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1.
Behav Brain Res ; 377: 112249, 2020 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-31541673

RESUMO

Negative mood states are hypothesized to increase the risk for overeating among restrained eaters. However, neurophysiological features of inhibitory control during negative mood states among restrained eaters are poorly understood. To address this gap, we aimed to assess event related potentials (ERPs) associated with food-specific inhibitory control among successful restrained eaters (SREs, n = 17), unsuccessful restrained eaters (UREs, n = 18), and non-restrained eaters (NREs, n = 18) engaged in a food-related go/no-go task during negative versus neutral mood states. Compared to neutral mood states, negative mood states were related to comparatively greater no-go N2a amplitudes in the entire sample. Regarding group differences, no-go N2a amplitudes of SREs were greater than those of UREs across negative and neutral mood states. However, no-go P3 amplitudes of SREs decreased significantly during the negative mood state while no change was observed for other groups. Results suggest that negative mood states may interfere with conflict monitoring and behavioral inhibition in general, but inhibitory control over food may be enhanced among SREs during negative mood states relative to UREs. Results may provide psychophysiological bases for understanding why SREs succeed and UREs fail in controlling appetitive responses to external food cues.

2.
Mater Sci Eng C Mater Biol Appl ; 106: 110158, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31753379

RESUMO

In the current work, an intestinal anastomosis ring made of magnesium­zinc­strontium (Mg-Zn-Sr) alloy was developed and fabricated in order to take advantages of the appropriate biocompatibility and degradability of Mg-based alloys. As-fabricated anastomosis rings were implanted into the intestinal tracts of Bama miniature pigs to evaluate their biological performance in vivo. At the injury site, the formation of edema and granulation tissue was observed for 2 weeks after surgery. Till week 4, the edema transformed to firm scar tissue, which reached the healing standard of intestinal tissue. The levels of biochemical indicators such as blood routine, liver and kidney functions as well as electrolytes were all under normal conditions, indicating that the implantation of Mg alloy did not have remarkable influence on the blood system as well as liver and kidney functions. Pathological results revealed that no obvious abnormality was found in heart, liver, spleen, lung, kidney and brain tissues. The Mg ions were found to be excreted from the body through urine. The intestinal anastomosis ring could be discharged through excretion around 2 weeks after surgery, of which the surface was corroded and covered by a layer of Ca- and P-containing minerals. According to histological images, a mild inflammatory response was noticed on week 2. At this stage, dilated and congested capillaries were found in the muscular layer. Moreover, the mucosal layer and villi at the injury site were disordered. Till week 4, the muscular and mucosal layer were similar to their healthy counterparts even though the villi were slightly shorter than normal ones. Together, the results indicate that Mg-Zn-Sr alloy is a promising candidate for the fabrication of biodegradable intestinal anastomosis ring.

3.
J Phys Condens Matter ; 32(1): 015701, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31509817

RESUMO

Recently, two-dimensional ferromagnetic semiconductors have been an important class of materials for many potential applications in spintronic devices. Based on density functional theory, we systematically explore the magnetic and electronic properties of CrGeS3 with the monolayer structures. It is found that the bandgap of spin-up state is 1.01 eV when it is 1.07 eV in spin-down state. The exchange splitting is calculated as 0.67 eV (2.21 eV by HSE06 functional), which originates from bonding [Formula: see text] hybridized states of Cr e g -S p  and unoccupied Cr t 2g -Ge p  hybridization. After that, the comparison of total energy between different magnetic states ensures the ferromagnetic ground state of monolayer CrGeS3. The reason of the magnetic states originates mainly from the competition between antiferromagnetic direct neighboring Cr-Cr exchange and ferromagnetic superexchange mediated by S atom. And the results also show the magnetic moment of 6 [Formula: see text] per unit cell, including two Cr atoms. Besides, we estimate that the monolayer CrGeS3 possesses the Curie temperature of 161 K by mean-field theory. The results suggest that monolayer CrGeS3 crystals will possess potential applications in nanoscale spintronics.

