Plexin D1 negatively regulates macrophage-derived foam cell migration via the focal adhesion kinase/Paxillin pathway.

BACKGROUND: Macrophage-derived foam cell formation is a hallmark of atherosclerosis and is retained during plaque formation. Strategies to inhibit the accumulation of these cells hold promise as viable options for treating atherosclerosis. Plexin D1 (PLXND1), a member of the Plexin family, has elevated expression in atherosclerotic plaques and correlates with cell migration; however, its role in macrophages remains unclear. We hypothesize that the guidance receptor PLXND1 negatively regulating macrophage mobility to promote the progression of atherosclerosis. METHODS: We utilized a mouse model of atherosclerosis based on a high-fat diet and an ox-LDL- induced foam cell model to assess PLXND1 levels and their impact on cell migration. Through western blotting, Transwell assays, and immunofluorescence staining, we explored the potential mechanism by which PLXND1 mediates foam cell motility in atherosclerosis. RESULTS: Our study identifies a critical role for PLXND1 in atherosclerosis plaques and in a low-migration capacity foam cell model induced by ox-LDL. In the aortic sinus plaques of ApoE-/- mice, immunofluorescence staining revealed significant upregulation of PLXND1 and Sema3E, with colocalization in macrophages. In macrophages treated with ox-LDL, increased expression of PLXND1 led to reduced pseudopodia formation and decreased migratory capacity. PLXND1 is involved in regulating macrophage migration by modulating the phosphorylation levels of FAK/Paxillin and downstream CDC42/PAK. Additionally, FAK inhibitors counteract the ox-LDL-induced migration suppression by modulating the phosphorylation states of FAK, Paxillin and their downstream effectors CDC42 and PAK. CONCLUSION: Our findings indicate that PLXND1 plays a role in regulating macrophage migration by modulating the phosphorylation levels of FAK/Paxillin and downstream CDC42/PAK to promoting atherosclerosis.

Emodin relieves morphine-stimulated BV2 microglial activation and inflammation through the TLR4/NF-κB/NLRP3 pathway.

The objective of this study is to disclose the role of emodin, a natural anthraquinone derivative that has been proposed to suppress microglial activation and inflammation, in morphine tolerance. Here, cell counting kit-8 method assayed the viability of BV2 microglial cells treated by ascending concentrations of emodin. In emodin-pretreated BV2 microglial cells challenged with morphine with or without transfection of toll-like receptor 4 (TLR4) overexpression plasmids, transwell assay measured cell migration. Immunofluorescence staining and western blot detected the expression of microglial markers. Inflammatory levels were subjected to ELISA and western blot. BODIPY 581/591 C11 assay estimated lipid reactive oxygen species activity. Iron assay kit examined total iron content. Western blot tested the expression of ferroptosis- and TLR4/nuclear factor-kappaB (NF-κB)/NOD-like receptor 3 (NLRP3) pathway-associated proteins. Molecular docking predicted the binding affinity of emodin to TLR4. Emodin was noted to obstruct the migration, activation, inflammatory response, and ferroptosis of BV2 microglial cells induced by morphine. In addition, emodin had a high binding affinity with TLR4 and inactivated TLR4/NF-κB/NLRP3 pathway in morphine-challenged BV2 microglial cells. Upregulation of TLR4 partially countervailed the protective role of emodin against morphine-elicited BV2 microglial cell migration, activation, inflammation, and ferroptosis. Accordingly, emodin might target TLR4 and act as an inactivator of TLR4/NF-κB/NLRP3 pathway, thus inhibiting BV2 microglial activation and inflammation to mitigate morphine tolerance.

Al@SiO2 Core-Shell Fillers Enhance Dielectric Properties of Silicone Composites.

