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Background: Vascular smooth muscle cell (VSMC) contractility is critical for blood pressure regulation and vascular homeostasis. Identifying the key molecule that maintains VSMC contractility may provide a novel therapeutic target for vascular remodeling. ALK3 (activin receptor-like kinase 3) is a serine/threonine kinase receptor, and deletion of ALK3 causes embryonic lethality. However, little is known about the role of ALK3 in postnatal arterial function and homeostasis. Methods: We conducted in vivo studies in a tamoxifen-induced postnatal VSMC-specific ALK3 deletion mice suitable for analysis of blood pressure and vascular contractility. Additionally, the role of ALK3 on VSMC was determined using Western blot, collagen-based contraction assay and traction force microscopy. Furthermore, interactome analysis were performed to identify the ALK3-associated proteins and bioluminescence resonance energy transfer assay was used to characterize Gαq activation. Results: ALK3 deficiency in VSMC led to spontaneous hypotension and impaired response to angiotensin II in mice. In vivo and in vitro data revealed that ALK3 deficiency impaired contraction force generation by VSMCs, repressed the expression of contractile proteins, and inhibited the phosphorylation of myosin light chain. Mechanistically, Smad1/5/8 signaling mediated the ALK3-modulated contractile protein expressions but not myosin light chain phosphorylation. Furthermore, interactome analysis revealed that ALK3 directly interacted with and activated Gαq (guanine nucleotide-binding protein subunit αq)/Gα11 (guanine nucleotide-binding protein subunit α11), thereby stimulating myosin light chain phosphorylation and VSMC contraction. Conclusions: Our study revealed that in addition to canonical Smad1/5/8 signaling, ALK3 modulates VSMC contractility through direct interaction with Gαq/Gα11, and therefore, might serve as a potential target for modulating aortic wall homeostasis.

Spinal muscular atrophy (SMA) is a neurodegenerative disease characterized by the selective loss of spinal motor neurons (MNs) and concomitant muscle weakness. Mutation of SMN1 is known to cause SMA, and restoring SMN protein levels via antisense oligonucleotide treatment is effective for ameliorating symptoms. However, this approach is hindered by exorbitant costs, invasive procedures, and poor treatment responses of some patients. Here, we seek to circumvent these hurdles by identifying reliable biomarkers that could predict treatment efficacy. We uncovered that MiR34 exhibits consistent downregulation during SMA progression in both human and rodent contexts. Importantly, Mir34 family-knockout mice display axon swelling and reduced neuromuscular junction (NMJ) endplates, recapitulating SMA pathology. Introducing MiR34a via scAAV9 improved the motor ability of SMNΔ7 mice, possibly by restoring NMJ endplate size. Finally, we observed a consistent decreasing trend in MiR34 family expression in the cerebrospinal fluid (CSF) of type I SMA patients during the loading phase of nusinersen treatment. Baseline CSF MiR34 levels before nusinersen injection proved predictive of patient motor skills 1 year later. Thus, we propose that MiR34 may serve as a biomarker of SMA since it is associated with the pathology and can help evaluate the therapeutic effects of nusinersen.

Vapor deposition of polymers is known to result in densified thin films, and recent developments have advanced these polymers with interesting fabrication techniques to a variety of controlled structures other than thin films. With the advantages of chemical modification and functionalization of these polymers, advancements have combined both the physical and chemical properties of these vapor-deposited polymers to obtain controlled anisotropic polymers, including layer-by-layer, gradient, hierarchical, porosity, and the combination of the above, meaning that the produced polymers are functional and are addressed in devised physical configurations and chemical compositions. The main purpose of using polymer coatings as a tool for surface modification is to provide additional properties that decouple the natural properties of the underlying materials (including metals, polymers, oxides/ceramics, glass, silicon, etc.), and recent advancements have rendered novel insights into combined physical and chemical properties to fulfill the increasing needs of sophisticated requirements of materials for users. The review herein intends to deliver messages of recent progress of the advancements of vapor-deposited polymers, with discussions of the variations of the physical structures and chemical functionalities, and how these two aspects are integrated with novel fabrication techniques. The advanced vapor polymers now have the capability of controlled anisotropy in the physical structure and chemical composition and are expected to pave the way for interface engineering toward prospective material designs.Table of content:1. Introduction2. Fabrication and Materials3. Controls of Anisotropy4. Applications5. Conclusions and Outlook