4.
Mater Sci Eng C Mater Biol Appl ; 107: 110327, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31761167

RESUMO

In this work, Ti and Mg alloys were made into composites by using spark plasma sintering, in order to combine the high stiffness of Ti and biodegradability of Mg alloys. The effect of the content of Mg-3Zn alloy on the degradability was studied by using electrochemical corrosion, immersion in simulated body fluid and incubation of Saos-2 cells. The results show that Ti-Mg metal-metal composites (MMCs) exhibit low Young's modulus (31-48 GPa) and good yield strength (616.5-642.8 MPa). The modulus and strength both decrease with the content of Mg-3Zn alloy increasing. The electrochemical corrosion rates of Ti-Mg MMCs were much lower than that of monolithic Mg-3Zn alloys. After an immersion for 21 days, Ti matrix remained integrity and Mg phase dissolved in the solution, inducing the formation of apatite layer on the MMCs. The precipitation of apatite increased with the content of Mg-3Zn alloy increasing. In vitro study indicated that Ti-10Mg and Ti-20 Mg MMCs possessed good biocompatibility to Saos-2 cells, while the biocompatibility of Ti-30 Mg MMCs decreased slightly. In summary, Ti-Mg MMCs are promising implant materials with adjustable degradation rates and improved biocompatibility for orthopedic applications.

5.
Chemosphere ; 238: 124645, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31472352

RESUMO

Bromophenols (BPs) are important organic compounds which have become dominant pollutants during these years. Our present study investigated the potential inhibition behaviour of BPs on the activity of one of the most important phase II drug-metabolizing enzymes (DMEs), UDP-glucuronosyltransferases (UGTs). Recombinant UDP-glucuronosyltransferases (UGTs)-catalyzed glucuronidation of 4-methylumbelliferone (4-MU) was utilized as the probe reaction. 100 µM of BPs was utilized as the inhibition screening concentrations, and the complete inhibition profile of UGT isoforms by BPs was obtained. UGT1A7 was the most vulnerable UGT isoform towards BPs. Some structure-activity relationship for the inhibition of UGTs by BPs was found, and this relationship can be furtherly explained by the hydrophobic contacts of BPs with the activity cavity of UGTs using in silico docking method. The inhibition kinetics determination showed that the inhibition kinetic parameter Ki value was calculated to be 2.85, 3.99 and 31.00 µM for the inhibition of UGT1A3, UGT1A7, and UGT2B7 by representative BPs, 2,4,6-TBP. Combined with in vivo exposure concentration of 2,4,6-TBP, in vitro-in vivo extrapolation (IVIVE) was employed to demonstrate the moderate possibility for the inhibition of UGT1A3 and UGT1A7 by 2,4,6-TBP. In conclusion, our study gave the full description towards the inhibition of BPs towards UGT isoforms, which will provide a new perspective for elucidating the toxicity mechanism of bromophenols (BPs).

6.
J Hazard Mater ; 383: 121126, 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-31505428

RESUMO

Here, TPB (triphenylbenzene) - DMTP (dimethoxyterephthaldehyde) -COF was prepared, characterized and used as effective adsorbent for the removal of sulfamerazine (SMT) from aqueous solution. Its adsorption characteristics and mechanism were explored. With large channel (∼3.3 nm), high specific surface area (2115 m2/g), and high crystallite, TPB-DMTP-COF showed high adsorption capacity (209 mg/g), fast adsorption equilibrium (80 min), and good reusability. Natural pH condition was optimal for its adsorption capacity, while electrostatic repulsion between TPB-DMTP-COF and SMT accounted for the low adsorption performance at acidic or alkaline conditions. According to the DFT method, SMT molecules adsorbed in the pore-sites of COFs via C-H···π interaction was the predominant and stable adsorption configuration accounting for the efficient removal of SMT in large quantity. This study revealed the great adsorption potential of COFs skeleton itself in the application of environmental remediation.