Over the past decade, there has been significant interest in polysiloxane-based dielectric elastomers as promising soft electroactive materials. Nevertheless, the natural low permittivity of polydimethylsiloxane has limited its practical applications. In this study, we have developed silicone rubber/Al@SiO2 composites with a high dielectric constant, low dielectric loss, and high electrical breakdown strength by controlling the shell layer thickness and the content of the core-shell filler. We also investigated the dielectric behavior of the composites. The use of core-shell fillers has increased the Maxwell-Wagner-Sillars (MWS) relaxation process while reducing the dielectric loss of direct current conductance in silicone rubber composites. Moreover, the temperature dependence of the MWS relaxation time in the composites follows the Arrhenius equation. This strategy of increasing the permittivity of silicone composites through core-shell structural fillers can inspire the preparation of other high dielectric constant composites.

Identification of Potential Differentially-Methylated/Expressed Genes in Chronic Obstructive Pulmonary Disease.

Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory lung disease that causes obstructed airflow from the lungs. DNA methylation can regulate gene expression. Understanding the potential molecular mechanism of COPD is of great importance. The aim of this study was to find differentially methylated/expressed genes in COPD. DNA methylation and gene expression profiles in COPD were downloaded from the dataset, followed by functional analysis of differentially-methylated/expressed genes. The potential diagnostic value of these differentially-methylated/expressed genes was determined by receiver operating characteristic (ROC) analysis. Expression validation of differentially-methylated/expressed genes was performed by in vitro experiment and extra online datasets. Totally, 81 hypermethylated-low expression genes and 121 hypomethylated-high expression genes were found in COPD. Among which, 9 core hypermethylated-low expression genes (CD247, CCR7, CD5, IKZF1, SLAMF1, IL2RB, CD3E, CD7 and IL7R) and 8 core hypomethylated-high expression genes (TREM1, AQP9, CD300LF, CLEC12A, NOD2, IRAK3, NLRP3 and LYZ) were identified in the protein-protein interaction (PPI) network. Moreover, these genes had a potential diagnostic utility for COPD. Some signaling pathways were identified in COPD, including T cell receptor signaling pathway, cytokine-cytokine receptor interaction, hematopoietic cell lineage, HTLV-I infection, endocytosis and Jak-STAT signaling pathway. In conclusion, differentially-methylated/expressed genes and involved signaling pathways are likely to be associated with the process of COPD.

Identification of candidate aberrant differentially methylated/expressed genes in asthma.

BACKGROUND: Asthma is an important non-communicable disease worldwide. DNA methylation is associated with the occurrence and development of asthma. We are aimed at assuring differential expressed genes (DEGs) modified by aberrantly methylated genes (DMGs) and pathways related to asthma by integrating bioinformatics analysis. METHODS: One mRNA dataset (GSE64913) and one gene methylation dataset (GSE137716) were selected from the Gene Expression Omnibus (GEO) database. Functional enrichment analysis was performed using GeneCodies 4.0 database. All gene expression matrices were analyzed by Gene set enrichment analysis (GSEA) software. STRING was applied to construct a protein-protein interaction (PPI) network to find the hub genes. Then, electronic validation was performed to verify the hub genes, followed by the evaluation of diagnostic value. Eventually, quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to detect the expression of hub genes. RESULTS: In total, 14 hypomethylated/high-expression genes and 10 hypermethylated/low-expression genes were obtained in asthma. Among them, 10 hub genes were identified in the PPI network. Functional analysis demonstrated that the differentially methylated/expressed genes were primarily associated with the lung development, cytosol and protein binding. Notably, HLA-DOA was enriched in asthma. FKBP5, WNT5A, TM4SF1, PDK4, EPAS1 and GMPR had potential diagnostic value for asthma. CONCLUSION: The project explored the pathogenesis of asthma, which may provide a research basis for the prediction and the drug development of asthma.

Perceived impact of the COVID-19 pandemic on infection containment training and mental state of dental residents in China: A longitudinal study.