This study aimed to evaluate the shape-memory effect (SME) of wood (Populus x beijingensis W. Y. Hsu) and identify the net-points and switches in its molecular and morphological structures. During several cycles of deformation and subsequent recovery, a high shape recovery rate and ratio were maintained. The transverse compression tests of wet and dry wood reveal that the hydrothermal coupling stimulation can considerably reduce the strength of wood. The X-ray diffraction characterization of wood under hydrothermal stimulation shows that the role of network nodes in the SME of wood is influenced by temperature. The wavenumber shifting and changes in the intensity ratio of the characteristic Fourier transform infrared peaks showed that hydrogen bonds acted as switches for the water-stimulated shape-memory behavior. By taking into account viscoelastic relaxation, a kinetic model derived from nonequilibrium thermodynamic fluctuation theory was used to describe the shape recovery process. The effects of hydration on recovery kinetics, activation, and dynamic mechanical behaviors were also studied. To explain the shape-memory mechanism of wood under hydrothermal stimulation, a hybrid-structure network model based on a single three-dimensional switch network was proposed in this study.

Patients with Graves' disease who remain hyperthyroid under the treatment of antithyroid drugs (ATD) or cannot tolerate ATD usually receive radioactive iodine (RAI) to control disease activity. This pilot study aimed to identify predictors of prolonged euthyroidism12 months after receiving RAI.Demographic, clinical, and laboratory data from 117 patients receiving RAI were retrospectively collected, including age, gender, body surface area, smoking status, free thyroxine, thyrotropin, thyrotropin binding inhibiting immunoglobulin (TBII), microsomal antibody (MA), thyroglobulin antibody (TA), medication history, and thyroid volume. Only 85 patients without missing values were included in statistical analysis. The calculated RAI dose was the estimated thyroid volume × 0.4. The difference and ratio between the actual and calculated RAI doses were examined. A stepwise logistic regression analysis was conducted to identify important predictors of prolonged euthyroidism12 months. The cut-off values for discretizing continuous covariates were estimated by fitting generalized additive models.Among the 85 patients on RAI, 18 (21.2%) achieved prolonged euthyroidism12 months, 38 (44.7%) remained hyperthyroid with decreased ATD doses, but 29 (34.1%) suffered permanent hypothyroidism and needed long-term levothyroxine. Logistic regression analysis revealed that patients with age66 years, 33age ≤66 years, quitting smoking vs. non-smoking or current smoking, 600MA ≤1729 IU/mL, 47%TBII ≤81%, 7TA ≤162 IU/mL, 0.63ratio between actual and calculated RAI doses ≤1.96, or taking hydroxychloroquine (HCQ) would have a higher chance of reaching prolonged euthyroidism12 months after receiving RAI. Its area under the Receiver Operating Characteristic curve was 0.932.Patients with Graves' disease who received an actual RAI dose close to the calculated RAI dose achieved prolonged euthyroidism12 months if they also took HCQ during RAI treatment.

Background: Alzheimer’s disease (AD) is the most common form of neurodegenerative dementia among the elderly. Excitotoxicity has been implicated as playing a dominant role in AD, especially related to the hyperactivation of excitatory neurons. Death-associated protein kinase 1 (DAPK1) is a calcium/calmodulin-dependent kinase and involved in the pathogenesis of AD, but the roles and mechanisms of DAPK1 in excitotoxicity in AD are still uncertain. Objective: We mainly explored the underlying mechanisms of DAPK1 involved in the excitotoxicity of AD and its clinical relevance. Methods: Differentiated SH-SY5Y human neuroblastoma cells, PS1 V97 L transgenic mice, and human plasma samples were used. Protein expression was assayed by immunoblotting, and intracellular calcium and neuronal damage were analyzed by flow cytometry. Plasma DAPK1 was measured by ELISA. Results: We found that DAPK1 was activated after amyloid-β oligomers (AβOs) exposure in differentiated SH-SY5Y cells. Besides, we found the phosphorylation of GluN2B subunit at Ser1303 was increased, which contributing to excitotoxicity and Ca2+ overload in SH-SY5Y cells. Inhibiting DAPK1 activity, knockdown of DAPK1 expression, and antagonizing GluN2B subunits could effectively prevent AβOs-induced activation of GluN2B subunit, Ca2+ overload, and neuronal apoptosis. Additionally, we found that DAPK1 was elevated in the brain of AD transgenic mouse and in the plasma of AD patients. Conclusion: Our finding will help to understand the mechanism of DAPK1 in the excitotoxicity in AD and provide a reference for the diagnosis and therapy of AD.

