFE and reducing the leakage current, and then the energy storage density is increased. After introducing the PTFE insulation coating, the high-field leakage current in the PVDF film shows an order of magnitude reduction. Moreover, the composite film presents a 30.8% improvement in breakdown strength, and a 70% enhancement in energy storage density is simultaneously achieved. The all-organic structure design provides a new idea for the application of PVDF in electrostatic capacitors.

Research on the application of the blended BOPPPS based on an online and offline mixed teaching model in the course of fermentation engineering in applied universities.

This study aimed to investigate the teaching effect of the blended BOPPPS based on an online and offline mixed teaching model ("B + BOPPPS") in the course of fermentation engineering in applied universities. The participants were 142 undergraduates majoring from the course of fermentation engineering in Food Science and Engineering in 2019 and 2020 in Huanghuai University, Zhumadian city, Henan province, China. The students in the control group (68 students) were taught in 2019, and the students in the experimental group (74 students) were taught in 2020. The traditional teaching method and "B + BOPPPS" were implemented, respectively. The teaching effect was evaluated using the questionnaire survey of course satisfaction and theoretical knowledge test. The results showed that the scores of the theoretical knowledge test in the experimental group adopting "B + BOPPPS" were significantly higher than those in the control group, and the difference was statistically significant (p < 0.01). The students had a good evaluation of the "B + BOPPPS" in many aspects, which included achieving learning goals, providing in-depth understanding of knowledge points, stimulating interest in learning, training in the ability to analyze and think about problems, and so on. The results suggested that "B + BOPPPS" could stimulate students' interest in learning and improve their subjective initiative. They could also improve students' ability to master and apply knowledge, which was conducive to improving the theoretical teaching quality of the course of fermentation engineering.

Predictive plasma biomarker for gestational diabetes: A case-control study in China.

OBJECTIVE: This study aims to find new plasma biomarkers in early pregnancy. DESIGN: The original study enrolled 1219 pregnant women. We investigated protein expression profiles of placental tissues from women with GDM (n = 89) and normal glucose tolerance (NGT) (n = 83). Maternal plasma samples between two groups in early and middle pregnancy were used for validation of candidate biomarkers. METHODS: Differentially expressed proteins (DEPs) were identified by label-free quantitative proteomics from human placenta samples between two groups. Several DEPs were validated in plasma by Luminex assays. An automatic biochemical analyzer was used to detect blood lipid indexes. The associations of GAL-3BP with biochemical indicators were demonstrated by Pearson's correlation analysis. Binary logistic regression was used to model potential predictive indicators in early pregnancy of GDM. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic accuracy of the predictive model and the value of GAL-3BP. RESULTS: 123 DEPs were found in placenta involved in ribosomal function, pancreatic secretion, oxidative phosphorylation, and inflammatory signaling pathway. Plasma GAL-3BP are significantly higher in women with GDM than NGT in the first (p = 0.008) and second (p = 0.026) trimester, but C9 and VWF have no difference. The predictive value of GAL-3BP in the first trimester of pregnancy (AUC 0.64) is better than that in the second trimester (AUC 0.61), and combined predictive model of TG and GAL-3BP at early pregnancy has greater predictive and diagnostic value for GDM (AUC 0.69) than individual GAL-3BP (AUC 0.64). CONCLUSIONS: Plasma TG and GAL-3BP has good predictive and diagnostic value at early pregnancy, suggesting that these two indicators may be used as biomarkers for early prediction and diagnosis of GDM. SIGNIFICANCE: The advantage of this study is that circulating TG and GAL-3BP might differentiate the progress of women with GDM and normal glucose tolerance (NGT) at the early stage of pregnancy. It is the first study to consider the role of GAL-3BP as an early predictive biomarker in the development of GDM during the whole pregnancy. Another advantage is that volunteers in this study were recruited from two provinces in China to eliminate the impacts of environmental confounders. The similar changes of blood glucose/lipid indicators for women with GDM and NGT in both regions was found in the first and second trimester of pregnancy, which added to the reliability of analytical results.

Oyster Peptide-Zinc Complex Ameliorates Di-(2-ethylhexyl) Phthalate-Induced Testis Injury in Male Mice and Improving Gut Microbiota.