Background: COVID-19 has presented a challenge for dental settings and dental schools: how to continue providing dental care and maintain education during the pandemic while remaining healthy. We highlight the necessity of infection containment control training for dental residents and rethink the tasks of safeguarding trainees' health and cultivating their abilities to deal with public health crises in the future. This paper may also serve as a health policy reference for policy makers. Objective: The study aimed to compare the formats, frequency, contents, emphasis, and test scores of infection containment control training pre- and post-pandemic. Besides, after the COVID-19 outbreak, we assessed the increased anxiety level, communication difficulties, and confidence of dental residents impacted by the pandemic. Methods: A total of 251 dental residents in Stomatological Hospital of Chongqing Medical University were recruited to complete a questionnaire of their routine involvement in infection control training before and after the COVID-19 outbreak. A self-designed 10-point Likert scale was used to assess the increased anxiety level, communication difficulties, and confidence in facing with the future public health crisis impacted by the pandemic. Results: After the outbreak, although more trainees chose online assessment than offline assessment, most of them (74.90%) still preferred in-person training rather than online training. Contents that trainees had been focusing on were affected by the COVID-19 outbreak. Thereafter, they were more inclined to learn crisis management. Over half of the participants (56.17%) participated in training more frequently after the outbreak. However, postgraduate students participated in training less frequently than others after the outbreak (p < 0.01). First-year trainees accounted for the majority in the population who emphasized considerably on infection control training and whose test scores had increased after the outbreak. In addition, the percentage of women scoring increasingly in post-pandemic assessment was significantly higher than that of men. In this study, the average increased anxiety level caused by COVID-19 was 5.51 ± 2.984, which was positively related to communication difficulties with patients caused by the pandemic. The trainees whose homes were located in Hubei Province showed higher increased anxiety levels (8.29 ± 2.93) impacted by the pandemic than the trainees from other provinces (p < 0.05). However, the former's confidence in coping with future public health crises was not significantly different from that of others (p > 0.05). Conclusions: Owing to the impact of COVID-19, the contents that the trainees focused on, frequency, emphasis, and test scores of infection containment control training were changed. Some recommendations have been provided for policy makers to attach importance to crisis-based training to cultivate dental residents in the post-pandemic era.

Analytical method for detection and quantification of new emerging drug etaqualone in human blood and urine by gas chromatography tandem mass spectrometry.

Analytical procedure for detection and quantification of etaqualone in human blood and urine using GC-MS/MS was established and applied to authentic human samples obtained from volunteers. A liquid-liquid extraction method was employed. Each 1.0 mL of blood or urine was alkalized and extracted with diethyl ether. The solvent layer was evaporated to dryness and reconstituted with methanol then analyzed by GC-MS/MS. linear relationships within the concentration range of 1-100 ng/mL were obtained in calibrators for both blood and urine, demonstrating correlation coefficients values being>0.999. For blood and urine samples, the intra-day assay precision and accuracy values are each less than 3.65%, 7.13%, and 6.02%, 9.12%; those values of the inter-day assay are each less than 1.82%, 6.74%, and 3.99%, 7.41%. The extraction recovery rates for etaqualone ranged from 98.7% to 106%. The lower limit of quantifications was 1.0 ng/mL in both blood and urine. Stabilities of etaqualone in blood and urine were satisfactory under various temperatures within 15 days. 8.51 and 2.06 ng/mL of etaqualone in blood and urine were detected at 4 h later oral ingestion; 6.91 and 3.94 ng/mL of etaqualone were also detected 30 min and 2 h later smoking from blood and urine.

Propofol inhibits the malignant development of osteosarcoma U2OS cells via AMPK/FΟΧO1-mediated autophagy.

