Publications by authors named "Yanyan Zhao"

328 Publications

Conjugated linoleic acid prompts bone formation in ovariectomized osteoporotic rats and weakens osteoclast formation after treatment with ultraviolet B.

Ann Transl Med 2021 Mar;9(6):503

Department of Orthopedic Surgery, the 1st Affiliated Hospital of Soochow University, Suzhou, China.

Background: Ultraviolet B (UVB) has been reported to prevent bone loss by promoting the synthesis of vitamin D. However, UVB can also enhance osteoclastic differentiation, inhibit osteogenic differentiation, and cause oxidative damage. The present study aimed to analyze the osteoprotective effects of UVB and conjugated linoleic acid (CLA) in rats with ovariectomy-induced osteoporosis, and to determine the interactions between UVB and CLA and their effects on bone mesenchymal stem cells (BMSCs) and bone marrow mononuclear cells (BMMCs).

Methods: , the distance of UVB irradiation and the dose of CLA were selected by immunofluorescence assays and Cytotoxicity assay. BMSCs and BMMCs were detected by immunohistochemical and immunofluorescence assays. , three-month-old female Sprague-Dawley rats that had undergone ovariectomy were treated with UVB and CLA. After 8 weeks of therapy, the femurs of the rats were examined by micro-computed tomography (CT) and immunohistochemical detection to assess the therapeutic efficacy.

Results: The least inhibitive UVB distance and dosage of CLA were selected for the experiments. CLA effectively weakened the osteogenic inhibitory effect of UVB (72 cm), significantly improved the activity of alkaline phosphatase (ALP), promoted the formation of mineralized nodules, and alleviated the oxidative damage induced by UVB. CLA also effectively weakened the osteoclast-promoting effect of UVB (72 cm), inhibited osteoclast formation, and inhibited the inflammatory damage to BMMCs caused by UVB (72 cm) irradiation. Micro-CT results showed that UVB irradiation could promote bone formation in ovariectomized Sprague-Dawley rats, while CLA could significantly promote bone regeneration. Immunofluorescence assays results showed that CLA alleviated UVB-induced oxidative damage to osteoblasts. The ROS detection results demonstrated that CLA effectively alleviated UVB-induced oxidative damage to BMSCs. Furthermore, Immunohistochemical assays showed that UVB and CLA treatment increased bone density, inhibited osteolytic osteolysis, and enhanced osteogenic activity.

Conclusions: CLA can effectively weaken osteoclast promotion, osteogenic inhibition, and oxidative damage caused by UVB. Combination treatment of UVB and CLA exerts an osteoprotective effect on ovariectomized osteoporotic rats and stimulates osteogenesis. The molecular mechanism of this interaction requires further investigation.
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http://dx.doi.org/10.21037/atm-21-934DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8039685PMC
March 2021

Long-term trends in the phylogenetic and functional diversity of Anatidae in South China coastal wetlands.

Ecol Appl 2021 Apr 4:e2344. Epub 2021 Apr 4.

Guangdong Key Laboratory of Animal Conservation and Resource Utilization, Guangdong Public Laboratory of Wild Animal Conservation and Utilization, Institute of Zoology, Guangdong Academy of Sciences, Guangzhou, China.

Species loss has attracted much attention among scientists for more than half a century. However, we have little information on the trends in phylogenetic and functional changes behind the species loss although this information is always asynchronous and important for conservation and management. We measured community trends in Anatidae (ducks and geese) for the last 50 years to quantify trends in phylogenetic and functional diversity patterns coinciding with taxonomic historical dynamics. We used one-way ANOVAs to test if there was a significant historical trend in communities of Anatidae. We characterized taxonomic, phylogenetic and functional diversity of communities. For taxonomic diversity, we used species richness (SR). For phylogenetic diversity, we calculated the standardized effect size of mean pairwise distances (ses.MPD) and the standard effect size of mean nearest taxon distances (ses.MNTD) in communities. For functional diversity, we calculated functional richness (FRic), functional evenness (FEve), functional divergence (FDiv) and the community-level weighted means (CWM) of trait values for diet, foraging stratum and body mass, separately. From the 1950's to 2010's, species richness declined without significant trends. The ses.MNTD of Anatidae communities showed no clear trends. However, ses.MPD of Anatidae communities declined dramatically during this period. For functional diversity, functional evenness of diet, foraging stratum, body mass and functional dispersion of diet, foraging stratum did not increase or decline significantly. However, functional evenness of all traits, functional richness and functional dispersion of body mass showed declined trends. The basic phylogenetic diversity and species body mass of Anatidae communities declined significantly because of a declining trend in the relative independent branch of geese. This makes it more challenging for implement community recovery in the future. More attention in conservation biology should consider taxonomic diversity and asynchrony in phylogenetic and functional diversity.
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http://dx.doi.org/10.1002/eap.2344DOI Listing
April 2021

Role of TLR4/MyD88/NF-κB signaling in heart and liver-related complications in a rat model of type 2 diabetes mellitus.

J Int Med Res 2021 Mar;49(3):300060521997590

The PLA Rocket Force Characteristic Medical Center, Beijing, P.R. China.

Aims: To analyze expression of members of the Toll-like receptor (TLR)4/myeloid differentiation primary response 88 (MyD88)/nuclear factor (NF)-κB signaling pathway in the heart and liver in a rat model of type 2 diabetes mellitus (T2DM). Our overall goal was to understand the underlying pathophysiological mechanisms.

Methods: We measured fasting blood glucose (FBG) and insulin (FINS) in a rat model of T2DM. Expression of members of the TLR4/MyD88/NF-κB signaling pathway as well as downstream cytokines was investigated. Levels of mRNA and protein were assessed using quantitative real-time polymerase chain reaction and western blotting, respectively. Protein content of tissue homogenates was assessed using enzyme-linked immunosorbent assays.

Results: Diabetic rats had lower body weights, higher FBG, higher FINS, and higher intraperitoneal glucose tolerance than normal rats. In addition, biochemical indicators related to heart and liver function were elevated in diabetic rats compared with normal rats. TLR4 and MyD88 were involved in the occurrence of T2DM as well as T2DM-related heart and liver complications. TLR4 caused T2DM-related heart and liver complications through activation of NF-κB.