Di-(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer, which can cause damage to male reproductive organs, especially the atrophy of the testis. Meanwhile, DEHP can also lead to a decrease in testicular zinc content, but the role of zinc remains unclear. This study aims to prepare oyster peptide-zinc complex (OPZC) to alleviate DEHP-induced reproductive damage in mice. OPZC was successfully obtained through electron microscopy, X-ray diffraction, and thermogravimetric analysis, with stable structure and high water-solubility. Low dose oyster peptide-zinc complex (OPZCL) significantly reduced the reproductive damage caused by DEHP in mice. Further research had shown that OPZCL restored the content of serum hormones and the activity of oxidative stress kinases to normal, while also normalizing testicular zinc and selenium levels. In addition, it also recovered the disorder of gut microbiota, reduced the proportion of Bacteroides, increased the abundance of Ligilactobacillus, and restored the proportion of Acidobacteriota, Chloroflexi, and Proteobacteria. Therefore, OPZCL can relieve the reproductive damage caused by DEHP in mice by restoring testicular zinc homeostasis and the composition of intestinal microbiota, indicating that OPZCL has a potential protective effect on male reproductive health.

Discovery of a maximally charged Weyl point.

The hypothetical Weyl particles in high-energy physics have been discovered in three-dimensional crystals as collective quasiparticle excitations near two-fold degenerate Weyl points. Such momentum-space Weyl particles carry quantised chiral charges, which can be measured by counting the number of Fermi arcs emanating from the corresponding Weyl points. It is known that merging unit-charged Weyl particles can create new ones with more charges. However, only very recently has it been realised that there is an upper limit - the maximal charge number that a two-fold Weyl point can host is four - achievable only in crystals without spin-orbit coupling. Here, we report the experimental realisation of such a maximally charged Weyl point in a three-dimensional photonic crystal. The four charges support quadruple-helicoid Fermi arcs, forming an unprecedented topology of two non-contractible loops in the surface Brillouin zone. The helicoid Fermi arcs also exhibit the long-pursued type-II van Hove singularities that can reside at arbitrary momenta. This discovery reveals a type of maximally charged Weyl particles beyond conventional topological particles in crystals.

Rhamnose-Containing Compounds: Biosynthesis and Applications.

Rhamnose-associated molecules are attracting attention because they are present in bacteria but not mammals, making them potentially useful as antibacterial agents. Additionally, they are also valuable for tumor immunotherapy. Thus, studies on the functions and biosynthetic pathways of rhamnose-containing compounds are in progress. In this paper, studies on the biosynthetic pathways of three rhamnose donors, i.e., deoxythymidinediphosphate-L-rhamnose (dTDP-Rha), uridine diphosphate-rhamnose (UDP-Rha), and guanosine diphosphate rhamnose (GDP-Rha), are firstly reviewed, together with the functions and crystal structures of those associated enzymes. Among them, dTDP-Rha is the most common rhamnose donor, and four enzymes, including glucose-1-phosphate thymidylyltransferase RmlA, dTDP-Glc-4,6-dehydratase RmlB, dTDP-4-keto-6-deoxy-Glc-3,5-epimerase RmlC, and dTDP-4-keto-Rha reductase RmlD, are involved in its biosynthesis. Secondly, several known rhamnosyltransferases from Geobacillus stearothermophilus, Saccharopolyspora spinosa, Mycobacterium tuberculosis, Pseudomonas aeruginosa, and Streptococcus pneumoniae are discussed. In these studies, however, the functions of rhamnosyltransferases were verified by employing gene knockout and radiolabeled substrates, which were almost impossible to obtain and characterize the products of enzymatic reactions. Finally, the application of rhamnose-containing compounds in disease treatments is briefly described.

Optically reconfigurable higher-order valley photonic crystals based on enhanced Kerr effect.

The reconfigurable higher-order topological states are realized in valley photonic crystals with enhanced optical Kerr nonlinearity. The inversion symmetry of the designed valley photonic crystal is broken due to the difference in optical responses between adjacent elements rather than their geometry structures. Therefore, by constructing photonic crystals with distinct topological phases, valley-dependent topological states can be realized, and their reconfigurability is demonstrated based on the Kerr effect. The investigated higher-order topological photonic crystals exhibit great robustness against the structural defects and inferior quality of pump introduced around the corner. Our work provides a new, to the best of our knowledge, platform for studying optical field manipulation and optical devices fabrication in the context of nonlinear higher-order topology.