It has previously been reported that propofol regulates the development of human osteosarcoma (OS). However, the specific molecular mechanisms underlying the effect of propofol on OS remain poorly understood. Therefore, the aim of the present study was to explore the effects of propofol on OS U2OS cells and the potential underlying mechanism. The Cell Counting Kit-8 and colony formation assays were performed to assess cell viability and proliferation. Furthermore, cell apoptosis was assessed using the TUNEL assay and western blotting. Wound healing and Transwell assays were performed to evaluate OS cell migration and invasion abilities, respectively. The protein expression levels of epithelial-mesenchymal transition (EMT)-, autophagy- and adenosine monophosphate-activated protein kinase (AMPK)/FOXO1 signaling pathway-related proteins were also determined using western blotting. The results demonstrated that propofol significantly reduced the viability of OS cells and promoted autophagy in a dose-dependent manner. Moreover, cell treatment with propofol significantly enhanced the protein expression levels of phosphorylated (p)-AMPK and FOXO1, while decreasing the protein levels of p-FOXO1. Furthermore, treatment with propofol significantly suppressed cell viability, migration and invasion abilities and the EMT of OS cells, and potentially promoted cell apoptosis via inducing autophagy via the AMPK/FOXO1 signaling pathway. In summary, the present study indicated that propofol potentially had an inhibitory effect on the development of OS cells via AMPK/FOXO1-mediated autophagy. These results have therefore provided an experimental basis for further studies into the therapeutic effect of propofol on OS.

Screening Potential Drugs for the Development of NAFLD Based on Drug Perturbation Gene Set.

Nonalcoholic fatty liver disease (NAFLD) has become one of the problems affecting the health of the population worldwide. The progressive disease includes nonalcoholic steatohepatitis (NASH) and fibrosis, which with no approved therapy, system identification of effective drugs remains challenging. In this work, we applicated drug perturbation gene set enrichment analysis to screen drugs for the development of NAFLD. A total 15490 small-molecule compounds were analyzed in our study; based on the p value of enrichment score, 7 small-molecule compounds were found to have a potential role in NASH and fibrosis. After pathway analyses, we found indoximod had effects on nonalcoholic fatty liver disease through regulated TNFa, AP-1, AKT, PI3K, etc. Furthermore, we established the NAFLD cell model with LO2 cells induced using PA; ELISA showed that the levels of TG, ALT, and AST were significantly improved by indoximod. In summary, our study offers optimal therapeutic drugs, which may provide novel insight into the precise treatment of NAFLD and promote researches.

Changes in the intestinal expression of drug metabolism-related genes in a piglet model of parenteral nutrition.

BACKGROUND: Parenteral nutrition (PN) may serve as a nutritional supportive therapy accompanied by oral medication, but the effect of PN on intestinal expression of drug metabolism-related genes remains unknown. METHODS: Twelve Bama piglets receiving PN for 14 days were used as in vivo model. Changes in intestinal drug metabolism-related genes were examined by proteomic analysis. Serum levels of fibroblast growth factor 19 (FGF19) were determined by ELISA, and the effect of FGF19 on the expression of drug metabolism-related genes was examined using murine ileum organoids. RESULTS: A total of 1063 differentially expressed proteins were identified in PN group. Of note, two drug transporters (Abcb1 and Abcc2) were significantly decreased in PN group, along with two glutathione-related drug-metabolizing enzymes, glutathione peroxidase (Gpx2) and glutathione S-transferase (Gsta1). Serum FGF19 levels were dramatically reduced in PN group. Treatment with recombinant FGF19 in vitro dose-dependently up-regulated the expression of Abcb1, Abcc2, Gpx2 and Gsta1 in organoids. CONCLUSION: Our data indicated that intestinal drug metabolism-related genes were significantly dysregulated under PN, and some of the changed genes were attributed to gut-derived FGF19.

Does the Efficacy of Behavior Management Techniques Differ Between Children From Single-Child and Multi-Child Families?: A Quasi-Experimental Study.

