Publications by authors named "Zhe Han"

147 Publications

Inactivating histone deacetylase HDA promotes longevity by mobilizing trehalose metabolism.

Nat Commun 2021 03 31;12(1):1981. Epub 2021 Mar 31.

Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.

Histone acetylations are important epigenetic markers for transcriptional activation in response to metabolic changes and various stresses. Using the high-throughput SEquencing-Based Yeast replicative Lifespan screen method and the yeast knockout collection, we demonstrate that the HDA complex, a class-II histone deacetylase (HDAC), regulates aging through its target of acetylated H3K18 at storage carbohydrate genes. We find that, in addition to longer lifespan, disruption of HDA results in resistance to DNA damage and osmotic stresses. We show that these effects are due to increased promoter H3K18 acetylation and transcriptional activation in the trehalose metabolic pathway in the absence of HDA. Furthermore, we determine that the longevity effect of HDA is independent of the Cyc8-Tup1 repressor complex known to interact with HDA and coordinate transcriptional repression. Silencing the HDA homologs in C. elegans and Drosophila increases their lifespan and delays aging-associated physical declines in adult flies. Hence, we demonstrate that this HDAC controls an evolutionarily conserved longevity pathway.
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http://dx.doi.org/10.1038/s41467-021-22257-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8012573PMC
March 2021

Functional analysis of SARS-CoV-2 proteins in Drosophila identifies Orf6-induced pathogenic effects with Selinexor as an effective treatment.

Cell Biosci 2021 Mar 25;11(1):59. Epub 2021 Mar 25.

Center for Precision Disease Modeling, Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, USA.

Background: SARS-CoV-2 causes COVID-19 with a widely diverse disease profile that affects many different tissues. The mechanisms underlying its pathogenicity in host organisms remain unclear. Animal models for studying the pathogenicity of SARS-CoV-2 proteins are lacking.

Methods: Using bioinformatic analysis, we found that 90% of the virus-host interactions involve human proteins conserved in Drosophila. Therefore, we generated a series of transgenic fly lines for individual SARS-CoV-2 genes, and used the Gal4-UAS system to express these viral genes in Drosophila to study their pathogenicity.

Results: We found that the ubiquitous expression of Orf6, Nsp6 or Orf7a in Drosophila led to reduced viability and tissue defects, including reduced trachea branching as well as muscle deficits resulting in a "held-up" wing phenotype and poor climbing ability. Furthermore, muscles in these flies showed dramatically reduced mitochondria. Since Orf6 was found to interact with nucleopore proteins XPO1, we tested Selinexor, a drug that inhibits XPO1, and found that it could attenuate the Orf6-induced lethality and tissue-specific phenotypes observed in flies.

Conclusions: Our study established Drosophila as a model for studying the function of SARS-CoV2 genes, identified Orf6 as a highly pathogenic protein in various tissues, and demonstrated the potential of Selinexor for inhibiting Orf6 toxicity using an in vivo animal model system.
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http://dx.doi.org/10.1186/s13578-021-00567-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7992514PMC
March 2021

Characterization of SARS-CoV-2 proteins reveals Orf6 pathogenicity, subcellular localization, host interactions and attenuation by Selinexor.

Cell Biosci 2021 Mar 25;11(1):58. Epub 2021 Mar 25.

Center for Precision Disease Modeling, Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, USA.

Background: SARS-CoV-2 causes COVID-19 which has a widely diverse disease profile. The mechanisms underlying its pathogenicity remain unclear. We set out to identify the SARS-CoV-2 pathogenic proteins that through host interactions cause the cellular damages underlying COVID-19 symptomatology.

Methods: We examined each of the individual SARS-CoV-2 proteins for their cytotoxicity in HEK 293 T cells and their subcellular localization in COS-7 cells. We also used Mass-Spec Affinity purification to identify the host proteins interacting with SARS-CoV-2 Orf6 protein and tested a drug that could inhibit a specific Orf6 and host protein interaction.

Results: We found that Orf6, Nsp6 and Orf7a induced the highest toxicity when over-expressed in human 293 T cells. All three proteins showed membrane localization in COS-7 cells. We focused on Orf6, which was most cytotoxic and localized to the endoplasmic reticulum, autophagosome and lysosomal membranes. Proteomics revealed Orf6 interacts with nucleopore proteins (RAE1, XPO1, RANBP2 and nucleoporins). Treatment with Selinexor, an FDA-approved inhibitor for XPO1, attenuated Orf6-induced cellular toxicity in human 293 T cells.

Conclusions: Our study revealed Orf6 as a highly pathogenic protein from the SARS-CoV-2 genome, identified its key host interacting proteins, and Selinexor as a drug candidate for directly targeting Orf6 host protein interaction that leads to cytotoxicity.
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http://dx.doi.org/10.1186/s13578-021-00568-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7993076PMC
March 2021

Clathrin Heavy Chain 1 Plays Essential Roles During Oocyte Meiotic Spindle Formation and Early Embryonic Development in Sheep.

Front Cell Dev Biol 2021 25;9:609311. Epub 2021 Feb 25.

State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, China.

As a major protein of the polyhedral coat of coated pits and vesicles, clathrin molecules have been shown to play a stabilization role for kinetochore fibers of the mitotic spindle by acting as inter-microtubule bridges. Clathrin heavy chain 1 (CLTC), the basic subunit of the clathrin coat, plays vital roles in both spindle assembly and chromosome congression during somatic-cell mitosis. However, its function in oocyte meiotic maturation and early embryo development in mammals, especially in domesticated animals, has not been fully investigated. In this study, the expression profiles and functional roles of CLTC in sheep oocytes were investigated. Our results showed that the expression of CLTC was maintained at a high level from the germinal vesicle (GV) stage to metaphase II stage and that CLTC was distributed diffusely in the cytoplasm of cells at interphase, from the GV stage to the blastocyst stage. After GV breakdown (GVBD), CLTC co-localized with beta-tubulin during metaphase. Oocyte treatments with taxol, nocodazole, or cold did not affect CLTC expression levels but led to disorders of its distribution. Functional impairment of CLTC by specific morpholino injections in GV-stage oocytes led to disruptions in spindle assembly and chromosomal alignment, accompanied by impaired first polar body (PB1) emissions. In addition, knockdown of CLTC before parthenogenetic activation disrupted spindle formation and impaired early embryo development. Taken together, the results demonstrate that CLTC plays a vital role in sheep oocyte maturation via the regulation of spindle dynamics and an essential role during early embryo development.
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http://dx.doi.org/10.3389/fcell.2021.609311DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7946971PMC
February 2021

Heterozygosity for a Pathogenic Variant in That Causes Autosomal Recessive Gitelman Syndrome Is Associated with Lower Serum Potassium.

