Early brain cognitive development in late preterm infants: an event-related potential and resting EEG study.

BACKGROUND: Late preterm infants (LPIs) are at risk of neurodevelopmental delay. Research on their cognitive development is helpful for early intervention and follow-up. METHODS: Event-related potential (ERP) and resting electroencephalography (RS-EEG) were used to study the brain cognitive function of LPIs in the early stage of life. The Gesell Developmental Scale (GDS) was used to track the neurodevelopmental status at the age of 1 year after correction, and to explore the neurophysiological indicators that could predict the outcome of cognitive development in the early stage. RESULTS: The results showed that mismatch response (MMR) amplitude, RS-EEG power spectrum and functional connectivity all suggested that LPIs were lagging behind. At the age of 1 year after correction, high-risk LPIs showed no significant delay in gross motor function, but lagged behind in fine motor function, language, personal social interaction and adaptability. The ROC curve was used to evaluate the predictive role of MMR amplitude in the brain cognitive development prognosis at 1 year, showing a sensitivity of 80.00% and a specificity of 90.57%. The area under the curve (AUC) was 0.788, with a P-value of 0.007. CONCLUSIONS: Based on our findings we supposed that the cognitive function of LPI lags behind that of full-term infants in early life. Preterm birth and perinatal diseases or high risk factors affected brain cognitive function in LPIs. MMR amplitude can be used as an early predictor of brain cognitive development in LPIs. TRIAL REGISTRATION: This clinical trial is registered with the Chinese Clinical Trial Registry (ChiCTR). TRIAL REGISTRATION NUMBER: ChiCTR2100041929. Date of registration: 2021-01-10. URL of the trial registry record: https://www.chictr.org.cn/ .

Architecture of the baculovirus nucleocapsid revealed by cryo-EM.

Baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) has been widely used as a bioinsecticide and a protein expression vector. Despite their importance, very little is known about the structure of most baculovirus proteins. Here, we show a 3.2 Å resolution structure of helical cylindrical body of the AcMNPV nucleocapsid, composed of VP39, as well as 4.3 Å resolution structures of both the head and the base of the nucleocapsid composed of over 100 protein subunits. AcMNPV VP39 demonstrates some features of the HK97-like fold and utilizes disulfide-bonds and a set of interactions at its C-termini to mediate nucleocapsid assembly and stability. At both ends of the nucleocapsid, the VP39 cylinder is constricted by an outer shell ring composed of proteins AC104, AC142 and AC109. AC101(BV/ODV-C42) and AC144(ODV-EC27) form a C14 symmetric inner layer at both capsid head and base. In the base, these proteins interact with a 7-fold symmetric capsid plug, while a portal-like structure is seen in the central portion of head. Additionally, we propose an application of AlphaFold2 for model building in intermediate resolution density.

STT3A-mediated viral N-glycosylation underlies the tumor selectivity of oncolytic virus M1.

Oncolytic viruses are emerging as promising anticancer agents. Although the essential biological function of N-glycosylation on viruses are widely accepted, roles of N-glycan and glycan-processing enzyme in oncolytic viral therapy are remain elusive. Here, via cryo-EM analysis, we identified three distinct N-glycans on the envelope of oncolytic virus M1 (OVM) as being necessary for efficient receptor binding. E1-N141-glycan has immediate impact on the binding of MXRA8 receptor, E2-N200-glycan mediates the maturation of E2 from its precursor PE2 which is unable to bind with MXRA8, and E2-N262-glycan slightly promotes receptor binding. The necessity of OVM N-glycans in receptor binding make them indispensable for oncolysis in vitro and in vivo. Further investigations identified STT3A, a key catalytic subunit of oligosaccharyltransferase (OST), as the determinant of OVM N-glycosylation, and STT3A expression in tumor cells is positively correlated with OVM-induced oncolysis. Increased STT3A expression was observed in various solid tumors, pointing to a broad-spectrum anticancer potential of OVM. Collectively, our research supports the importance of STT3A-mediated N-glycosylation in receptor binding and oncolysis of OVM, thus providing a novel predictive biomarker for OVM.

Directed natural evolution generates a next-generation oncolytic virus with a high potency and safety profile.