Aims Whether changes in endothelial tight junctions (TJs) lead to the formation of thoracic aortic aneurysm and dissection (TAAD) and serve as an early indicator and therapeutic target remains elusive. Methods and results Single-cell RNA sequencing analysis showed aberrant endothelial TJ expressions in the thoracic aortas of patients with TAAD. In a β-aminopropionitrile (BAPN)-induced TAAD mouse model, endothelial TJ function was disrupted in the thoracic aortas at an early stage (5 and 10 days) as observed by a vascular permeability assay, while the intercellular distribution of crucial TJ components was significantly decreased by en face staining. For the non-invasive detection of endothelial TJ function, two dextrans of molecular weights 4 and 70 kDa were conjugated with the magnetic resonance imaging (MRI) contrast agent Gd-DOTA to synthesize FITC-dextran-DOTA-Gd and rhodamine B-dextran-DOTA-Gd. MRI images showed that both probes accumulated in the thoracic aortas of the BAPN-fed mice. Particularly, the mice with increased accumulated signals from 5 to 10 days developed TAAD at 14 days, whereas the mice with similar signals between the two time points did not. Furthermore, the protease-activated receptor 2 inhibitor AT-1001, which seals TJs, alleviated the BAPN-induced impairment of endothelial TJ function and expression and subsequently reduced TAAD incidence. Notably, endothelial-targeted ZO-1 conditional knockout increased TAAD incidence. Mechanistically, vascular inflammation and edema were observed in the thoracic aortas of the BAPN-fed mice, whereas these phenomena were attenuated by AT-1001. Conclusion The disruption of endothelial TJ function is an early event prior to TAAD formation, herein serving as a potential indicator and a promising target for TAAD.

The in-situ formed Li–B alloy provides abundant nucleation sites for inducing uniform Li deposition and inhibiting Li dendrite formation. The 3D porous Ni foam can provide enough space for relieving volume change.

A star-like CoIII2DyIII2 complex (1) and a cross-shaped CoII/III–Dy complex (2) have been obtained, in which 1 exhibits double magnetic relaxation behavior, while 2 only shows slow magnetic relaxation signals.

A novel separator of Co-doped MnO2 with abundant oxygen vacancies exhibits a high reversible capacity of 902.0 mA h g−1 at 0.5 C after 200 cycles for Li–S batteries.

Background: Few randomized controlled trials (RCTs) investigated the effects of mindfulness intervention on affective and cognitive symptoms in older adults with mild cognitive impairment (MCI). Furthermore, no RCTs on mindfulness followed participants beyond two years. Objective: To examine the longitudinal effects of a mindful awareness practice (MAP) intervention on depressive, anxiety, and cognitive symptoms in MCI. Methods: In this parallel-arm and assessor-blinded RCT, 55 community-dwelling older adults with MCI were randomized into the MAP or active control, i.e., health education program (HEP). Intervention sessions were conducted weekly for three months and monthly for the subsequent six months. Assessments and follow-up were conducted at baseline, 3-month, 9-month, and 5-year time points. Depressive, anxiety, and cognitive symptoms were measured using the Geriatric Depression Scale-15 (GDS-15), Geriatric Anxiety Inventory-20 (GAI-20), and Mini-Mental State Examination (MMSE), respectively. Linear-mixed models, following the intention-to-treat principle, were used for data analyses. Results: A total of 55 participants aged 60 to 86 (Mean age: 71.3±6 years old) was recruited, with n = 28 allocated to the MAP arm and n = 27 allocated to the HEP arm. Compared to HEP, GDS-15, GAI-20, and MMSE scores did not differ significantly in MAP during follow-ups. Conclusion: Compared to HEP, MAP did not improve affective symptoms nor delay deteriorations in general cognition in community-dwelling older adults with MCI. Compared to our previous findings showing domain-specific improvements in MAP over HEP in attention and memory up to 9 months, this study highlights the importance of examining domain-specificity using detailed cognitive measures in non-pharmacological intervention with MCI.