Conclusions: TLR4/MyD88/NF-κB signaling induces production of tumor necrosis factor-α, interleukin-6, and monocyte chemoattractant protein-1, leading to the heart- and liver-related complications of T2DM.
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http://dx.doi.org/10.1177/0300060521997590DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8020098PMC
March 2021

Comparative Transcriptome Analysis of Early- and Late-Bolting Traits in Chinese Cabbage ().

Front Genet 2021 4;12:590830. Epub 2021 Mar 4.

Institute of Horticulture, Henan Academy of Agricultural Sciences, Zhengzhou, China.

Chinese cabbage is one of the most important and widely consumed vegetables in China. The developmental transition from the vegetative to reproductive phase is a crucial process in the life cycle of flowering plants. In spring-sown Chinese cabbage, late bolting is desirable over early bolting. In this study, we analyzed double haploid (DH) lines of late bolting ("Y410-1" and "SY2004") heading Chinese cabbage ( var. ) and early-bolting Chinese cabbage ("CX14-1") ( ssp. var. ) by comparative transcriptome profiling using the Illumina RNA-seq platform. We assembled 721.49 million clean high-quality paired-end reads into 47,363 transcripts and 47,363 genes, including 3,144 novel unigenes. There were 12,932, 4,732, and 4,732 differentially expressed genes (DEGs) in pairwise comparisons of Y410-1 vs. CX14-1, SY2004 vs. CX14-1, and Y410-1 vs. SY2004, respectively. The RNA-seq results were confirmed by reverse transcription quantitative real-time PCR (RT-qPCR). A Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of DEGs revealed significant enrichment for plant hormone and signal transduction as well as starch and sucrose metabolism pathways. Among DEGs related to plant hormone and signal transduction, six unigenes encoding the indole-3-acetic acid-induced protein ARG7 (BraA02g009130), auxin-responsive protein SAUR41 (BraA09g058230), serine/threonine-protein kinase BSK11 (BraA07g032960), auxin-induced protein 15A (BraA10g019860), and abscisic acid receptor PYR1 (BraA08g012630 and BraA01g009450), were upregulated in both late bolting Chinese cabbage lines (Y410-1 and SY2004) and were identified as putative candidates for the trait. These results improve our understanding of the molecular mechanisms underlying flowering in Chinese cabbage and provide a foundation for studies of this key trait in related species.
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http://dx.doi.org/10.3389/fgene.2021.590830DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7969806PMC
March 2021

Nedd4-2 haploinsufficiency in mice causes increased seizure susceptibility and impaired Kir4.1 ubiquitination.

Biochim Biophys Acta Mol Basis Dis 2021 Jun 13;1867(6):166128. Epub 2021 Mar 13.

Department of Clinical Genetics, Shengjing Hospital of China Medical University, Shenyang, China. Electronic address:

Neural precursor cell expressed developmentally down-regulated gene 4-like (NEDD4-2) encodes a ubiquitin E3 ligase that is involved in epileptogenesis with mechanisms needing further investigation. We constructed a novel Nedd4-2 mouse model with half level of both Nedd4-2 long and short isoforms in the brain. Nedd4-2 haploinsufficiency caused increased susceptibility and severity of pentylenetetrazole (PTZ)-induced seizures. Of the 3379 proteins identified by the hippocampal proteomic analysis, 55 were considered altered in Nedd4-2 mice compared with wild-type control, among which the inwardly rectifying K channel Kir4.1 was up-regulated by 1.83-fold. Kir4.1 was subsequently confirmed to be less ubiquitinated in response to comprised Nedd4-2 in mouse brains and C6 cells. Kir4.1 associated with Nedd4-2 through the threonine-proline motif in the intracellular domain by target mutagenesis. Adaptor protein 14-3-3 facilitated Nedd4-2-mediated ubiquitination of Kir4.1. Our data consolidate the detailed molecular mechanism of Nedd4-2-mediated Kir4.1 ubiquitination, and provide a possible relationship between increased seizure susceptibility and impaired Kir4.1 ubiquitination in the brain.
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http://dx.doi.org/10.1016/j.bbadis.2021.166128DOI Listing
June 2021

Synthesis and Assessment of Novel Probes for Imaging Tau Pathology in Transgenic Mouse and Rat Models.

ACS Chem Neurosci 2021 Mar 10. Epub 2021 Mar 10.

Molecular Imaging Chemistry Laboratory, Wolfson Brain Imaging Centre, University of Cambridge, Cambridge CB2 0QQ, United Kingdom.

Aggregated tau protein is a core pathology present in several neurodegenerative diseases. Therefore, the development and application of positron emission tomography (PET) imaging radiotracers that selectively bind to aggregated tau in fibril form is of importance in furthering the understanding of these disorders. While radiotracers used in human PET studies offer invaluable insight, radiotracers that are also capable of visualizing tau fibrils in animal models are important tools for translational research into these diseases. Herein, we report the synthesis and characterization of a novel library of compounds based on the phenyl/pyridinylbutadienylbenzothiazoles/benzothiazolium (PBB3) backbone developed for this application. From this library, we selected the compound LM229, which binds to recombinant tau fibrils with high affinity ( = 3.6 nM) and detects with high specificity (a) pathological 4R tau aggregates in living cultured neurons and mouse brain sections from transgenic human P301S tau mice, (b) truncated human 151-351 3R (SHR24) and 4R (SHR72) tau aggregates in transgenic rat brain sections, and (c) tau neurofibrillary tangles in brain sections from Alzheimer's disease (3R/4R tau) and progressive supranuclear palsy (4R tau). With LM229 also shown to cross the blood-brain barrier and its effective radiolabeling with the radioisotope carbon-11, we have established a novel platform for PET translational studies using rodent transgenic tau models.
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http://dx.doi.org/10.1021/acschemneuro.0c00790DOI Listing
March 2021

3D printing of chemical-empowered tendon stem/progenitor cells for functional tissue repair.

Biomaterials 2021 Apr 15;271:120722. Epub 2021 Feb 15.