7.
Appl Opt ; 58(30): 8295-8301, 2019 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-31674504

RESUMO

A grating projection shape measurement system has been a commonly used method in the field of three-dimensional (3D) reconstruction in recent years, and global point cloud registration is a key step in this method. However, in the registration process, a large amount of low-precision overlapping redundant data (ORD) is generated between adjacent camera stations, which will seriously affect the speed and accuracy of later modeling. Therefore, how to eliminate these low-precision ORD is a major problem to be solved at present. Determining all overlapping 3D point pairs between two adjacent stations and deleting the points with low precision in the point pairs is the key to solving this problem. Therefore, based on an omnidirectional rotation measurement system, combined with the constraint relationships between the projection space and the acquisition space in the global registration process and the stereo-matching method of space conversion, an elimination algorithm for ORD with a two-dimensional (2D) phase constraint and a 2D pixel constraint is proposed. The experimental results show that the proposed algorithm can faster locate overlapping 3D point pairs between adjacent stations, with a higher elimination rate, and the accuracy of the overall point cloud is higher after the redundancy elimination.

8.
Opt Express ; 27(19): 27046-27061, 2019 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-31674573

RESUMO

We present an efficient and faithful hyperentanglement purification protocol (hyper-EPP) for three-photon system in mixed hyperentangled Greenberger-Horne-Zeilinger states with bit-flip errors in both spatial-mode and polarization degrees of freedom (DOFs), resorting to the fidelity-robust quantum gates and hyperentanglement link. Our high-efficiency hyper-EPP comes from two aspects. One is to pump the higher-fidelity hyperentanglement from different three-photon systems into the same three-photon system with fidelity-robust swap gates, the other is to reproduce some hyperentangled three-photon systems from hyperentangled two-photon subsystems based on hyperentanglement link. Moreover, as the infidelity originating from imperfect single-photon scattering can be heralded as a failure by triggering a detector, our hyper-EPP operates faithfully with the present quantum circuits. Furthermore, our hyper-EPP can be directly extended to purify multiple photon systems entangled in one DOF or hyperentangled in multiple DOFs.

9.
Opt Express ; 27(19): 27151-27158, 2019 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-31674581

RESUMO

We demonstrate a tunable gain-switched Dy3+-doped ZBLAN fiber laser around 3 µm, for the first time, to the best of our knowledge. The pump source is a homemade actively Q-switched Yb3+-doped silica fiber laser at 1.1 µm, yielding a repetition rate of 80 kHz and a pulse width of 300 ns. A plane-ruled grating in a Littrow configuration functions as the tuning element. At the launched pump power of 2.78 W, stable gain-switched pulses tunable within a ~300 nm range of 2800~3095 nm are achieved. When tuning the wavelength to 2946.5 nm, a maximum output power of 218.6 mW with a repetition rate of 80 kHz is obtained. The corresponding pulse width and energy are 530 ns and 2.73 µJ, respectively. The results imply that gain-switching of Dy3+-doped ZBLAN fiber laser is an alternative way for mid-infrared nanosecond pulse generation around 3 µm.

10.
Nurs Res ; 68(6): 433-438, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31693548

RESUMO

BACKGROUND: The relationship between an increasing elderly population and wide-ranging neurological conditions has led to heightened rates of cognitive function impairment. Some researchers have found that health literacy risk may be associated with cognitive impairment in older adults. OBJECTIVES: The purpose of this study was to delineate the difference in health literacy risk between older adults with mild cognitive impairment and those with normal cognitive function. METHODS: We conducted a survey study to explore the health literacy risk in older adults with and without mild cognitive impairment. Data were collected from 412 subjects between 60 and 91 years of age (260 without and 152 with mild cognitive impairment) between June and December 2016 in China. Cognitive function was measured using the Mini-Mental Status Examination, Montreal Cognitive Assessment, Global Deterioration Scale, Activities of Daily Living, and Hamilton Depression Rating Scale. Health literacy was measured using the Chinese Citizen Health Literacy Questionnaire. RESULTS: The scores of total health literacy and its three dimensions (basic health knowledge and mind, basic skills, and health lifestyle and behavior) were lower in those with mild cognitive impairment. Older adults with normal cognitive function had adequate or marginal health literacy levels, whereas those with mild cognitive impairment had marginal or inadequate health literacy levels. Using multiple logistic regression analysis, we found that health literacy risk, education level, age, marital status, and body mass index were independent risk factors for mild cognitive impairment. DISCUSSION: These results suggest that low health literacy may be a predictor of mild cognitive impairment. Screening for lower health literacy risk should be included in multidimensional geriatric evaluation.