Oncolytic viruses (OVs) represent a type of encouraging multi-mechanistic drug for the treatment of cancer. However, attenuation of virulence, which is generally required for the development of OVs based on pathogenic viral backbones, is frequently accompanied by a compromised killing effect on tumor cells. By exploiting the property of viruses to evolve and adapt in cancer cells, we perform directed natural evolution on refractory colorectal cancer cell HCT-116 and generate a next-generation oncolytic virus M1 (NGOVM) with an increase in the oncolytic effect of up to 9690-fold. The NGOVM has a broader antitumor spectrum and a more robust oncolytic effect in a range of solid tumors. Mechanistically, two critical mutations are identified in the E2 and nsP3 genes, which accelerate the entry of M1 virus by increasing its binding to the Mxra8 receptor and antagonize antiviral responses by inhibiting the activation of PKR and STAT1 in tumor cells, respectively. Importantly, the NGOVM is well tolerated in both rodents and nonhuman primates. This study implies that directed natural evolution is a generalizable approach for developing next-generation OVs with an expanded scope of application and high safety.

The structure of a 12-segmented dsRNA reovirus: New insights into capsid stabilization and organization.

Infecting a wide range of hosts, members of Reovirales (formerly Reoviridae) consist of a genome with different numbers of segmented double stranded RNAs (dsRNA) encapsulated by a proteinaceous shell and carry out genome replication and transcription inside the virion. Several cryo-electron microscopy (cryo-EM) structures of reoviruses with 9, 10 or 11 segmented dsRNA genomes have revealed insights into genome arrangement and transcription. However, the structure and genome arrangement of 12-segmented Reovirales members remain poorly understood. Using cryo-EM, we determined the structure of mud crab reovirus (MCRV), a 12-segmented dsRNA virus that is a putative member of Reovirales in the non-turreted Sedoreoviridae family, to near-atomic resolutions with icosahedral symmetry (3.1 Å) and without imposing icosahedral symmetry (3.4 Å). These structures revealed the organization of the major capsid proteins in two layers: an outer T = 13 layer consisting of VP12 trimers and unique VP11 clamps, and an inner T = 1 layer consisting of VP3 dimers. Additionally, ten RNA dependent RNA polymerases (RdRp) were well resolved just below the VP3 layer but were offset from the 5-fold axes and arranged with D5 symmetry, which has not previously been seen in other members of Reovirales. The N-termini of VP3 were shown to adopt four unique conformations; two of which anchor the RdRps, while the other two conformations are likely involved in genome organization and capsid stability. Taken together, these structures provide a new level of understanding for capsid stabilization and genome organization of segmented dsRNA viruses.

Applications and prospects of cryo-EM in drug discovery.

Drug discovery is a crucial part of human healthcare and has dramatically benefited human lifespan and life quality in recent centuries, however, it is usually time- and effort-consuming. Structural biology has been demonstrated as a powerful tool to accelerate drug development. Among different techniques, cryo-electron microscopy (cryo-EM) is emerging as the mainstream of structure determination of biomacromolecules in the past decade and has received increasing attention from the pharmaceutical industry. Although cryo-EM still has limitations in resolution, speed and throughput, a growing number of innovative drugs are being developed with the help of cryo-EM. Here, we aim to provide an overview of how cryo-EM techniques are applied to facilitate drug discovery. The development and typical workflow of cryo-EM technique will be briefly introduced, followed by its specific applications in structure-based drug design, fragment-based drug discovery, proteolysis targeting chimeras, antibody drug development and drug repurposing. Besides cryo-EM, drug discovery innovation usually involves other state-of-the-art techniques such as artificial intelligence (AI), which is increasingly active in diverse areas. The combination of cryo-EM and AI provides an opportunity to minimize limitations of cryo-EM such as automation, throughput and interpretation of medium-resolution maps, and tends to be the new direction of future development of cryo-EM. The rapid development of cryo-EM will make it as an indispensable part of modern drug discovery.

Improvement of semantic processing ability of Chinese characters in school children: A comparative study based on 2009 and 2019 data.

