Evaluation of prenatal and postnatal ultrasonography for the diagnosis of fetal double bubble sign.

Background: Congenital duodenal obstruction (CDO) is commonly detected antenatally through the presence of the "double bubble" sign on prenatal ultrasound, denoting dilatation of the stomach and duodenum. Subsequent postnatal ultrasonography plays a pivotal role in determining the causes of obstruction, thereby informing surgical strategies and neonatal management. The aim of this study was to investigate the diagnostic accuracy of postnatal ultrasonography in comparison to that of prenatal ultrasound and surgical findings in a cohort of 43 patients with fetal double bubble sign. Methods: A total of 43 patients, comprising 24 males and 19 females, who exhibited double bubble sign on prenatal ultrasound were subjected to postnatal ultrasound assessment at a tertiary care facility during the 2018-2023 period. The accuracy of both pre-and postnatal ultrasonography in the identification and diagnosis of CDO, as well as its underlying causes, was compared to that of the established gold standard of surgical findings. Results: The accuracy rates for prenatal and postnatal ultrasonic diagnosis of CDO were 97.7% (42/43) and 100% (42/42), respectively. In terms of etiological diagnosis, prenatal and postnatal ultrasound correctly identified the causes of obstruction in 45.2% (19/42) and 81.0% (34/42) of cases, respectively, as confirmed by surgical intervention. Conclusions: The presence of the prenatal double bubble sign serves as a highly reliable indicator for CDO. Additionally, postnatal ultrasonography proved to be a valuable tool in refining the diagnosis and determining the underlying causes of obstruction in neonates.

Analysis of risk factors associated with incision complications in modified "L" approach for calcaneal fracture.

OBJECTIVE: This study aimed to identify risk factors associated with incision complications following the modified "L" approach for calcaneal fractures. METHODS: Data from 100 patients treated with the modified "L" approach for calcaneal fractures between January 2018 and December 2021 were analyzed. These included 52 cases in the poorly healing group and 48 in the well-healing group. Variables such as patient age, sex, body mass index, fracture type (Sanders classification), smoking history, alcohol consumption, diabetes status, timing of surgery, tourniquet use, bone grafting, suture method, and postoperative incision care were evaluated. A nomogram was developed using R software to predict the risk of incision complications, validated through the area under the ROC curve, C-index, and decision curve analysis. RESULTS: Both univariate and multivariate regression analyses identified fracture type, smoking history, diabetes, timing of surgery, and duration of tourniquet application as significant predictors of incision complications. These factors were incorporated into a clinical predictive nomogram. The nomogram's calibration curves demonstrated high accuracy, both internally and externally. The unadjusted concordance indes (C-index) was 0.793 [95% confidence interval (CI), 0.825-0.995], and the area under the curve for the nomogram was 0.7875882. Decision curve analysis confirmed the clinical applicability of the model at a threshold probability of 20-60%. CONCLUSION: We have developed a reliable clinical nomogram to predict the risk factors for incision complications in the modified "L" approach for calcaneal fractures, enhancing decision-making in clinical settings.

Transcription-coupled DNA repair protects genome stability upon oxidative stress-derived DNA strand breaks.

Elevated oxidative stress, which threatens genome stability, has been detected in almost all types of cancers. Cells employ various DNA repair pathways to cope with DNA damage induced by oxidative stress. Recently, a lot of studies have provided insights into DNA damage response upon oxidative stress, specifically in the context of transcriptionally active genomes. Here, we summarize recent studies to help understand how the transcription is regulated upon DNA double strand breaks (DSB) and how DNA repair pathways are selectively activated at the damage sites coupling with transcription. The role of RNA molecules, especially R-loops and RNA modifications during the DNA repair process, is critical for protecting genome stability. This review provides an update on how cells protect transcribed genome loci via transcription-coupled repair pathways.

Naphthalene Diimide-Based Metallacage as an Artificial Ion Channel for Chloride Ion Transport.