Assuntos
Disfunção Cognitiva/psicologia , Alfabetização em Saúde , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Estudos de Casos e Controles , China , Escolaridade , Feminino , Humanos , Modelos Logísticos , Masculino , Estado Civil , Pessoa de Meia-Idade , Fatores de Risco
11.
Stem Cell Res ; 41: 101635, 2019 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-31707215

RESUMO

Autism spectrum disorder (ASD) is a childhood-onset neurodevelopmental disorder challenged in social reciprocity and restrictive repetitive behaviors. Here, we generated an induced pluripotent stem cell (iPSC) line SDQLCHi014-A from a patient with ASD and hyperactivity, carrying a 303 kb de novo deletion at chr3p26.1 implicating GRM7 gene by reprogramming urine cells with non-integrating vectors. SDQLCHi014-A have shown full pluripotency, differentiation capacity and genetic stability. This iPSC line provides a valuable resource to study the molecular mechanisms underlying ASD.

12.
J Fluoresc ; 2019 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-31773380

RESUMO

Herein, we develop a novel mitochondria-targetable fluorescence probe (FVPI) based on fluorene cation derivative. This mitochondria-targetable fluorescence probe exhibits large Stokes shift in various solutions. In addition, FVPI displays numerous advantages, including high photostability, low cytotoxicity, and two-photon properties. View of the above features, FVPI is successfully capable of imaging mitochondria in biological systems with three different sets of signals. This finding will provide a new platform for the constructing mitochondria-targetable fluorescent probes with excellent optical properties.

13.
Cancer Sci ; 2019 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-31778279

RESUMO

Metastasis is a critical determinant for the treatment strategy and prognosis in patient with squamous cell carcinoma of the head and neck (SCCHN). However, mechanisms underlying SCCHN metastasis are poorly understood. Our study sought to find out the key microRNAs and their functional mechanisms involved in SCCHN metastasis. The Cancer Genome Atlas (TCGA) data analysis, qPCR was used to quantify the level of miR-30e-5p in SCCHN and its clinical significance was further analyzed. A series of in vitro and in vivo experiments were applied to determine the effects of miR-30e-5p and its target AEG-1 on SCCHN metastasis. Mechanism investigation further revealed that AEG-1 was implicated in the angiogenesis and metastasis mediated by miR-30e-5p. Overall, our study validates that miR-30e-5p is a valuable predictive biomarker and potential therapeutic target in SCCHN metastasis.

14.
Water Res ; 169: 115279, 2019 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-31734392

RESUMO

Nitrate production during anammox can decrease total nitrogen removal efficiency, which will negatively impact its usefulness for the removal of nitrogen from waste streams. However, neither the performance characteristics nor physiological shifts associated with nitrate accumulation in anammox reactors under different nitrogen loading rates (NLRs) is well understood. Consequently, these parameters were studied in a lower NLR anammox reactor, termed R1, producing higher than expected levels of nitrate and compared with a higher NLR reactor, termed R2, showing no excess nitrate production. While both reactors showed high NH4+-N removal efficiencies (>90%), the total nitrogen removal efficiency (<60%) was much lower in R1 due to higher nitrate production. Metagenomic analysis found that the number of reads derived from anammox bacteria were significantly higher in R2. Another notable trend in reads occurrence was the relatively higher levels of reads from genes predicted to be nitrite oxidoreductases (nxr) in R1. Binning yielded 27 high quality draft genomes from the two reactors. Analysis of bin occurrence found that R1 showing both a decrease in anammox bacteria and an unexpected increase in nxr. In-situ assays confirmed that R1 had higher rates of nitrite oxidation to nitrate and suggested that it was not solely due to obligate NOB, but other nxr-containing bacteria are important contributors as well. Our results demonstrate that nitrate accumulation can be a serious operational concern for the application of anammox technology to low-strength wastewater treatment and provide insight into the community changes leading to this outcome.