To explore the characteristics of semantic cognitive development of school children by observing the development changes over 10 years, a retrospective event-related potential (ERP) study was conducted on the semantic processing characteristics of Chinese characters in children aged 7-11 years with the same study design in 2009 and 2019. For the EEGs recorded in 2009, the N400 amplitude of semantic processing in children aged 7-11 years showed an approximately inverted U-shaped development trend with a slow rise at the age of 7-9, a peak at the age of 10, then a rapid decline at the age of 11. However, for the EEGs recorded in 2019, the N400 amplitude showed a gradually decreasing development trend with a slow decline for the 7-11 years class. Our data suggested that the semantic processing of Chinese characters in children aged 7-11 years in 2019 was one age stage earlier than that in 2009. The children's brain cognition is in the process of development and change with high plasticity. 10 years of favorable social and educational environmental factors have significantly improved children's semantic processing ability of Chinese characters.

Metabolic risk score as a predictor in a nomogram for assessing myometrial invasion for endometrial cancer.

The purpose of the present study was to investigate the predictive value of metabolic syndrome in evaluating myometrial invasion (MI) in patients with endometrial cancer (EC). The study retrospectively included patients with EC who were diagnosed between January 2006 and December 2020 at the Department of Gynecology of Nanjing First Hospital (Nanjing, China). The metabolic risk score (MRS) was calculated using multiple metabolic indicators. Univariate and multivariate logistic regression analyses were performed to determine significant predictive factors for MI. A nomogram was then constructed based on the independent risk factors identified. A calibration curve, a receiver operating characteristic (ROC) curve and decision curve analysis (DCA) were used to evaluate the effectiveness of the nomogram. A total of 549 patients were randomly assigned to a training or validation cohort, with a 2:1 ratio. Data was then gathered on significant predictors of MI in the training cohort, including MRS [odds ratio (OR), 1.06; 95% confidence interval (CI), 1.01-1.11; P=0.023], histological type (OR, 1.98; 95% CI, 1.11-3.53; P=0.023), lymph node metastasis (OR, 3.15; 95% CI, 1.61-6.15; P<0.001) and tumor grade (grade 2: OR, 1.71; 95% CI, 1.23-2.39; P=0.002; Grade 3: OR, 2.10; 95% CI, 1.53-2.88; P<0.001). Multivariate analysis indicated that MRS was an independent risk factor for MI in both cohorts. A nomogram was generated to predict a patient's probability of MI based on the four independent risk factors. ROC curve analysis showed that, compared with the clinical model (model 1), the combined model with MRS (model 2) significantly improved the diagnostic accuracy of MI in patients with EC (area under the curve in model 1 vs. model 2: 0.737 vs. 0.828 in the training cohort and 0.713 vs. 0.759 in the validation cohort). Calibration plots showed that the training and validation cohorts were well calibrated. DCA showed that a net benefit is obtained from the application of the nomogram. Overall, the present study developed and validated a MRS-based nomogram predicting MI in patients with EC preoperatively. The establishment of this model may promote the use of precision medicine and targeted therapy in EC and has the potential to improve the prognosis of patients affected by EC.

Identification of the receptor of oncolytic virus M1 as a therapeutic predictor for multiple solid tumors.

Over the last decade, oncolytic virus (OV) therapy has shown its promising potential in tumor treatment. The fact that not every patient can benefit from it highlights the importance for defining biomarkers that help predict patients' responses. As particular self-amplifying biotherapeutics, the anti-tumor effects of OVs are highly dependent on the host factors for viral infection and replication. By using weighted gene co-expression network analysis (WGCNA), we found matrix remodeling associated 8 (MXRA8) is positively correlated with the oncolysis induced by oncolytic virus M1 (OVM). Consistently, MXRA8 promotes the oncolytic efficacy of OVM in vitro and in vivo. Moreover, the interaction of MXRA8 and OVM studied by single-particle cryo-electron microscopy (cryo-EM) showed that MXRA8 directly binds to this virus. Therefore, MXRA8 acts as the entry receptor of OVM. Pan-cancer analysis showed that MXRA8 is abundant in most solid tumors and is highly expressed in tumor tissues compared with adjacent normal ones. Further study in cancer cell lines and patient-derived tumor tissues revealed that the tumor selectivity of OVM is predominantly determined by a combinational effect of the cell membrane receptor MXRA8 and the intracellular factor, zinc-finger antiviral protein (ZAP). Taken together, our study may provide a novel dual-biomarker for precision medicine in OVM therapy.

Cryo-electron Tomography Reveals the Roles of FliY in Helicobacter pylori Flagellar Motor Assembly.

