Publications by authors named "Yuxiang Zhang"

137 Publications

Molecular basis and homeostatic regulation of Zinc taste.

Protein Cell 2021 Apr 23. Epub 2021 Apr 23.

School of Life Sciences, IDG/McGovern Institute for Brain Research, Tsinghua University, Beijing, 100084, China.

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http://dx.doi.org/10.1007/s13238-021-00845-8DOI Listing
April 2021

Dynamic Changes of Cytoskeleton-Related Proteins Within Reward-Related Brain Regions in Morphine-Associated Memory.

Front Neurosci 2020 28;14:626348. Epub 2021 Jan 28.

College of Forensic Medicine, Xi'an Jiaotong University Health Science Center, Xi'an, China.

Drug-induced memory engages complex and dynamic processes and is coordinated at multiple reward-related brain regions. The spatiotemporal molecular mechanisms underlying different addiction phases remain unknown. We investigated the role of β-actin, as well as its potential modulatory protein activity-regulated cytoskeletal-associated protein (Arc/Arg3.1) and extracellular signal-regulated kinase (ERK), in reward-related associative learning and memory using morphine-induced conditioned place preference (CPP) in mice. CPP was established by alternate morphine (10 mg/kg) injections and extinguished after a 10-day extinction training, while the withdrawal group failed to extinguish without training. In the nucleus accumbens (NAc), morphine enhanced the level of β-actin and Arc only during extinction, while p-ERK1/2 was increased during both CPP acquisition and extinction phases. In the dorsal hippocampus, morphine induced an upregulation of p-ERK only during extinction, while p-β-actin was elevated during both CPP establishment and extinction. In the dorsal hippocampus, Arc was elevated during CPP formation and suppressed during extinction. Compared with the NAc and dorsal hippocampus, dynamic changes in the medial prefrontal cortex (mPFC) and caudate putamen (CPu) were not very significant. These results suggested region-specific changes of p-β-actin, Arc/Arg3.1, and p-ERK1/2 protein during establishment and extinction phases of morphine-induced CPP. These findings unveiled a spatiotemporal molecular regulation in opiate-induced plasticity.
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http://dx.doi.org/10.3389/fnins.2020.626348DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7876246PMC
January 2021

[Experience of treating batches of exertional heat stroke patients in military training].

Zhonghua Wei Zhong Bing Ji Jiu Yi Xue 2020 Dec;32(12):1522-1525

Department of Critical Care Medicine, the Eighth Medical Center of Chinese PLA General Hospital, Beijing 100091, China. Corresponding author: Zhang Yuxiang, Email:

Objective: To explore the process and segments of intense treatment of patients suffered from exertional heat stroke (EHS), and to provide the experience and guidance for treating batches of EHS patients.

Methods: A retrospective study of the curing process of eight EHS patients admitted to the Eighth Medical Center of Chinese PLA General Hospital on May 18th, 2019 was conducted, including onsite treatment and subsequent transshipment to hospital for treatment.

Results: Based on the medical history with high-intensity exercise and the clinical manifestation of central nervous system dysfunction, the eight patients were quickly diagnosed as EHS on the site, and were immediately rehydrated and attained the systematic cooling. An emergency plan for the treatment of batches of patients was initiated instantly for quick medical evacuation and care, and the patients were quickly transferred to the Eighth Medical Center of Chinese PLA General Hospital. A therapeutic principle was strictly followed, concerning the transshipment accompanied by systematic cooling while the medical evacuation and care, therapeutic preparation and immediate opening of the green channel were started up in the hospital. Sticking to the plan of allocating patients vs. nurses vs. doctor by 1:3:1, enough persons were dispatched. Ventilators, hemofiltration machines and ice blanket machines were prepared. Through the professional diagnosis and treatment in the emergency department, the patients were transferred to the ICU department. The relevant medical departments were coordinated with the centralization of the ICU department. The vital signs of patients were monitored closely. The titration and precise treatment were implemented, based on the changes in blood routine examination, blood biochemical indexes, blood clotting function, and blood gas analysis. The continuous renal replacement therapy (CRRT) was performed for four patients, the ice blanket machine cooling therapy for two patients, and the tracheal intubation and mechanical ventilation for one patient. Low molecular weight heparin sodium anticoagulant treatment was used in one patient with the diffuse intravascular coagulation (DIC) on the basis of coagulation factors such as plasmas and fibrinogens supplementation. The function of the damaged organs was significantly improved in 5-11 days. Eight patients were finally transferred out from ICU department. All patients were recovered and discharged after 11-20 days.

Conclusions: A normative treatment process (rapid recognition on the site, rapid cooling, rapid and safe medical evacuation, in-hospital multidisciplinary collaboration) combined bundles of therapeutic strategy such as systematic cooling and rehydration, CRRT, and supplying the coagulation factors while anticoagulation treatment can provide a strong guarantee for rescuing batches of EHS patients and achieve satisfactory treatment outcomes.
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http://dx.doi.org/10.3760/cma.j.cn121430-20200303-00236DOI Listing
December 2020

A short ORF-encoded transcriptional regulator.

Proc Natl Acad Sci U S A 2021 Jan;118(4)

Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037;

Recent technological advances have expanded the annotated protein coding content of mammalian genomes, as hundreds of previously unidentified, short open reading frame (ORF)-encoded peptides (SEPs) have now been found to be translated. Although several studies have identified important physiological roles for this emerging protein class, a general method to define their interactomes is lacking. Here, we demonstrate that genetic incorporation of the photo-crosslinking noncanonical amino acid AbK into SEP transgenes allows for the facile identification of SEP cellular interaction partners using affinity-based methods. From a survey of seven SEPs, we report the discovery of short ORF-encoded histone binding protein (SEHBP), a conserved microprotein that interacts with chromatin-associated proteins, localizes to discrete genomic loci, and induces a robust transcriptional program when overexpressed in human cells. This work affords a straightforward method to help define the physiological roles of SEPs and demonstrates its utility by identifying SEHBP as a short ORF-encoded transcription factor.
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http://dx.doi.org/10.1073/pnas.2021943118DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7848545PMC
January 2021

Adaptive Decision-Making for Automated Vehicles Under Roundabout Scenarios Using Optimization Embedded Reinforcement Learning.

IEEE Trans Neural Netw Learn Syst 2020 Dec 30;PP. Epub 2020 Dec 30.

