Deletion of BTB and CNC homology 1 protects against Staphylococcus aureus-induced acute lung injury.

BTB and CNC homology 1 (BACH1) plays a crucial role in the pathogenesis of acute lung injury (ALI) caused by gram-negative bacteria. However, its exact mechanisms and roles in Staphylococcus aureus (SA)-induced ALI, a gram-positive bacterial infection, remain incompletely understood. In this study, we generated a BACH1-knockout mouse model (BACH1-/-) to investigate the role of BACH1 and its underlying mechanisms in regulating the development of sepsis-induced acute lung injury (ALI). Elevated levels of BACH1 were observed in both serum samples from septic patients and mouse models. Deletion of BACH1 alleviated ALI symptoms induced by sepsis. In bone marrow-derived macrophages, BACH1 deletion or knockdown suppressed NF-κB p65 phosphorylation and the induction of pro-inflammatory cytokines. Mechanistic studies demonstrated that BACH1 downregulated tumor necrosis factor-alpha-induced protein 3 (TNFAIP3) mRNA expression by binding to its promoter region. These findings uncover inhibiting BACH1 may be a promising therapeutic strategy for treating gram-positive bacteria-induced ALI.

Self-assembled ordered AuNRs-modified electrodes for simultaneous determination of dopamine and topotecan with improved data reproducibility.

Gold nanomaterials have been widely explored in electrochemical sensors due to their high catalytic property and good stability in multi-medium. In this paper, the reproducibility of the signal among batches of gold nanorods (AuNRs)-modified electrodes was investigated to improve the data stabilization and repeatability. Ordered and random self-assembled AuNRs-modified electrodes were used as electrochemical sensors for the simultaneous determination of dopamine (DA) and topotecan (TPC), with the aim of obtaining an improved signal stability in batches of electrodes and realizing the simultaneous determination of both substances. The morphology and structure of the assemblies were analyzed and characterized by UV-Vis spectra, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X-ray powder diffraction (XRD). Electrochemical studies showed that the ordered AuNRs/ITO electrodes have excellent signal reproducibility among several individuals due to the homogeneous mass transfer in the ordered arrangement of the AuNRs. Under the optimized conditions, the simultaneous detection results of DA and TPC showed good linearity in the ranges 1.75-45 µM and 1.5-40 µM, and the detection limits of DA and TPC were 0.06 µM and 0.17 µM, respectively. The results showed that the prepared ordered AuNR/ITO electrode had high sensitivity, long-term stability, and reproducibility for the simultaneous determination of DA and TPC, and it was expected to be applicable for real sample testing.

Carbon quantum dots as immune modulatory therapy in a Sjögren's syndrome mouse model.

OBJECTIVES: The objective of the study was to evaluate the therapeutic effects of carbon quantum dots (CQDs) in immunomodulation on non-obese diabetic (NOD) mice, as the model for Sjögren's syndrome (SS). METHODS: Carbon quantum dots were generated from Setaria viridis via a hydrothermal process. Their toxic effects were tested by cell viability and blood chemistry analysis, meanwhile therapeutic effects were investigated in NOD mice in the aspects of saliva flow, histology, and immune cell distribution. RESULTS: Carbon quantum dots, with rich surface chemistry and unique optical properties, showed non-cytotoxicity in vitro or no damage in vivo. Intravenously applied CQDs alleviated inflammation in the submandibular glands in NOD mice after 6-week treatments. The inflammatory area index and focus score were significantly decreased in CQD-treated mice. Besides, the levels of anti-SSA and anti-SSB were decreased in the presence of CQDs. The stimulated saliva flow rates and weight of submandibular glands were significantly increased in CQD-treated mice by reducing the apoptosis of cells. The CD3+ and CD4+ T cells distributed around the ducts of submandibular glands were significantly decreased, while the percentage of Foxp3+ cells was higher in CQD-treated mice than that in the control group. CONCLUSIONS: Our findings suggest that CQDs may ameliorate the dysregulated immune processes in NOD mice.

Age at adiposity rebound and the relevance for obesity: a systematic review and meta-analysis.