J Am Soc Nephrol 2021 Mar 4;32(3):756-765. Epub 2021 Feb 4.

Program in Personalized and Genomic Medicine, Division of Endocrinology, Diabetes and Nutrition, Department of Medicine, University of Maryland School of Medicine, Baltimore, Maryland

Background: Potassium levels regulate multiple physiologic processes. The heritability of serum potassium level is moderate, with published estimates varying from 17% to 60%, suggesting genetic influences. However, the genetic determinants of potassium levels are not generally known.

Methods: A whole-exome sequencing association study of serum potassium levels in 5812 subjects of the Old Order Amish was performed. A dietary salt intervention in 533 Amish subjects estimated interaction between p.R642G and sodium intake.

Results: A cluster of variants, spanning approximately 537 kb on chromosome 16q13, was significantly associated with serum potassium levels. Among the associated variants, a known pathogenic variant of autosomal recessive Gitelman syndrome (p.R642G ) was most likely causal; there were no homozygotes in our sample. Heterozygosity for p.R642G was also associated with lower chloride levels, but not with sodium levels. Notably, p.R642G showed a novel association with lower serum BUN levels. Heterozygotes for p.R642G had a two-fold higher rate of self-reported bone fractures and had higher resting heart rates on a low-salt diet compared with noncarriers.

Conclusions: This study provides evidence that heterozygosity for a pathogenic variant in causing Gitelman syndrome, a canonically recessive disorder, contributes to serum potassium concentration. The findings provide insights into biology and the effects of heterozygosity on electrolyte homeostasis and related subclinical phenotypes that may have implications for personalized medicine and nutrition.
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http://dx.doi.org/10.1681/ASN.2020071030DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7920171PMC
March 2021

CircSND1 Regulated by TNF-α Promotes the Migration and Invasion of Cervical Cancer Cells.

Cancer Manag Res 2021 12;13:259-275. Epub 2021 Jan 12.

Tianjin Life Science Research Center, Tianjin Key Laboratory of Inflammation Biology, Collaborative Innovation Center of Tianjin for Medical Epigenetics, Department of Pathogen Biology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin, People's Republic of China.

Aim: To explore the role and potential mechanism of circSND1 in cervical cancer (CC).

Main Methods: qRT-PCR was used to determine the expression of circSND1 in tumor necrosis factor-α (TNF-α)-treated HeLa cells. CircSND1 overexpression and knockdown were performed to indicate the functional role of circSND1 in vitro and in vivo. Luciferase assay was used to analyze promoter activity. The expression and regulation of circSND1, miR-125a-3p and FUT6 were evaluated using EGFP fluorescent reporter assay and rescue experiments. Immunofluorescence and Western blot assays were used to analyze the activation of nuclear factor-κB (NF-κB).

Results: In HeLa cells, TNF-α up-regulated the expression of circSND1 by activating the NF-κB signaling pathway. Overexpression of circSND1 significantly increased the migration and invasion and the epithelial-mesenchymal transition (EMT) process of CC cells, and promoted tumor metastasis in xenograft nude mouse model, whereas down-regulation of circSND1 exerted opposite effects. Furthermore, circSND1 enhanced the expression of FUT6 via sponging miR-125a-3p, and FUT6 activated NF-κB signaling pathway.

Conclusion: We found that circSND1 promoted the expression of FUT6 and the malignant behavior of cervical cancer through the ceRNA mechanism, and there was a TNF-α/NF-κB/circSND1/miR-125a-3p/FUT6/NF-κB positive feedback pathway between them, which suggests that circSND1 can be a promising prognostic marker and therapeutical target for cervical cancer.
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http://dx.doi.org/10.2147/CMAR.S289032DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7811455PMC
January 2021

Exome Sequencing and Congenital Heart Disease in Sub-Saharan Africa.

Circ Genom Precis Med 2021 Feb 15;14(1):e003108. Epub 2021 Jan 15.

Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda (A.F.M., B.O., C.T.-N., Y.A.A., M.M., P.K.).

Background: Congenital heart disease (CHD) is the most common birth defect and affects roughly 1% of the global population. There have been many large CHD sequencing projects in developing countries but none in sub-Saharan Africa. In this exome sequencing study, we recruited families from Lagos, Nigeria, affected by structural heart disease.

Methods: Ninety-eight participants with CHD and an average age of 3.6 years were recruited from Lagos, Nigeria. Exome sequencing was performed on probands and parents when available. For genes of high interest, we conducted functional studies in using a cardiac-specific RNA interference-based gene silencing system.

Results: The 3 most common CHDs were tetralogy of Fallot (20%), isolated ventricular septal defect (14%), and transposition of the great arteries (8%). Ten percent of the cohort had pathogenic or likely pathogenic variants in genes known to cause CHD. In 64 complete trios, we found 34 de novo variants that were not present in the African population in the Genome Aggregation Database (v3). Nineteen loss of function variants were identified using the genome-wide distribution of selection effects for heterozygous protein-truncating variants (s). Nine genes caused a significant mortality when silenced in the heart, including 4 novel disease genes not previously associated with CHD (, and ).

Conclusions: This study identifies novel candidate genes and variants for CHD and facilitates comparisons with previous CHD sequencing studies in predominantly European cohorts. The study represents an important first step in genomic studies of CHD in understudied populations. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT01952171.
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http://dx.doi.org/10.1161/CIRCGEN.120.003108DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7887052PMC
February 2021

Autophagy inhibition rescues structural and functional defects caused by the loss of mitochondrial chaperone in .

Autophagy 2021 Jan 25:1-15. Epub 2021 Jan 25.

Research Group Synaptic Plasticity, Hertie Institute for Clinical Brain Research, University of Tübingen , Tübingen, Germany.