A pair of undescribed dihydrobenzofuran neolignan enantiomers, (+/−)-phybrenan A (1a/1b), two new benzofuran neolignans, phybrenan B and C (2 and 3), along with four known neolignans (4 − 7) were obtained from the plants of Phyllanthodendron breynioides P. T. Li. The planar structures of all isolates were demonstrated by the analysis of detailed spectroscopic evidence (NMR, HRMS, and IR), and the absolute configurations of novel neolignans were elucidated by combined calculated and experimental ECD data analysis. The neuroprotective activities of all benzofuran neolignans against sodium nitroprusside (SNP)-induced cell death were examined in rat pheochromocytoma PC12 cells. The results exhibited that three compounds (4 − 6) possessed remarkable neuroprotective activities at 10 µM, better than the positive drug edaravone.

To explore the clinical and radiological effects of the free vascularized iliac bone flap (FVIBF) based on deep iliac circumflex vessels for Association Research Circulation Osseous (ARCO) stage 3 traumatic osteonecrosis of the femoral head (TONFH) in young adults.From February 2017 to August 2020, a total of 29 (29 hips) TONFH patients with ARCO stage 3 were enrolled in this study. Following the FVIBF standard procedure, the necrotic area of the femoral head was removed and a free vascularized iliac bone flap was harvested and grafted. X-rays were performed at 1, 3, 6, 9, and 12 months and every 6 months thereafter postoperatively for evaluation of the degree of femoral head collapse. The Beijing University of Chinese Medicine X-ray Evaluation Method (BUCMXE) was used to evaluate the hip preservation effects based on anteroposterior hip radiographs. Hip functions were evaluated using the Harris hip score (HHS) every 6 months and annually after 24 months, and the quality of life was assessed using the SF-36 questionnaire at 18 months postoperatively.The mean follow-up time was 30 months (range 12-54 months). None of the patients was lost to follow-up, and the wounds healed primarily without infection and deep venous thrombosis of lower limbs. The HHS at 12 months postoperatively was significantly improved than that before surgery (83.4 ± 7.4 vs. 64.8 ± 14.1; p 0.05). The BUCMXE radiographic score at 12 months postoperatively did not reveal any significant differences in femoral head morphology and osteonecrotic lesion, when compared to preoperation. The SF-36 questionnaire showed significant improvements in social functioning, role physical, bodily pain, and general health (p 0.05). However, eight patients had varying degrees of femoral head collapse after surgery, and only one of them underwent THA due to pain and collapse aggravation. BUCMXE-Osteoarthritis scores showed that 23 hips were stable and six hips were advanced, among which five hips staged ARCO 3B and one staged ARCO 3A preoperatively, with statistically significant differences (p 0.05).Although some patients showed no improvement of radiological outcomes, most patients with severe TONFH could still benefit from the FVIBF procedure. Further studies should attempt to improve this surgical procedure and explore its long-term efficacy.