OBJECTIVES: To understand the sex difference in age at adiposity rebound (AR), integrate the prevalence of early AR (EAR), and provide a quantitative association between early age at AR and overweight/obesity. METHODS: Literature review was conducted in different databases, including the Web of Science, PubMed, EMBASE, Wiley, Chinese National Knowledge Infrastructure, and ScienceDirect databases up to August 2021. Studies that reported data related to AR were considered for inclusion. Pooled effect sizes and their respective 95% confidence intervals (CIs) were calculated using random effects models, depending on the size of heterogeneity. Heterogeneity was tested by using the I2 statistics. RESULTS: 28 studies with a combined sample size of 106,397 people were included in the final meta-analysis. Girls had a significantly earlier age of AR than boys (mean difference = 3.38 months; 95% CI 2.14-4.63). The overall prevalence of EAR was 40% (95% CI 31% to 50%), and the prevalence in girls was 5% higher than that in boys based on the definition of age at AR < 5.0-5.1 years. The overall pooled prevalence of EAR showed an increasing trend by child's birth year [1934-1973]: 29% (95% CI 22% to 37%), 1991-2001: 35% (95% CI 26% to 44%), and 2002-2009: 52% (95% CI 40-63%). Early age at AR (age at AR < 5.0-5.1 years) was associated with a significantly increased risk of overweight/obesity (OR = 5.07; 95% CI 3.60-7.12), overweight (OR = 3.10; 95% CI 1.69-5.70), and obesity (OR = 6.97; 95% CI 4.32-11.26) from the preschool period to adulthood. CONCLUSIONS: The overall prevalence of EAR is increasing, and girls experience AR earlier than boys. The early age at AR in children may be an early and effective marker of obesity.

Proteomic characteristics of PM2.5-induced differentially expressed proteins in k-ras-silenced HBE cells.

The human bronchial epithelial cells (HBE) and K-ras-silenced HBE cells were treated with fine particulate matter (PM2.5) samples from Taiyuan for 24 h. To screen the proteomic characteristics of PM2.5-induced differentially expressed proteins (DEPs), the Q Exactive mass spectrometer was used. Gene ontology (GO) analysis, Kyoto encyclopedia of genes and genomes (KEGG) analysis, functional prediction, protein-protein interaction (PPI) network analysis, and visualization of differential protein interactions were performed. 251 DEPs in K-ras silenced cells and 535 DEPs in normal HBE cells were identified, respectively. KEGG analysis showed that the differentially expressed proteins of PM2.5-treated cells were related to the biosynthesis of ribosomes, antibiotics, and amino acids. On the other hand, K-ras silenced cells were related to metabolic pathways, RNA transport, and DNA replication. Through the construction of a PPI network, the top 10 hub proteins were screened from the two cell groups, among which MRPL13, RPS20, and EIF1AX were of great significance. Our results indicated that the K-ras gene plays an important role in PM2.5-induced DEPs, and the findings provide a scientific basis for the further study of PM2.5 toxic mechanisms and biomarkers.

[Determination of lipophilic marine biotoxins in shellfish foods by ultra performance liquid chromatography-hybrid triple quadrupole linear ion trap-mass spectrometry].

OBJECTIVE: An ultra performance liquid chromatography-hybrid triple quadrupole linear ion trap-mass spectrometry(UPLC-QqLIT-MS) was established for determination of lipophilic marine biotoxins in shellfish. And the 12 lipophilic marine biotoxins in shellfish were surveyed. METHODS: The lipophilic marine biotoxins in homogenized shellfish were ultrasonically extracted by methanol in super-sonic instrument, and cleaned up by solid phase extraction of Strata-X column, and eluted with methanol(containing 0.3% ammonia water). The elution was diluted with water, and cleaned by 0.22 µm millipore filter. The filtrate was separated on a Waters ACQUITY UPLC BEH C_(18) column(150 mm×2.1 mm, 1.7 µm)by gradient elution in 12 minutes with acetronitrile-water(containing 0.01%(V/V) ammonia and 2 mmol/L ammonium formate) as mobile phase, and detected by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS), identified by electrospray ionization(ESI) in simultaneous scanning mode of positive and negative ions using multiple reaction monitoring, and quantified with external standards. Information dependent acquisition scan function(IDA) combined with enhanced production scan(EPI) was used to confirm the 12 lipophilic marine biotoxins. RESULTS: The calibration curves of 12 lipophilic marine biotoxins showed good linearity in the range of 0.5-50 µg/L with correlation coefficients were 0.9984-0.9999.The detection limits of the method were 0.15-0.29 µg/kg. The recoveries of three spiking levels ranged from 80.0% to 116.0%, and the relative standard deviation(RSD) were 0.6%-6.4%(n=7). CONCLUSION: The method for determination of 12 lipophilic marine biotoxins in shellfish by UPLC-QqLIT-MS was of operation convenience, less interference from impurities and good accuracy, which could meet the requirements for the determination of 12 lipophilic marine biotoxins residues in sea foods.