Dr. Li Dak Sum & Yip Yio Chin Center for Stem Cells and Regenerative Medicine, Department of Orthopedic Surgery of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China; Key Laboratory of Tissue Engineering and Regenerative Medicine of Zhejiang Province, School of Medicine, Zhejiang University, Hangzhou, China; China Orthopaedic Regenerative Medicine (CORMed), Hangzhou, China. Electronic address:

Tendon injuries are the leading cause of chronic debilitation to patients. Tendon stem/progenitor cells (TSPCs) are potential seed cells for tendon tissue engineering and regeneration, but TSPCs are prone to lose their distinct phenotype in vitro and specific differentiation into the tenocyte lineage is challenging. Utilizing small molecules in an ex vivo culture system may be a promising solution and can significantly improve the therapeutic applications of these cells. Here, by using an image-based, high-throughput screening platform on small molecule libraries, this study established an effective stepwise culture strategy for TSPCs application. The study formulated a cocktail of small molecules which effected proliferation, tenogenesis initiation and maturation phases, and significantly upregulated expression of various tendon-related genes and proteins in TSPCs, which were demonstrated by high-throughput PCR, ScxGFP reporter assay and immunocytochemistry. Furthermore, by combining small molecule-based culture system with 3D printing technology, we embedded living, chemical-empowered TSPCs within a biocompatible hydrogel to engineer tendon grafts, and verified their enhanced ability in promoting functional tendon repair and regeneration both in vivo and in situ. The stepwise culture system for TSPCs and construction of engineered tendon grafts can not only serve as a platform for further studies of underlying molecular mechanisms of tenogenic differentiation, but also provide a new strategy for tissue engineering and development of novel therapeutics for clinical applications.
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http://dx.doi.org/10.1016/j.biomaterials.2021.120722DOI Listing
April 2021

Feeding Value Assessment of Substituting Cassava () Residue for Concentrate of Dairy Cows Using an In Vitro Gas Test.

Animals (Basel) 2021 Jan 26;11(2). Epub 2021 Jan 26.

State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.

The feeding value of replacing concentrate with cassava () residue in the feed of Holstein cows was confirmed using an in vitro gas test. The treatments consisted of 0% (control, CON), 5%, 10%, 15%, 20%, 25%, and 30% inclusion of cassava residue in fermentation culture medium composed of buffer solution (50 mL) and filtrated rumen fluid (25 mL). The parameters analyzed included the kinetics of gas production and fermentation indexes. Forty-eight hours later, there were no significant differences on in vitro dry matter disappearance (IVDMD), pH, and microbial crude protein (MCP) content among treatments ( > 0.05). However, the "cumulative gas production at 48 h" (GP), the "asymptotic gas production" (A), and the "maximum gas production rate" (RmaxG) all increased linearly or quadratically ( < 0.01). The GP was significantly higher in the 25% treatment compared to the other treatments, except for the 30% ( < 0.01). The A was significantly larger in the 25% treatment compared to the other treatments, except for the 20% and 30% ( < 0.01). The RmaxG was distinctly larger in the 25% treatment compared to other treatments ( < 0.01); moreover, the "time at which RmaxG is reached" (TRmaxG) and the "time at which the maximum rate of substrate degradation is reached" (TRmaxS) were significantly higher in the 25% treatment than the CON, 20%, and 30% treatments ( < 0.01). Additionally, the content of ammonia-N (NH-N) in all treatments showed linearly and quadratically decreases ( < 0.01), whereas total volatile fatty acid (VFA), iso-butyrate, butyrate, and iso-valerate contents changed quadratically ( = 0.02, = 0.05, = 0.01, and = 0.02, respectively); all of these values peaked in the 25% treatment. In summary, the 25% treatment was associated with more in vitro gas and VFA production, indicating that this cassava residue inclusion level may be used to replace concentrate in the feed of Holstein cows. However, these results need to be verified in vivo.
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http://dx.doi.org/10.3390/ani11020307DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7912291PMC
January 2021

Identifying of 22q11.2 variations in Chinese patients with development delay.

BMC Med Genomics 2021 Jan 22;14(1):26. Epub 2021 Jan 22.

Department of Clinical Genetics, Shengjing Hospital of China Medical University, Shenyang, China.

Background: 22q11.2 variation is a significant genetic factor relating to development delay and/or intellectual disability. However, the prevalence, genetic characteristics and clinical phenotype in Chinese patients are unknown.

Methods: In total 6034 patients with development delay and/or intellectual disability were screened by multiplex ligation-dependent probe amplification (MLPA) P245 and G-band karyotyping. The positive patients with 22q11.2 imbalance were confirmed by MLPA P250 assay.

Results: 52 (0.86%) patients were found to carry different levels of 22q11.2 variations, in which 37 cases (71.2%) had heterozygous deletions, whereas 15 (28.8%) had heterogeneous duplications. 34 cases (65.4%) carried typical imbalance from low copy repeat (LCR) 22 A to D. The other cases had atypical variations, relating to LCR22 A-B, LCR22 C-D, LCR22 B-D, LCR22 D-E, LCR22 E-F and LCR22 B-F region. The phenotypes of these 52 patients were variable, including development delay, language delay, facial anomalies, heart defects, psychiatric/behavior problems, epilepsy, periventricular leukomalacia, hearing impairment, growth delay etc. CONCLUSION: These data revealed the prevalence and variability of 22q11.2 genomic imbalance in Chinese patients with development delay and/or intellectual disability. It suggested that genetic detection of 22q11.2 is necessary, especially for the patients with mental retardation and development disorders, which deserves the attention of all pediatricians in their daily work.
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http://dx.doi.org/10.1186/s12920-020-00849-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7821542PMC
January 2021

rGO/BiWO composite as a highly efficient and stable visible-light photocatalyst for norfloxacin degradation in aqueous environment.

J Colloid Interface Sci 2021 May 9;589:336-346. Epub 2021 Jan 9.

College of Food Science and Technology, Northwest University, Xi'an 710069, PR China. Electronic address:

Fabrication of binary composite has proved to be an efficient approach to improve the photocatalytic activity of monomer photocatalyst. In this contribution, an effective Reduced Graphene Oxide/Bismuth Tungsten Oxide (rGO/BiWO) composite with outstanding photocatalytic activity was designed by employing BiWO as a primary photocatalyst and rGO as an electron acceptor and transporter for norfloxacin degradation in aquatic environment. The rGO/BiWO composite displayed higher photocatalytic activity compare with pure BiWO, which could degrade about 87.49% of norfloxacin with 180 min under visible light irradiation. The results of the UV-vis diffuse reflection spectrum, photoluminescence spectra and transient photocurrent response implied that the enhanced photocatalytic activity of the rGO/BiWO composite could be attributed to the improved visible light-harvesting ability and the efficient charge separation ability. Additionally, the reactive-species-trapping experiments indicated that ⋅OH and e played dominant roles during the photocatalytic degradation process. Four possible intermediates and two possible transformation pathways of norfloxacin degradation were detected by LC-MS. This present work provided a low-cost and facile green method to design of Bi-based composite.
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http://dx.doi.org/10.1016/j.jcis.2021.01.016DOI Listing
May 2021

Electrochemically Triggered Chain Reactions for the Conversion of Furan Derivatives.

