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1.
Reprod Fertil Dev ; 28(3): 302-9, 2016 Mar.
Article in English | MEDLINE | ID: mdl-25145348

ABSTRACT

During reprogramming, there is exchange of histone H1c and the oocyte-specific linker histone, and H1c may play a critically important role in the reprogramming process of somatic cell nuclear transfer (SCNT). The aim of the present study was to investigate the role of the H1c gene in SCNT reprogramming in Chinese swamp buffalo (Bubalus bubalis) using RNA interference (RNAi). Chinese swamp buffalo H1c gene sequences were obtained and H1c-RNAi vectors were designed, synthesised and then transfected into a buffalo fetal skin fibroblast cell line. Expression of H1c was determined by real-time polymerase chain reaction to examine the efficiency of vector interference. These cells were then used as a nuclear donor for SCNT so as to observe the further development of SCNT embryos. Inhibition of H1c gene expression in donor cells significantly improved the developmental speed of embryos from the 1-cell to 8-cell stage. Furthermore, compared with the control group, inhibition of H1c gene expression significantly reduced the blastocyst formation rate. It is concluded that linker histone H1c is very important in SCNT reprogramming in Chinese swamp buffalo. Correct expression of the H1c gene plays a significant role in preimplantation embryonic development in B. bubalis.


Subject(s)
Blastocyst/metabolism , Buffaloes/metabolism , Cellular Reprogramming Techniques/veterinary , Cellular Reprogramming , Histones/metabolism , Nuclear Transfer Techniques/veterinary , Reproductive Techniques, Assisted/veterinary , Animals , Buffaloes/embryology , Buffaloes/genetics , Cell Line , Embryo Culture Techniques/veterinary , Embryonic Development , Fertilization in Vitro/veterinary , Gene Expression Regulation, Developmental , Histones/genetics , In Vitro Oocyte Maturation Techniques/veterinary , RNA Interference , Time Factors , Transfection
2.
Yi Chuan ; 34(3): 342-7, 2012 Mar.
Article in Zh | MEDLINE | ID: mdl-22425953

ABSTRACT

Long-tailed macaque-pig interspecies somatic cell nuclear transfer (iSCNT) is beneficial to yield embryonic stem cells from iSCNT embryos with similar genetic background as human, which can be used as materials for medical and basic research. The primary objective of this study was to investigate the effects of concentrations and treatment duration of two histone deacetylase inhibitors-Trichostatin A (TSA) and Valproic acid (VPA) and two different embryo culture media (PZM-3 and HECM-10) on the in vitro development of iSCNT embryos. The results suggested that when PZM-3 was used as the embryo culture medium, the blastocyst rate of 10 nmol/L TSA treatment for 48 h was significantly higher than the control group (22.78% vs 9.86%, P< 0.05). However, neither in PZM-3 nor in HECM-10, 2-10 mmol/L VPA treatment did not increase the in vitro developmental potential of iSCNT embryos. It was concluded that TSA treatment could enhance the in vitro developmental potential of long-tailed macaque-pig iSCNT embryos.


Subject(s)
Hydroxamic Acids/pharmacology , Macaca fascicularis , Nuclear Transfer Techniques , Swine , Valproic Acid/pharmacology , Animals , Culture Media/pharmacology , Dose-Response Relationship, Drug , Embryonic Development/drug effects , Female , Male , Time Factors
3.
Front Immunol ; 12: 641562, 2021.
Article in English | MEDLINE | ID: mdl-33679805

ABSTRACT

Natural killer-like B (NKB) cells, which are newly identified immune subsets, reveal a critical immunoregulatory property in the eradication of microbial infection via the secretion of interleukin (IL)-18. For the first time, this study investigated the role of NKB cells in secreting IL-18 in the pathogenesis of periodontitis. In this study, NKB cells' percentage and IL-18 concentration in peripheral blood and periodontium in periodontitis patients was measured using flow cytometry and ELISA. The role of IL-18 in regulating periodontal inflammation was examined in a Porphyromonas gingivalis (P. gingivalis)-induced periodontitis murine model. Peripheral and periodontal-infiltrating CD3-CD19+NKp46+ NKB cells, which were the main source of IL-18, were elevated and correlated with attachment loss in periodontitis patients. In vitro IL-18 stimulation promoted proinflammatory cytokine production in periodontal ligament cells. P. gingivalis infection induced elevation of IL-18 receptor in periodontium in a periodontitis murine model. IL-18 neutralization not only suppressed P. gingivalis-induced alveolar bone resorption, but also inhibited recruitment of antigen-non-specific inflammatory cells into the periodontium, probably via dampening expressions of cytokines, chemokines, and matrix metalloproteinases. NKB cells secreting IL-18 appeared to be an important mediator in the inflammatory response following intraoral P. gingivalis infection. These findings might be relevant to the development of immunotherapies for periodontitis.


Subject(s)
B-Lymphocyte Subsets/immunology , Interleukin-18/immunology , Periodontitis/immunology , Adult , Animals , Bacteroidaceae Infections/immunology , Female , Humans , Inflammation/immunology , Male , Mice , Mice, Inbred C57BL , Middle Aged , Porphyromonas gingivalis
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