Developing synthetic molecular devices for controlling ion transmembrane transport is a promising research field in supramolecular chemistry. These artificial ion channels provide models to study ion channel diseases and have huge potential for therapeutic applications. Compared with self-assembled ion channels constructed by intermolecular weak interactions between smaller molecules or cyclic compounds, metallacage-based ion channels have well-defined structures and can exist as single components in the phospholipid bilayer. A naphthalene diimide-based artificial chloride ion channel is constructed through efficient subcomponent self-assembly and its selective ion transport activity in large unilamellar vesicles and the planar lipid bilayer membrane by fluorescence and ion-current measurements is investigated. Molecular dynamics simulations and density functional theory calculations show that the metallacage spans the entire phospholipid bilayer as an unimolecular ion transport channel. This channel transports chloride ions across the cell membrane, which disturbs the ion balance of cancer cells and inhibits the growth of cancer cells at low concentrations.

Meiotic protein SYCP2 confers resistance to DNA-damaging agents through R-loop-mediated DNA repair.

Drugs targeting the DNA damage response (DDR) are widely used in cancer therapy, but resistance to these drugs remains a major clinical challenge. Here, we show that SYCP2, a meiotic protein in the synaptonemal complex, is aberrantly and commonly expressed in breast and ovarian cancers and associated with broad resistance to DDR drugs. Mechanistically, SYCP2 enhances the repair of DNA double-strand breaks (DSBs) through transcription-coupled homologous recombination (TC-HR). SYCP2 promotes R-loop formation at DSBs and facilitates RAD51 recruitment independently of BRCA1. SYCP2 loss impairs RAD51 localization, reduces TC-HR, and renders tumors sensitive to PARP and topoisomerase I (TOP1) inhibitors. Furthermore, our studies of two clinical cohorts find that SYCP2 overexpression correlates with breast cancer resistance to antibody-conjugated TOP1 inhibitor and ovarian cancer resistance to platinum treatment. Collectively, our data suggest that SYCP2 confers cancer cell resistance to DNA-damaging agents by stimulating R-loop-mediated DSB repair, offering opportunities to improve DDR therapy.

Combined exposure to lead and microplastics increased risk of glucose metabolism in mice via the Nrf2/NF-κB pathway.

The purpose of this study was to explore the effects of combined lead (Pb) and two types of microplastic (MP) (polyvinyl chloride [PVC] and polyethylene [PE]) exposure on glucose metabolism and investigate the role of the nuclear factor erythroid 2-related factor 2 (Nrf2)/nuclear factor-kappa B (NF-κB) signaling pathway in mediating these effects in mice. Adult C57BL/6J mice were randomly divided into four groups: control, Pb (100 mg/L), MPs (containing 10 mg/L PE and PVC), and Pb + MPs, each of which was treated with drinking water. Treatments were conducted for 6 weeks. Co-exposure to Pb + MPs exhibited increase glycosylated serum protein levels, insulin resistance, and damaged glucose tolerance compared with the control mice. Additionally, treatment with Pb + MPs caused more severe damage to hepatocytes than when exposed to them alone concomitantly, exposed to Pb + MPs exhibited improved the levels of interleukin-6, tumor necrosis factor-alpha, and malondialdehyde, but reduced superoxide dismutase, glutathione peroxidase, and catalase assay in livers. Furthermore, they increase the Kelch-like ECH-associated protein 1 (Keap1) and phosphorylated p-NF-κB protein levels but reduced the protein levels of heme oxygenase-1 and Nrf2, as well as increased Keap1 mRNA and Nrf2 mRNA. Co-exposure to Pb + MP impacts glucose metabolism via the Nrf2 /NF-κB pathway.

The inhibition of ß-catenin activity by luteolin isolated from Paulownia flowers leads to growth arrest and apoptosis in cholangiocarcinoma.