The roundabout is a typical changeable, interactive scenario in which automated vehicles should make adaptive and safe decisions. In this article, an optimization embedded reinforcement learning (OERL) is proposed to achieve adaptive decision-making under the roundabout. The promotion is the modified actor of the Actor-Critic framework, which embeds the model-based optimization method in reinforcement learning to explore continuous behaviors in action space directly. Therefore, the proposed method can determine the macroscale behavior (change lane or not) and medium-scale behaviors of desired acceleration and action time simultaneously with high sample efficiency. When scenarios change, medium-scale behaviors can be adjusted timely by the embedded direct search method, promoting the adaptability of decision-making. More notably, the modified actor matches human drivers' behaviors, macroscale behavior captures the human mind's jump, and medium-scale behaviors are preferentially adjusted through driving skills. To enable the agent adapts to different types of the roundabout, task representation is designed to restructure the policy network. In experiments, the algorithm efficiency and the learned driving strategy are compared with decision-making containing macroscale behavior and constant medium-scale behaviors of the desired acceleration and action time. To investigate the adaptability, the performance under an untrained type of roundabout and two more dangerous situations are simulated to verify that the proposed method changes the decisions with changeable scenarios accordingly. The results show that the proposed method has high algorithm efficiency and better system performance.
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http://dx.doi.org/10.1109/TNNLS.2020.3042981DOI Listing
December 2020

Synthesis and discovery of ω-3 polyunsaturated fatty acid- alkanolamine (PUFA-AA) derivatives as anti-inflammatory agents targeting Nur77.

Bioorg Chem 2020 12 8;105:104456. Epub 2020 Nov 8.

Fujian Provincial Key Laboratory of Innovative Drug Target Research and State Key Laboratory of Cellular Stress Biology, School of Pharmaceutical Sciences, Xiamen University, Xiamen 361102, China. Electronic address:

In this work, three series of ω-3 polyunsaturated fatty acid-alkanolamine derivatives (PUFA-AAs) were synthesized, characterized and their anti-inflammatory activity in vivo was evaluated. Compounds 4a, 4f, and 4k exhibited marked anti-inflammatory activity in LPS-stimulated RAW 264.7 cells. The most promising compound 4k dose-dependently suppressed the cytokines with IC values in the low micromolar range. Further, 4k exhibited potential in vitro Nur77-binding affinity (K = 6.99 × 10 M) which is consistent with the result of docking studies. Next, the anti-inflammatory mechanism of 4k was found to be through NF-κB signal pathway in a Nur77-dependent manner. Moreover, we also observed 4k significantly inhibited LPS-induced expression of cytokines (IL-6, TNF-α, and IL-1β) through suppressing NF-κB activation and attenuated LPS-induced inflammation in mouse acute lung injury (ALI) model. In conclusion, the study strongly suggests that the PUFA-AA derivatives can be particularly as new Nur77 mediators for further treatment in inflammatory diseases.
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http://dx.doi.org/10.1016/j.bioorg.2020.104456DOI Listing
December 2020

Water extract of sporoderm-broken spores of Ganoderma lucidum enhanced pd-l1 antibody efficiency through downregulation and relieved complications of pd-l1 monoclonal antibody.

Biomed Pharmacother 2020 Nov 18;131:110541. Epub 2020 Sep 18.

The Second Affiliated Hospital Zhejiang University School of Medicine, Jiefang Campus, 88 Jiefang Road, Shangcheng District, Hangzhou, 310009, China; Zhejiang University School of Medicine, Zhejiang University Huajiachi Campus, 268 Kaixuan Road, Jianggan District, Hangzhou, 310029, China. Electronic address:

Purpose: Osteosarcoma is a malignant musculoskeletal tumor with early metastasis and a poor prognosis, especially in adolescents. Ganoderma lucidum (Leyss. Ex Fr.) Karst (G. lucidum), a traditional East Asian medicine, has been reported to play a critical role in antitumor and immunomodulatory activity. The aim of this study was to investigate the effects and molecular mechanisms of water extract of sporoderm-broken spores of G. lucidum (BSGWE) on osteosarcoma PD-L1 (programmed cell death-ligand 1) transcriptional regulation, efficacy enhancement, and side effect remission.

Methods: The antitumor effects on cell proliferation of BSGWE in osteosarcoma cells were detected by apoptosis flow cytometry, and the migration ability of HOS and K7M2 cells were evaluated by cell scratch assay. Potential signaling regulation of PD-L1 was detected by western blotting. To confirm the signaling pathway of BSGWE-related PD-L1 downregulation, a pho-STAT3 turnover experiment was carried out. Colivelin was administered as a pho-STAT3 activator to rescue the BSGWE-induced PD-L1 inhibition. To further study in vivo signaling, in a Balb/c osteosarcoma allograft model, tumor volume was measured using an in vivo bioluminescence imaging system. The body weight curve and tumor volume curve were analyzed to reveal the remission effects of BSGWE on PD-L1 antibody-related body weight loss and its immunomodulatory effects on the osteosarcoma and spleen. The PD-L1 expression level and expression of related transcription-factor pho-STAT3 in tumor cells and spleens were assessed by IHC analysis.

Results: BSGWE suppressed the proliferation and migration of osteosarcoma cells in vitro via induction of apoptosis. In addition, BSGWE downregulated PD-L1 expression and related STAT3 (signal transducers and activators of transcription) phosphorylation levels in a dose-dependent manner. Western blotting and qRT-PCR assay revealed that BSGWE downregulated PD-L1 expression by inhibiting STAT3 phosphorylation. A turnover experiment showed that colivelin administration could rescue PD-L1 inhibition via pho-STAT3 activation. BSGWE not only downregulated PD-L1 expression via the STAT3 pathway in an allograft Balb/c mouse model, but also relieved complications including weight loss and spleen atrophy in a mouse monoclonal antibody therapy model on the basis of its traditional advantages in immune enhancement.

Conclusion: BSGWE downregulated PD-L1 expression via pho-STAT3 inhibition of protein and RNA levels. BSGWE enhanced PD-L1 antibody efficacy via phosphorylated STAT3 downregulation in vitro and in vivo. BSGWE also relieved complications of weight loss and spleen atrophy in a murine allograft osteosarcoma immune checkpoint blockade therapy model.
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http://dx.doi.org/10.1016/j.biopha.2020.110541DOI Listing
November 2020

The 5-HT Receptors in the Ventrolateral Orbital Cortex Attenuate Allodynia in a Rodent Model of Neuropathic Pain.

Front Neurosci 2020 18;14:884. Epub 2020 Aug 18.

College of Forensic Medicine, Xi'an Jiaotong University Health Science Center, Xi'an, China.