Endoplasmic reticulum stress (ERS) is involved in the pathological process of vascular dementia (VD). GJ-4 is extracted from Gardenia jasminoides J. Ellis and has been reported to have protective roles in ischemia-related brain damage. However, the role of GJ-4 in ERS has not been elucidated. We established a VD rat model through bilateral common carotid arteries occlusion (2-VO). The rats were intragastrically administrated with GJ-4 (10, 25, and 50[Formula: see text]mg/kg) and nimodipine (10[Formula: see text]mg/kg). Data from a Morris water maze test showed that GJ-4 could significantly alleviate learning and memory deficits in VD rats. Nissl and cleaved caspase-3 staining revealed that GJ-4 can inhibit apoptosis and thus exert a protective role in the brain of 2-VO rats. Western blot results suggested that GJ-4 significantly reduced ERS-related protein expression and inhibited apoptosis through suppression of the PERK/eIF2[Formula: see text]/ATF4/CHOP signaling pathway. For in vitro studies, the oxygen-glucose deprivation (OGD) SH-SY5Y model was employed. Western blot and Hoechst 33342/PI double staining were utilized to explore the effects of crocetin, the main active metabolite of GJ-4. Like GJ-4 in vivo, crocetin in vitro also decreased ERS-related protein expression and inhibited the activation of the PERK/eIF2[Formula: see text]/ATF4/CHOP signaling pathway. Thus, crocetin exerted similar protective roles on OGD challenged SH-SY5Y cells in vitro. In summary, GJ-4 and crocetin reduce the ERS in the brain of VD rats and SY5Y cells subjected to OGD and inhibit neuronal apoptosis through suppression of the PERK/eIF2[Formula: see text]/ATF4/CHOP pathway, suggesting that GJ-4 may be useful for the treatment of VD.

The assignment of functions based on homology has recently been challenged by the frequent discovery of functional divergence among homologous gene family members of enzymes involved in plant secondary metabolism. Secologanin synthase (SLS) is the key CYP450 enzyme that acts critically in the biosynthesis of Strychnos alkaloid scaffold. In this study, to fully elucidate the mechanism that underlies metabolic variation, the CYP450 paralogs that participate in oxidative transformation of the secoiridoid pathway were functionally characterized by combining multitiered strategies of metabolite profiling, phylogenetic analyses, biochemistry assays and reverse genetics techniques. Five CaSLSs-like homologous genes were mined and isolated from an integrative multi-omics database of Camptotheca acuminata. Protein sequences, structural comparisons, and phylogenetic analyses confirmed that CaSLS1-2 and CaSLS4-5 were grouped into the SLS clade, and only CaSLS3 clustered into the 7DLH clade. Five homologs, including two previously identified enzymatic genes, were thus designated as CaSLAS1, CaSLAS2, Ca7DLH, CaSLS4 and CaSLS5. Enzymatic assays of the recombinant proteins in yeast showed that CaSLAS1 and CaSLAS2 displayed multi-catalytic activities of SLS, secologanic acid synthase (SLAS) and secoxyloganin synthase (SXS). Additionally, the reactions of CaSLASs enzymes generated stereospecific isomers of secoiridoid products, and a new product of secoxyloganin was observed. CaSLS5, a third SLS enzyme isoform that catalyzes the formation of secologanin, was reported for the first time. However, CaSXS enzymatic activities in vitro had little physiological impact on the biosynthesis of camptothecin (CPT) in Camptotheca acuminata. The primary and secondary roles of CaSLSs-like genes in secoiridoid metabolism were confirmed by virus-induced gene silencing (VIGS) in plant leaves. Efficient silencing and transcriptional downregulation of CaSLAS2, compared with the CaSLAS1 homologs, resulted in a greater reduction of the accumulation of CPT within silenced plants, and CaSLS5 had barely any effect on the contents of metabolites in planta. Thus, CaSLAS2, rather than CaSLAS1, appeared to function as a major participant in the biosynthesis of CPT, and there were redundant functions in the CaSLSs-like enzymes. Consistent with such roles, CaSLAS2 was ubiquitously expressed at very high levels in Camptotheca tissues, and CaSLAS2 was specifically expressed in young leaves. In contrast, CaSLS5 was poorly expressed in every tissue tested. Our findings demonstrate that homologs that belong to the CYP72 gene family are functionally diverse and exhibit divergence and thereby uncover an expanding group of enzymatic genes that determine the chemo-diversity of the iridoid pathway.