Survey on the Mechanical Properties of Lamellar Ag-MXA Supercluster Architectures.

Lamellar nanomaterials with specific architectures and novel properties have received increasing attention from both scientific and technological fields in recent years because of their potential applications in catalysis, energy conversion, and storage devices. Bulk supercluster pellets with well-defined lamellar structures were fabricated by assembling silver clusters and mercaptoalkyl acids (MXA) to investigate the mechanical properties. The relationship between the assembled structure and pressure resistance was surveyed for the first time. The enhanced interlayer interactions were found to increase the elastic modulus of the Ag-MXA supercluster architectures.

Serum metabolomics profiling and potential biomarkers of myopia using LC-QTOF/MS.

Myopia is the most common form of refractive eye disease, and the prevalence is increasing rapidly worldwide. However, the key metabolic alterations in individuals with high myopia are not understood clearly, and serum biomarkers remain to be determined. The objectives of this study were to identify serum biomarkers and investigate the metabolic alterations of myopia. The serum metabolomics profiling was investigated on 30 high myopia cases and 30 controls (without myopia) using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF/MS), and an independent additional cohort including 20 cases and 19 controls were investigated to validate potential metabolite candidates for biomarkers. According to the metabolic differences, the myopia patients and controls could be divided into different clusters and nine metabolites were found to be closely correlated with myopia. In the cohort of validation, eight metabolites were confirmed. Metabolic pathway analyses of these metabolites of high myopia involved abnormal phospholipid, diacylglycerol, amino acid, and vitamin metabolism, which were closely correlated with oxidative stress and inflammation. Multiple logistic regression analyses showed that γ-glutamyltyrosine and 12-oxo-20-trihydroxy-leukotriene B4 were potential biomarkers of myopia with a combined high sensitivity (97%), specificity (90%), and area under the curve value (0.983). These findings may contribute to an understanding of the pathophysiological changes and pathogenesis of myopia, and provide novel insight into the early prevention and control of high myopia.

Correction for color artifacts using the RGB intersection and the weighted bilinear interpolation.

Due to the defects of optical systems, image sensors, and imperfect algorithms for image acquisition, compression, and restoration, color artifacts often appear in images obtained by imaging devices such as digital cameras and scanners. Moreover, color artifacts are difficult to eliminate because of technical limitations, even in some mature commercial cameras. On the basis of red, green, and blue (RGB) intersection (RGBI), a correction method for color artifacts is proposed in this paper, where the RGB intersection-based method can effectively detect various types of color artifacts. Also, by combining the object information with weighted bilinear interpolation, the continuity of the image is kept while restoring the real color. Experiments demonstrate that the RGBI method, which is applicable to all color images, can eliminate various types of color artifacts with accurate detection and less artifact residue, even if the image has severe color distortion or the areas of the color artifacts are small and discrete.

Biocompatibility of Magnetic Resonance Imaging Nanoprobes Improved by Transformable Gadolinium Oxide Nanocoils.

To design functional nanomaterials for biomedical applications, the challenge for scientists is to gain further understanding of their unique toxicological properties. Nonspecific adhesion of proteins and endocytosis are considered to be the major biotoxic sources of imaging nanoprobes. Here, we fabricated ultrathin gadolinium oxide (Gd2O3) nanocoils with a low Young's modulus, which provides transformable properties in solution. The spatial configurational freedom of ultrathin nanocoils induces the steric repulsion to the nonspecific adsorption of proteins that, in turn, suppresses cellular uptake and thus improves their biocompatibility. The larger number of exposed surface gadolinium atoms of the ultrathin nanocoils provided enhanced T1 magnetic resonance (MR) imaging contrast with high signal activation. Such nanocontrast agents were applied to in vivo MR bioimaging to achieve prolonged circulation lifetime. The improved biocompatibility by transformable Gd2O3 nanocoils could open up a new perspective toward the design and construction of various nano-biomedicines in the future.