We investigated in larval and adult models whether loss of the mitochondrial chaperone is sufficient to cause pathological alterations commonly observed in Parkinson disease. At affected larval neuromuscular junctions, no effects on terminal size, bouton size or number, synapse size, or number were observed, suggesting that we studied an early stage of pathogenesis. At this stage, we noted a loss of synaptic vesicle proteins and active zone components, delayed synapse maturation, reduced evoked and spontaneous excitatory junctional potentials, increased synaptic fatigue, and cytoskeleton rearrangements. The adult model displayed ATP depletion, altered body posture, and susceptibility to heat-induced paralysis. Adult phenotypes could be suppressed by knockdown of , and . The knockdown of components of the macroautophagy/autophagy machinery or overexpression of human broadly rescued larval and adult phenotypes, while disease-associated variants did not. Overexpression of or promotion of autophagy exacerbated defects. AEL: after egg laying; AZ: active zone; brp: bruchpilot; Csp: cysteine string protein; dlg: discs large; eEJPs: evoked excitatory junctional potentials; GluR: glutamate receptor; HO: hydrogen peroxide; mEJP: miniature excitatory junctional potentials; MT: microtubule; NMJ: neuromuscular junction; PD: Parkinson disease; : PTEN-induced putative kinase 1; PSD: postsynaptic density; SSR: subsynaptic reticulum; SV: synaptic vesicle; VGlut: vesicular glutamate transporter.
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http://dx.doi.org/10.1080/15548627.2020.1871211DOI Listing
January 2021

Phycocyanin Improves Reproductive Ability in Obese Female Mice by Restoring Ovary and Oocyte Quality.

Front Cell Dev Biol 2020 13;8:595373. Epub 2020 Nov 13.

State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, China.

Reproductive dysfunction associated with obesity is increasing among women of childbearing age. Emerging evidence indicates that maternal obesity impairs embryo development and offspring health, and these defects are linked to oxidative stress in the ovary and in oocytes. Phycocyanin (PC) is a biliprotein from that possesses antioxidant, anti-inflammatory, and radical-scavenging properties. Our previous studies have shown that PC can reduce reactive oxygen species (ROS) accumulation in oocytes in D-gal-induced aging mice. Here, at the Institute of Cancer Research (ICR) mice fed a high-fat diet (HFD) to model obesity were used to test the effect of PC on reversing the fertility decline caused by obesity. We observed a significant increase in litter size and offspring survival rates after PC administration to obese mice. Further, we found that PC not only ameliorated the level of ovarian antioxidant enzymes, but also reduced the occurrence of follicular atresia in obese female mice. In addition, the abnormal morphology of the spindle-chromosome complex (SCC), and the abnormal mitochondrial distribution pattern in oocytes both recovered. The obesity-related accumulation of ROS, increased number of early apoptotic cells, and the abnormal expression of H3K9me3 in oocytes were all partially reversed after PC administration. In summary, this is the first demonstration that PC can improve fertility by partially increasing ovarian and oocyte quality in obese female mice and provides a new strategy for clinically treating obesity-related infertility in females.
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http://dx.doi.org/10.3389/fcell.2020.595373DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7691388PMC
November 2020

Superfine pulverisation pretreatment to enhance crystallinity of cellulose from Lycium barbarum L. leaves.

Carbohydr Polym 2021 Feb 13;253:117207. Epub 2020 Oct 13.

Department of Food Science and Engineering, College of Biological Sciences and Technology, Beijing Key Laboratory of Forest Food Processing and Safety, Beijing Forestry University, Beijing, 100083, China.

Superfine pulverisation (SFP) pretreatment of Lycium barbarum L. leaves was performed to obtain highly crystalline cellulose. Compared with other common pulverisation methods, SFP enhanced cellulosic crystallinity by 18.3 % and 8.4 %, with and without post-acid treatments, respectively. XRD and solid-state NMR analyses showed that SFP facilitated the exposure of amorphous substances (i.e., hemicellulose and lignin) to NaOH and HO. Large amounts of silicon (5.5 %) and aluminium (2.1 %) were found to incorporate into the crystalline regions of SFP-produced cellulose. Further FTIR and thermogravimetric analyses revealed that SFP-produced cellulose contained large amounts of hydroxyl groups, affecting the cellulosic crystallinity and thermal stability. These findings demonstrate the potential for SFP to serve as a green technology for production of highly crystalline and mineral-rich cellulose.
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http://dx.doi.org/10.1016/j.carbpol.2020.117207DOI Listing
February 2021

Mechanistic Insight into Palladium-Catalyzed γ-C(sp)-H Arylation of Alkylamines with 2-Iodobenzoic Acid: Role of the -Carboxylate Group.

Inorg Chem 2020 Dec 30;59(24):18295-18304. Epub 2020 Nov 30.

Key Lab of Colloid and Interface Chemistry, Ministry of Education, Institute of Theoretical Chemistry, School of Chemistry and Chemical engineering, Shandong University, Jinan 250100, People's Republic of China.

Density functional theory calculations were performed to understand the distinctly different reactivities of carboxylate-substituted aryl halides and pristine aryl halides toward the Pd-catalyzed γ-C(sp)-H arylation of secondary alkylamines. It is found that, when 2-iodobenzoic acid (a representative of -carboxylate-substituted aryl halides) is used as an aryl transfer agent, the arylation reaction is energetically favorable, while when the pristine aryl halide iodobenzene is used as the aryl transfer reagent, the reaction is kinetically difficult. Our calculations showed an operative Pd/Pd/Pd redox cycle, which differs in the mechanistic details from the cycle proposed by the experimental authors. The improved mechanism emphasizes that (i) the intrinsic role of the -carboxylate group is facilitating the C(sp)C(sp) bond reductive elimination from Pd rather than facilitating the oxidative addition of the aryl iodide on Pd, (ii) the decarboxylation occurs at the Pd species instead of the Pd species, and (iii) the 1,2-arylpalladium migration proceeds via a stepwise mechanism where the reductive elimination occurs before decarboxylation, not via a concerted mechanism that merges the three processes decarboxylation, 1,2-arylpalladium migration, and C(sp)-C(sp) reductive elimination into one. The experimentally observed exclusive site selectivity of the reaction was also rationalized well.
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http://dx.doi.org/10.1021/acs.inorgchem.0c02895DOI Listing
December 2020

Effects of Atorvastatin Combined with Nano-Selenium on Blood Lipids and Oxidative Stress in Atherosclerotic Rats.

J Nanosci Nanotechnol 2021 Feb;21(2):1331-1337

The Second Department of Cardiology, Affiliated Hospital of Hebei University, Baoding City, 071000, Hebei Province, China.