Mechanical allodynia, characterized by a painful sensation induced by innocuous stimuli, is thought to be caused by disruption in pain-related regions. Identification and reversal of this pathologic neuroadaptation are therefore beneficial for clinical treatment. Previous evidence suggests that 5-HT receptors in the ventrolateral orbital cortex (VLO) are involved in neuropathic pain, but their function is poorly understood. The aim of the present study is to unveil the role of 5-HT receptors in the VLO and the underlying mechanisms in pain modulation. Here, by using the spared nerve injury (SNI) pain model, first, we report that 5-HT receptor protein decreased in the contralateral VLO compared with the ipsilateral VLO in rats with allodynia. Second, microinjection of the selective 5-HT receptor agonists EMD-386088 and WAY-208466 into the contralateral VLO consistently and significantly depressed allodynia. Third, microinjection of the selective antagonist SB-258585 blocked the agonist-induced anti-allodynic effect, while the antagonist applied alone to the VLO had no effect. Furthermore, the anti-nociceptive effect of EMD-386088 on neuropathic pain was prevented by the adenylate cyclase (AC) inhibitor SQ-22536, and protein kinase A (PKA) inhibitor H89, suggesting that AC/PKA signaling might underlie the antinociception of agonists. Finally, the 5-HT receptors were found to be colocalized with a glutamate transporter (EAAC1) by immunofluorescent staining, and the glutamate receptor antagonist kynurenic acid was found to completely block antinociception. These findings indicated that the antinociceptive effect of 5-HT receptor agonists might occur via interaction with the glutamatergic system. Altogether, the agonists activated 5-HT receptors present in the glutamatergic neurons in the VLO to facilitate the AC/PKA cascade, which subsequently might evoke glutamate release, thus depressing allodynia. These findings suggest a potential therapeutic role of 5-HT receptor agonists in treating neuropathic pain.
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http://dx.doi.org/10.3389/fnins.2020.00884DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7461796PMC
August 2020

The neurophysiology and seizure outcomes of late onset unexplained epilepsy.

Clin Neurophysiol 2020 11 19;131(11):2667-2672. Epub 2020 Sep 19.

Department of Neurology, Massachusetts General Hospital, Harvard Medical School, MA, USA; Department of Neurology, Center for Alzheimer Research and Treatment, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.

Objective: To investigate neurophysiologic and neuroimaging characteristics of patients with late onset unexplained epilepsy (LOUE).

Methods: We performed a retrospective chart review of elderly patients with ICD9 diagnosis codes consistent with epilepsy/seizures. Inclusion criteria included unprovoked seizures, and absence of cortical lesions on magnetic resonance imaging (MRI). Electroencephalograms (EEGs) findings were also analyzed. MRI images were scored for degree of white matter hyperintensities (Fazekas Scale) and mesial temporal atrophy (MTA). Vascular risk factors, and Framingham Heart Study general cardiovascular disease (FHS-CVD) risk scores were compared to controls from the Harvard Aging Brain study (HABS).

Results: We identified 224 LOUE patients and 8% were drug resistant. Epileptiform abnormalities were captured on EEG in 35%. The location was temporal with left sided predominance in 49%. Fazekas scale consisted of 25% beginning of confluent lesions, and 10% large confluent lesions. MTA scores consisted of 21% moderate-severe hippocampal atrophy. LOUE patients had on average a 2.3% (adjusted), 7.4% (unadjusted) increased FHS-CVD score.

Conclusions: Our findings highlight LOUE as pharmacosensitive and left temporal predominant. Given the higher prevalence of vascular risk factors, investigations are needed to study their role in pathophysiology.

Significance: Physicians caring for patients with LOUE should evaluate for vascular risk factors and investigate the presence of hippocampal atrophy.
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http://dx.doi.org/10.1016/j.clinph.2020.08.014DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7644268PMC
November 2020

Spectral-Spatial Weighted Kernel Manifold Embedded Distribution Alignment for Remote Sensing Image Classification.

IEEE Trans Cybern 2020 Jul 10;PP. Epub 2020 Jul 10.

Feature distortions of data are a typical problem in remote sensing image classification, especially in the area of transfer learning. In addition, many transfer learning-based methods only focus on spectral information and fail to utilize spatial information of remote sensing images. To tackle these problems, we propose spectral-spatial weighted kernel manifold embedded distribution alignment (SSWK-MEDA) for remote sensing image classification. The proposed method applies a novel spatial information filter to effectively use similarity between nearby sample pixels and avoid the influence of nonsample pixels. Then, a complex kernel combining spatial kernel and spectral kernel with different weights is constructed to adaptively balance the relative importance of spectral and spatial information of the remote sensing image. Finally, we utilize the geometric structure of features in manifold space to solve the problem of feature distortions of remote sensing data in transfer learning scenarios. SSWK-MEDA provides a novel approach for the combination of transfer learning and remote sensing image characteristics. Extensive experiments have demonstrated that the proposed method is more effective than several state-of-the-art methods.
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http://dx.doi.org/10.1109/TCYB.2020.3004263DOI Listing
July 2020

An oriented-collagen scaffold including Wnt5a promotes osteochondral regeneration and cartilage interface integration in a rabbit model.

FASEB J 2020 08 6;34(8):11115-11132. Epub 2020 Jul 6.

Department of Orthopedic Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, P.R. China.

Articular cartilage regeneration remains a major challenge in orthopedics. Noncanonical Wnt5a is a particularly attractive growth factor in this context; Wnt5a inhibits chondrocyte hypertrophy but maintains chondrogenesis. We designed a novel, vertically oriented-collagen scaffold. The effect of Wnt5a on MSCs and chondrocytes and the therapeutic effects of the Wnt5a/oriented-collagen scaffold in terms of osteochondral repair and cartilage integration were evaluated. In vitro, the proliferation, migration, and differentiation of MSCs and chondrocytes treated with Wnt5a, and the mechanisms thereof, were assessed. mRNA microarray analysis was performed to compare the expression profiles of MSCs before and after Wnt5a treatment. In vivo, full-thickness cylindrical osteochondral defects (4 mm in diameter, 3 mm in depth) were created in the patellar grooves of 24 New Zealand white rabbits and implanted with oriented-collagen scaffolds (n = 8), Wnt5a/oriented-collagen scaffolds (n = 8), or nothing (n = 8). After 6 and 12 weeks, integration and tissue responses were evaluated. The proliferation, migration, chondrogenic differentiation, and extracellular matrix formation of/by MSCs and chondrocytes improved greatly after treatment with Wnt5a. Western blotting showed that the PI3K/AKT/JNK signaling pathway was activated. Microarray analysis revealed that the Wnt5a group exhibited a significant upregulation of the PI3K pathway. Reactome GSEA pathway interaction analysis revealed that such upregulation was associated with collagen and extracellular matrix formation. In vivo, the Wnt5a/oriented-collagen scaffold group exhibited optimal interface integration, cartilage regeneration, and collagenous fiber arrangement, accompanied by significantly increased glycosaminoglycan and collagen accumulations in the zones of regeneration and integration, compared to the other groups. Gene expression analysis showed that the levels of mRNAs encoding genes involved in cartilage formation were significantly increased in the Wnt5a/oriented, collagen scaffold group (all P < .05). Wnt5a promoted the proliferation, migration, and chondrogenic differentiation of MSCs and chondrocytes via the activation of the PI3K/AKT/JNK signaling pathway. The Wnt5a/oriented-collagen constructs enhanced the structure-specific regeneration of hyaline cartilage in a rabbit model and may be a promising treatment for the repair of human cartilage defects.
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http://dx.doi.org/10.1096/fj.202000280RDOI Listing
August 2020