Global mRNA and Long Non-Coding RNA Expression in the Placenta and White Adipose Tissue of Mice Fed a High-Fat Diet During Pregnancy.

BACKGROUND/AIMS: Gestational diabetes mellitus (GDM) is a common complication of pregnancy, but the mechanisms underlying the disorders remain unclear. The study aimed to identify mRNA and long non-coding RNA (lncRNA) profiles in placenta and gonadal fat of pregnant mice fed a high-fat diet and to investigate the transcripts and pathways involved in the development of gestational diabetes mellitus. METHODS: Deep and broad transcriptome profiling was performed to assess the expression of mRNAs and lncRNAs in placenta and gonadal fat from 3 mice fed an HFD and chow during pregnancy. Then, differentially expressed mRNAs and lncRNAs were validated by quantitative real-time PCR. The function of the differentially expressed mRNAs was determined by pathway and Gene Ontology (GO) analyses, and the physical or functional relationships between the lncRNAs and the corresponding mRNAs were determined. RESULTS: Our study revealed that 82 mRNAs and 52 lncRNAs were differentially expressed in the placenta of mice fed an HFD during pregnancy, and 202 mRNAs and 120 lncRNAs were differentially expressed in gonadal fat. GO and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed differentially expressed mRNAs of placenta were closely related to extracellular matrix interactions, digestion, adhesion, and metabolism, whereas the differentially expressed mRNAs in adipose tissue were related to metabolic and insulin signalling pathways. The gene network demonstrated that Actg2, Cnfn, Muc16, Serpina3k, NONMMUT068202, and NONMMUT068203, were the core of the network in placental tissue, and the genes Tkt, Acss2, and Elovl6 served as the core of the network in gonadal fat tissue. CONCLUSION: These newly identified key genes and pathways in mice might provide valuable information regarding the pathogenesis of GDM and might be used to improve early diagnosis, prevention, drug design, and clinical treatment.

Ultrasensitive Surface-Enhanced Raman Scattering Sensor of Gaseous Aldehydes as Biomarkers of Lung Cancer on Dendritic Ag Nanocrystals.

Surface-enhanced Raman scattering (SERS) is expected as a technique that even theoretically detected chemicals at the single molecule level by surface plasmon phenomena of noble metal nanostructures. Insensitivity of detecting Raman weak-intensity molecules and low adsorptivity of gaseous molecules on solid substrates are two main factors hindering the application of SERS in gas detectors. In this manuscript, we demonstrated an operational SERS strategy to detect gaseous Raman weak-intensity aldehydes that have been considered as a biomarker of lung cancer for abnormal content was measured in volatile organic compounds (VOCs) of lung cancer patients. To enhance the adsorption of gaseous molecules, dendritic Ag nanocrystals mimicking the structural feature (dendritic) of moth's antennae were formed, wherein the existence of numerous cavity traps in Ag dendritic nanocrystals prolonged reaction time of the gaseous molecules on the surface of solid surface through the "cavity-vortex" effect. By the nucleophilic addition reaction with the Raman-active probe molecule p-aminothiophenol (4-ATP) pregrafted on dendritic Ag nanocrystals, the gaseous aldehyde molecules were sensitively captured to detect at the ppb (parts per billion) level. Additionally, the sensitivity of this operational SERS strategy to detection of lung cancer biomarkers was not affected by the humidity, which represented a great potential in fast, easy, cost-effective, and noninvasive recognition of lung malignancies.

Understanding the Selective Detection of Fe3+ Based on Graphene Quantum Dots as Fluorescent Probes: The Ksp of a Metal Hydroxide-Assisted Mechanism.

Graphene quantum dots (GQDs) have been widely used as fluorescence probes to detect metal ions with satisfactory selectivity. However, the diverse chemical structures of GQDs lead to selectivity for multiple metal ions, and this can lead to trouble in the interpretation of selectivity due to the lack of an in depth and systematic analysis. Herein, bare GQDs were synthesized by oxidizing carbon black with nitric acid and used as fluorescent probes to detect metal ions. We found that the specific ability of GQDs to recognize ferric ions relates to the acidity of the medium. Specifically, we demonstrated that the coordination between GQDs and Fe3+ is regulated by the pH of the aqueous GQDs solution. Dissociative Fe3+ can coordinate with the hydroxyl groups on the surface of the GQDs to form aggregates (such as iron hydroxide), which induces fluorescence quenching. A satisfactory selectivity for Fe3+ ions was achieved under relatively acidic conditions; this is because of the extremely small Ksp of ferric hydroxide compared to those of other common metal hydroxides. To directly survey the key parameter for Fe3+ ion specificity, we performed the detection experiment in an environment free of interference from the buffer solution, noninherent groups, and other complex factors. This study will help researchers understand the selectivity mechanisms of GQDs as fluorescence probes for metal ions, which could guide the design of other GQD-based sensor platforms.