Aim: Behavior management techniques (BMTs) efficiently deliver dental treatment to children with dental anxiety. The objective of this quasi-experimental study was to examine whether the efficacy of BMTs applied for the improvement of compliance in pediatric patients differs between children 3-10-year-olds from single-child and multi-child families. Materials and Methods: In this quasi-experimental, 197 caregiver-child couples were divided into two groups: single-child group (116 couples) and multi-child group (81 couples). Children's pre- and post-treatment anxiety levels were measured by facial mood scale (FMS) and Frankl Behavior Rating Scale (FBRS), respectively.Caregivers' dental anxiety was measured by the Chinese version of the Modified Dental Anxiety Scale (MDAS), which was included in the self-designed questionnaire. Data were analyzed by using the Mann-Whitney U-test, chi-square tests, and binary multivariate regression analysis. Results: There was no statistically significant difference in the demographic characteristics of the children between the two groups. BMTs were found to be capable of reducing children's dental anxiety (CDA): the compliance rate was 45.69-88.79% in the single-child group and 44.44-85.79% in the multi-child group pre- and post-BMTs, but there was no significant difference in the change of compliance between the two groups (p > 0.05). In the subgroup analysis, parenting style (odds ratio [OR] = 0.054, p < 0.05) and father's education (OR = 8.19, p < 0.05) affected the varies of children's compliance in the single-child group. In contrast, in the multi-child group, gender (OR = 8.004, p < 0.05) and mother's occupation (OR = 0.017, p < 0.05) were associated with these changes in compliance. Conclusions: In this study, BMTs were proved to be beneficial in improving compliance in 3- to 10-year-olds children in dental treatment. Though there was no significant difference in the change of compliance between children from single-child and multi-child families, different associated factors may affect the two groups. Therefore, the related family factors should be taken into account when professionals manage each child's behavior in dental practice.

Three-dimensional digital measurement of the facet joint in normal individual and lumbar intervertebral disc herniation patients aged 13 - 18 years.

BACKGROUND: Lumbar facet joint is an important element of spinal "three-joint complex". Whether there is a relationship between strange structure of facet joint and adolescent lumbar disc herniation (ALDH) is nonetheless controversial, and the current research is mainly centered on adults. OBJECTIVE: To find out the normal lumbar facet joints between 13 and 18 years old to provide anatomical basis for early diagnosis and therapy of lumbar disc herniation. METHODS: CT imaging information of 32 sufferers with lumbar disc herniation aged from 13 to 18 years old in Inner Mongolia have been collected as the ALDH group, and 62 wholesome subjects in the equal period had been chosen as the normal group. Uncooked records of continuous scanning lumbar tomography pix were imported into MIMICS 21.0 for evaluation and size in DICOM format. The parameters include facet joint height, facet joint width, et al. RESULTS: 1. The left and right transverse angle of L5S1 segment in the ALDH group were (52.41 ± 9.2) ° and (55.99 ± 10.91) ° (P < 0.05), and the differences were statistically significant. The right side is larger than the left side. 2. Facet joint thickness in L3-L5 segment of the normal group was significantly higher than that of male (1.63 ± 0.32) mm than that of female (1.38 ± 0.25) mm; In 16-18 years old group, comparison of facet joint cross-sectional area was statistically significant (22.1 ± 3.04) mm2 in male than (18.92 ± 3.71) mm2 in female. 3. In comparison between normal and ALDH group, there was significant difference in L3-4 transverse angle (P < 0.05), L4-5 facet joint height and facet joint thickness (P < 0.05), L5S1 facet joint thickness and transverse angle (P < 0.05). CONCLUSION: When ALDH occurs in the L5S1 segment, there is a substantial difference between the left and right sides of the transverse angle, and there is a difference in the thickness and the facet joint cross-sectional area between males and females, which is generally larger in males than in females. Facet joint height is larger, transverse angle of left and right is asymmetric, inferior articular process is larger, and facet joint thickness is smaller can indicate that lumbar disc herniation is effortless to occur.

Analysis of sagittal curvature and its influencing factors in adolescent idiopathic scoliosis.