Helicobacter pylori plays a causative role in gastric diseases. The pathogenicity of H. pylori depends on its ability to colonize the stomach guided by motility. FliY is a unique flagellar motor switch component coexisting with the classical FliG, FliM, and FliN switch proteins in some bacteria and has been shown to be essential for flagellation. However, the functional importance of FliY in H. pylori flagellar motor assembly is not well understood. Here, we applied cryo-electron tomography and subtomogram averaging to analyze the in situ structures of flagellar motors from wild-type strain, fliY-null mutant and complementation mutants expressing the N-terminal or C-terminal domain of FliY. Loss of full-length FliY or its C-terminal domain interrupted the formation of an intact C ring and soluble export apparatus, as well as the hook and flagellar filaments. Complementation with FliY C-terminal domain restored all these missing components of flagellar motor. Taken together, these results provide structural insights into the roles of FliY, especially its C-terminal domain in flagellar motor assembly in H. pylori. IMPORTANCE Helicobacter pylori is the major risk factor related with gastric diseases. Flagellar motor is one of the most important virulence factors in H. pylori. However, the assembly mechanism of H. pylori flagellar motor is not fully understood yet. Previous report mainly described the overall structures of flagellum but had not focused on its specific components. Here, we focus on H. pylori flagellar C-ring protein FliY. We directly visualize the flagellar structures of H. pylori wild-type and FliY N-/C-terminal complementary strains by cryo-electron tomography and subtomogram averaging. Our results show that deletion of FliY or its C-terminal domain causes the loss of C ring, whereas deletion of FliY N-terminal does not affect C-ring assembly and flagellar structures. Our results provide direct evidence that C-ring protein FliY, especially its C-terminal domain, plays an indispensable role in H. pylori motor assembly and flagellar formation. This study will deepen our understanding about H. pylori pathogenesis.

Correction: Evaluation of auditory perception development in neonates by quantitative electroencephalography and auditory event-related potentials.

[This corrects the article DOI: 10.1371/journal.pone.0183728.].

Predictive value of electroencephalogram, event-related potential, and general movements quality assessment in neurodevelopmental outcome of high-risk infants.

OBJECTIVE: The objective of the study is to investigate the predictive value of electroencephalogram (EEG), event-related potential (ERP), and general movements (GMs) quality assessment in the neurodevelopmental outcome of high-risk infants at one year old. METHODS: EEG and ERP were performed in high-risk infants at four weeks old, and GMs quality was evaluated once at 4 weeks and once at 12 weeks. The Gesell score was used to assess neurodevelopment outcome at one year old. A comparative analysis of the effects of EEG, GMs, EEG + ERP, and EEG + ERP + GMs was used to predict high-risk neonatal neurodevelopmental outcome. RESULTS: Of 71 high-risk infants at the age of one year, 3 (4.23%) had cerebral palsy, 14 (19.72%) had psychomotor retardation, and 54 (76.05%) were normal. The sensitivity, specificity, positive predictive value, and negative predictive value of EEG + ERP + GMs method were 90.00%, 95.08%, 75.00%, and 98.31%, respectively, and these indexes were the highest among the four methods (EEG, GMs, EEG + ERP, and EEG + ERP + GMs). The kappa statistic for the reliability of predicting neurodevelopmental outcome of high risk newborns by the EEG + ERP + GMs method was substantial at 0.785, while the other three methods obtained relatively low Kappa values (0.599, 0.586, and 0.712, respectively). CONCLUSIONS: The combination of EEG, ERP, and GMs quality assessment can greatly improve the prediction of neurodevelopmental outcome of high-risk newborns.

Cryo-EM reveals a previously unrecognized structural protein of a dsRNA virus implicated in its extracellular transmission.

Mosquito viruses cause unpredictable outbreaks of disease. Recently, several unassigned viruses isolated from mosquitoes, including the Omono River virus (OmRV), were identified as totivirus-like viruses, with features similar to those of the Totiviridae family. Most reported members of this family infect fungi or protozoans and lack an extracellular life cycle stage. Here, we identified a new strain of OmRV and determined high-resolution structures for this virus using single-particle cryo-electron microscopy. The structures feature an unexpected protrusion at the five-fold vertex of the capsid. Disassociation of the protrusion could result in several conformational changes in the major capsid. All these structures, together with some biological results, suggest the protrusions' associations with the extracellular transmission of OmRV.

Identification of an intraocular microbiota.