Angew Chem Int Ed Engl 2021 03 24;60(14):7534-7539. Epub 2021 Feb 24.

Department of Chemistry, Boston College, Merkert Chemistry Center, 2609 Beacon St., Chestnut Hill, MA, 02467, USA.

We report an electrochemical method for coupling biomass-derived C5/C6 compounds to value-added fuel precursors. Using only 2 % of equivalent charges, 2-methylfuran (2-MF) was oxidized to yield a cation radical, which readily reacted with 3-hexene-2,5-dione, a derivate of 2,5-dimethylfuran, to produce 3-(5-methylfuran-2-yl)hexane-2,5-dione. The product was converted to 4-ethylnonane (a component of biodiesel/jet fuel) in a single step in excellent yield. Importantly, the reaction was not sensitive to oxygen, and a trace amount of water was found to promote the reaction. Detailed mechanistic studies confirmed the proposed reaction pathways. Key to the mechanism is the radical generation that is enabled by electrochemistry. The radical is regenerated at the end of a reaction cycle to ensure chain propagation for an average of ca. 47 times, resulting in an apparent Faradaic efficiency of 4700 %.
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http://dx.doi.org/10.1002/anie.202016601DOI Listing
March 2021

The impact of urbanization on body size of Barn Swallows .

Ecol Evol 2021 Jan 19;11(1):612-625. Epub 2020 Dec 19.

Department of Health and Environmental Sciences Xi'an Jiaotong-Liverpool University Suzhou China.

Urbanization implies a dramatic impact on ecosystems, which may lead to drastic phenotypic differences between urban and nonurban individuals. For instance, urbanization is associated with increased metabolic costs, which may constrain body size, but urbanization also leads to habitat fragmentation, which may favor increases in body mass when for instance it correlates with dispersal capacity. However, this apparent contradiction has rarely been studied. This is particularly evident in China where the urbanization process is currently occurring at an unprecedented scale. Moreover, no study has addressed this issue across large geographical areas encompassing locations in different climates. In this regard, Barn Swallows () are a suitable model to study the impact of urbanization on wild animals because they are a widely distributed species tightly associated with humans. Here, we collected body mass and wing length data for 359 breeding individuals of Barn Swallow () from 128 sites showing different levels of urbanization around the whole China. Using a set of linear mixed-effects models, we assessed how urbanization and geography influenced body size measured using body mass, wing length, and their regression residuals. Interestingly, we found that the impact of urbanization was sex-dependent, negatively affecting males' body mass, its regression residuals, and females' wing length. We also found that northern and western individuals were larger, regarding both body mass and wing length, than southern and eastern individuals. Females were heavier than males, yet males had slightly longer wings than females. Overall, our results showed that body mass of males was particularly sensitive trait to urbanization, latitude, and longitude, while it only showed a weak response to latitude in females. Conversely, while wing length showed a similar geographical pattern, it was only affected by urbanization in the case of females. Further research is needed to determine whether these phenotypic differences are associated with negative effects of urbanization or potential selective advantages.
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http://dx.doi.org/10.1002/ece3.7088DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7790637PMC
January 2021

Molecular and behavioral responses of zebrafish embryos/larvae after sertraline exposure.

Ecotoxicol Environ Saf 2021 Jan 23;208:111700. Epub 2020 Nov 23.

Department of Physiological Sciences and Center for Environmental and Human Toxicology, University of Florida Genetics Institute, Interdisciplinary Program in Biomedical Sciences Neuroscience, College of Veterinary Medicine, University of Florida, Gainesville, FL, 32611 USA.

Sertraline (SER) is one of the most frequently detected antidepressant drugs in aquatic environments. However, knowledge regarding SER-induced behavioral alterations in fish is insufficient, as well as the mechanisms underlying SER-induced toxicity. The present study aimed to determine behavioral and molecular responses in larval fish following SER exposure with a focus on its mode of action. Zebrafish embryos (~6 h-post-fertilization, hpf) were exposed to one of three concentrations of SER (1, 10, 100 μg/L) for 6 days, respectively. Evaluated parameters included development, behavior, transcripts related to serotonin signaling, serotonin levels, and acetylcholinesterase activity. Accelerated hatching of zebrafish embryos was observed for those fish exposed to 100 μg/L SER at 54 hpf. Locomotor activity (e.g. distance moved and mobile cumulative duration) was significantly reduced in larval zebrafish following exposure to 10 and 100 μg/L SER. Conversely, larval fish showed increased dark-avoidance after exposure to 1-100 μg/L SER. Of the measured transcripts related to serotonin signaling, only serotonin transporter (serta) and serotonin receptor 2c (5-ht2c) mRNA levels were increased in fish in response to 10 μg/L SER treatment. However, serotonin levels were unaltered in larvae exposed to SER. There were no differences among groups in acetylcholinesterase activity at any concentration tested. Taking together, the results evidenced that exposure to SER alters behavioral responses in early-staged zebrafish, which may be related to the abnormal expression of 5-ht2c. This study elucidates molecular responses to SER and characterizes targets that may be sensitive to antidepressant pharmaceuticals in larval fish.
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http://dx.doi.org/10.1016/j.ecoenv.2020.111700DOI Listing
January 2021

Protective effects of dioscin on vascular remodeling in pulmonary arterial hypertension via adjusting GRB2/ERK/PI3K-AKT signal.

Biomed Pharmacother 2021 Jan 28;133:111056. Epub 2020 Nov 28.