15.
Opt Lett ; 44(23): 5772-5775, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31774776

RESUMO

The nanoscale wavelength and polarization router, which can simultaneously separate wavelength and polarization modes, is an essential component of on-chip nanophotonic devices. Here, an on-chip wavelength and polarization router is realized experimentally based on a three-layer hybrid waveguide of Au-SiO2-LiNbO3 etched with asymmetric nano-cavities. The central area size of the device is only 1.60×1.96 µm2. A broad operation band covers from 500 nm to 1150 nm with low cross talk of under 10 dB. The monolithic-LiNbO3is introduced for the first time, to the best of our knowledge, to on-chip multichannel wavelength and polarization routers. This work plays a key role for dense chip integration, visible light displays, and communications, and can inspire LiNbO3-based nanophotonic devices.

16.
J Zhejiang Univ Sci B ; 20(12): 995-1002, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31749346

RESUMO

OBJECTIVE: This study aimed to clone and characterize the oxiranedicarboxylate hydrolase (ORCH) from Labrys sp. WH-1. METHODS: Purification by column chromatography, characterization of enzymatic properties, gene cloning by protein terminal sequencing and polymerase chain reaction (PCR), and sequence analysis by secondary structure prediction and multiple sequence alignment were performed. RESULTS: The ORCH from Labrys sp. WH-1 was purified 26-fold with a yield of 12.7%. It is a monomer with an isoelectric point (pI) of 8.57 and molecular mass of 30.2 kDa. It was stable up to 55 °C with temperature at which the activity of the enzyme decreased by 50% in 15 min (T5015) of 61 °C and the half-life at 50 °C (t1/2, 50 °C) of 51 min and was also stable from pH 4 to 10, with maximum activity at 55 °C and pH 8.5. It is a metal-independent enzyme and strongly inhibited by Cu2+, Ag+, and anionic surfactants. Its kinetic parameters (Km, kcat, and kcat/Km) were 18.7 mmol/L, 222.3 s-1, and 11.9 mmol/(L·s), respectively. The ORCH gene, which contained an open reading frame (ORF) of 825 bp encoding 274 amino acid residues, was overexpressed in Escherichia coli and the enzyme activity was 33 times higher than that of the wild strain. CONCLUSIONS: The catalytic efficiency and thermal stability of the ORCH from Labrys sp. WH-1 were the best among the reported ORCHs, and it provides an alternative catalyst for preparation of L(+)-2,3-dihydrobutanedioic acid.

17.
Mol Vis ; 25: 654-662, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31741654

RESUMO

Purpose: To identify any novel mutations in CYP4V2 in 85 Chinese families with Bietti corneoretinal crystalline dystrophy (BCD) by using next-generation sequencing, and to summarize the mutation spectrum in this population, along with any genotype-phenotype correlations. Methods: A total of 90 patients with BCD from 85 unrelated Chinese families were recruited. All probands were analyzed by using gene chip-based next-generation sequencing, to capture and sequence all the exons of 57 known hereditary retinal degeneration-associated genes. The candidate variants were validated with PCR and Sanger sequencing. Results: Twenty-eight mutations were detected in all patients, including thirteen novel mutations (five missense, six deletions, one splicing and one frame-shift mutations) and 15 previously reported mutations. Mutations in 64 patients were inherited from their parents, while three patients had de novo mutations. c.802-8_810del17insGC was the most common mutation, accounting for 78% of the mutations. Although 16 patients were homozygous at this site, the clinical features of all 16 patients were highly heterogeneous. Conclusions: These results expand the spectrum of mutations in CYP4V2, and suggest that mutations in CYP4V2 may be common in the Chinese population. The phenotype of patients with the homozygous mutation (hom.c.802-8_810del17insGC) is highly heterogeneous.