BCR selection and affinity maturation in Peyer's patch germinal centres.

Nature 2020 06 6;582(7812):421-425. Epub 2020 May 6.

Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.

The antigen-binding variable regions of the B cell receptor (BCR) and of antibodies are encoded by exons that are assembled in developing B cells by V(D)J recombination. The BCR repertoires of primary B cells are vast owing to mechanisms that create diversity at the junctions of V(D)J gene segments that contribute to complementarity-determining region 3 (CDR3), the region that binds antigen. Primary B cells undergo antigen-driven BCR affinity maturation through somatic hypermutation and cellular selection in germinal centres (GCs). Although most GCs are transient, those in intestinal Peyer's patches (PPs)-which depend on the gut microbiota-are chronic, and little is known about their BCR repertoires or patterns of somatic hypermutation. Here, using a high-throughput assay that analyses both V(D)J segment usage and somatic hypermutation profiles, we elucidate physiological BCR repertoires in mouse PP GCs. PP GCs from different mice expand public BCR clonotypes (clonotypes that are shared between many mice) that often have canonical CDR3s in the immunoglobulin heavy chain that, owing to junctional biases during V(D)J recombination, appear much more frequently than predicted in naive B cell repertoires. Some public clonotypes are dependent on the gut microbiota and encode antibodies that are reactive to bacterial glycans, whereas others are independent of gut bacteria. Transfer of faeces from specific-pathogen-free mice to germ-free mice restored germ-dependent clonotypes, directly implicating BCR selection. We identified somatic hypermutations that were recurrently selected in such public clonotypes, indicating that affinity maturation occurs in mouse PP GCs under homeostatic conditions. Thus, persistent gut antigens select recurrent BCR clonotypes to seed chronic PP GC responses.
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http://dx.doi.org/10.1038/s41586-020-2262-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7478071PMC
June 2020

Near-infrared-excited upconversion photodynamic therapy of extensively drug-resistant Acinetobacter baumannii based on lanthanide nanoparticles.

Nanoscale 2020 Jul 12;12(26):13948-13957. Epub 2020 May 12.

State Key Laboratory of Structural Chemistry, CAS Key Laboratory of Design and Assembly of Functional Nanostructures, and Fujian Key Laboratory of Nanomaterials, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fujian College, University of Chinese Academy of Sciences, Fuzhou, Fujian 350002, China.

Extensively drug-resistant Acinetobacter baumannii (XDR-AB) has raised considerable concerns due to its mortal damage to humans and its high transmission rate of infections in hospitals. However, current antibiotics not only show poor anti-infection effects in vivo but also frequently cause high nephrotoxicity and neurotoxicity. Herein, we report a near-infrared (NIR) light-initiated antimicrobial photodynamic therapy (aPDT) to effectively treat in vivo XDR-AB infections based on photosensitizer (PS) loaded upconversion nanoparticles (UCNPs, LiYF:Yb/Er). Such nanoagents feature robust NIR triggered UC luminescence and high-efficiency energy transfer from UCNPs to the loaded PS, thereby allowing NIR-triggered generation of reactive oxygen species (ROS) for destroying the bacterial cell membrane. This strategy permits a high antibacterial activity against XDR-AB, resulting in a decline of 4.72 log in viability at a dose of 50 μg mL UCNPs-PVP-RB with 980 nm laser irradiation (1 W cm). More significantly, we can achieve excellent therapeutic efficacy against deep-tissue (about 5 mm) XDR-AB infections without causing any side effects in the murine model. In brief, such NIR-activated aPDT may open up new avenues for treating various deep-tissue intractable infections.
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http://dx.doi.org/10.1039/d0nr01073aDOI Listing
July 2020

Induction of recurrent break cluster genes in neural progenitor cells differentiated from embryonic stem cells in culture.

Proc Natl Acad Sci U S A 2020 05 24;117(19):10541-10546. Epub 2020 Apr 24.

Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115;

Mild replication stress enhances appearance of dozens of robust recurrent genomic break clusters, termed RDCs, in cultured primary mouse neural stem and progenitor cells (NSPCs). Robust RDCs occur within genes ("RDC-genes") that are long and have roles in neural cell communications and/or have been implicated in neuropsychiatric diseases or cancer. We sought to develop an in vitro approach to determine whether specific RDC formation is associated with neural development. For this purpose, we adapted a system to induce neural progenitor cell (NPC) development from mouse embryonic stem cell (ESC) lines deficient for XRCC4 plus p53, a genotype that enhances DNA double-strand break (DSB) persistence to enhance detection. We tested for RDCs by our genome-wide DSB identification approach that captures DSBs via their ability to join to specific genomic Cas9/single-guide RNA-generated bait DSBs. In XRCC4/p53-deficient ESCs, we detected seven RDCs, all of which were in genes and two of which were robust. In contrast, in NPCs derived from these ESC lines we detected 29 RDCs, a large fraction of which were robust and associated with long, transcribed neural genes that were also robust RDC-genes in primary NSPCs. These studies suggest that many RDCs present in NSPCs are developmentally influenced to occur in this cell type and indicate that induced development of NPCs from ESCs provides an approach to rapidly elucidate mechanistic aspects of NPC RDC formation.
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http://dx.doi.org/10.1073/pnas.1922299117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7229678PMC
May 2020

Telomere length heterogeneity in ALT cells is maintained by PML-dependent localization of the BTR complex to telomeres.

Genes Dev 2020 05 26;34(9-10):650-662. Epub 2020 Mar 26.

Laboratory of Genome Integrity, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.