College of Pharmacy, Dalian Medical University, Western 9 Lvshunnan Road, Dalian 116044, China; Key Laboratory for Basic and Applied Research on Pharmacodynamic Substances of Traditional Chinese Medicine of Liaoning Province, Dalian Medical University, Dalian 116044, China. Electronic address:

Pulmonary arterial hypertension (PAH) is a progressive and lethal cardiopulmonary. Pulmonary vascular remodeling (PVR) caused by excessive proliferation and apoptosis resistance of pulmonary artery smooth muscle cells (PASMCs) is the chief pathological feature of PAH. Dioscin is a natural product that possesses multiple pharmacological activities, but its effect on PAH remains unclear. In this study, effect of dioscin on vascular remodeling in PAH was assessed in hypoxia-induced PASMCs, hypoxia-induced and monocrotaline (MCT)-induced rats. Western blot, Real-time PCR and siRNA transfection tests were applied to evaluate the possible mechanisms of dioscin. In vitro experiments, results showed dioscin markedly inhibited the proliferation and migration, and promoted apoptosis of hypoxic PASMCs. In vivo, dioscin significantly decreased the right ventricular systolic pressure (RVSP) and right ventricular hypertrophy index (RVHI), and improved pulmonary vascular stenosis in rats induced by hypoxia or MCT. Molecular mechanism studies showed that dioscin significantly reduced the expression of growth factor receptor-bound protein 2 (GRB2). Subsequently, dioscin reduced the expressions of Ras, Cyclin D1, CDK4, c-Fos, PCNA and p-ERK to inhibit proliferation and migration of PASMCs, inhibited p-PI3K and p-AKT levels and increased Bax/Bcl2 ratio to promote cell apoptosis. GRB2 siRNA transfection in PASMCs further confirmed that the inhibitory action of dioscin in PAH was evoked by adjusting GRB2/ERK/PI3K-AKT signal. Taken together, our study indicated that dioscin attenuates PAH through adjusting GRB2/ERK/PI3K-AKT signal to inhibit PASMCs proliferation and migration, and promote apoptosis, and dioscin may be developed as a therapeutic strategy for treating PAH in the future.
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http://dx.doi.org/10.1016/j.biopha.2020.111056DOI Listing
January 2021

Crosstalk between phosphorylation and ubiquitination is involved in high salt-induced WNK4 expression.

Exp Ther Med 2021 Feb 10;21(2):133. Epub 2020 Dec 10.

Department of Clinical Genetics, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110003, P.R. China.

With no lysine 4 (WNK4) is a serine/threonine kinase, which is expressed in the kidney and associated with salt-sensitive hypertension. However, how salt regulates WNK4 remains unclear. In the present study, the C57BL/6 mice and HEK293 cells were treated with high salt and the expression of WNK4 protein and its ubiquitination and phosphorylation levels were detected. Western blotting demonstrated that WNK4 expression was significantly increased in high salt-treated mice and cells. Meanwhile, co-immunoprecipitation analysis demonstrated that the ubiquitination of WNK4 was decreased under high-salt simulation. It was also identified that the Lys-1023 site was the most important ubiquitination site for WNK4, and it was found that phosphorylation at the Ser-1022 site was a prerequisite for ubiquitination. These results suggested that there was crosstalk between phosphorylation and ubiquitination in the WNK4 protein, and high salt may downregulate its phosphorylation and, in turn, decrease its ubiquitination, leading to a decrease in WNK4 degradation. This eventually resulted in an increase in the abundance of WNK4 protein.
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http://dx.doi.org/10.3892/etm.2020.9565DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7751476PMC
February 2021

Programmed Cell Death in Stigmatic Papilla Cells Is Associated With Senescence-Induced Self-Incompatibility Breakdown in Chinese Cabbage and Radish.

Front Plant Sci 2020 7;11:586901. Epub 2020 Dec 7.

State Key Laboratory of Crop Biology, College of Horticulture Science and Engineering, Shandong Agricultural University, Tai'an, China.

Self-incompatibility (SI) is a genetic mechanism flowering plants adopted to reject self-pollen and promote outcrossing. In the Brassicaceae family plants, the stigma tissue plays a key role in self-pollen recognition and rejection. We reported earlier in Chinese cabbage () that stigma tissue showed upregulated ethylene responses and programmed cell death (PCD) upon compatible pollination, but not in SI responses. Here, we show that SI is significantly compromised or completely lost in senescent flowers and young flowers of senescent plants. Senescence upregulates senescence-associated genes in . Suppressing their expression in young stigmas by antisense oligodeoxyribonucleotide abolishes compatible pollination-triggered PCD and inhibits the growth of compatible pollen tubes. Furthermore, ethylene biosynthesis genes and response genes are upregulated in senescent stigmas, and increasing the level of ethylene or inhibiting its response increases or decreases the expression of senescence-associated genes, respectively. Our results show that senescence causes PCD in stigmatic papilla cells and is associated with the breakdown of SI in Chinese cabbage and in radish.
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http://dx.doi.org/10.3389/fpls.2020.586901DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7750362PMC
December 2020

Development and Molecular Investigation into the Effects of Carbamazepine Exposure in the Zebrafish ().

Int J Environ Res Public Health 2020 11 29;17(23). Epub 2020 Nov 29.

Department of Physiological Sciences and Center for Environmental and Human Toxicology, University of Florida Genetics Institute, Interdisciplinary Program in Biomedical Sciences Neuroscience, College of Veterinary Medicine, University of Florida, Gainesville, FL 32611, USA.

Concerns regarding environmental exposures and the impacts of pharmaceuticals on non-target aquatic organisms continue to increase. The antiepileptic drug carbamazepine (CBZ) is often detected as an aquatic contaminant and can disrupt various behaviors of fishes. However, there are few reports which investigate the mechanism of CBZ action in fish. The aim of the current study was to evaluate the effects of CBZ on embryonic development (i.e., hatching rate, heart rate, and body length) and early spontaneous movement. Moreover, we sought to investigate potential mechanisms by focusing on the gamma-aminobutyric acid (GABA) neurotransmitter system in zebrafish 6 days after of exposure. The results show that CBZ exposure did not cause significant effects on embryo development (hatching rate, heart rate, nor body length) at the test concentrations. However, the early spontaneous movement of embryos was inhibited following 10 μg/L CBZ exposure at 28-29 h post-fertilization (hpf). In addition, acetylcholinesterase (AChE) activity and GABA concentrations were increased with exposure, whereas glutamate (Glu) concentrations were decreased in larval zebrafish. Gene expression analysis revealed that GABA and glutamate metabolic pathways in zebrafish larvae were altered following exposure to CBZ. GABA transaminase () and glutamic acid decarboxylase () decreased to 100 µg/L, and glutamate receptor, ionotropic, N-methyl D-aspartate 1b () as well as the glutamate receptor, ionotropic, α-amino-3hydroxy-5methylisoxazole-4propionic 2b () were down-regulated with exposure to 1 µg/L CBZ. Our study suggests that CBZ, which can act as an agonist of the GABA receptor in humans, can also induce alterations in the GABAergic system in fish. Overall, this study improves understanding of the neurotoxicity and behavioral toxicity of zebrafish exposed to CBZ and generates data to be used to understand mechanisms of action that may underlie antiepileptic drug exposures.
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http://dx.doi.org/10.3390/ijerph17238882DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7731368PMC
November 2020

20-HETE downregulates Na/K-ATPase α1 expression via the ubiquitination pathway.

