1.Fucoidan sulfate from Sargassum fusiforme regulates the SARS-CoV-2 receptor AXL expression in human embryonic lung diploid fibroblast cells.
Xuqiang ZHOU ; Weihua JIN ; Di JIANG ; Yipeng XU ; Sanying WANG ; Xinna WU ; Yunchuang CHANG ; Huili SU ; Tianjun ZHU ; Xiaogang XU ; Genxiang MAO
Journal of Zhejiang University. Science. B 2023;24(11):1047-1052
新冠病毒感染疫情严重威胁着世界各国人民的生命健康。目前,对病毒感染的防治研究主要集中在抑制病毒与分子受体的结合上。AXL作为新发现的严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)受体,在协助病毒感染人体呼吸系统中发挥着重要作用,是未来临床干预的潜在靶点。本研究对已发表的单细胞测序数据进行整理和分析,发现AXL在年轻人肺细胞中的表达水平明显高于老年人。人胚肺二倍体成纤维细胞(2BS)是衰老研究的公认细胞株。本文采用2BS细胞构建复制性细胞衰老模型,发现年轻细胞中AXL的蛋白水平明显高于衰老细胞,据此推测年轻人感染的风险可能更高,需要注意防护。我们发现一种羊栖菜褐藻多糖硫酸酯组分(SFW-3)可显著下调年轻2BS细胞中AXL的表达水平,表明SFW-3具有一定的抗SARS-CoV-2感染的研究价值,同时表明2BS细胞株也可作为潜在的SARS-CoV-2体外感染模型。
Humans
;
SARS-CoV-2
;
Sargassum/metabolism*
;
Diploidy
;
Sulfates/metabolism*
;
COVID-19
;
Polysaccharides/pharmacology*
;
Lung
2.Distinct mononuclear diploid cardiac subpopulation with minimal cell-cell communications persists in embryonic and adult mammalian heart.
Miaomiao ZHU ; Huamin LIANG ; Zhe ZHANG ; Hao JIANG ; Jingwen PU ; Xiaoyi HANG ; Qian ZHOU ; Jiacheng XIANG ; Ximiao HE
Frontiers of Medicine 2023;17(5):939-956
A small proportion of mononuclear diploid cardiomyocytes (MNDCMs), with regeneration potential, could persist in adult mammalian heart. However, the heterogeneity of MNDCMs and changes during development remains to be illuminated. To this end, 12 645 cardiac cells were generated from embryonic day 17.5 and postnatal days 2 and 8 mice by single-cell RNA sequencing. Three cardiac developmental paths were identified: two switching to cardiomyocytes (CM) maturation with close CM-fibroblast (FB) communications and one maintaining MNDCM status with least CM-FB communications. Proliferative MNDCMs having interactions with macrophages and non-proliferative MNDCMs (non-pMNDCMs) with minimal cell-cell communications were identified in the third path. The non-pMNDCMs possessed distinct properties: the lowest mitochondrial metabolisms, the highest glycolysis, and high expression of Myl4 and Tnni1. Single-nucleus RNA sequencing and immunohistochemical staining further proved that the Myl4+Tnni1+ MNDCMs persisted in embryonic and adult hearts. These MNDCMs were mapped to the heart by integrating the spatial and single-cell transcriptomic data. In conclusion, a novel non-pMNDCM subpopulation with minimal cell-cell communications was unveiled, highlighting the importance of microenvironment contribution to CM fate during maturation. These findings could improve the understanding of MNDCM heterogeneity and cardiac development, thus providing new clues for approaches to effective cardiac regeneration.
Animals
;
Mice
;
Diploidy
;
Heart
;
Myocytes, Cardiac/metabolism*
;
Cell Communication
;
Gene Expression Profiling
;
Mitochondria
;
Regeneration
;
Mammals/genetics*
3.Comparative analysis of active components and transcriptome between autotetraploid and diploid of Dendrobium huoshanense.