To develop an inhibitor targeting the Wnt/ß-catenin signaling pathway, flavonoid monomer that can interact with ß-catenin was isolated from Paulownia flowers. Luteolin may form stable hydrogen bonds with ß-catenin by molecular docking. Fluorescence quenching analysis determined the physical interaction between luteolin and ß-catenin. The binding of luteolin to ß-catenin caused a loss of α-helical structure and induced a conformational change through circular dichroism spectroscopy. Luteolin inhibits the activity of the Wnt signaling, causing cholangiocarcinoma (CCA) cell cycle arrest in the G2/M phase, leading to cell apoptosis and inhibition of cell migration. In addition, transcriptome and proteomics analysis showed that the differentially expressed proteins were significantly enriched in the Wnt/ß-catenin pathway. ß-catenin protein in the nucleus was significantly decreased, while C-Myc and cyclin D1 in the CCA cells were significantly decreased after luteolin treatment. Additionally, activation of the Wnt/ß-catenin signaling reversed the inhibitory effect of luteolin on the migration of CCA cells. Therefore, luteolin can directly interact with ß-catenin and act as an inhibitor of ß-catenin, inhibiting proliferation and reducing the migration ability of CCA cells by inhibiting the Wnt/ß-catenin pathway. This study provides a scientific basis for the development of Wnt/ß-catenin pathway inhibitors and the prevention and treatment of CCA.

Impact of neoadjuvant chemotherapy on breast cancer-related lymphedema after axillary lymph node dissection: a retrospective cohort study.

PURPOSE: We aimed to evaluate whether neoadjuvant chemotherapy (NAC) could be a risk factor for breast cancer-related lymphedema (BCRL) associated with axillary lymph node dissection (ALND). PATIENTS AND METHODS: A total of 596 patients with cT0-4N0-3M0 breast cancer who underwent ALND and chemotherapy were retrospectively analyzed between March 2012 and March 2022. NAC was administered in 188 patients (31.5%), while up-front surgery in 408 (68.5%). Univariate and multivariable Cox regression analyses were performed to determine whether NAC was an independent risk factor for BCRL. With propensity score matching (PSM), the NAC group and up-front surgery group were matched 1:1 by age, body mass index (BMI), molecular subtypes, type of breast surgery, and the number of positive lymph nodes. Kaplan-Meier survival analyses were performed for BCRL between groups before and after PSM. Subgroup analyses were conducted to explore whether NAC differed for BCRL occurrence in people with different characteristics. RESULTS: At a median follow-up of 36.3 months, 130 patients (21.8%) experienced BCRL [NAC, 50/188 (26.60%) vs. up-front surgery, 80/408 (19.61%); P = 0.030]. Multivariable analysis identified that NAC [hazard ratio, 1.503; 95% CI (1.03, 2.19); P = 0.033] was an independent risk factor for BCRL. In addition, the hormone receptor-negative/human epidermal growth factor receptor 2-negative (HR-/HER2-) subtype, breast-conserving surgery (BCS), and increased positive lymph nodes significantly increased BCRL risk. After PSM, NAC remained a risk factor for BCRL [hazard ratio, 1.896; 95% CI (1.18, 3.04); P = 0.007]. Subgroup analyses showed that NAC had a consistent BCRL risk in most clinical subgroups. CONCLUSION: NAC receipt has a statistically significant increase in BCRL risk in patients with ALND. These patients should be closely monitored and may benefit from early BCRL intervention.

The Safety of CT-Guided Percutaneous Lung Biopsy in Elderly Patients With Solitary Pulmonary Nodules.