Prostaglandins Other Lipid Mediat 2021 Feb 24;152:106503. Epub 2020 Oct 24.

Department of Clinical Genetics, Shengjing Hospital of China Medical University, Shenyang, China. Electronic address:

In this article, we found that 20-Hydroxyeicosatetraenoic acid (20-HETE) reduced Na/K-ATPase α1 expression via the ubiquitin-proteasome pathway. The ubiquitination level of Na/K-ATPase α1 protein was increased in 20-HETE-treated mouse cortical collecting duct cells and the kidney tissues of CYP4F2 transgenic mice. We also demonstrated that 20-HETE-induced high level phosphorylation of Na/K-ATPase α1 was necessary for its ubiquitination.The protein kinase C inhibitor sotrastaurin significantly reduced the phosphorylation of Na/K-ATPase α1 and increased the expression of Na/K-ATPase α1 although 20-HETE stimulus being applied at the same time. Moreover, high level of 20-HETE increased the expression and neddylation of Cullin3,which is an important ubiquitin E3 ligase in kidney. MLN4924, an inhibitor of NEDD8-activating enzyme, inhibited neddylation of Cullin3 and reversed the reduction of Na/K-ATPase α1 expression caused by 20-HETE. Thus, 20-HETE downregulates Na/K-ATPase α1 via the ubiquitination pathway, and phosphorylation of Na/K-ATPase α1 is a prerequisite to ubiquitination. Additionally, 20-HETE regulates Cullin3 expression via neddylation.
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http://dx.doi.org/10.1016/j.prostaglandins.2020.106503DOI Listing
February 2021

Discovery of driver non-coding splice-site-creating mutations in cancer.

Nat Commun 2020 11 4;11(1):5573. Epub 2020 Nov 4.

Department of Medicine, Washington University in St. Louis, St. Louis, MO, 63110, USA.

Non-coding mutations can create splice sites, however the true extent of how such somatic non-coding mutations affect RNA splicing are largely unexplored. Here we use the MiSplice pipeline to analyze 783 cancer cases with WGS data and 9494 cases with WES data, discovering 562 non-coding mutations that lead to splicing alterations. Notably, most of these mutations create new exons. Introns associated with new exon creation are significantly larger than the genome-wide average intron size. We find that some mutation-induced splicing alterations are located in genes important in tumorigenesis (ATRX, BCOR, CDKN2B, MAP3K1, MAP3K4, MDM2, SMAD4, STK11, TP53 etc.), often leading to truncated proteins and affecting gene expression. The pattern emerging from these exon-creating mutations suggests that splice sites created by non-coding mutations interact with pre-existing potential splice sites that originally lacked a suitable splicing pair to induce new exon formation. Our study suggests the importance of investigating biological and clinical consequences of noncoding splice-inducing mutations that were previously neglected by conventional annotation pipelines. MiSplice will be useful for automatically annotating the splicing impact of coding and non-coding mutations in future large-scale analyses.
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http://dx.doi.org/10.1038/s41467-020-19307-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7642382PMC
November 2020

Validation of the DAPT score in large-scale consecutive and contemporary patients population in the real world.

Platelets 2020 Nov 3:1-8. Epub 2020 Nov 3.

From State Key Laboratory of Cardiovascular Disease, Department of Cardiology, Cardiovascular Institute, Fuwai Hospital and National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College , Beijing, China.

Dual antiplatelet therapy (DAPT) score emerged as a tool for quantification of ischemia and bleeding risks. However, there was discrepancy of the prediction ability of DAPT score in previous studies. We aimed to assess the utility of DAPT score in a large-scale cohort of consecutive percutaneous coronary intervention (PCI) patients. This study enrolled 9,114 patients who had undergone PCI at Fuwai Hospital in 2013, adhered to DAPT and were event-free within the first 12 months following PCI. The endpoints included primary ischemic endpoints (major adverse cardiovascular and cerebrovascular events, and myocardial infarction and/or stent thrombosis), and bleeding endpoint from 12 through 24 months after PCI. Patients were classified into low (score <2, n = 3,989) and high (score ≥2, n = 5,125) DAPT score groups. The incidence rates of primary ischemic endpoints and bleeding endpoint were similar between the two groups. Multivariable analysis demonstrated DAPT score not to be an independent predictor of primary ischemic endpoints or bleeding endpoint. Based on receiver operating characteristic curves analysis, the C-statistic of DAPT score for primary ischemic endpoints or bleeding endpoint did not achieve a significant extent. In this large-scale cohort of PCI patients, DAPT score did not discriminate the risks of ischemic and bleeding events.
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http://dx.doi.org/10.1080/09537104.2020.1841894DOI Listing
November 2020

A 2.3-ps RMS Resolution Time-to-Digital Converter Implemented in a Low-Cost Cyclone V FPGA.

IEEE Trans Instrum Meas 2019 Oct 13;68(10):3647-3660. Epub 2018 Dec 13.

Department of Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA 94720 USA.