Placental DNA methylation changes in gestational diabetes mellitus.

In this study, we investigated the association between altered methylation in the maternal placenta and hyperglycaemia and explored the epigenetic mechanisms underlying gestational diabetes mellitus (GDM). Reduced representation bisulphite sequencing (RRBS) and RNA sequencing (RNA-seq) were performed on placental tissues obtained from women with GDM and healthy controls. Further, pyrosequencing, correlation analyses, and linear regression analyses were performed to valuate relationships between aberrantly methylated-differentially expressed genes and clinical parameters. The EMBOSS and JASPAR databases were used for a computational analysis of CpG islands and transcription factor-binding sites in the TRIM67 promoter region. A CpG island with a length of 264 bp in the placental TRIM67 promoter region in the GDM group exhibited significant hypermethylation at four CpG sites. The hypermethylation of the TRIM67 promoter region in the maternal placenta showed a significant, positive correlation with the 1 h and 2 h oral glucose tolerance test (OGTT) values and a negative correlation with lipoprotein(a). Placental DNA methylation levels in the TRIM67 promoter region were markedly elevated in GDM and were associated with blood glucose and lipid levels during healthy pregnancy.

Integrated single-cell RNA-seq and DNA methylation reveal the effects of air pollution in patients with recurrent spontaneous abortion.

BACKGROUND: Maternal air pollutants exposure is associated with a number of adverse pregnancy outcomes, including recurrent spontaneous abortion (RSA). However, the underlying mechanisms are still unknown. The present study aimed to understand the mechanism of RSA and its relationship with air pollution exposure. We compared data of decidual tissue from individuals with induced abortions and those with RSA by bulk RNA sequencing (RNA-seq), reduced representation bisulfite sequencing (RRBS), and single-cell RNA sequencing (scRNA-seq). Differentially expressed genes (DEGs) were verified using RT-qPCR and pyrosequencing. A logistic regression model was used to investigate the association between air pollutants exposure and RSA. RESULTS: We identified 98 DEGs with aberrant methylation by overlapping the RRBS and RNA-seq data. Nineteen immune cell subsets were identified. Compared with normal controls, NK cells and macrophages accounted for different proportions in the decidua of patients with RSA. We observed that the methylation and expression of IGF2BP1 were different between patients with RSA and controls. Furthermore, we observed significant positive associations between maternal air pollutants exposure during the year prior to pregnancy and in early pregnancy and the risk of RSA. Mediation analyses suggested that 24.5% of the effects of air pollution on the risk of RSA were mediated through IGF2BP1 methylation. CONCLUSION: These findings reveal a comprehensive cellular and molecular mechanism of RSA and suggest that air pollution might cause pregnancy loss by affecting the methylation level of the IGF2BP1 promoter.

Establishment of DNA Methylation Profile Associated with TCM Syndrome in Endometriosis.

Objectives: To screen the potential epigenetic biomarkers associated with endometriosis (EMS) and traditional Chinese medicine (TCM) syndrome EMS types. Methods: A cohort of 99 participants comprising 42 EMS patients with cold coagulation blood stasis (CCBS) syndrome, 35 EMS patients with Qi stagnation blood stasis (QSBS) syndrome, and 22 women of childbearing age without EMS were recruited. Reduced representation bisulfite sequencing (RRBS) was used to establish the differential DNA methylation profiles in human peripheral blood samples obtained from four non-EMS and four EMS patients with CCBS or QSBS syndrome, respectively. Differentially expressed genes (DEGs) were verified in 18 non-EMS, 38 CCBS-EMS, and 31 QSBS-EMS using pyrosequencing. Results: Methylation sites of 123942, 127229, and 115961 were found in peripheral blood DNA of non-EMS, CCBS-EMS, and QSBS-EMS patients, respectively. GO and KEGG analyses showed that the pathological process of EMS may be closely related to the nervous system development, cell junctions, GABA-gated chloride ion channel activity, nicotine addiction, Hippo signaling pathway, mRNA surveillance pathway, and Wnt signaling pathway. The methylation level at CpG site within HDAC6 gene in QSBS-EMS patients was significantly different from that in control women. Conclusions: The changes in DNA methylation in peripheral blood samples may be associated with EMS and TCM syndrome EMS types. The methylation level of HDAC6 gene may be used to distinguish QSBS-EMS patients from women without EMS.