OBJECTIVE: Computed tomography (CT)-guided percutaneous lung biopsy is an effective diagnostic procedure for patients with solitary pulmonary nodules (SPN). The aim of this study is to evaluate the safety of this procedure for elderly patients with SPN. METHODS: A total of 125 patients with SPN who received a CT-guided percutaneous lung biopsy were retrospectively analyzed. Patients were divided into elderly (age 65 and above) and non-elderly groups. The patients' characteristics and procedure-related complications were compared between the two groups. RESULTS: The elderly and non-elderly groups included 74 and 51 patients, respectively. The success rate of a CT-guided percutaneous lung biopsy was 100%. The diagnosis rate of lung cancer in the elderly group was significantly higher than that in the non-elderly group (83.78% vs. 64.70%, p = 0.014). The incidence of pulmonary hemorrhage after lung biopsy in the elderly group (44, 59.45%) was significantly higher than that in the non-elderly group (21, 41.17%, p = 0.044), and moderate hemorrhage was the main contributor. The incidence rate of pneumothorax in the elderly group numerically increased, but the difference did not reach statistical significance. CONCLUSION:  Computed tomography-guided percutaneous lung biopsy was an efficient procedure for diagnosing SPN in elderly patients. Although complication rates were relatively higher in elderly patients, the safety of this procedure was acceptable.

The RNA m5C modification in R-loops as an off switch of Alt-NHEJ.

The roles of R-loops and RNA modifications in homologous recombination (HR) and other DNA double-stranded break (DSB) repair pathways remain poorly understood. Here, we find that DNA damage-induced RNA methyl-5-cytosine (m5C) modification in R-loops plays a crucial role to regulate PARP1-mediated poly ADP-ribosylation (PARylation) and the choice of DSB repair pathways at sites of R-loops. Through bisulfite sequencing, we discover that the methyltransferase TRDMT1 preferentially generates m5C after DNA damage in R-loops across the genome. In the absence of m5C, R-loops activate PARP1-mediated PARylation both in vitro and in cells. Concurrently, m5C promotes transcription-coupled HR (TC-HR) while suppressing PARP1-dependent alternative non-homologous end joining (Alt-NHEJ), favoring TC-HR over Alt-NHEJ in transcribed regions as the preferred repair pathway. Importantly, simultaneous disruption of both TC-HR and Alt-NHEJ with TRDMT1 and PARP or Polymerase θ inhibitors prevents alternative DSB repair and exhibits synergistic cytotoxic effects on cancer cells, suggesting an effective strategy to exploit genomic instability in cancer therapy.

Cluster-Locating Algorithm Based on Deep Learning for Silicon Pixel Sensors.

The application of silicon pixel sensors provides an excellent signal-to-noise ratio, spatial resolution, and readout speed in particle physics experiments. Therefore, high-performance cluster-locating technology is highly required in CMOS-sensor-based systems to compress the data volume and improve the accuracy and speed of particle detection. Object detection techniques using deep learning technology demonstrate significant potential for achieving high-performance particle cluster location. In this study, we constructed and compared the performance of one-stage detection algorithms with the representative YOLO (You Only Look Once) framework and two-stage detection algorithms with an RCNN (region-based convolutional neural network). In addition, we also compared transformer-based backbones and CNN-based backbones. The dataset was obtained from a heavy-ion test on a Topmetal-M silicon pixel sensor at HIRFL. Heavy-ion tests were performed on the Topmetal-M silicon pixel sensor to establish the dataset for training and validation. In general, we achieved state-of-the-art results: 68.0% AP (average precision) at a speed of 10.04 FPS (Frames Per Second) on Tesla V100. In addition, the detection efficiency is on the same level as that of the traditional Selective Search approach, but the speed is higher.

The correlation between prenatal maternal active smoking and neurodevelopmental disorders in children: a systematic review and meta-analysis.

OBJECTIVE: To systematically evaluate the association between maternal active smoking during pregnancy and Tourette syndrome (TS), chronic tic disorder (CTD), and developmental coordination disorder (DCD) in children, and to provide evidence-based medical references to reduce the incidence of neurodevelopmental disorders in children. METHOD: We searched PubMed, Web of Science, Embase, and Cochrane Library to obtain relevant articles published before 4 August 2021. Two reviewers independently assessed the articles for eligibility and extracted data. RESULTS: We included eight studies involving a total of 50,317 participants (3 cohort, 3 case-control, and 2 cross-sectional studies). The pooled effect estimates suggested that prenatal maternal active smoking is related to an increased risk of neurodevelopmental disorders (OR = 1.91, 95% CI: 1.30-2.80), especially DCD (OR = 2.25, 95% CI: 1.35-3.75). Maternal active smoking during pregnancy is not associated with TS (OR = 1.07, 95% CI: 0.66-1.73) in children. CONCLUSION: In this meta-analysis, we found evidence for a correlation between active smoking exposure in pregnant women and neurodevelopmental disorders in children. Owing to the differences in sample size, smoking categories and diagnostic methods, further research is needed to validate our results.