ABSTRACT: This study aimed to explore the characteristics of changes in the sagittal arrangement of the spine between adolescent patients with idiopathic scoliosis (AIS) and normal adolescents, the risk factors for AIS and the factors affecting the progress of AIS.X-ray images of the full length of the spine in standing position were taken in AIS patients and normal adolescents. Radiographic measurements made at intermediate follow-up included the following:C1 and C2 cervical lordosis and C2 - C7 curvature of cervical lordosis, C2-C7sagittal horizontal distance (C2-C7SagittalVerticalAxis, C2-C7SVA), TS-CL, after thoracic lobe (Thoracic Kyphosis, TK), thoracic lumbar segment Angle (thoracolumbar kyphosis, [TLK]), lumbar lordosis Angle (Lumbar Lordosis, LL), sacral slope Angle (Sacrum Slope, SS), pelvic tilt Angle (Pelvic Tilt, PT), pelvic incidence (PI), L5 Incidence (Lumbar5 Slope (L5S), L5 incidence (Lumbar5 Incidence (L5I), sagittal horizontal distance (CSVA), lower depression Angle of the 2nd cervical spine. The difference of sagittal plane parameters between AIS group and normal adolescent group was compared. To evaluate the progress of AIS, correlation analysis was conducted between diagonal 2 and other parameters. The main risk factors of AIS were determined by binary Logistic analysis.The CSVA of AIS patients was higher than that of healthy adolescents (AIS: 27.64 ±â€Š19.56) mm. Healthy adolescents: (17.74 ±â€Š12.8) mm), L5S (AIS: 19.93°= 7.07° and healthy adolescents: 15.38°= 7.78°, P = .024 < .05), C2 downward sag Angle (AIS: 15.12°= 2.7°;Healthy adolescents: 12.97°= 4.56°); AIS patients had lower TS-CL (AIS: 22.48 ±â€Š6.09 and healthy adolescents: 28.26°= 10.32°), PT (AIS: 10.42°= 4.53° and healthy adolescents: 15.80°=7.68°), (AIS: 41.87°=9.72° and healthy adolescents: 48.75°= 8.22°). The main risk factor for idiopathic scoliosis in adolescents was L5 (OR = 1.239, 95%CI = 1.049-1.463, P = .012 < .05).L5S is a major risk factor for idiopathic scoliosis in adolescents. The larger PI is, the higher the risk of scoliosis progression is. In AIS patients, lumbar lordosis is increased, cervical lordosis is reduced, and even cervical kyphosis occurs.

Surgical treatment for prostatic utricle cyst in children: A single-center report of 15 patients.

OBJECTIVES: To report our single-center experience of the management of children with prostatic utricle cysts. METHODS: We retrospectively analyzed 15 children who were incidentally found to have a prostatic utricle cyst and were admitted to our department between October 2013 and August 2020. Clinical characteristics and management were collected and catalogued. RESULTS: Recurrent genitourinary tract infections were the most frequent complaint, and two-thirds of patients also had hypospadias. A connection between the posterior urethra and the prostatic utricle cyst was found in all cases. Two patients directly had their progressively enlarging prostatic utricle cyst resected laparoscopically. Endoscopic techniques were used in 13 patients, two of whom underwent laparoscopic excision for repeated symptoms. The mean (range) follow-up period was 34.9 (2-82) months. No recurrences were observed in four patients who underwent prostatic utricle cyst excision and eight patients who received endoscopic treatment. Three patients had recurrent symptoms after endoscopic treatment and were managed by nonsurgical treatment. CONCLUSIONS: Prostatic utricle cyst is a rare disease which can cause recurrent genitourinary tract infections. Extra attention should be paid to evaluation for prostatic utricle cyst in children with external genital anomalies. Retrograde urethrogram and magnetic resonance imaging are useful tools with which to distinguish prostatic utricle cyst from other cystic lesions that are located in the midline pelvis in male patients. Individualized treatment is appropriate when considering fertility preservation, recurrences and malignancy. Laparoscopic excision is feasible for symptomatic and large prostatic utricle cyst. Regular long-term monitoring is recommended for all patients with prostatic utricle cyst.

The Interplay Between Language Form and Concept During Language Switching: A Behavioral Investigation.