Dyslipidemia and oxidative stress injury of blood vessel walls play important roles in the formation of atherosclerosis (AS) and plaque progression. This is also the main pathological basis for atherosclerosis. Statins, as inhibitors of HMG-CoA reductase in the process of cholesterol biosynthesis, have become key drugs for lipid-lowering treatment. Many studies have found the anti-atherosclerotic effect of atorvastatin is far beyond the lipid-lowering effect. Its lipid-lowering effects are also involved, such as anti-inflammatory, inhibiting endothelial cell ROS production, and improving endothelial cell damage. Nano selenium (Nano-Se) shows stronger anti-oxidation ability, lower toxicity, high efficiency absorption and strong immune regulation ability. Because of the unique biological effects of Nano-Se, it has broad prospects in the field of human health care. Therefore, in this study, by constructing a rat model of abnormal lipid metabolism, we observed changes in parameters such as serum peroxidase (MPO), propylene glycol (MDA), superoxide dismutase (SOD), and blood lipid levels in atherosclerotic rats Happening, furthermore, the effects of atorvastatin+nano-selenium on lipid metabolism disorders and the protective effects and mechanisms of oxidative stress injury in rats were investigated and with a view to providing new targets for the treatment of arteriosclerosis. The results of this study demonstrated that contrast to the AS rat, the combined use of atorvastatin+nano-selenium group could significantly reduce serum TC, TG, and LDL-C contents, and declined tissue lesions such as aortic arch and liver; Significantly enhanced the activities of GPx-1 and SOD in serum, decreased MDA content, and increased the SOD activity in rat aorta. These results suggested that the combined use of atorvastatin+nano-selenium has good protection against oxidative stress caused by disorders of lipid metabolism.
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http://dx.doi.org/10.1166/jnn.2021.18633DOI Listing
February 2021

Hydrogenation of Carbon Dioxide to Methanol over Non-Cu-based Heterogeneous Catalysts.

ChemSusChem 2020 Dec 17;13(23):6160-6181. Epub 2020 Nov 17.

State Key Laboratory of Catalysis, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, P.R. China.

The increasing atmospheric CO level makes CO reduction an urgent challenge facing the world. Catalytic transformation of CO into chemicals and fuels utilizing renewable energy is one of the promising approaches toward alleviating CO emissions. In particular, the selective hydrogenation of CO to methanol utilizing renewable hydrogen potentially enables large scale transformation of CO . The Cu-based catalysts have been extensively investigated in CO hydrogenation. However, it is not only limited by long-term instability but also displays unsatisfactory catalytic performance. The supported metal-based catalysts (Pd, Pt, Au, and Ag) can achieve high methanol selectivity at low temperatures. The mixed oxide catalysts represented by M ZrO (M =Zn, Ga, and Cd) solid solution catalysts present high methanol selectivity and catalytic activity as well as excellent stability. This Review focuses on the recent advances in developing Non-Cu-based heterogeneous catalysts and current understandings of catalyst design and catalytic performance. First, the thermodynamics of CO hydrogenation to methanol is discussed. Then, the progress in supported metal-based catalysts, bimetallic alloys or intermetallic compounds catalysts, and mixed oxide catalysts is discussed. Finally, a summary and a perspective are presented.
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http://dx.doi.org/10.1002/cssc.202002054DOI Listing
December 2020

Master regulator genes and their impact on major diseases.

PeerJ 2020 6;8:e9952. Epub 2020 Oct 6.

The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, Hunan, China.

Master regulator genes (MRGs) have become a hot topic in recent decades. They not only affect the development of tissue and organ systems but also play a role in other signal pathways by regulating additional MRGs. Because a MRG can regulate the concurrent expression of several genes, its mutation often leads to major diseases. Moreover, the occurrence of many tumors and cardiovascular and nervous system diseases are closely related to MRG changes. With the development in omics technology, an increasing amount of investigations will be directed toward MRGs because their regulation involves all aspects of an organism's development. This review focuses on the definition and classification of MRGs as well as their influence on disease regulation.
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http://dx.doi.org/10.7717/peerj.9952DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7546222PMC
October 2020

MicroRNA-363-3p downregulation in papillary thyroid cancer inhibits tumor progression by targeting NOB1.

J Investig Med 2021 01 19;69(1):66-74. Epub 2020 Oct 19.

Thyroid Surgery, First Hospital of Jilin University, Changchun, China

MicroRNA-363-3 p (miR-363-3 p) has been reported to play a crucial role in tumor development and progression, and function as a tumor suppressor in many types of cancer. In our previous studies, we found that miRNA-363-3 p inhibited papillary thyroid carcinoma (PTC) progression by targeting PIK3CA. Meanwhile, we found that NIN1/RPN12 binding protein 1 (NOB1) was significantly upregulated in thyroid carcinoma tissue and downregulation of NOB1 expression significantly inhibited cell proliferation, migration and invasion in PTC. However, the correlation of NOB1 and miR-363-3 p has not been investigated. Here, we performed bioinformatic analysis to explore miRNA targeting NOB1. We found that NOB1 was a target of miR-363-3 p and miR-363-3 p regulated NOB1 expression at the translational and transcriptional levels by targeting its 3' untranslated region (3'-UTR). Further, we showed that miR-363-3 p inhibited tumor progression by targeting NOB1 in vitro and in vivo. We found that overexpression miR-363-3 p or silencing NOB1 significantly increased G0/G1-phase and decreased S-phase in the human papillary thyroid cells, which led to a significant delay in cell proliferation, indicating miR-363-3 p and NOB1 are crucial for human papillary thyroid cancer tumorigenesis. Collectively, our data unveil that miR-363-3 p negatively regulates NOB1 activity by reducing its stability. This study provides a new therapeutic target for regulation of NOB1 stability to modulate human papillary thyroid cancer progression.
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http://dx.doi.org/10.1136/jim-2020-001562DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7803892PMC
January 2021

Intracystic Hemorrhage and Its Management During Ultrasound-Guided Percutaneous Microwave Ablation for Cystic Thyroid Nodules.

Front Endocrinol (Lausanne) 2020 8;11:477. Epub 2020 Sep 8.

Department of Thyroid Surgery, The First Hospital of Jilin University, Changchun, China.

Intracystic hemorrhage can present occasionally during ultrasound-guided percutaneous microwave ablation (PMWA) for cystic thyroid nodules. It can affect treatment outcome, can lead to ablation failure, and even conversion to open surgery. We aim to avoid such cases in the future by exploring their causes and management. From March 2017 to December 2019, we retrospectively studied 87 cystic thyroid nodules in 59 patients who underwent PMWA in the First Hospital of Jilin University. All patients were followed at 1, 3, 6, and 12 months after treatment. All patients completed the treatment successfully. Nine cystic thyroid nodules presented with intracystic hemorrhage during the ultrasound-guided PMWA, giving an incidence of 10.3% (9/87 cysts). Larger cystic thyroid nodules were more likely to develop intracystic bleeding during ultrasound-guided PMWA. Intracystic hemorrhage resulted in significantly prolonged ablation time and had a negative effect on treatment outcome. No patients had other complications, but temporary post-operative pain and local swelling were more obvious in patients with intracystic hemorrhage. Intracystic hemorrhage is not rare during ultrasound-guided PMWA for cystic thyroid nodules. Doctors should pay more attention to it, learn to manage it and try to avoid it in clinical practice.
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http://dx.doi.org/10.3389/fendo.2020.00477DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7506029PMC
September 2020

The Visual Acuity Outcome and Relevant Factors Affecting Visual Improvement in Pediatric Sporadic Chiasmatic-Hypothalamic Glioma Patients Who Received Surgery.