Synchronous multiple primary malignant neoplasms in breast, kidney, and bilateral thyroid: A case report.

BACKGROUND: Multiple primary malignant neoplasms (MPMNs) are rare, while synchronous MPMNs (SMPMNs) are even less common. Owing to the progression of medical technology and the extension of life expectancy, its incidence is gradually increasing. CASE SUMMARY: Although reports of breast and thyroid dual cancers are common, cases of an additional diagnosis of kidney primary cancer within the same individual are rare. CONCLUSION: We present a case of simultaneous MPMN of three endocrine organs, reviewing the relevant literature to enhance our understanding of SMPMNs while emphasizing the increasingly important need for accurate diagnosis and multidisciplinary management whenever this challenging situation arises.

The GLIM criteria as an effective tool for survival prediction in gastric cancer patients.

BACKGROUND: The Global Leadership Initiative on Malnutrition released a new version of the malnutrition criteria (GLIM criteria). To investigate the influence of the GLIM criteria on the long-term efficacy of radical gastric cancer surgery and establish a nomogram to predict the long-term prognosis of patients with gastric cancer. METHODS: A retrospective analysis of 1121 patients with gastric cancer in our department from 2010 to 2013 was performed. A nomogram was established to predict overall survival (OS) based on the GLIM criteria. Patients were divided into the low-risk group (LRG) and high-risk group (HRG) based on the established nomogram. RESULTS: Multivariate Cox regression analyses showed that GLIM criteria was an independent risk factor for the 5-year OS (HR = 1.768, Cl:1.341-2.329, p < 0.001). The C index, AUC and Time-ROC of the nomogram were significantly better than that of GLIM criteria and traditional criteria. The 5-year OS of patients receiving adjuvant chemotherapy in the high-risk group was significantly higher than that of patients without chemotherapy (45.77% vs. 24.73%,p < 0.001). CONCLUSIONS: The GLIM criteria independently influence the long-term outcome of patients after radical gastric cancer surgery. The established nomogram can predict the long-term survival of patients with gastric cancer, and postoperative adjuvant chemotherapy for HRG can significantly improve the 5-year OS of patients.

SAFB2 Inhibits the Progression of Breast Cancer by Suppressing the Wnt/ß-Catenin Signaling Pathway via NFAT5.

Aberrant scaffold attachment factor-B2 (SAFB2) expression is associated with several malignant tumors. In this study, we investigated how SAFB2 worked in the process of breast cancer as well as the underlying mechanism. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting analysis were used to investigate the expression of SAFB2 and nuclear factor of activated T cells 5 (NFAT5). Cellular proliferative ability was detected with cell counting kit 8 (CCK8), colony formation and 5-Ethynyl-2'-deoxyuridine (EdU) staining assays. Cell apoptosis was measured via flow cytometry and western blotting analysis. Wound healing, transwell assays, and western blotting analysis were executed to estimate cell migration and invasion. The relationship between SAFB2 and NFAT5 was verified by RNA immunoprecipitation (RIP) assay and NFAT5 mRNA stability was examined with actinomycin (Act) D assay. Western blotting analysis also tested the expression of Wnt/ß-catenin signaling-associated proteins. As a result, SAFB2 was downregulated in breast cancer cell lines, while NFAT5 was highly expressed in most breast cancer cell lines. Overexpression of SAFB2 suppressed the proliferation, migration, and invasion while exacerbated the apoptosis of breast cancer cells. SAFB2 interacted with NFAT5 mRNA and declined the stability of NFAT5 mRNA. Overexpression of NFAT5 counteracted anti-proliferative, anti-metastatic and pro-apoptotic effects of SAFB2 in breast cancer cells. Mechanistically, SAFB2 overexpression inhibited the Wnt/ß-catenin signaling pathway, while this effect was partially eliminated by NFAT5. Collectively, SAFB2 hindered breast cancer development and inactivated Wnt/ß-catenin signaling via regulation of NFAT5, suggesting that SAFB2 might be a promising therapeutic target for breast cancer.