Cheng-Cai ZHANG ; Zhen GAO ; Li-Na LUO ; Hui-Hui LIANG ; Zeng-Xu XIANG
China Journal of Chinese Materia Medica 2020;45(23):5669-5676
In this study, the roots, stems and leaves of diploid and autotetraploid Dendrobium huoshanense were used as materials to compare their contents of polysaccharides and alkaloids, and the transcriptome sequencing analysis was carried out. The results showed that the contents of polysaccharides and alkaloids in the roots, stems and leaves of tetraploid were 7.6%, 34.5%, 17.2%, 0.01%, 0.024% and 0.035% higher than those of diploid D. huoshanense, respectively. The contents of active components in different tissues were significantly different. There were 3 687 differentially expressed genes in diploid and tetraploid D. huoshanense, of which 2 346 genes were up-regulated and 1 341 down regulated. Go functional analysis showed that these genes were mainly involved in growth and development, stress resistance and other related functions. KEGG pathway analysis showed that most of the differential genes were concentrated in the processes of carbon metabolism, signal transduction, carbohydrate metabolism, amino acid metabolism and energy metabolism. The differential expression of key genes involved in the metabolism of polysaccharides, terpenes and polyketones, amino acid metabolism, hormone synthesis and signal transduction in diploid and tetraploid plants may be the main reason for the high energy content, the increase of active components and the growth potential of tetraploid plants.
Alkaloids
;
Dendrobium/genetics*
;
Diploidy
;
Plant Roots
;
Polysaccharides
;
Transcriptome
4.MIME-Mitosis instead of meiosis and its application in crop apomixis.
Yanhong HOU ; Guizhi GONG ; Zhuchun PENG ; Qianqian DONG ; Ai LUO ; Qibin HONG
Chinese Journal of Biotechnology 2020;36(4):612-621
Apomixis has been widely concerned because of its great potential in heterosis fixation. Artificial apomixis is an important direction of current apomixis research. Mitosis instead of Meiosis (MIME) produces diploid gametes that is identical with the maternal genetic composition and is a key step in the artificial creation of apomixes. This paper reviews the occurrence of MIME and its application in crop apomixis and the problems encountered, in an aim to provide reference for expanding the application of MIME in crop apomixis.
Apomixis
;
Crops, Agricultural
;
genetics
;
Diploidy
;
Germ Cells
;
Meiosis
;
Mitosis
5.Inhibitory effect of the aqueous extract of a tetraploid ‘etteum’ variety in Platycodon grandiflorum on degranulation and inflammatory mediator release in RBL-2H3 cells.
Jae In JUNG ; Hyung Seo KIM ; Han Kyeol JI ; Hyun Sook LEE ; Jae Yong LEE ; Eun Ji KIM
Journal of Nutrition and Health 2018;51(3):208-214
PURPOSE: Platycodon grandiflorum (a domestic diploid variety, DV-PG) has been used as a food and component of various traditional oriental medicines. Although DV-PG is known to have an anti-allergic effect, little is known about the beneficial health effects of the tetraploid ‘Etteum’ variety in the Platycodon grandiflorum (TV-PG), which is a recently developed variety. In this study, we investigated the effect of TV-PG on the rat basophilic leukemia mast cell (RBL-2H3)-mediated allergic response. METHODS: To examine the effects of TV-PG on the allergic response, RBL-2H3 cells were sensitized with dinitropheny (DNP)-immunoglobin E, treated with various concentrations of TV-PG, and challenged with DNP-human serum albumin. We estimated cell granulation by measuring the release of β-hexosaminidase and production of inflammatory mediators by ELISA. RESULTS: TV-PG had no effect on the proliferation or cytotoxicity of RBL-2H3 cells within the concentration range of 0 to 200 µg/mL. TV-PG inhibited degranulation of RBL-2H3 cells by antigen stimulation in a dose-dependent manner. TV-PG also suppressed the production of inflammatory cytokines and mediators such as interleukin-4, tumor necrosis factor-α, prostagladin E2, and leukotriene B4 in RBL-2H3 cells by antigen stimulation. CONCLUSION: These results indicate that TV-PG exhibits anti-allergic activity via inhibition of degranulation as well as suppression of inflammatory mediators and cytokine release. These findings suggest that TV-PG may have potential as a preventive and therapeutic agent for the treatment of various allergic diseases.
Animals
;
Basophils
;
Cytokines
;
Diploidy
;
Enzyme-Linked Immunosorbent Assay
;
Functional Food
;
Hypersensitivity
;
Inflammation Mediators
;
Interleukin-4
;
Leukemia
;
Leukotriene B4
;
Mast Cells
;
Medicine, East Asian Traditional
;
Necrosis
;
Platycodon*
;
Rats
;
Serum Albumin
;
Tetraploidy*
6.Pathogenic variant in NLRP7 (19q13.42) associated with recurrent gestational trophoblastic disease: Data from early embryo development observed during in vitro fertilization.