This study addresses well-known shortcomings of poly(ethylene glycol) (PEG)-based conjugates. PEGylation is by far the most common method employed to overcome immunogenicity and suboptimal pharmacokinetics of, for example, therapeutic proteins but has significant drawbacks. First, PEG offers no protection from denaturation during lyophilization, storage, or oxidation (e.g., by biological oxidants, reactive oxygen species); second, PEG's inherent immunogenicity, leading to hypersensitivity and accelerated blood clearance (ABC), is a growing concern. We have here developed an active-stealth' polymer, poly(thioglycidyl glycerol)(PTGG), which in human plasma is less immunogenic than PEG (35% less complement activation) and features a reactive oxygen species-scavenging and anti-inflammatory action (50% less TNF-α in LPS-stimulated macrophages at only 0.1 mg/mL). PTGG was conjugated to proteins via a one-pot process; molar mass- and grafting density-matched PTGG-lysozyme conjugates were superior to their PEG analogues in terms of enzyme activity and stability against freeze-drying or oxidation; the latter is due to sacrificial oxidation of methionine-mimetic PTGG chains. Both in mice and rats, PTGG-ovalbumin displayed circulation half-lives up to twice as long as PEG-ovalbumin, but most importantly and differently from PEG without any associated ABC effect seen either in the time dependency of blood concentration, in the liver/splenic accumulation, or in antipolymer IgM/IgG titers. Furthermore, similar pharmacokinetic results were obtained with PTGGylated/PEGylated liposomal nanocarriers. PTGG's active-stealth' character therefore makes it a highly promising alternative to PEG for conjugation to biologics or nanocarriers.

To assess how age-friendly deprescribing trials are regarding intervention design and outcome assessment. Reduced use of potentially inappropriate medications (PIMs) can be addressed by deprescribing-a systematic process of discontinuing and/or reducing the use of PIMs. The 4Ms-"Medication", "Mentation", "Mobility", and "What Matters Most" to the person-can be used to guide assessment of age-friendliness of deprescribing trials.Published literature.Scoping review.The literature was identified using keywords related to deprescribing and polypharmacy in PubMed, EMBASE, Web of Science, ProQuest, CINAHL, and Cochrane and snowballing. Study characteristics were extracted and evaluated for consideration of 4Ms.Thirty-seven of the 564 trials identified met the review eligibility criteria. Intervention design: "Medication" was considered in the intervention design of all trials; "Mentation" was considered in eight trials; "Mobility" (n = 2) and "What Matters Most" (n = 6) were less often considered in the design of intervention. Most trials targeted providers without specifying how matters important to older adults and their families were aligned with deprescribing decisions."Medication" was the most commonly assessed outcome (n = 33), followed by "Mobility" (n = 13) and "Mentation" (n = 10) outcomes, with no study examining "What Matters Most" outcomes."Mentation" and "Mobility", and "What Matters Most" have been considered to varying degrees in deprescribing trials, limiting the potential of deprescribing evidence to contribute to improved clinical practice in building an age-friendly health care system.

To establish and validate a radiomics model based on multiparametric magnetic resonance imaging (MRI), and to predict microsatellite instability (MSI) status in rectal cancer patients.A total of 199 patients with pathologically confirmed rectal cancer were included. The MSI status was confirmed by immunohistochemistry (IHC) staining. Clinical factors and laboratory data associated with MSI status were analyzed. The imaging data of 100 patients from one of the hospitals were used as the training set. The remaining 99 patients from the other two hospitals were used as the external validation set. The regions of interest (ROIs) were delineated from T1-weighted imaging (T1WI), T2-weighted imaging (T2WI), diffusion-weighted imaging (DWI), and contrast-enhanced T1WI (CE-T1WI) sequence to extract the radiomics features. The Tree-based approach was used for feature selection. The models were constructed based on the four single sequences and a combination of the four sequences using the random forest (RF) algorithm. The external validation set was used to verify the generalization ability of each model. The receiver operating characteristic (ROC) curves and the area under the curve (AUC) were plotted to evaluate and compare the predictive performance of each model.In the four single-series models, the CE-T1WI model performed the best. The AUCs of the T1WI, T2WI, DWI, and CE-T1WI prediction models in the training set were 0.74, 0.71, 0.71, and 0.78, respectively, while in the external validation set, the corresponding AUCs were 0.67, 0.66, 0.70, and 0.77. The prediction and generalization performance of the combined model of multi-sequences was comparable to that of the CE-T1WI model and it was better than that of the remaining three single-series models, with AUC values of 0.78 and 0.78 in the training and validation sets, respectively.The established radiomics models based on CE-T1WI or multiparametric MRI have similar predictive performance. They have the potential to predict MSI status in rectal cancer patients.• A radiomics model for the prediction of MSI status in patients with rectal cancer was established and validated using external validation. • The models based on CE-T1WI or multiparametric MRI have better predictive performance than those based on single unenhanced sequence images. • The radiomics model has the potential to suggest MSI status in rectal cancer patients; however, it is not yet a substitute for histological confirmation.