Telomeres consist of TTAGGG repeats bound by protein complexes that serve to protect the natural end of linear chromosomes. Most cells maintain telomere repeat lengths by using the enzyme telomerase, although there are some cancer cells that use a telomerase-independent mechanism of telomere extension, termed alternative lengthening of telomeres (ALT). Cells that use ALT are characterized, in part, by the presence of specialized PML nuclear bodies called ALT-associated PML bodies (APBs). APBs localize to and cluster telomeric ends together with telomeric and DNA damage factors, which led to the proposal that these bodies act as a platform on which ALT can occur. However, the necessity of APBs and their function in the ALT pathway has remained unclear. Here, we used CRISPR/Cas9 to delete PML and APB components from ALT-positive cells to cleanly define the function of APBs in ALT. We found that PML is required for the ALT mechanism, and that this necessity stems from APBs' role in localizing the BLM-TOP3A-RMI (BTR) complex to ALT telomere ends. Strikingly, recruitment of the BTR complex to telomeres in a PML-independent manner bypasses the need for PML in the ALT pathway, suggesting that BTR localization to telomeres is sufficient to sustain ALT activity.
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http://dx.doi.org/10.1101/gad.333963.119DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7197349PMC
May 2020

Cell-Based Ligand Discovery for the ENL YEATS Domain.

ACS Chem Biol 2020 04 19;15(4):895-903. Epub 2020 Mar 19.

Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States.

ENL is a transcriptional coactivator that recruits elongation machinery to active -regulatory elements upon binding of its YEATS domain-a chromatin reader module-to acylated lysine side chains. Discovery chemistry for the ENL YEATS domain is highly motivated by its significance in acute leukemia pathophysiology, but cell-based assays able to support large-scale screening or hit validation efforts do not presently exist. Here, we report on the discovery of a target engagement assay that allows for high-throughput ligand discovery in living cells. This assay is based on the cellular thermal shift assay (CETSA) but does not require exposing cells to elevated temperatures, as small-molecule ligands are able to stabilize the ENL YEATS domain at 37 °C. By eliminating temperature shifts, we developed a simplified target engagement assay that requires just two steps: drug treatment and luminescence detection. To demonstrate its value for higher throughput applications, we miniaturized the assay to a 1536-well format and screened 37 120 small molecules, ultimately identifying an acyl-lysine-competitive ENL/AF9 YEATS domain inhibitor.
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http://dx.doi.org/10.1021/acschembio.0c00124DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7384521PMC
April 2020

Genomic Insights Into Sugar Adaptation in an Extremophile Yeast .

Front Microbiol 2019 11;10:3157. Epub 2020 Feb 11.

College of Food Science and Engineering, Northwest University, Xi'an, China.

The osmotolerant is known for its trait to survive in extreme high sugar environments. This ability determines its role in the fermentation process and leads to yeast spoilage in the food industry. However, our knowledge of the gene expression in response to high sugar stress remains limited. Here, we conducted RNA-sequencing (RNA-seq) under different sugar concentrations of the spoilage yeast, , which exhibit extremely high tolerance to sugar stress. The obtained differentially expressed genes (DEGs) are significantly different to that of the , which is sensitive to extreme high sugar stress. Most of the DEGs participated in the "glucan synthesis," "transmembrane transport," "ribosome," etc. In this work, we also demonstrated that the gene () encoding Kar2p can significantly affect the growth of under high sugar stress. In addition, we combined with a previous study on the genome sequence of , indicating that several gene families contain significantly more gene copies in the lineage, which involved in tolerance to sugar stress. Our results provide a gene insight for understanding the high sugar tolerance trait, which may impact food and biotechnological industries and improve the osmotolerance in other organisms.
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http://dx.doi.org/10.3389/fmicb.2019.03157DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026193PMC
February 2020

Influencing factors and correlation of anxiety, psychological stress sources, and psychological capital among women pregnant with a second child in Guangdong and Shandong Province.

J Affect Disord 2020 03 30;264:115-122. Epub 2019 Nov 30.

The Affiliated Brain Hospital of Guangzhou Medical University (Guangzhou Huiai Hospital), Guangzhou, 510370, China; School of Health Management, Guangzhou Medical University, Guangzhou, 510182, China. Electronic address:

Background: After the one-child policy has been effective for over 30 years, China implemented the universal two-child policy in 2015. The number of high-risk pregnant women had increased dramatically ever since, increasing negative health outcomes for both mothers and children. Our study aims to investigate the status of anxiety, psychological stress sources, and psychological capital among women pregnant with a second child, and to assess influencing factors and correlations, providing scientific basis for promoting women's health during pregnancy.

Methods: We recruited 513 participants from maternity hospitals in Guangdong and Shandong Province. All participants completed the Zung Self-Rating Anxiety Scale (SAS), Pregnant women psychological stress sources questionnaire and the psychological capital questionnaire (PCQ-24).

Results: The prevalence of anxiety of women pregnant with a second child was 27.5%. The score of psychological stress sources was 74.93 ± 16.07. The risk factors for the anxiety of the women pregnant with a second child included low education level of the husband, gender expectation and the sources of psychological stress of the pregnant woman. The risk factors for psychological stress were dissatisfaction of residence and unplanned pregnancy. The main factors influencing psychological capital were the participants' education level, whether they have learned antenatal care knowledge and satisfaction with the residence.

Limitations: All participants were recruited only from tertiary hospitals in Guangdong and Shandong province.

Conclusions: Anxiety and psychological stress is prevalent during pregnancy among women pregnant with a second child in Guangdong and Shandong. Psychological capital was a protective factor for anxiety.
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http://dx.doi.org/10.1016/j.jad.2019.11.148DOI Listing
March 2020

Water Extract of Sporoderm-Broken Spores of Induces Osteosarcoma Apoptosis and Restricts Autophagic Flux.

Onco Targets Ther 2019 31;12:11651-11665. Epub 2019 Dec 31.

Department of Orthopedics Research Institute, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, People's Republic of China.

Purpose: Osteosarcoma (OS) is a malignant bone tumor with easy metastasis and poor prognosis. (), a traditional Chinese medicine, was reported playing a critical role in suppressing multiple tumor progress. So we wanted to investigate the effects and molecular mechanisms of water extract of sporoderm-broken spores of (BSGLWE) on osteosarcoma.

Methods: In vitro, the effects on cell proliferation of BSGLWE in osteosarcoma cells were detected by CCK-8, colony formation assay and flow cytometry; migration ability of osteosarcoma cells was evaluated by cell scratch and transwell assays. Cell apoptosis and autophagy were tested by transmission electron microscopy (TEM). Potential signaling pathways were detected by Western blotting and immunofluorescence. In xenograft orthotopic model, the luminescence intensity measured by an in vivo bioluminescence imaging system, and the expression of related proteins in tumor cells were assessed by IHC analysis.