Front Neurol 2020 19;11:766. Epub 2020 Aug 19.

Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.

The role and effectiveness of primary surgical treatment for sporadic chiasmatic-hypothalamic glioma (CHG) are not clear. The present study was to describe sporadic CHG visual acuity (VA) outcomes after surgery and to analyze the relevant factors affecting VA improvement. Forty-five pediatric sporadic CHG patients who met the inclusion criteria were included in a retrospective study. All patients received primary intratumor partial resection. Disease characteristics, treatment strategies, complications, and VA outcome were analyzed. Univariate and multivariate analyses were performed to identify relevant factors of VA improvement. Receiver operating characteristic (ROC) analysis was performed to evaluate the predictive accuracy of measurement indexes. There were 77 eyes of 45 children suffering from various levels of VA impairment before surgical treatment, and only 13 eyes had normal vision. Patients with resection extents >70, 50-70, and <50% accounted for 26.67, 24.44, and 48.89%, respectively. The percentages of VA maintained and deteriorated in normal vision eyes were 61.54 and 38.46%. The percentages of VA improved, maintained, and deteriorated in visually impaired eyes after surgery were 29.87, 45.45, and 24.68%, respectively. There was a positive correlation between the IVA level and VA improvement. There was no significant difference in VA improvement between the different resection extents. Blindness occurred in ~4.4%. Approximately 11.1% of the children had complications that affected quality of life, which correlated with resection extent. IVA and tumor size were correlated with VA improvement. The AUC for IVA + tumor size predicting VA improvement was 0.831. The cutoff points for IVA level and tumor volume were 4.5 and 43.50 cm, respectively. IVA and tumor size were correlated with VA improvement after primary intratumor partial resection. Children with IVA ≥ level 5 were more likely to achieve visual improvement after decompression surgery, but decompression had limited effectiveness on vision improvement in patients with tumor volumes ≥ 43.50 cm. Performing resections < 50% was safe and did not reduce the effect of decompression to improve VA.
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http://dx.doi.org/10.3389/fneur.2020.00766DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466562PMC
August 2020

Protein regulator of cytokinesis 1 regulates chromosome dynamics and cytoplasmic division during mouse oocyte meiotic maturation and early embryonic development.

FEBS J 2020 Dec;287(23):5130-5147

State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Science, Inner Mongolia University, Hohhot, China.

In contrast to the homeokinesis of mitosis, asymmetric division of cytoplasm is the conspicuous feature of meiosis in mammalian oocytes. Protein regulator of cytokinesis 1 (PRC1) is an important regulator during mitotic spindle assembly and cytoplasmic division, but its functions in oocyte meiosis and early embryo development have not been fully elucidated. In this study, we detected PRC1 expression and localization and revealed a nuclear, spindle midzone-related dynamic pattern throughout meiotic and mitotic progressions. Treatment of oocytes with the reagents taxol or nocodazole disturbed the distribution of PRC1 in metaphase II oocytes. Further, PRC1 depletion led to failure of first polar body (PB1) extrusion and spindle migration, aneuploidy and defective kinetochore-microtubule attachment and spindle assembly. Overexpression of PRC1 resulted in PB1 extrusion failure, aneuploidy and serious defects of spindle assembly. To investigate PRC1 function in early embryos, we injected Prc1 morpholino into zygotes and 2-cell stage embryos. Depletion of PRC1 in zygotes impaired 4-cell, morula and blastocyst formation. Loss of PRC1 in single or double blastomeres in 2-cell stage embryos significantly impaired cell division, indicating its indispensable role in early embryo development. Co-immunoprecipitation showed that PRC1 interacts with polo-like kinase 1 (PLK1), and functional knockdown and rescue experiments demonstrated that PRC1 recruits PLK1 to the spindle midzone to regulate cytoplasmic division during meiosis. Finally, kinesin family member 4 knockdown downregulates PRC1 expression and leads to PRC1 localization failure. Taken together, our data suggest PRC1 plays an important role during oocyte maturation and early embryonic development by regulating chromosome dynamics and cytoplasmic division.
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http://dx.doi.org/10.1111/febs.15458DOI Listing
December 2020

Oral administration of olaquindox negatively affects oocytes quality and reproductive ability in female mice.

Ecotoxicol Environ Saf 2020 Sep 7;201:110826. Epub 2020 Jun 7.

State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, People's Republic of China. Electronic address:

As an effective feed additive in the livestock industry, olaquindox (OLA) has been widely used in domestic animal production. However, it is unclear whether OLA has negative effects on mammalian oocyte quality and fetal development. In this study, toxic effects of OLA were tested by intragastric gavage ICR mice with water, low-dose OLA (5 mg/kg/day), or high-dose OLA (60 mg/kg/day) for continuous 45 days. Results showed that high-dose OLA gavage severely affected the offspring birth and growth. Significantly, high-dose OLA impaired oocyte maturation and early embryo development, indicated by the decreased percentage of germinal vesicle breakdown, first polar body extrusion and blastocyst formation. Meanwhile, oxidative stress levels were increased in oocytes or ovaries, indexed by the increased levels of ROS, MDA, HO, NO, and decreased levels of GSH, SOD, CAT, GSH-Px and GSH-Rd. Furthermore, aberrant mitochondria distribution, defective spindle assembly, abnormal H3K4me2/H3K9me3 levels, increased DNA double-strand breaks and early apoptosis rate, were observed after high-dose OLA gavage. Taken together, our results for the first time illustrated that high-dose OLA gavage led to sub-fertility of females, which means that restricted utilization of OLA as feed additive should be considered.
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http://dx.doi.org/10.1016/j.ecoenv.2020.110826DOI Listing
September 2020

Single-cell RNA sequencing identifies novel cell types in Drosophila blood.

J Genet Genomics 2020 04 9;47(4):175-186. Epub 2020 Mar 9.