Background Pulmonary fibrosis is considered to be an irreversible lung injury, which can be caused by paraquat (PQ) poisoning. MiRNAs have been demonstrated crucial roles in pulmonary fibrosis caused by numerous approaches including PQ induction. The purpose of this study was to investigate the role and the underlying mechanism of miR-215 in PQ-induced pulmonary fibrosis. Methods The cell and animal models of pulmonary fibrosis were established through PQ intervention. Cell viability was performed to test by MTT assay. Immunofluorescence assay was used to detect COL1A1 expression and its location. The relationships among E2F1, miR-215-5p, and BMPR2 were validated by dual luciferase reporter gene assay, chromatin immunoprecipitation and RNA-binding protein immunoprecipitation. Lung morphology was evaluated by hematoxylin and eosin staining. Results MiR-215-5p was upregulated in PQ-induced pulmonary fibrosis in vitro and in vivo. MiR-215-5p silencing relieved PQ-induced pulmonary fibrosis progression by enhancing cell viability and reducing the expression of fibrosis-related markers (COL1A1, COL3A1, and α-SMA). Mechanistically, miR-215-5p directly targeted BMRP2. BMPR2 knockdown abolished the suppressive effects of miR-215-5p knockdown on PQ-induced pulmonary fibrosis. In addition, E2F1 interacted with miR-215-5p promoter and positively regulated miR-215-5p expression. E2F1 downregulation reduced miR-215-5p level and promoted BMPR2 level via regulating TGF-β/Smad3 pathway, and then suppressed PQ-induced pulmonary fibrosis, whereas these effects were compromised by miR-215-5p sufficiency. Conclusion MiR-215-5p was activated by E2F1 to repress BMPR2 expression and activate TGF-β/Smad3 pathway, which aggravated PQ-induced pulmonary fibrosis progression. Targeting the E2F1/miR-215-5p/BMPR2 axis might be a new approach to alleviate PQ-induced pulmonary fibrosis.

This study aims to explore the molecular mechanism of N6-methyladenosine (m6A) modification-related long noncoding RNA (lncRNA)-microRNA (miRNA)-messenger RNA (mRNA) network in regulating autophagy and affecting the occurrence and development of acute pancreatitis (AP). RNA-seq datasets related to AP were obtained from Gene Expression Omnibus (GEO) database and merged after batch effect removal. lncRNAs significantly related to m6A in AP, namely candidate lncRNA, were screened by correlation analysis and differential expression analysis. In addition, candidate autophagy genes were screened through the multiple databases. Furthermore, the key pathways for autophagy to play a role in AP were determined by functional enrichment analysis. Finally, we predicted the miRNAs binding to genes and lncRNAs through TargetScan, miRDB and DIANA TOOLS databases and constructed two types of lncRNA-miRNA-mRNA regulatory networks mediated by upregulated and downregulated lncRNAs in AP. Nine lncRNAs related to m6A were differentially expressed in AP, and 21 candidate autophagy genes were obtained. Phosphoinositide 3-kinase (PI3K)-Akt signaling pathway and Forkhead box O (FoxO) signaling pathway might be the key pathways for autophagy to play a role in AP. Finally, we constructed a lncRNA-miRNA-mRNA regulatory network. An upregulated lncRNA competitively binds to 13 miRNAs to regulate 6 autophagy genes, and a lncRNA-miRNA-mRNA regulatory network in which 2 downregulated lncRNAs competitively bind to 7 miRNAs to regulate 2 autophagy genes. m6A modification-related lncRNA Pvt1, lncRNA Meg3 and lncRNA AW112010 may mediate the lncRNA-miRNA-mRNA network, thereby regulating autophagy to affect the development of AP.