Results: BSGLWE suppressed the proliferation and migration of osteosarcoma cells in a dose-dependent manner, and osteosarcoma cell cycle progression at the G2/M phase was arrested by the BSGLWE. In addition, increased apoptosis-related protein expression meant BSFLWE induced caspase-dependent apoptosis of osteosarcoma cells. TEM results indicated that BSGLWE promoted the formation of apoptotic bodies and autophagosomes in HOS and U2 cells. Western blotting or immunofluorescence and rescue assay revealed that BSGLWE blocked autophagic flux by inducing initiation of autophagy and increasing autophagosome accumulation of osteosarcoma cells. BSGLWE not only repressed the angiogenesis in the mouse model, but also induced apoptosis and blocked autophagy in vivo.

Conclusion: BSGLWE inhibits osteosarcoma progression.
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http://dx.doi.org/10.2147/OTT.S226850DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6942530PMC
December 2019

Assessment of chemical composition and sensorial properties of ciders fermented with different non-Saccharomyces yeasts in pure and mixed fermentations.

Int J Food Microbiol 2020 Apr 10;318:108471. Epub 2019 Dec 10.

College of Food Science and Engineering, Northwest A&F University, Yangling, 712100, China; Laboratory of Quality & Safety Risk Assessment for Agro-products (Yangling), Ministry of Agriculture, Yangling, 712100, China; National Engineering Research Center of Agriculture Integration Test (Yangling), Yangling 712100, China; College of Food Science and Technology, Northwest University, Xi'an, China. Electronic address:

This work presents the attempt to enhance the flavor complexity of cider fermented by different non-Saccharomyces species. Pichia kluyveri and Hanseniaspora vineae pure cultures were used as reference ciders. Mixed cultures between all 4 species gave 5 fermentations, where Hanseniaspora uvarum or Torulaspora quercuum were included for apple juice fermentation. Chemical composition and sensorial properties of all ciders were studied. The results indicated that the growth of P. kluyveri and H. vineae were interreacted and also affected by H. uvarum and T. quercuum. H. vineae was more capable of consuming sugar than P. kluyveri. Ciders from the single culture fermentation with P. kluyveri (Pk), as well as from mixed fermentation with P. kluyveri and H. uvarum (Pk-Hu), had high residual sugar, sugar/acid ratio, and glucose-fructose consumption ratio. Large shifts in the consumption and production of organic acids and polyphenols among all ciders were observed. The calculation of the relative odor activity value (rOAV) showed that 17 volatile compounds had an rOAV >1 in at least one sample, and acetate esters and ethyl esters were the groups with the highest number of volatile compounds of importance to the cider aroma. Among these 17 compounds, 3-methylbutyl acetate, 2-methylbutyl acetate, ethyl hexanoate, ethyl octanoate, and β-damascenone exhibited high rOAVs in some ciders and might contribute fruity, floral, and sweet features to the cider aroma. Besides, the tropical fruity aroma from 3-methylbutyl acetate was only perceived in Pk and Pk-Hu. The partial least squares regression (PLSR) analysis revealed that acetate esters contributed positively to the roasted and cooked odor of all ciders. This is the first study evaluating simultaneous fermentation of two non-Saccharomyces yeasts to produce cider, which provides new insights into cider production.
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http://dx.doi.org/10.1016/j.ijfoodmicro.2019.108471DOI Listing
April 2020

Thiol-functionalized inactivated yeast embedded in agar aerogel for highly efficient adsorption of patulin in apple juice.

J Hazard Mater 2020 04 30;388:121802. Epub 2019 Nov 30.

College of Food Science and Engineering, Northwest A&F University, Yangling, 712100, China; Laboratory of Quality & Safety Risk Assessment for Agro-products (Yangling), Ministry of Agriculture, Yangling, 712100, China; National Engineering Research Center of Agriculture Integration Test (Yangling), Yangling, 712100, China. Electronic address:

The issue of patulin (PAT) contamination in apple juice has attracted widespread concern. Recently, inactivated yeast based biosorbents have shown great advantages in the removal of toxic contaminants. However, the traditional yeast adsorbents have disadvantages of a limited adsorption capacity in juice and separation difficulty. In the present work, five chemical thiol-functionalization methods were used to increase the PAT adsorption efficiency of yeast cells in apple juice. Thereinto, glutaraldehyde cross-linking increased the thiol (-SH) content of yeast cells to 1.26 mmol g and improved the PAT adsorption capacity of inactivated yeast in apple juice by 150 times. The covalent bonding of -SH and PAT played an important role in the improvement of adsorption capacity. The as-prepared thiol-modification yeast (Y-SH(Gl)) was then embedded in the agar aerogel to obtain Y-SH(Gl)@Agar free of separation. PAT adsorption of Y-SH(Gl)@Agar was consistent with the Freundlich model and the pseudo-second-order kinetic model. Moreover, Y-SH(Gl)@Agar was competent for PAT removal in apple juice and manifested negligible effects on juice quality. Cytotoxicity investigation indicated its good biocompatibility and ignorable food safety risk, thereby demonstrating that Y-SH(Gl)@Agar may be a promising adsorbent material for the control of PAT contaminant in juice.
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http://dx.doi.org/10.1016/j.jhazmat.2019.121802DOI Listing
April 2020

Structure-Dependent Inhibition of by Polyphenol and Its Impact on Cell Membrane.

Front Microbiol 2019 13;10:2646. Epub 2019 Nov 13.

College of Food Science and Engineering, Northwest A&F University, Yangling, China.

As natural occurring antimicrobial substances, phenolic compounds have been used to inhibit various bacteria. 4-1, a strain isolated from food, exhibited spoilage potential with proteolysis and lipolysis at 25°C. The present study evaluated the antibacterial properties of 13 polyphenols on 4-1, and selected 6 compounds (ferulic acid, -coumaric acid, caffeic acid, chlorogenic acid, (-)-epigallocatechin, and phloretin) for binary combination treatments. The results revealed that antibacterial activities of polyphenols were structure-dependent, and cinnamic acid showed strong inhibitory effects, with a minimum inhibitory concentration (MIC) of 0.125 mg/mL. Importantly, we did not observe any obvious synergistic effects across all binary combinations. The antibacterial mechanism of cinnamic acid was related to membrane damage, caused by the loss of cell membrane integrity and alteration of cell morphology. These findings suggest that cinnamic acid is a promising candidate for the control of spoilage bacteria in food.
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http://dx.doi.org/10.3389/fmicb.2019.02646DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6863799PMC
November 2019

Laboratory study on the hydraulic characteristics of mechanically and biologically treated waste in China.