E Scott SILLS ; Alexandra J OBREGON-TITO ; Harry GAO ; Thomas K MCWILLIAMS ; Anthony T GORDON ; Catharine A ADAMS ; Rima SLIM
Clinical and Experimental Reproductive Medicine 2017;44(1):40-46
OBJECTIVE: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. METHODS: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. RESULTS: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. CONCLUSION: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.
Abortion, Habitual
;
Aneuploidy
;
Biopsy
;
Blastocyst
;
Diploidy
;
Embryonic Development*
;
Embryonic Structures*
;
Female
;
Fertilization
;
Fertilization in Vitro*
;
Genetic Testing
;
Gestational Trophoblastic Disease*
;
Humans
;
Hydatidiform Mole
;
In Vitro Techniques*
;
Infertility
;
Oocytes
;
Parents
;
Ploidies
;
Pregnancy
;
Sperm Injections, Intracytoplasmic
7.Malvidin Protects WI-38 Human Fibroblast Cells Against Stress-induced Premature Senescence.
Hye Rin SEO ; Mi Jin CHOI ; Ji Myung CHOI ; Jong Cheol KO ; Jee Yeon KO ; Eun Ju CHO
Journal of Cancer Prevention 2016;21(1):32-40
BACKGROUND: Malvidin is one of the most abundant components in red wines and black rice. The effects of malvidin on aging and lifespan under oxidative stress have not been fully understood. This study focused on the anti-aging effect of malvidin on stress-induced premature senescence (SIPS) in WI-38 human lung-derived diploid fibroblasts. METHODS: In order to determine the viability of WI-38 cells, MTT assay was conducted, and malondialdehyde level was determined using thiobarbituric acid-reactive substance assay. Protein expression of inflammation-related factors was also evaluated by Western blot analysis. RESULTS: Acute and chronic oxidative stress via hydrogen peroxide (H2O2) treatment led to SIPS in WI-38 cells, which showed decreased cell viability, increased lipid peroxidation, and a shortened lifespan in comparison with non-H2O2-treated WI-38 cells. However, malvidin treatment significantly attenuated H2O2-induced oxidative stress by inhibiting lipid peroxidation and increasing cell viability. Furthermore, the lifespan of WI-38 cells was prolonged by malvidin treatment. In addition, malvidin downregulated the expression of oxidative stress-related proteins, including NF-κB, COX-2, and inducible nitric oxide synthase. Furthermore, protein expression levels of p53, p21, and Bax were also regulated by malvidin treatment in WI-38 cells undergoing SIPS. CONCLUSIONS: Malvidin may potentially inhibit the aging process by controlling oxidative stress.
Aging*
;
Blotting, Western
;
Cell Survival
;
Diploidy
;
Fibroblasts*
;
Humans*
;
Hydrogen Peroxide
;
Lipid Peroxidation
;
Malondialdehyde
;
Nitric Oxide Synthase Type II
;
Oxidative Stress
;
Wine
8.Utility of p57KIP2 and Her-2 fluorescence in situ hybridization in differentiating partial from complete hydatidiform mole.
Diwa Michele H. ; Kim Min-A ; Avila Jose Maria C. ; Pedroza David G. ; Encinas-Latoy Michelle Anne M.
Acta Medica Philippina 2016;50(4):318-325
INTRODUCTION: Hydatidiform mole (HM) is an abnormal gestation characterized by significant hydropic enlargement, trophoblastic proliferation and atypia involving part or all of the chorionic villi. The diagnosis and classification of hydatidiform moles is subject to great inter-observer variability due to significant morphologic overlaps. This study aims to evaluate the utility of p57KIP2 immunohistochemistry and ploidy by Her-2 FISH in refining the diagnosis of molar tissues.
METHOD: 113 and 78 molar cases were retrieved from the archives of the Histopathology Section of the Philippine General Hospital and Pathology Department of Seoul National University Hospital, respectively. TMA sections were submitted for immunohistochemical analysis for p57KIP2. Ploidy was determined by fluorescence in situ hybridization using Her-2 probe. An interrater reliability analysis was done using the Kappa statistics with 95% confidence interval.