Replication Origin Deletion Enhances Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate) Synthesis in Haloarchaea.

Although the use of multiple replication origins for chromosome replication has been widely characterized in haloarchaea, whether it is possible to manipulate the chromosome copy number by their genetic engineering is not known, and how it would affect the cell functioning is poorly understood. Here, we demonstrate that deletion of the three active chromosomal origins in Haloferax mediterranei remarkably reduces its DNA amounts and ploidy numbers. Consequently, the mutant strain H. mediterranei Δ123 is more sensitive to UV and mitomycin C. Surprisingly, the cell size increases by 21.2%, and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) production in shake flask culture enhances from 7.23 to 8.11 g/L in ΔEPSΔ123, although there is also a decrease in cell growth. In this mutant, the chromosomal copy number decreases, whereas the pha-encoding pHM300 megaplasmid copy number increases. Moreover, our transcriptome analysis reveals that the genes involved in primary metabolisms are significantly downregulated in ΔEPSΔ123, whereas those responsible for starch utilization and precursor supplying for PHBV monomers are upregulated. This indicates that more energy and carbon flux is redirected from primary metabolism to PHBV synthesis, thereby enhancing its PHBV accumulation. These findings may therefore provide a rational design to enhance PHBV synthesis by simply tuning the replication origins to modulate the chromosome/megaplasmid copy number ratio and subsequently influence cellular metabolism and physiological functions. IMPORTANCE The haloarchaeon Haloferax mediterranei is a potential producer of PHBV (100% biodegradable plastic) from inexpensive carbon sources. We previously reported that H. mediterranei possessed three active chromosomal origins and, when these origins were deleted, a dormant origin was activated to initiate the replication of chromosome. In this context, in the present study, we first found a close connection between replication initiation and PHBV accumulation. We describe the potential industrial advantages of the strain H. mediterranei ΔEPSΔ123, which includes the enlargement of cell volume by 21.2% and enhancement of PHBV production by 11.2%. We further reveal the possible mechanism that contributes to the greater PHBV production in the ΔEPSΔ123 strain. Overall, we provide here a conceptual advance in the field of synthetic biology by modulating chromosome replication to improve the production of bio-based chemicals.

Effectiveness of Perioperative Comprehensive Evaluation of Hip Fracture in the Elderly.

Objective: The objective is to observe the effect of Comprehensive Geriatric Assessment (CGA) in the perioperative period of hip fracture. Methods: From October 2018 to October 2021, 155 patients over the age of 65 diagnosed with hip fracture and treated with surgery at the Department of Trauma Orthopaedics of General Hospital of Ningxia Medical University were randomly divided into two groups using a prospective research method. A total of 70 cases in the CGA group received a perioperative comprehensive assessment of the geriatric, and 85 cases in the control group received routine medical consultation. Results: Elderly patients with hip fractures have a high comorbidity index. Patients with abnormal daily activity before injury accounted for 55%, the abnormal rate of nutrition was 58.1%, the abnormal rate of cognition, anxiety, and depression was 81.8%, and 77.3% of the patients were in a weak state. There was no significant difference in age, gender, ASA grade, fracture type, and operation mode between the two groups, but there were significant differences in operation rate at 48 h (χ 2 = 22.153; P ≤ 0.001), preoperative waiting time (Z = -6.387; P ≤ 0.001), total hospital stay (Z = -11.756; P ≤ 0.001), and incidence of postoperative delirium (χ 2 = 23.897; P ≤ 0.001). Conclusions: The implementation of CGA shortened the preoperative waiting time and total hospital stay, increased the 48 h operation rate, and reduced the incidence of postoperative delirium.