Language switching involves multiple processing stages. Previous studies have not dissociated the cognitive process underlying language form switches and concept switches. Here, we examined the two factors using a novel language-switching paradigm. Chinese-English bilinguals named individually presented pictures in either Chinese or English according to a language cue. Pictures in two consecutive trials represented either identical, semantically related, or unrelated concepts. Results showed both language (form) switch costs and concept switch costs. The interaction between these two factors suggested that the effects were additive, with the longest naming response times observed when two pictures were semantically unrelated and involved a switch between languages. These findings suggest that the functional loci of the language control mechanism occur at multiple processing stages. Implications of the findings are discussed within current models of language processing in bilinguals.

Aligned electrospun cellulose scaffolds coated with rhBMP-2 for both in vitro and in vivo bone tissue engineering.

Physical properties of scaffolds such as nanofibers and aligned structures have been reported to exert profound effects on the growth and differentiation of stem cells due to their homing-effect features and contact guidance. However, the biological function of aligned nanofibers utilized as bone-scaffold has not been rigorously characterized. In the present study, aligned electrospun cellulose/CNCs nanocomposite nanofibers (ECCNNs) loaded with bone morphogenic protein-2 (BMP-2) were used for the first time to investigate (1) in vitro osteogenic differentiation of human mesenchymal stem cells (BMSCs) and (2) in vivo collagen assembly direction and cortical bone regeneration. Aligned ECCNNs scaffolds loaded with BMP-2 possess good biological compatibility. The growth orientation of BMSCs followed the underlying aligned nanofiber morphology, accompanied with increased alizarin red stain, alkaline phosphatase (ALP) activity and calcium content in vitro while, a rabbit calvaria bone defect model was used in an in vivo study with micro CT and histology analyses.

Butyrate promotes the adaptation of intestinal smooth muscle cells through the yes-associated protein (YAP) pathway in a rat model of short bowel syndrome.

Our previous study demonstrated that the proliferation of human intestinal smooth muscle (ISM) cells was stimulated by butyrate through the yes-associated protein (YAP) pathway in vitro, suggesting a valuable approach for intestinal adaption of short bowel syndrome (SBS). This study was conducted to confirm these findings in vivo. Three-week-old Sprague-Dawley rats were randomly divided into the following groups: Sham group (bowel transection and reanastomosis), SB W group (80% small bowel resection/water ad libitum), and SB Bu group (80% small bowel resection/50 mM sodium butyrate ad libitum). Morphological changes were determined by hematoxylin and eosin staining; the proliferation rate of ISM cells was examined by Ki67 staining, and apoptosis was determined in the TUNEL assay. Changes in the expression of YAP and its downstream genes were evaluated by quantitative-polymerase chain reaction and western blotting. Fourteen days post-operation, a significant increase in ISM thickness was observed in the SB Bu group compared to the SB W group, accompanied by enhanced proliferation of ISM cells and suppression of apoptosis. Notably, YAP expression was also significantly increased in the SB Bu group, with a 6.5-fold increase in the proportion of YAP-positive ISM cells, 2.2-fold increase in YAP mRNA expression, and 3.4-fold increase in protein expression. In conclusion, our results suggest that butyrate promotes ISM adaption through YAP in vivo, which may be a potential therapeutic approach for SBS patients.

Autophagy May Protect Against Parenteral Nutrition-Associated Liver Disease by Suppressing Endoplasmic Reticulum Stress.

BACKGROUND: The objectives of this study were to address the role of autophagy in the pathogenesis of parenteral nutrition (PN)-associated liver disease (PNALD) and its possible mechanism in vivo. METHODS: Five-week-old male Sprague Dawley rats were fed Shoobree chow (Xietong Organism, Jiangsu, China) and administered intravenous 0.9% saline (sham group), PN (PN group), PN plus rapamycin (1 mg/kg; PN + Rapa group), or rapamycin (Rapa group) for 7 days. Before and after study, body weight, biochemical indicators, hepatic histology, level of autophagy, hepatocyte apoptosis, reactive oxygen species (ROS), and endoplasmic reticulum (ER) stress indicators including binding immunoglobulin protein (BIP), spliced X-box-binding protein-1 (sXBP1), and CCAAT-enhancer-binding protein homologous protein (CHOP) were measured. RESULTS: Autophagy was suppressed in the PNALD model, which was demonstrated by less light chain 3 fluorescence (LC3) puncta and lower LC3II expression. Rapamycin effectively induced hepatic autophagy in PN rats. The PN + Rapa group presented improved hepatic function, decreased pathology scores, and less steatosis than the PN group. In addition, rapamycin treatment decreased terminal deoxynucleotidyl transferase dUTP nick end labeling and cleaved-caspase 3 expression, indicating a lower level of hepatocyte apoptosis. Compared with the PN group, the PN + Rapa group had lower levels of ROS and reduced expression of ER stress-related protein markers, such as BIP, sXBP1 and CHOP. CONCLUSIONS: Autophagy was suppressed in the PNALD model. Rapamycin treatment induced autophagy and protected against PNALD, possibly by suppressing ROS-induced ER stress.