Center for Precision Disease Modeling, Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, 21201, USA; Division of Endocrinology, Diabetes and Nutrition, Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, 21201, USA. Electronic address:

Drosophila has been extensively used to model the human blood-immune system, as both systems share many developmental and immune response mechanisms. However, while many human blood cell types have been identified, only three were found in flies: plasmatocytes, crystal cells and lamellocytes. To better understand the complexity of fly blood system, we used single-cell RNA sequencing technology to generate comprehensive gene expression profiles for Drosophila circulating blood cells. In addition to the known cell types, we identified two new Drosophila blood cell types: thanacytes and primocytes. Thanacytes, which express many stimulus response genes, are involved in distinct responses to different types of bacteria. Primocytes, which express cell fate commitment and signaling genes, appear to be involved in keeping stem cells in the circulating blood. Furthermore, our data revealed four novel plasmatocyte subtypes (Ppn, CAH7, Lsp and reservoir plasmatocytes), each with unique molecular identities and distinct predicted functions. We also identified cross-species markers from Drosophila hemocytes to human blood cells. Our analysis unveiled a more complex Drosophila blood system and broadened the scope of using Drosophila to model human blood system in development and disease.
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http://dx.doi.org/10.1016/j.jgg.2020.02.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7321924PMC
April 2020

Novel frameshift variant in MYL2 reveals molecular differences between dominant and recessive forms of hypertrophic cardiomyopathy.

PLoS Genet 2020 05 26;16(5):e1008639. Epub 2020 May 26.

Center for Cardiovascular Research, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States of America.

Hypertrophic cardiomyopathy (HCM) is characterized by thickening of the ventricular muscle without dilation and is often associated with dominant pathogenic variants in cardiac sarcomeric protein genes. Here, we report a family with two infants diagnosed with infantile-onset HCM and mitral valve dysplasia that led to death before one year of age. Using exome sequencing, we discovered that one of the affected children had a homozygous frameshift variant in Myosin light chain 2 (MYL2:NM_000432.3:c.431_432delCT: p.Pro144Argfs*57;MYL2-fs), which alters the last 20 amino acids of the protein and is predicted to impact the most C-terminal of the three EF-hand domains in MYL2. The parents are unaffected heterozygous carriers of the variant and the variant is absent in control cohorts from gnomAD. The absence of the phenotype in carriers and the infantile presentation of severe HCM is in contrast to HCM associated with dominant MYL2 variants. Immunohistochemical analysis of the ventricular muscle of the deceased patient with the MYL2-fs variant showed a marked reduction of MYL2 expression compared to an unaffected control. In vitro overexpression studies further indicate that the MYL2-fs variant is actively degraded. In contrast, an HCM-associated missense variant (MYL2:p.Gly162Arg) and three other MYL2 stop-gain variants (p.E22*, p.K62*, p.E97*) that result in loss of the EF domains are stably expressed but show impaired localization. The degradation of the MYL2-fs can be rescued by inhibiting the cell's proteasome function supporting a post-translational effect of the variant. In vivo rescue experiments with a Drosophila MYL2-homolog (Mlc2) knockdown model indicate that neither the MYL2-fs nor the MYL2:p.Gly162Arg variant supports normal cardiac function. The tools that we have generated provide a rapid screening platform for functional assessment of variants of unknown significance in MYL2. Our study supports an autosomal recessive model of inheritance for MYL2 loss-of-function variants in infantile HCM and highlights the variant-specific molecular differences found in MYL2-associated cardiomyopathy.
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http://dx.doi.org/10.1371/journal.pgen.1008639DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7274480PMC
May 2020

Clinical and Ultrasonic Risk Factors for Lateral Lymph Node Metastasis in Papillary Thyroid Microcarcinoma: A Systematic Review and Meta-Analysis.

Front Oncol 2020 3;10:436. Epub 2020 Apr 3.

Department of Thyroid Surgery, The 1st Hospital of Jilin University, Changchun, China.

Clinical and ultrasonic risk factors for lateral lymph node metastasis (LLNM) in papillary thyroid microcarcinoma (PTMC) are not well-defined. Herein, a systematic review and meta-analysis was conducted to investigate clinicopathologic and ultrasonic risk features for LLNM in PTMC. A systematic search of electronic databases (PubMed, Embase, Cochrane Library, and Web of Science) for studies published until April 2019 was performed. Case-control studies and randomized controlled trials that studied clinical and ultrasonic risk factors of LLNM in PTMC were included. Fourteen studies (all retrospective studies) involving 43,750 patients met final inclusion criteria. From the pooled analyses, younger age<45 (OR, 1.55; 95% CI, 1.16-2.07; = 0.003), male patients (OR, 1.94; 95% CI, 1.55-2.42; < 0.00), extrathyroidal extension (OR, 3.63; 95% CI, 2.28-5.77; <0.00), tumor multifocality (OR, 2.24; 95% CI, 1.53-3.28; <0.00), tumor > 0.5 cm (OR, 2.24; 95% CI, 1.53-3.28; < 0.00), central lymph node metastasis (OR, 5.61; 95% CI, 4.64-6.79; < 0.00), >25% tumor contact with thyroid capsule (OR, 6.66; 95% CI, 1.96-22.65; = 0.002), tumor calcification (OR, 2.90; 95% CI, 1.71-4.93; < 0.00), upper tumor (OR, 3.18; 95% CI, 2.23-4.55; < 0.00) were significantly associated with increased risk of LLNM in PTMC, while Hashimoto's thyroiditis and other ultrasonic features (solid tumor, hypoechoic tumor, smooth margin, and taller than wide tumor) were not significantly associated with LLNM in PTMC. Our analysis identified several clinicopathologic and ultrasonic factors associated with LLNM in PTMC. This finding highlights the need for a cautious and frequent postoperative surveillance of the lateral neck in high-risk PTMC patients. Moreover, high-risk ultrasonic features also need to be considered during selection of PTMC for active surveillance.
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http://dx.doi.org/10.3389/fonc.2020.00436DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7145902PMC
April 2020

Exocyst Genes Are Essential for Recycling Membrane Proteins and Maintaining Slit Diaphragm in Nephrocytes.

J Am Soc Nephrol 2020 05 1;31(5):1024-1034. Epub 2020 Apr 1.

Center for Precision Disease Modeling, Department of Medicine, University of Maryland School of Medicine, Baltimore, Maryland

Background: Studies have linked mutations in genes encoding the eight-protein exocyst protein complex to kidney disease, but the underlying mechanism is unclear. Because nephrocytes share molecular and structural features with mammalian podocytes, they provide an efficient model for studying this issue.

Methods: We silenced genes encoding exocyst complex proteins specifically in nephrocytes and studied the effects on protein reabsorption by lacuna channels and filtration by the slit diaphragm. We performed nephrocyte functional assays, carried out super-resolution confocal microscopy of slit diaphragm proteins, and used transmission electron microscopy to analyze ultrastructural changes. We also examined the colocalization of slit diaphragm proteins with exocyst protein Sec15 and with endocytosis and recycling regulators Rab5, Rab7, and Rab11.