RESULTS: All 68 (100%) cases diagnosed as CH were negative for p57KIP2 staining and are diploid. Among the 54 cases of PH, only 1 (2%) is positive for p57KIP2 and is diploid. The interrater reliability between p57KIP2 and Her-2 FISH ploidy results is 0.66 (p <.0.001), 95% CI (0.02, 1.00) which is considered "fair to good." The kappa value between review diagnosis and p57KIP2 is 0.024 while the kappa between review diagnosis and Her-2 FISH ploidy is 0.050 both signifying poor agreement beyond chance.
CONCLUSION: Morphologic assessment alone may not be sufficient in problematic cases. p57KIP2 in conjunction with by Her-2 FISH are good adjuncts in the diagnosis and classification of hydatidiform mole.
Human ; Male ; Female ; Pregnancy ; Chorionic Villi ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Diploidy ; Reproducibility Of Results ; Hydatidiform Mole ; Trophoblasts ; Ploidies ; Molar
9.The parental origin correlates with the karyotype of human embryos developing from tripronuclear zygotes.
Mette Warming JOERGENSEN ; Rodrigo LABOURIAU ; Johnny HINDKJAER ; Magnus STOUGAARD ; Steen KOLEVRAA ; Lars BOLUND ; Inge Errebo AGERHOLM ; Lone SUNDE
Clinical and Experimental Reproductive Medicine 2015;42(1):14-21
OBJECTIVE: It has previously been suggested that embryos developing from intracytoplasmic sperm-injected (ICSI) zygotes with three pronuclei (3PN) are endowed with a mechanism for self-correction of triploidy to diploidy. 3PN are also observed in zygotes after conventional in vitro fertilization (IVF). The parental origin, however, differs between the two fertilization methods. Whereas the vast majority of 3PN IVF zygotes are of dispermic origin and thus more likely to have two centrioles, the 3PN ICSI zygotes are digynic in origin and therefore, more likely to have one centriole. In the present study, we examine whether the parental origin of 3PN embryos correlates with the karyotype. METHODS: The karyotype of each nucleus was estimated using four sequential fluorescence in situ hybridizations-each with two probes-resulting in quantitative information of 8 different chromosomes. The karyotypes were then compared and correlated to the parental origin. RESULTS: 3PN ICSI embryos displayed a significantly larger and more coordinated reduction from the assumed initial 3 sets of chromosomes than 3PN IVF embryos. CONCLUSION: The differences in the parental origin-and hence the number of centrioles-between the 3PN IVF and the 3PN ICSI zygotes are likely to be the cause of the differences in karyotypes.
Centrioles
;
Diploidy
;
Embryonic Structures*
;
Fertilization
;
Fertilization in Vitro
;
Fluorescence
;
Humans
;
Karyotype*
;
Parents*
;
Ploidies
;
Sperm Injections, Intracytoplasmic
;
Triploidy
;
Zygote*
10.Study on exogenous hormones inducing parthenocarpy fruit growth and development and quality of Siraitia grosvenorii.
Jie HUANG ; Dong-ping TU ; Xiao-jun MA ; Chang-ming MO ; Li-mei PAN ; Long-hua BAI ; Shi-xin FENG
China Journal of Chinese Materia Medica 2015;40(18):3567-3572
To explore the growth and development and analyze the quality of the parthenocarpy fruit induced by exogenous hormones of Siraitia grosvenorii. the horizontal and vertical diameter, volume of the fruit were respectively measured by morphological and the content of endogenous hormones were determined by ELISA. The size and seed and content of mogrosides of mature fruit were determined. The results showed that the fruit of parthenocarpy was seedless and its growth and development is similar to the diploid fruit by hand pollination and triploid fruit by hand pollination or hormones. But the absolute value of horizontal and vertical diameter, volume of parthenocarpy fruit was less than those of fruit by hand pollination, while triploid was opposite. The content of IAA, ABA and ratio of ABA/GA was obviously wavy. At 0-30 d the content of IAA and ABA of parthenocarpy fruit first reduced then increased, content of IAA and GA parthenocarpy fruit was higher than that of fruit by hand pollination. Mogrosides of parthenocarpy fruit was close to pollination fruit. Hormones can induce S. grosvenorii parthenocarpy to get seedless fruit and the fruit shape and size and quality is close to normal diploid fruit by hand pollination and better than triploid fruit by hormone or hand pollination.
Cucurbitaceae
;
chemistry
;
drug effects
;
genetics
;
growth & development
;
Diploidy
;
Fruit
;
chemistry
;
genetics
;
growth & development
;
Plant Growth Regulators
;
pharmacology

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