We present a nonuniform multiphase (NUMP) method to construct a high-resolution time-to-digital converter (TDC) for low-cost field-programmable gate array (FPGA) devices. The NUMP method involves a system clock being passed through a series of delay elements to generate multiple clocks with different phase shifts. The phases of the rising and falling edges of all the clocks are sorted in order and the states of all the clocks are latched when a hit signal arrives. The sizes of the time bins (and precision) of the NUMP method are not limited by the uniformity and minimum value of the time delays of the delay lines. In theory, any delay sources with small jitters in an FPGA, not just very fine carry chains, can be used in the NUMP method to delay and randomize the clocks. Thus, the NUMP method can achieve excellent TDC timing resolutions in low-cost FPGAs without very fine delay lines. We implemented four NUMP TDC channels in a low-cost FPGA device (an Altera Cyclone V 5CEBA4F23C7N). The performance of the four NUMP TDCs was evaluated using both internal and external pulses. The root mean square (rms) for the timing resolution measured using the internal and the external pulses with short-time intervals (less than 1 ns) was 2.3 and 5.2 ps, respectively. A 14.1-ps rms timing resolution was measured at a time interval of 517 ns. The NUMP method is suitable for applications that require a number of high-performance TDC channels in a low-cost FPGA.
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http://dx.doi.org/10.1109/tim.2018.2880940DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7597900PMC
October 2019

Skp2 regulates DNA damage repair and apoptosis via interaction with Ku70.

Exp Cell Res 2020 12 22;397(1):112335. Epub 2020 Oct 22.

Center for Molecular Medicine, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang, 310013, PR China. Electronic address:

Purpose: Skp2, an oncoprotein, regulates tumor proliferation, invasion and metastasis. Ku70 is a critical component of the non-homologous end-joining (NHEJ) process. Both Skp2 and Ku70 are positively associated in multiple cancers. However, there is no report about the relationship between Skp2 and Ku70 proteins.

Methods: In this study, we carried out Bioinformatics and molecular biological methods to investigate the relationship between Skp2 and Ku70 proteins.

Results: We first observed Skp2 and Ku70 mRNAs were significantly increased in cervical cancer tissues. And we identified Ku70 as a Skp2-binding protein and the binding site located in the C-terminal of Ku70 protein. We further found that Skp2 knockdown decreased the Ku70 protein level in cells, and increase the cellular apoptosis and DNA damage, suggesting Skp2 mediates the Ku70 protein stability and function via post-translational modification.

Conclusion: The direct interaction between Skp2 and Ku70 proteins mediates the DNA damage repair and cellular apoptosis by regulating Ku70 stability and function via post-translational modification. The molecular mechanisms how Skp2 stabilize Ku70 would be clarified in our following research work.
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http://dx.doi.org/10.1016/j.yexcr.2020.112335DOI Listing
December 2020

Early prediction and identification for severe patients during the pandemic of COVID-19: A severe COVID-19 risk model constructed by multivariate logistic regression analysis.

J Glob Health 2020 Dec;10(2):020510

Center for Infectious Diseases, Second Affiliated Hospital of Air Force Medical University, Xi'an, Shaanxi, China.

Background: As an emergent and fulminant infectious disease, Corona Virus Disease 2019 (COVID-19) has caused a worldwide pandemic. The early identification and timely treatment of severe patients are crucial to reducing the mortality of COVID-19. This study aimed to investigate the clinical characteristics and early predictors for severe COVID-19, and to establish a prediction model for the identification and triage of severe patients.

Methods: All confirmed patients with COVID-19 admitted by the Second Affiliated Hospital of Air Force Medical University were enrolled in this retrospective non-interventional study. The patients were divided into a mild group and a severe group, and the clinical data were compared between the two groups. Univariate and multivariate analysis were used to identify the independent early predictors for severe COVID-19, and the prediction model was constructed by multivariate logistic regression analysis. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of the prediction model and each early predictor.

Results: A total of 40 patients were enrolled in this study, of whom 19 were mild and 21 were severe. The proportions of patients with venerable age (≥60 years old), comorbidities, and hypertension in severe patients were higher than that of the mild ( < 0.05). The duration of fever and respiratory symptoms, and the interval from illness onset to viral clearance were longer in severe patients ( < 0.05). Most patients received at least one form of oxygen treatments, while severe patients required more mechanical ventilation ( < 0.05). Univariate and multivariate analysis showed that venerable age, hypertension, lymphopenia, hypoalbuminemia and elevated neutrophil lymphocyte ratio (NLR) were the independent high-risk factors for severe COVID-19. ROC curves demonstrated significant predictive value of age, lymphocyte count, albumin and NLR for severe COVID-19. The sensitivity and specificity of the newly constructed prediction model for predicting severe COVID-19 was 90.5% and 84.2%, respectively, and whose positive predictive value, negative predictive value and crude agreement were all over 85%.

Conclusions: The severe COVID-19 risk model might help clinicians quickly identify severe patients at an early stage and timely take optimal therapeutic schedule for them.
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http://dx.doi.org/10.7189/jogh.10.020510DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7567445PMC
December 2020

Water flow inside various geometric nano-confinement channels.

Phys Chem Chem Phys 2020 Nov 23;22(42):24633-24639. Epub 2020 Oct 23.

College of Water Resources and Architectural Engineering, Northwest A&F University, 712100 Yangling, China.

In nano-confined systems, the properties of a fluid are different from those of macroscopic systems, and the properties of a nanotube can significantly affect water transport. However, our knowledge of the effects of nanotube shape is far from adequate. In the present work, we study the properties of a fluid transporting in different nano-confined configurations by molecular dynamics simulations. This study is aimed at gaining insight into the transport of water molecules in carbon nanotubes with different configurations. We find that the closer of channel shape to the circular nanotube (more sides of the channel), the lower friction coefficient of the solid-liquid interface has and the friction coefficient of nanochannels increases with R when R < 1.0 nm. The friction coefficient converges to a stable value (close to the friction coefficient of graphene/water) when R > 1.0 nm. A variety of configurations leads to the variation of the fluid properties in nanotubes. Our results can be applied to the nanofluid properties of a complex channel structure and water nanochannel microscopic design.
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http://dx.doi.org/10.1039/d0cp04491aDOI Listing
November 2020

Author Correction: Dioscin alleviates alcoholic liver fibrosis by attenuating hepatic stellate cell activation via the TLR4/MyD88/NF-κB signaling pathway.

Sci Rep 2020 Oct 22;10(1):18384. Epub 2020 Oct 22.

College of Pharmacy, Dalian Medical University, No. 9 West Part of Lvshunnan Road, Dalian, 116044, China.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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http://dx.doi.org/10.1038/s41598-020-74987-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7582170PMC
October 2020

Author Correction: Potent effects of dioscin against obesity in mice.

Sci Rep 2020 Oct 22;10(1):18382. Epub 2020 Oct 22.