[Effects of trichloroethylene toxicity on normal human liver cells and hepatocytes with CYP2E1 gene overexpression].

OBJECTIVE: To investigate the effects of trichloroethylene (TCE) toxicity on the normal human liver cells (L02 cells) and hepatocytes with CYP2E1 gene overexpression which was constructed through molecular cloning technology in our laboratory, then to explore the roles of CYP2E1 gene in TCE toxicity. METHODS: L02 cells and hepatocytes with CYP2E1 overexpression were treated with various doses of TCE (0,0.25, 0.5, 1.0, 2.0, 4.0 mmol/L) for 12h, the expression of apoptosis genes (Bcl-2, Caspase-3, Caspase-8, Caspase-9) and oncogenes (c-fos, c-myc, k-ras, p53) were determined by real-time fluorescent PCR. RESULTS: Bcl-2 mRNA expression levels increased significantly in normal liver cells and CYP2E1-overexpressing cells after TCE treatment, Bcl-2 levels were 20%∼50%higher in CYP2E1-overexpressing cells than in L02 liver cells at doses of 0.25∼2.0 mmol/L TCE. Caspase-3, Caspase-8 and caspase-9 mRNA expression increased by 30%∼600% in CYP2E1-overexpressing cells at doses of 0.5∼4.0 mmol/L TCE when compared with L02 cells (P < 0.01). Additionally, c-fos, k-ras and c-myc mRNA expression levels were 25%∼120% higher in CYP2E1-overexpressing cells than in L02 cells (P < 0.01), p53 mRNA expression levels were lower 10%∼50% in CYP2E1-overexpressing cells than in L02 cells (P < 0.05 or P < 0.01). CONCLUSIONS: There were significant differences for apoptosis gene and oncogene expression levels between normal liver cells and CYP2E1-overexpressing cells after they were treated with TCE, these findings indicated that CYP2E1 might play an important role in TCE metabolism in vivo.

A Label-Free Electrochemical Aptamer Sensor for Sensitive Detection of Cardiac Troponin I Based on AuNPs/PB/PS/GCE.

Cardiac troponin I (cTnI) monitoring is of great value in the clinical diagnosis of acute myocardial infarction (AMI). In this paper, a highly sensitive electrochemical aptamer sensor using polystyrene (PS) microspheres as the electrode substrate material in combination with Prussian blue (PB) and gold nanoparticles (AuNPs) was demonstrated for the sensitive and label-free determination of cTnI. PS microspheres were synthesized by emulsion polymerization and then dropped onto the glassy carbon electrode (GCE); PB and AuNPs were electrodeposited on the electrode in corresponding electrolyte solutions step by step. The PS microsphere substrate provided a large surface area for the loading mass of the biological affinity aptamers, while the PB layer improved the electrical conductivity of the modified electrode, and the electroactive AuNPs exhibited excellent catalytic performance for the subsequent electrochemical measurements. In view of the above mentioned AuNPs/PB/PS/GCE sensing platform, the fabricated label-free electrochemical aptamer sensor exhibited a wide detection range of 10 fg/mL~1.0 µg/mL and a low detection limit of 2.03 fg/mL under the optimal conditions. Furthermore, this biosensor provided an effective detection platform for the analysis of cTnI in serum samples. The introduction of this sensitive electrochemical aptamer sensor provides a reference for clinically sensitive detection of cTnI.

In Situ Synthesis of Hierarchical Co(CO3)0.5(OH)·0.11H2O@ZIF-67/WO3 With High Humidity Immunity and Response to H2S Sensing.