The current dogma in ophthalmology and vision research presumes the intraocular environment to be sterile. However, recent evidence of intestinal bacterial translocation into the bloodstream and many other internal organs including the eyes, found in healthy and diseased animal models, suggests that the intraocular cavity may also be inhabited by a microbial community. Here, we tested intraocular samples from over 1000 human eyes. Using quantitative PCR, negative staining transmission electron microscopy, direct culture, and high-throughput sequencing technologies, we demonstrated the presence of intraocular bacteria. The possibility that the microbiome from these low-biomass communities could be a contamination from other tissues and reagents was carefully evaluated and excluded. We also provide preliminary evidence that a disease-specific microbial signature characterized the intraocular environment of patients with age-related macular degeneration and glaucoma, suggesting that either spontaneous or pathogenic bacterial translocation may be associated with these common sight-threatening conditions. Furthermore, we revealed the presence of an intraocular microbiome in normal eyes from non-human mammals and demonstrated that this varied across species (rat, rabbit, pig, and macaque) and was established after birth. These findings represent the first-ever evidence of intraocular microbiota in humans.

Visualization of the Oncolytic Alphavirus M1 Life Cycle in Cancer Cells.

Oncolytic alphavirus M1 has been shown to selectively target and kill cancer cells, but cytopathic morphologies induced by M1 virus and the life cycle of the M1 strain in cancer cells remain unclear. Here, we study the key stages of M1 virus infection and replication in the M1 virus-sensitive HepG2 liver cancer cell line by transmission electron microscopy, specifically examining viral entry, assembly, maturation and release. We found that M1 virus induces vacuolization of cancer cells during infection and ultimately nuclear marginalization, a typical indicator of apoptosis. Specifically, our results suggest that the endoplasmic reticulum participates in the assembly of nucleocapsids. In the early and late stage of infection, three kinds of special cytopathic vacuoles are formed and appear to be involved in the replication, maturation and release of the virus. Taken together, our data displayed the process of M1 virus infection of tumor cells and provide the structural basis for the study of M1 virus-host interactions.

Development of Biomarker Signatures Associated with Anoikis to Predict Prognosis in Endometrial Carcinoma Patients.

PURPOSE: To generate a signature based on anoikis-related genes (ARGs) for endometrial carcinoma (EC) patients and elucidate the molecular mechanisms in EC. METHODS: On the basis of TCGA-UCEC dataset, we identified specific anoikis-related genes (ARGs) in EC. Cox-relative regression methods were used to generate an anoikis-related signature (ARS). The possible biological pathways of ARS-related genes were analyzed by GSEA. The clinical potency and immune status of ARS were analyzed by CIBERSORT method, ssGSEA algorithm, Tumor Immune Dysfunction and Exclusion (TIDE) analysis. Moreover, the expression patterns of ARS genes were verified by HPA database. RESULTS: Seven anoikis genes (CDKN2A, E2F1, ENDOG, EZH2, HMGA1, PLK1, and SLC2A1) were determined to develop a prognostic ARS. Both genes of ARS were closely bound up with the prognosis of EC patients. The ARS could accurately classify EC cases with different clinical outcome and mirror the specific immune status of EC. We observed that ARS-high patients could not benefit from immunotherapy. Finally, all the hub genes of ARS were proved to be upregulated in EC tissues by immunohistology. CONCLUSION: ARS can be used to stratify the risk and forecast the survival outcome of EC patients and provide prominent reference for individualized treatment in EC.

Clinical significance of electroencephalography power spectrum density and functional connection analysis in neonates with hypoxic-ischemic encephalopathy.