College of Pharmacy, Dalian Medical University, Western 9 Lvshunnan Road, Dalian, 116044, China.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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http://dx.doi.org/10.1038/s41598-020-74985-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7581761PMC
October 2020

Control of Photocarrier Separation and Recombination at Bismuth Oxyhalide Interface for Nitrogen Fixation.

J Phys Chem Lett 2020 Nov 21;11(21):9304-9312. Epub 2020 Oct 21.

Institute for Superconducting and Electronic Materials (ISEM), Australian Institute for Innovative Materials (AIIM), University of Wollongong, Wollongong, New South Wales 2500, Australia.

Developing high-efficiency photocatalysts for clean energy generation is a grand challenge, which requires simultaneously steering photocarrier dynamics and chemical activity for a specific reaction. To this end, here for the first time, we explore the real-time photocarrier transport property and catalytic mechanism of nitrogen reduction reaction (NRR) at the interface of bismuth oxyhalides (BiOX, X = Cl, Br, and I), an inexpensive and green semiconductor. By time-dependent non-adiabatic molecular dynamics simulations, we show that the separation and recombination processes of excited carriers as well as the catalytic activity can be concurrently optimized by precise band structure engineering. The exact influence of impurity states and heterojunction on the reduction power and lifetime of photogenerated carriers, light absorbance, and NRR activity/selectivity of BiOX are clearly unveiled, to provide essential physical insights for improving the photocatalytic efficiency of semiconductors for practical solar energy conversion and hydrogen fuel storage.
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http://dx.doi.org/10.1021/acs.jpclett.0c02480DOI Listing
November 2020

Homocysteine downregulates cardiac homeobox transcription factor NKX2.5 via IGFBP5.

Am J Physiol Heart Circ Physiol 2020 12 9;319(6):H1380-H1386. Epub 2020 Oct 9.

Department of Clinical Genetics, Shengjing Hospital of China Medical University, Shenyang, Liaoning, People's Republic of China.

Homocysteine (Hcy) is an independent risk factor of congenital heart disease (CHD), but its exact underlying mechanism is unclear. In this study, we collected amniotic fluid (AF) supernatant samples from pregnant women carrying CHD-affected ( = 16) or normal ( = 16) fetuses. We found that Hcy concentrations were higher in the AF of the CHD group when compared with normal pregnancies. Also, Western blot showed that NK2 homeobox 5 (NKX2.5) was decreased and insulin-like growth factor binding protein 5 (IGFBP5) was increased in the AF of the CHD group. In the H9C2 cell culture experiment, 500 μmol/L Hcy downregulated and upregulated . Real-time PCR and Western blot showed that expression was reduced in H9C2 cells treated with IGFBP5. Luciferase reporter gene demonstrated that IGFBP5 decreased the transcription of the promoter. Chromatin immunoprecipitation and electrophoretic mobility shift assay suggested that IGFBP5 binds to the promoter region. Thus, the data indicated that one of the possible mechanisms by which Hcy is involved in CHD may be that Hcy inhibits expression partly through IGFBP5. We found that Hcy and IGFBP5 were increased, whereas NKX2.5 was decreased, in AF of CHD. Meanwhile, Hcy could upregulate IGFBP5 but downregulate NKX2.5, and IGFBP5 inhibited NKX2.5 expression in vitro. Moreover, IGFBP5 can bind to the promoter region and reduce transcriptional activity.
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http://dx.doi.org/10.1152/ajpheart.00347.2020DOI Listing
December 2020

Impact of Public Health Emergency Response to COVID-19 on Management and Outcome for STEMI Patients in Beijing-A Single-Center Historic Control Study.

Curr Probl Cardiol 2021 Mar 1;46(3):100693. Epub 2020 Sep 1.

Coronavirus disease 2019 (COVID-19) pandemic poses great challenge on public health globally. To clarify the impact of COVID-19 pandemic on in-hospital management and outcomes for ST-segment elevation myocardial infarction (STEMI) patients in the nonepicenter. We enrolled consecutive STEMI patients who visited Fuwai Hospital from January to March, 2020 (N = 73) and also established a historical control including all consecutive STEMI patients in the same period of 2019 (N = 95). The primary outcome was defined as a composite endpoint of all-cause death, heart failure, cardiac shock, and cardiac arrest during hospitalization. Emergency response for COVID-19 resulted in a significant 77.6% reduction in the number of primary percutaneous coronary intervention, and a trend toward higher rate of primary composite endpoint (15.1% vs 11.6%, P = 0.51). COVID-19 pandemic results in a significant reduction in emergent reperfusion therapy, and a trend toward higher in-hospital adverse events risk.
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http://dx.doi.org/10.1016/j.cpcardiol.2020.100693DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7462598PMC
March 2021

An extended KASP-SNP resource for molecular breeding in Chinese cabbage(Brassica rapa L. ssp. pekinensis).

PLoS One 2020 2;15(10):e0240042. Epub 2020 Oct 2.

Institute of Horticulture, Henan Academy of Agricultural Sciences, Zhengzhou, China.

Kompetitive allele-specific PCR (KASP) is a cost-effective single-step SNP genotyping technology, With an objective to enhance the marker repertoire and develop high efficient KASP-SNP markers in Chinese cabbage, we re-sequenced four Chinese cabbage doubled haploid (DH) lines, Y177-47, Y635-10, Y510-1 and Y510-9, and generated a total of more than 38.5 billion clean base pairs. A total of 827,720 SNP loci were identified with an estimated density of 3,217 SNPs/Mb. Further, a total of 387,354 SNPs with at least 30 bp to the next most adjacent SNPs on either side were selected as resource for KASP markers. From this resource, 258 (96.27%) of 268 SNP loci were successfully transformed into KASP-SNP markers using a Roche LightCycler 480-II instrument. Among these markers, 221 (85.66%) were co-dominant markers, 220 (85.27%) were non-synonymous SNPs, and 257 (99.6%) were newly developed markers. In addition, 53 markers were applied for genotyping of 34 Brassica rapa accessions. Cluster analysis separated these 34 accessions into three clusters based on heading types. The millions of SNP loci, a large set of resource for KASP markers, as well as the newly developed KASP markers in this study may facilitate further genetic and molecular breeding studies in Brassica rapa.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0240042PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7531813PMC
November 2020