H2S gas sensors with facile preparation, low detection limits, and high selectivity are crucial for environmental and human health monitoring. However, it is difficult to maintain a high response of H2S gas sensors under high humidity in practical applications. To face this dilemma, a layer-by-layer growth method is applied to in situ prepare a nanostructured Co(CO3)0.5(OH)·0.11H2O/WO3 coated by a hydrophobic hierarchical ZIF-67 as the H2S sensor. This novel composite exhibits excellent humidity immunity without sacrificing the excellent sensitivity and selectivity of H2S. At a low operating temperature of 90 °C, a remarkable response value of 1052.3 to 100 ppm H2S has been achieved, which is 779 and 9.36 times higher than that of pure WO3 and Co(CO3)0.5(OH)·0.11H2O/WO3, respectively. More importantly, an 82.2% relative response value remains at a high humidity of 75%RH. The sensing mechanisms are investigated using gas chromatography-mass spectrometry (GC-MS), which revealed that the reaction products are H2O and SO2. The high humidity immunity and fast response of the Co(CO3)0.5(OH)·0.11H2O@ZIF-67/WO3 demonstrate the layer-by-layer in situ synthesis method holds the potential application for the development of high-performance WO3-based H2S sensors.

Ni foam: a novel three-dimensional porous sensing platform for sensitive and selective nonenzymatic glucose detection.

The present communication reports on the first use of commercially available three-dimensional porous Ni foam (NF) as a novel electrochemical sensing platform for nonenzymatic glucose detection. NF not only acts as a working electrode, but also functions as an effective electrocatalyst for electrooxidation of glucose. The sensor exhibits high selectivity toward glucose. The linear range and limit of detection were 0.05-7.35 mM (R = 0.995) and 2.2 µM with a signal-to-noise ratio of 3, respectively. The application of this glucose sensor in human blood serum has also been demonstrated successfully.

Facile synthesis of novel Ni(II)-based metal-organic coordination polymer nanoparticle/reduced graphene oxide nanocomposites and their application for highly sensitive and selective nonenzymatic glucose sensing.

The present paper reports on the facile preparation of novel Ni(II)-based metal-organic coordination polymer nanoparticle/reduced graphene oxide (NiCPNP/rGO) nanocomposites for the first time. The formation of the nanocomposites occurs in a single step, carried out by hydrothermal treatment of the mixture of tannic acid functioned graphene oxide and NiCl(2) aqueous solution in N,N-dimethylformamide. It is found that the NiCPNP/rGO nanocomposite-modified electrode shows high electrocatalytic activity for glucose oxidation in alkaline medium. This nonenzymatic glucose sensor exhibits high selectivity toward glucose and the linear range and limit of detection are estimated to be from 0.01 to 8.75 mM (r: 0.997) and 0.14 µM with a signal-to-noise ratio of 3, respectively. The application of this glucose sensor in human blood serum has also been demonstrated successfully.

Impact of pregnancy-related anxiety on preschoolers' emotional and behavioral development: Gender specificity, critical time windows and cumulative effect.

BACKGROUND: Studies on the gender-specific effect of PrA on children's emotional and behavioral development are limited. Lack of PrA data on the entire pregnancy had caused difficulties in identifying the key time window and cumulative effects. METHODS: Based on Ma'anshan Birth Cohort in China, mothers at pregnancy and children followed up to 4 years of age were tested using the PrA questionnaire and the Achenbach Child Behavior Checklist (CBCL) 1.5-5. Finally, 1699 mother-child pairs were included in the study and regression models were developed for analysis. RESULTS: Children of mothers with PrA are at significantly elevated risk for abnormal mood/behavior at preschool age. Girls seemed to be more sensitive to maternal PrA than boys, mainly manifested in internalizing problems; The third trimester of pregnancy might be a critical time window when maternal PrA affected children's internalizing problems; Longer the duration women had PrA during pregnancy, more possibilities their children would have to develop internalizing and externalizing problems. LIMITATIONS: The PrA questionnaire may be different from PrA questionnaires in other countries such as due to different cultural contexts. Findings need to be interpreted with more caution. Factors such as maternal postpartum depression and care practices of nursing staff were not considered. No data were collected on disease as well as caregiver emotional status, which also impacts the reporting and identification of emotional/behavioral problems in children. CONCLUSIONS: Gender-specific and cumulative effect of PrA on preschoolers' emotional/behavioral development is observed. The third trimester of pregnancy might be the critical time window.