OBJECTIVE: To investigate the effects of electroencephalography (EEG) power and functional connectivity analysis on cerebral cortex function in neonates with hypoxic-ischemic encephalopathy (HIE), and to identify the neurobiological indicators of neural development in HIE. METHODS: We recruited 20 mild HIE neonates, 15 moderate HIE neonates, and 30 controls. EEG was performed about 72 hr after birth. The power spectral density (PSD) and imaginary part of coherency (ICOH) were analyzed. Gesell developmental schedule (GDS) was used to evaluate the neural development in the mild and moderate HIE groups at 1 year of age, and the correlation between the quantitative EEG results and the state of neural development was analyzed. RESULTS: Compared with the controls, FP1, FP2, C3, C4, Fz, Cz, Pz , F3, and P4 of moderate HIE neonates showed that the PSD of theta, alpha and beta bands decreased significantly. In terms of the mean whole-brain PSD, the moderate HIE group showed a significant decrease in all frequency bands. ICOH of the moderate HIE group showed that functional connectivity was significantly less than that in the controls mainly in the delta band, and the functional connectivity of the delta, theta, alpha1, and alpha2 bands was markedly reduced compared with the mild HIE. GDS test at 1 year old showed that two infants in the moderate HIE group had suspected neurological delay in gross motor and language. The developmental quotient(DQ) of gross motor, language, and personal-social ability in the moderate HIE group were significantly lower than in the mild HIE group. And there was a significant positive correlation between PSD in each EEG frequency band and GDS score in the moderate HIE group. CONCLUSION: PSD and ICOH can be used to evaluate brain function. PSD can detect the delayed neurological development in infants with moderate HIE, and can be a neurobiomarker of brain development in HIE.

Advanced glycation end products increased placental vascular permeability of human BeWo cells via RAGE/NF-kB signaling pathway.

OBJECTIVE: This study aimed to investigate the mechanisms of advanced glycation end products (AGEs) on cell tight conjunction and placental vascular permeability in BeWo cells. STUDY DESIGN: Monolayer permeability assay and transmission electron microscopy were employed to reveal the transformation of the placental vascular permeability and cell tight conjunction. Immunofluorescence, western blot and RT-qPCR were adopted to determine the protein and mRNA levels. Anti-RAGE and NF-kB inhibitor (PDTC) were used to inactivate the RAGE/NF-kB signaling pathway. RESULTS: AGEs significantly decreased trans-epithelial electrical resistance (TEER), while increased paracellular permeability (P < 0.05). TEM showed that AGEs made cell junction loose. AGEs inhibited ZO-1 and Occludin expressions, while anti-RAGE or PDTC partially restored their levels. AGEs also significantly increased mRNA RAGE and NF-kB expressions in BeWo cells (P < 0.05), and their expressions were inhibited by anti-RAGEy or PDTC. CONCLUSION: AGEs could reduce the expressions of ZO-1 and Occludin by activating RAGE/NF-kB signaling pathway, thus increasing placental vascular permeability.

Low molecular weight heparin (nadroparin) improves placental permeability in rats with gestational diabetes mellitus via reduction of tight junction factors.

Placental structural abnormalities and dysfunction in those with gestational diabetes mellitus (GDM) can lead to increased placental permeability, which is in turn related to a poorer maternal and fetal prognosis. The present study sought to assess whether increased placental permeability in rats with GDM was accompanied by alterations in tight junction (TJ) factors and to evaluate the impact of low molecular weight heparin (LMWH) on these factors. The present study was conducted using pregnant female rats that were randomized into control, GDM and GDM + LMWH groups. Diabetes was induced via intraperitoneal administration of streptozotocin to rats in the GDM and GDM + LMWH groups, whereas rats in the GDM + LMWH group received daily subcutaneous LMWH starting on day 5 of pregnancy. On gestational day 16, all rats were sacrificed and Evans Blue (EB) assay was used to gauge vascular permeability based on EB dye leakage. Transmission electron microscopy was further used to assess TJ structures, and the TJ proteins zonular occludens­1 (ZO­1) and occludin (OCLN) were assessed using immunohistochemistry and western blotting. Blood samples were obtained from the abdominal aorta for ELISA measurements of advanced glycation end products (AGEs) concentrations, and placental receptor for AGEs (RAGE) and vascular endothelial growth factor (VEGF) expression was assessed using reverse transcription­quantitative PCR. In addition, western blotting was used to measure placental NF­κB. Compared with in the control group, EB leakage was markedly increased in GDM group rats; this was associated with reduced ZO­1 and OCLN expression. Conversely, LMWH attenuated this increase in placental permeability in rats with GDM and also mediated a partial recovery of ZO­1 and OCLN expression. Blood glucose and serum AGEs concentrations did not differ between the GDM and GDM + LMWH groups. Furthermore, LMWH treatment resulted in decreases in RAGE and VEGF mRNA expression levels, which were upregulated in the GDM group, whereas it had the opposite effect on the expression of NF­κB. In conclusion, GDM was associated with increased placental permeability and this may be linked with changes in TJs. LMWH intervention mediated protection against this GDM­associated shift in placental permeability via the RAGE/NF­κB pathway.