Severity of Hospitalized Children with Anti-NMDAR Autoimmune Encephalitis.

Background: Information on the clinical characteristics and severity of autoimmune encephalitis with antibodies against the N-methyl-d-aspartate receptor (NMDAR) in children is attracting more and more attention in the field of pediatric research. Methods: In this retrospective cohort study, all cases (n = 67) were enrolled from a tertiary children's hospital, from 2017 to 2020. We compared severe cases that received intensive care unit (ICU) care with nonsevere cases that did not receive ICU care and used machine learning algorithm to predict the severity of children, as well as using immunologic and viral nucleic acid tests to identify possible pathogenic triggers. Results: Mean age of children was 8.29 (standard deviation 4.09) years, and 41 (61.19%) were girls. Eleven (16.42%) were admitted to the ICU, and 56 (83.58%) were admitted to neurology ward. Ten individual parameters were statistically significant differences between severe cases and nonsevere cases (P < .05), including headache, abnormal mental behavior or cognitive impairment, seizures, concomitant tumors, sputum/blood pathogens, blood globulin, blood urea nitrogen, blood immunoglobulin G, blood immunoglobulin M, and number of polynucleated cells in cerebrospinal fluid. Random forest regression model presented that the overall prediction power of severity reached 0.806, among which the number of polynucleated cells in cerebrospinal fluid contributed the most. Potential pathogenic causes exhibited that the proportion of mycoplasma was the highest, followed by Epstein-Barr virus. Conclusion: Our findings provided evidence for early identification of autoimmune encephalitis in children, especially in severe cases.

Leucine-rich repeat kinase-2 deficiency protected against cardiac remodelling in mice via regulating autophagy formation and degradation.

Introduction: Leucine-rich repetitive kinase-2 (LRRK2) is a Parkinson's disease-related gene that also participates in many inflammatory diseases. However, the functional role of LRRK2 in cardiovascular disease is not clear. Objective: In this study, we aimed to elucidate the role of LRRK2 in cardiac remodelling under pressure overload. Methods: Aortic banding surgery was performed to induce cardiac remodelling in a LRRK2 knockout mouse model. A cardiomyocyte remodelling model was established by phenylephrine (PE) stimulation in neonatal rat cardiomyocytes. Results: LRRK2 was upregulated in remodelled mouse hearts and cardiomyocytes. Cardiac hypertrophy, fibrosis and dysfunction were ameliorated in LRRK2 knockout mice. LRRK2 silencing protected against the PE-induced cardiomyocyte hypertrophic response, while LRRK2 over-expression worsened the PE-induced hypertrophic response in cardiomyocytes. Decreased autophagy was observed in remodelled cardiomyocytes, whereas LRRK2 silencing increased autophagy levels and LRRK2 overexpression reduced autophagy levels. The autophagy inhibitors 3-MA, bafilomycin and chloroquine reversed the protective effects of LRRK2 deficiency. The autophagy activator rapamycin reversed the deleterious effects of LRRK2 overexpression. We found that LRRK2 inhibited Bcl-2 phosphorylation, thus decreasing the phosphorylation of Beclin1. The protective effects of LRRK2 knockout were partly counteracted by Beclin1(+/-) in vivo and Beclin1 silencing in vitro. We also observed an interaction between LRRK2 and Rab7, an autolysosome degradation-associated protein, which caused Rab7 downregulation. Rab7 knockdown almost completely reversed LRRK2 silencing-induced protection of cardiomyocytes. Conclusion: LRRK2 deficiency protected against cardiac remodelling under pressure overload by increasing Bcl-2/Beclin1 and Rab7-regulated autophagy levels in the heart.