Results: Silencing exocyst genes in nephrocytes led to profound changes in structure and function. Abolition of cellular accumulation of hemolymph proteins with dramatically reduced lacuna channel membrane invaginations offered a strong indication of reabsorption defects. Moreover, the slit diaphragm's highly organized surface structure-essential for filtration-was disrupted, and key proteins were mislocalized. Ultrastructural analysis revealed that exocyst gene silencing led to the striking appearance of novel electron-dense structures that we named "exocyst rods," which likely represent accumulated membrane proteins following defective exocytosis or recycling. The slit diaphragm proteins partially colocalized with Sec15, Rab5, and Rab11.

Conclusions: Our findings suggest that the slit diaphragm of nephrocytes requires balanced endocytosis and recycling to maintain its structural integrity and that impairment of the exocyst complex leads to disruption of the slit diaphragm and nephrocyte malfunction. This model may help identify therapeutic targets for treating kidney diseases featuring molecular defects in vesicle endocytosis, exocytosis, and recycling.
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http://dx.doi.org/10.1681/ASN.2019060591DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7217423PMC
May 2020

Zika virus non-structural protein NS4A restricts eye growth in through regulation of JAK/STAT signaling.

Dis Model Mech 2020 04 30;13(4). Epub 2020 Apr 30.

Department of Biological Sciences, The George Washington University, Washington, DC 20052, USA

To gain a comprehensive view of the changes in host gene expression underlying Zika virus (ZIKV) pathogenesis, we performed whole-genome RNA sequencing (RNA-seq) of ZIKV-infected adult flies. RNA-seq analysis revealed that ZIKV infection alters several and diverse biological processes, including stress, locomotion, lipid metabolism, imaginal disc morphogenesis and regulation of JAK/STAT signaling. To explore the interaction between ZIKV infection and JAK/STAT signaling regulation, we generated genetic constructs overexpressing ZIKV-specific non-structural proteins NS2A, NS2B, NS4A and NS4B. We found that ectopic expression of non-structural proteins in the developing eye significantly restricts growth of the larval and adult eye and correlates with considerable repression of the JAK/STAT reporter, At the cellular level, eye growth defects are associated with reduced rate of proliferation without affecting the overall rate of apoptosis. In addition, ZIKV NS4A genetically interacts with the JAK/STAT signaling components; co-expression of along with the dominant-negative form of or results in aggravated reduction in eye size, while co-expression of in (also known as ) mutant background partially rescues the -induced eye overgrowth phenotype. The function of ZIKV NS4A in regulating growth is maintained in the wing, where ZIKV overexpression in the pouch domain results in reduced growth linked with diminished expression of Notch targets, Wingless (Wg) and Cut, and the Notch reporter, Thus, our study provides evidence that ZIKV infection in results in restricted growth of the developing eye and wing, wherein eye phenotype is induced through regulation of JAK/STAT signaling, whereas restricted wing growth is induced through regulation of Notch signaling. The interaction of ZIKV non-structural proteins with the conserved host signaling pathways further advance our understanding of ZIKV-induced pathogenesis.This article has an associated First Person interview with the first author of the paper.
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http://dx.doi.org/10.1242/dmm.040816DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7197722PMC
April 2020

Pharmacy students' perceptions toward peer assessment and its use in teaching patient presentation skills.

Curr Pharm Teach Learn 2020 02 25;12(2):228-236. Epub 2019 Nov 25.

Department of Pharmacy, National University of Singapore, Block S4A, Level 3, 18 Science Drive 4, 117543, Republic of Singapore. Electronic address:

Background And Purpose: Conducting peer assessment has been associated with positive learning outcomes in higher education. The primary objective was to evaluate pharmacy students' perceptions of using peer assessment as a pedagogical strategy in learning patient presentation skills. Secondary objectives were to determine helpful factors for providing and/or receiving peer assessment and to compare students' perceptions of peer assessment relative to receiving feedback from teaching assistants (TAs).

Educational Activity And Setting: Patient presentation skills were taught to third-year pharmacy students in three sessions (session 1: didactic lecture, session 2: faculty-led patient presentation workshops followed by peer assessment, session 3: one-on-one patient presentations to TAs). An anonymous survey instrument consisting of five-point Likert scale, yes/no, and open-ended questions was administered.

Findings: A total of 187 students (98%) completed the survey. Peer assessment was perceived as a useful way to obtain feedback on patient presentations (87%). It facilitated higher level thinking and a self-reflection of students' own patient presentations. Most students felt that they received constructive feedback from peers (82%) that helped them improve their patient presentation skills (72%). However, students were more trusting of TAs' skills in assessing patient presentations (76% versus 93%, p < 0.001). Some students were concerned about the specificity and criticalness of feedback they received from peers.

Summary: Peer assessment is a useful pedagogical strategy for providing formative feedback to students in learning patient presentations skills in the classroom setting. Students may benefit from additional training to improve the quality of feedback in peer assessment.
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http://dx.doi.org/10.1016/j.cptl.2019.10.012DOI Listing
February 2020

Transient Delay-Period Activity of Agranular Insular Cortex Controls Working Memory Maintenance in Learning Novel Tasks.

Neuron 2020 03;105(5):934-946.e5

Institute of Neuroscience, State Key Laboratory of Neuroscience, Chinese Academy of Sciences, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Center for Brain Science and Brain-Inspired Technology, Shanghai 200031, China; School of Future Technology, University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address:

Whether transient or sustained neuronal activity during the delay period underlies working memory (WM) has been debated. Here, we report that transient, but not sustained, delay-period activity in mouse anterior agranular insular cortex (aAIC) plays a dominant role in maintaining WM information during learning of novel olfactory tasks. By optogenetic screening over 12 brain regions, we found that suppressing aAIC activity markedly impaired olfactory WM maintenance during learning. Single-unit recording showed that odor-selective aAIC neurons with predominantly transient firing patterns encoded WM information. Both WM task performance and transient-neuron proportion were enhanced and reduced by activating and suppressing the delay-period activity of the projection from medial prefrontal cortex (mPFC) to aAIC. The ability of mice to resist delay-period distractors also correlated with an increased percentage of transient neurons. Therefore, transient, but not sustained, aAIC neuronal activity during the delay period is largely responsible for maintaining information while learning novel WM tasks.
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http://dx.doi.org/10.1016/j.neuron.2019.12.008DOI Listing
March 2020

[Methods used for gene manipulation in mammal female germ cells and early embryos].