Waste Manag 2020 Feb 29;102:686-697. Epub 2019 Nov 29.

Zhejiang Sci-tech University, Hangzhou 310018, China.

Mechanical biological treatment (MBT) can greatly reduce the amount of municipal solid waste (MSW) and has become a hot topic in environmental geotechnical engineering. To study the effects of factors such as the pressure, dry density, compression time under pressure, strain, and hydraulic gradient, permeability tests with saturated MBT waste were conducted by using a compression and permeability combined apparatus in an environmental geotechnical laboratory. The results showed that Darcy's law was applicable to the saturated MBT waste. The permeability coefficient was found to be inversely related to the pressure, dry density, compression time under pressure and strain. The logarithmic permeability coefficient and the variables (e.g., pressure) could largely be fitted to a linear function. Additionally, a prediction model for the permeability coefficient was established, and the permeability coefficient at different depths of MBT landfills was predicted. The results were then compared to previous studies on MSW and MBT waste to investigate the permeability mechanism.
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http://dx.doi.org/10.1016/j.wasman.2019.11.026DOI Listing
February 2020

Chemical composition, sensorial properties, and aroma-active compounds of ciders fermented with Hanseniaspora osmophila and Torulaspora quercuum in co- and sequential fermentations.

Food Chem 2020 Feb 3;306:125623. Epub 2019 Oct 3.

College of Food Science and Engineering, Northwest A&F University, Yangling 712100, China; Laboratory of Quality & Safety Risk Assessment for Agro-products (Yangling), Ministry of Agriculture, Yangling 712100, China; National Engineering Research Center of Agriculture Integration Test (Yangling), Yangling 712100, China; College of Food Science and Technology, Northwest University, Xi' an 710000, China. Electronic address:

In this study, apple juice was fermented using Hanseniaspora osmophila X25-5 in pure culture as well as mixed culture with Torulaspora quercuum X24-4, which was inoculated simultaneously or sequentially. H. osmophila inhibited the growth of T. quercuum, while T. quercuum had little effect on the growth of H. osmophila. The simultaneous fermentation consumed relatively more sugar and resulted in the highest ethanol content. The production of organic acids varied depending on the yeast species employed and inoculation modality. Esters and alcohols were the main volatile families produced during fermentation, while ethyl esters and terpenes contributed most to the temperate fruity aroma. Gas chromatography-olfactometry (GC-O) showed that 3-methyl-1-butanol, ethyl 2-methylbutanoate, phenylethyl alcohol, β-phenethyl acetate, and β-damascenone were the most potent odorants in all samples. This study suggested that simultaneous fermentation with H. osmophila and T. quercuum might represent a novel strategy for the future production of cider.
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http://dx.doi.org/10.1016/j.foodchem.2019.125623DOI Listing
February 2020

[Progress in the application of high-flow nasal cannula oxygenation in immunosuppressed patients with acute respiratory failure].

Zhonghua Wei Zhong Bing Ji Jiu Yi Xue 2019 Aug;31(8):1048-1050

Department of Critical Care Medicine, the Eighth Medical Center of Chinese PLA General Hospital, Beijing 100091, China. Corresponding author: Zhang Yuxiang, Email:

Objective: High-flow nasal cannula oxygenation (HFNC) characterized as a new non-invasive respiratory support technology, has been widely used in recent years. Compared with conventional oxygen therapy (COT), non-invasive ventilation (NIV), HFNC can offset patient discomforts, and effectively arrest the deterioration of acute respiratory failure (ARF) in immunosuppressed patients. Although there is no benefit of HFNC over COT on reducing mortality in immunocompromised patients with ARF, HFNC is associated with a lower intubation rate and the improved prognosis of transplant recipients and solid cancer patients. Although the association between the prognosis of HFNC treatment and the pathogenic differences of ARF patients remains unknown, HFNC has, as one of the alternative methods for treating ARF in immunosuppressed patients, outstanding clinical significance. The treatment of HFNC in immunosuppressed patients with ARF, such as transplantation, malignancy, pneumocystis pneumonia, and interstitial pneumonia are reviewed in this article, in order to guide the clinical application of HFNC in such patients.
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http://dx.doi.org/10.3760/cma.j.issn.2095-4352.2019.08.029DOI Listing
August 2019

Direct Rubidium-Strontium Dating of Hydrocarbon Charge Using Small Authigenic Illitic Clay Aliquots from the Silurian Bituminous Sandstone in the Tarim Basin, NW China.

Sci Rep 2019 Aug 29;9(1):12565. Epub 2019 Aug 29.

Australise Resources and Environment Solutions (ARES), Wattle Grove, Perth, WA, Australia.

Illitic clay is ubiquitous in clastic hydrocarbon reservoirs, and the host for several radiometric isotopes such as the potassium-argon (K-Ar) and rubidium-strontium (Rb-Sr) systems. This study applied the isotope-dilution thermal ionization mass spectrometry technique for small samples (3-4 mg) to conduct illite Rb-Sr isotope dating of five illitic clay samples from the Silurian bituminous sandstone (SBS) intersected by five drillholes in the Tarim Basin, NW China. The Rb/Sr ratio of clays is fractionated mainly by the addition of Rb during the illitization of mixed-layer illite/smectite (I/S), which is the dominant clay species in the Tarim Basin samples. The subsample-scale Rb/Sr isotope values suggest that each subsample may contain I/S particles of slightly variable degrees of illitization. Three of the analyzed samples (H6, KQ1 and TZ67) generated Rb-Sr isochron ages of 141 ± 61 Ma, 332 ± 32 Ma and 235 ± 8 Ma (errors quoted at 2σ), respectively. These results are similar to the corresponding K-Ar ages (125 Ma, 389 Ma and 234 Ma). The isotopic ages are consistent with the timing of hydrocarbon charge which varies in different drillholes as constrained by basin modelling, indicating that a closed-system behavior is attained by the hydrocarbon charge that inhibits the illitization of I/S. The Rb-Sr isotope analyses of the other two samples (YM35-1 and Q1) that did not yield isochron ages suggest the conditions for producing isochrons were not satisfied, which may be caused by disturbance of the isotope system by a post-charge hydrothermal event. The outcomes of this study show the robust potential of Rb-Sr clay subsample geochronology for cross-checking isotopic ages yielded by other systems (e.g. K-Ar system) and constraining the timing of hydrocarbon charge.
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http://dx.doi.org/10.1038/s41598-019-48988-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6715745PMC
August 2019

Nonlinear Coda Wave Interferometry: Sensitivity to wave-induced material property changes analyzed via numerical simulations in 2D.