18.
Adv Ther ; 2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31755037

RESUMO

INTRODUCTION: This study aimed to investigate the expression of a 70-kDa heat shock protein [heat shock 70-kDa protein 8 (HSPA8)/heat shock protein 70 (Hsc70)] in human degenerative lumbar intervertebral discs and its relationship with the degree of degeneration of human intervertebral discs. METHODS: A total of 72 cases of lumbar intervertebral disc nucleus pulposus tissues were collected. Among these, 18 cases of nucleus pulposus tissue were assigned to the control group, while 54 cases of nucleus pulposus tissues were assigned to the experimental group. According to the preoperative MRI, cases in the experimental group were further divided into three groups: protrusion group (n = 18), extrusion group (n = 18), and sequestration group (n = 18). Western blot was performed to determine the relative expression of HSPA8 in the nucleus pulposus in each group. Hematoxylin and eosin staining was performed to determine the number of nucleus pulposus cells, morphological differences, and cell densities of the degenerated intervertebral discs and normal intervertebral discs. Immunohistochemistry was performed to determine the expression of HSPA8 in nucleus pulposus tissues in each group. RESULTS: Hematoxylin and eosin staining results: There were significant differences in cell morphology and number between the control group and the experimental group. Furthermore, there were significant differences in cell density (F = 936.80, P < 0.01). Immunohistochemistry results: HSPA8 was expressed in lumbar intervertebral disc nucleus pulposus tissues, and its expression of gradually decreased with the severity of the disease, and the differences were significant (F = 2110.43, P < 0.01). Western blot results: The expression of HSPA8 in human degenerative nucleus pulposus tissues gradually decreased, and the differences were significant (F = 1841.72, P < 0.01). CONCLUSION: HSPA8 is stably expressed in human intervertebral disc nucleus pulposus tissues, and its expression is associated with the degree of intervertebral disc degeneration.

19.
Biosens Bioelectron ; 148: 111810, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31710960

RESUMO

Poly (ADP-ribose) polymerase-1 (PARP-1) was defined as a new biomarker, which has achieved wide attention in recent years. In this work, we designed a renewable electrochemical sensor based on host-guest recognition for the detection of PARP-1 activity. Mono-(6-Mercapto-6-deoxy)-beta-Cyclodextrin (SH-ß-CD) was modified on the electrode surface to recognize the trans-azobenzene labeled dsDNA (Abz-dsDNA). In the presence of PARP-1, PAR with abundant PO43- was generated and reacted with MoO42- to form PMo12O403-, producing strong current. The proposed method avoided the unspecific adsorption effectively and improved the detection accuracy. Under UV irradiation, Abz-dsDNA was removed from the electrode surface because of the configuration change of azobenzene from trans to cis structure, allowing the electrode to be recycled. The sensor realized the linear detection of PARP-1, ranging from 0.01 U to 1.0 U with a detection limit of 0.008 U, which is comparable to results from reported methods. It is expected to be a potential tool for clinical detection because of its high sensitivity and selectivity.

20.
Stem Cells ; 2019 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-31721342

RESUMO

The tunica adventitia ensheathes arteries and veins and contains presumptive mesenchymal stem cells (MSCs) involved in vascular remodeling. We show here that a subset of human adventitial cells express the CD10/CALLA cell surface metalloprotease. Both CD10+ and CD10- adventitial cells displayed phenotypic features of MSCs when expanded in culture. However, CD10+ adventitial cells exhibited higher proliferation, clonogenic and osteogenic potentials in comparison to their CD10- counterparts. CD10+ adventitial cells increased expression of the cell cycle protein CCND2 via ERK1/2 signaling and osteoblastogenic gene expression via NF-κB signaling. CD10 expression was upregulated in adventitial cells through Sonic hedgehog-mediated GLI1 signaling. These results suggest that CD10, which marks rapidly dividing cells in other normal and malignant cell lineages, plays a role in perivascular MSC function and cell fate specification. These findings also point to a role for CD10+ perivascular cells in vascular remodeling and calcification.

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