CELF1/p53 axis: a sustained antiproliferative signal leading to villus atrophy under total parenteral nutrition.

Intestinal villus atrophy is a major complication of total parenteral nutrition (TPN). Our previous study revealed that TPN-induced villus atrophy is accompanied by elevated expression of CUGBP, Elav-like family member 1 (CELF1); however, its mechanism of action has not been fully understood. Herein, we report a pivotal role of CELF1/p53 axis, which induces a sustained antiproliferative signal, leading to suppressed proliferation of intestinal epithelial cells (IECs). By using a rat model of TPN, we found synchronous upregulation of CELF1 and p53 in jejunum mucosa, accompanied by a 51% decrease in crypt cell proliferation rate. By using HCT-116 cells as an IEC model in vitro, we found that the expression of CELF1 altered dynamically in parallel to proliferation rate, suggesting a self-adaptive expression pattern in IECs in vitro. Furthermore, ectopic overexpression of CELF1 elicited a significant antiproliferative effect in HCT-116, Caco-2, and IEC-6 cells, whereas knockdown of CELF1 elicited a significant proproliferative effect. Moreover, cell-cycle assay revealed that ectopic overexpression of CELF1 induced sustained G2 arrest and G1 arrest in HCT-116 and IEC-6 cells, respectively, which could be abolished by p53 silencing. Mechanistically, polysomal profiling and nascent protein analysis revealed that regulation of p53 by CELF1 was mediated through accelerating its protein translation in polysomes. Taken together, our findings revealed a sustained suppression of IEC proliferation evoked by CELF1/p53 axis, which may be a potential therapeutic target for the treatment of TPN-induced villus atrophy.-Yan, J.-K., Zhang, T., Dai, L.-N., Gu, B.-L., Zhu, J., Yan, W.-H., Cai, W., Wang, Y. CELF1/p53 axis: a sustained antiproliferative signal leading to villus atrophy under total parenteral nutrition.

Butyrate stimulates the growth of human intestinal smooth muscle cells by activation of yes-associated protein.

Intestinal smooth muscle cells play a critical role in the remodeling of intestinal structure and functional adaptation after bowel resection. Recent studies have shown that supplementation of butyrate (Bu) contributes to the compensatory expansion of a muscular layer of the residual intestine in a rodent model of short-bowel syndrome (SBS). However, the underlying mechanism remains elusive. In this study, we found that the growth of human intestinal smooth muscle cells (HISMCs) was significantly stimulated by Bu via activation of Yes-Associated Protein (YAP). Incubation with 0.5 mM Bu induced a distinct proliferative effect on HISMCs, as indicated by the promotion of cell cycle progression and increased DNA replication. Notably, YAP silencing by RNA interference or its specific inhibitor significantly abolished the proliferative effect of Bu on HISMCs. Furthermore, Bu induced YAP expression and enhanced the translocation of YAP from the cytoplasm to the nucleus, which led to changes in the expression of mitogenesis genes, including TEAD1, TEAD4, CTGF, and Cyr61. These results provide evidence that Bu stimulates the growth of human intestinal muscle cells by activation of YAP, which may be a potential treatment for improving intestinal adaptation.