Ultrasonics 2019 Nov 26;99:105968. Epub 2019 Jul 26.

Laboratoire d'Acoustique de l'Universitédu Mans, LAUM, UMR 6613 CNRS, Le Mans Université, Avenue Olivier Messiaen, 72085 Le Mans Cedex 9, France.

The numerical studies conducted in this paper are based on our previous research (Chen et al., 2017); through use of the spectral element method, parametric sensitivity studies of Nonlinear Coda Wave Interferometry (NCWI) are established here and divided into two parts. In the first part, CWI observables are found to be proportional to the product of the changes in elastic modulus within the Effective Damaged Zone (EDZ) and the EDZ surface area. The modifications to intrinsic properties are quantified via an overall wave velocity variation, as probed by a reverberated coda wave. However, for high-level changes, CWI may fail due to meaningless decorrelation values. In this context, parametric studies are conducted to yield a maximum range for EDZ contrast and area. To further validate these observations using a more realistic numerical model, instead of introducing a homogeneous EDZ model, the second part of this paper adds random cracks with random orientations into the EDZ of a material sample. The influence of a strong pump wave on localized nonlinear damage is reestablished; results show that the cracks added into the EDZ reduce the property changes required to match the previous experimental dataset.
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http://dx.doi.org/10.1016/j.ultras.2019.105968DOI Listing
November 2019

Hybrid model for the prediction of municipal solid waste generation in Hangzhou, China.

Waste Manag Res 2019 Aug 2;37(8):781-792. Epub 2019 Jul 2.

Zhejiang Sci-tech University, Hangzhou, China.

Accurate prediction of municipal solid waste (MSW) generation is necessary for choosing appropriate waste treatment methods and for planning the distribution of disposal facilities. In this study, a hybrid model was established to forecast MSW generation through the combination of the ridge regression and GM(1,N) models. The hybrid model is multivariate and involves total urban population, total retail sales of social consumer goods, per capita consumption expenditure of urban areas, tourism, and college graduation. Compared with the constituent models alone, the hybrid model yields higher accuracy, with a mean absolute percentage error (MAPE) of only 2.59%. Through weight allocation and optimal treatment of residuals, the hybrid model also balances the growth trends of the individual models, making the prediction curve smoother. The model coefficients and correlation analysis show that population, economics, and educational factors are influential for waste generation. MSW output in Hangzhou will gradually increase in the future, and is expected to reach 5.12 million tons in 2021. Results can help decision makers to develop the measures and policies of waste management in the future.
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http://dx.doi.org/10.1177/0734242X19855434DOI Listing
August 2019

BZR1 Family Transcription Factors Function Redundantly and Indispensably in BR Signaling but Exhibit BRI1-Independent Function in Regulating Anther Development in Arabidopsis.

Mol Plant 2019 10 21;12(10):1408-1415. Epub 2019 Jun 21.

Ministry of Education Key Laboratory of Molecular and Cellular Biology, Hebei Key Laboratory of Molecular and Cellular Biology, College of Life Sciences, Hebei Normal University, Shijiazhuang 050024, China; Hebei Collaboration Innovation Center for Cell Signaling, Shijiazhuang 050024, China. Electronic address:

BRASSINAZOLE-RESISTANT 1 family proteins (BZRs) are central transcription factors that govern brassinosteroid (BR)-regulated gene expression and plant growth. However, it is unclear whether there exists a BZR-independent pathway that mediates BR signaling. In this study, we found that disruption of all BZRs in Arabidopsis generated a hextuple mutant (bzr-h) displaying vegetative growth phenotypes that were almost identical to those of the null mutant of three BR receptors, bri1brl1brl3 (bri-t). By RNA sequencing, we found that global gene expression in bzr-h was unaffected by 2 h of BR treatment. The anthers of bzr-h plants were loculeless, but a similar phenotype was not observed in bri-t, suggesting that BZRs have a BR signaling-independent regulatory role in anther development. By real-time PCR and in situ hybridization, we found that the expression of SPOROCYTELESS (SPL), which encodes a transcription factor essential for anther locule development, was barely detectable in bzr-h, suggesting that BZRs regulate locule development by affecting SPL expression. Our findings reveal that BZRs are indispensable transcription factors required for both BR signaling and anther locule development, providing new insight into the molecular mechanisms underlying the microsporogenesis in Arabidopsis.
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http://dx.doi.org/10.1016/j.molp.2019.06.006DOI Listing
October 2019

Microporous acellular extracellular matrix combined with adipose-derived stem cell sheets as a promising tissue patch promoting articular cartilage regeneration and interface integration.

Cytotherapy 2019 08 11;21(8):856-869. Epub 2019 Jun 11.

Department of Orthopedic Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, P.R. China. Electronic address:

Background: Acute or chronic injury of articular cartilage leads to localized destruction. Difficulties with interface integration between the implant and native cartilage tissue can lead to an undesirable outcome. To improve cartilage repair and interface integration, we explored the therapeutic efficacy of microporous acellular extracellular matrix (ECM) combined with adipose-derived stem cell (ASC) sheets.

Methods: Methods for fabricating ASC sheets and microporous acellular ECM were explored before transplanting the constructed ASC sheet/matrix in vivo and in vitro, respectively. After the operation, distal femur samples were collected at 6 and 12 weeks for further analysis.

Results: The decellularization process removed 90% of the DNA but retained 82.4% of glycosaminoglycans (GAGs) and 82.8% of collagen, which are the primary components of cartilage matrix. The acellular matrix/ASC sheet construct treatment in vivo showed better interface integration, cartilage regeneration, and collagenous fiber arrangement, which resembles the native structure. There was a significant increase in GAG and collagen accumulation at the zone of regeneration and integration compared to other groups. Gene expression analysis showed that the mRNA level associated with cartilage formation significantly increased in the acellular matrix/ASC sheet group (p<0.05), which is consistent with the histological analysis.

Discussion: ASC sheets promote interface integration between the implant and native tissue. This effect, together with the acellular matrix as a graft, is beneficial for cartilage defect repair, which suggests that acellular matrix/ASC sheet bioengineered cartilage implants may be a better approach for cartilage repair due to their enhanced integration.
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http://dx.doi.org/10.1016/j.jcyt.2019.02.005DOI Listing
August 2019