Sheng Li Xue Bao 2020 Feb;72(1):31-47

State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, The Research Center for Laboratory Animal Science, School of Life Sciences, Inner Mongolia University, Hohhot 010070, China.

For sexual reproduction, oocytes are mammalian female germ cells that provide the majority of maternal genetic material for early stage embryo production and development. Early stage embryos begin the process of multicellular organism formation through cell differentiation. Studies on mammalian female germ cells (oocytes) not only reveal its unique physiological characteristics, but also help understand the mechanism involved in cell differentiation of other cell types. However, because it is difficult to culture in vitro, our understanding of the function of oocytes and early stage embryos remains very limited. Gene editing or manipulation is one of the most commonly used method, which is also useful in the field of gametes study. In this review, we summarized the principles, advantages and disadvantages of techniques, which include conditional knockout, RNA interference, Morpholino, Trim-Away and antibody-mediated inhibition of protein function, currently used for gene manipulation in oocytes and early stage embryos. We also discuss the issues the investigators need to consider. Finally, we highlight the future directions for gene manipulation or editing in female germ cells and early stage embryos.
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February 2020

Blastocyst hatching site is regularly distributed and does not influence foetal development in mice.

Sci Rep 2020 02 12;10(1):2475. Epub 2020 Feb 12.

State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, Inner Mongolia, People's Republic of China.

Hatching out from the zona pellucida (ZP) is a crucial step for blastocyst implantation and development. However, it is still unknown whether the location of the hatching site relative to the inner cell mass (ICM) affects embryo implantation and foetal development. Here, we classified hatching blastocysts into three categories, 0° ≤ θ ≤ 30°, 30° < θ ≤ 60°, and 60° < θ ≤ 90°, in which θ is determined based on the relative position of the hatching site to the arc midpoint of the ICM. Non-surgical embryo transfer (NSET) devices were employed to evaluate blastocyst implantation and embryo development. Of 1,827 hatching blastocysts, 43.84%, 30.60%, and 21.67% were categorized as 30° < θ ≤ 60°, 0° ≤ θ ≤ 30°, and 60° < θ ≤ 90°, respectively. Embryos with different hatching sites showed no distinct differences in blastocyst implantation; surrogate female pregnancy; embryo development to term; litter size, or offspring survival, gender, or body weight. Our results indicate that mouse blastocyst hatching site is not randomly distributed. Embryo implantation and development are not correlated with the blastocyst hatching site in mice. Thus, assessment of the blastocyst hatching site should not be recommended to evaluate mouse blastocyst implantation and developmental potential.
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http://dx.doi.org/10.1038/s41598-020-59424-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7015891PMC
February 2020

Knockdown of MCM10 Gene Impairs Glioblastoma Cell Proliferation, Migration and Invasion and the Implications for the Regulation of Tumorigenesis.

J Mol Neurosci 2020 May 6;70(5):759-768. Epub 2020 Feb 6.

Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.

Minichromosome maintenance 10 (MCM10) plays an important role in DNA replication and is expressed in a variety of tumors, including glioma. However, its role and mechanism in glioma remain elusive. The purpose of this study was to examine the molecular function of MCM10 in glioblastoma cell lines in vitro and to further investigate the molecular mechanisms in the network mediated by MCM10. Cell proliferation, invasion, and migration were investigated in the absence of MCM10 mediated by RNA interference (RNAi) in U87 and U251 cell lines. Microarray data were obtained from U87 cells infected with a lentivirus expressing a small interfering RNA (siRNA) targeting MCM10, and ingenuity pathway analysis (IPA) was performed. Molecular signaling pathways, gene functions, and upstream and downstream regulatory genes and networks were analyzed. MCM10 was positively stained in human glioblastoma multiforme (GBM) samples according to immunohistochemistry. Silencing MCM10 in U87 and U251 cells significantly reduced cell proliferation, migration, and invasion. In U87 cells transfected with MCM10, 274 genes were significantly upregulated, while 313 genes were downregulated. IPA revealed that MCM10 is involved in the IGF-1 signaling pathway, and calcitriol appears to be a significant upstream regulator of MCM10. Other factors, such as TWIST1 and Stat3, also interact within the MCM10-mediated network. Our data indicate that MCM10 is involved in the regulation of GBM in vitro and may provide more evidence for understanding the molecular mechanisms of this fatal disease.
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http://dx.doi.org/10.1007/s12031-020-01486-yDOI Listing
May 2020

Empagliflozin Attenuates Hyperuricemia by Upregulation of ABCG2 via AMPK/AKT/CREB Signaling Pathway in Type 2 Diabetic Mice.

Int J Biol Sci 2020 1;16(3):529-542. Epub 2020 Jan 1.

NHC Key Laboratory of Hormones and Development (Tianjin Medical University), Tianjin Key Laboratory of Metabolic Diseases, Tianjin Medical University Chu Hsien-I Memorial Hospital & Tianjin Institute of Endocrinology, Tianjin 300134, China.

Hyperuricemia (HUA) is a metabolic disease characterized by elevated serum uric acid (SUA). Empagliflozin, a kind of sodium-glucose cotransporter 2 inhibitors, has recently emerged as a new antidiabetic agent by facilitating glucose excretion in urine. Moreover, there was evidence of SUA reduction following treatment with empagliflozin in addition to glycaemic control, while the molecular mechanisms remain unknown. To investigate the potential mechanisms, the model of type 2 diabetes (T2DM) with HUA was established by combination of peritoneal injection of potassium oxonate and intragastric administration of hypoxanthine in KK-Ay mice. A series of method such as RT-PCR, western blot, immunochemistry, immunofluorescence were conducted to explore the mechanism. Our results showed that empagliflozin significantly ameliorated the levels of SUA and blood glucose in T2DM mice with HUA. Furthermore, in both kidney and ileum, empagliflozin obviously promoted protein expression of uric acid (UA) transporter ABCG2, p-AMPK, p-AKT and p-CREB. The same trend was observed in human tubular epithelial (HK-2) cells. Additionally, through application of an AMPK inhibitor (Compound C), it was further confirmed empagliflozin exerted its anti-hyperuricemic effects in an AMPK dependent manner. Meanwhile, with the help of ChIP assay and luciferase reporter gene assay, we found that CREB further activated ABCG2 via binding to the promoter of ABCG2 to induce transcription. Taken together, our study demonstrated that empagliflozin treatment played an essential role in attenuating HUA by upregulation of ABCG2 via AMPK/AKT/CREB signaling pathway.
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http://dx.doi.org/10.7150/ijbs.33007DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6990905PMC
January 2021