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1.
J Biomed Res ; : 1-15, 2024 Apr 25.
Article in English | MEDLINE | ID: mdl-38808565

ABSTRACT

Microtubule-severing enzymes (MTSEs) play important roles in mitosis and meiosis of the primitive organisms. However, no studies have assessed their roles in mammalian meiosis of females, whose abnormality accounts for over 80% of the cases of gamete-originated human reproductive disease. In the current study, we reported that katanin-like 2 (KL2) was the only MTSE concentrating at chromosomes. Furthermore, the knockdown of KL2 significantly reduced chromosome-based increase in the microtubule (MT) polymer, increased aberrant kinetochore-MT (K-MT) attachment, delayed meiosis, and severely affected normal fertility. Importantly, we demonstrated that the inhibition of aurora B, a key kinase for correcting aberrant K-MT attachment, eliminated KL2 from chromosomes completely. KL2 also interacted with phosphorylated eukaryotic elongation factor-2 kinase; they competed for chromosome binding. We also observed that the phosphorylated KL2 was localized at spindle poles, and that KL2 phosphorylation was regulated by extracellular signal-regulated kinase 1/2. In summary, our study reveals a novel function of MTSEs in mammalian female meiosis and demonstrates that multiple kinases coordinate to regulate the levels of KL2 at chromosomes.

2.
Phytomedicine ; 129: 155713, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38735196

ABSTRACT

BACKGROUND: Oligoasthenozoospermia is the most common type of semen abnormality in male infertile patients. Betaine (BET) has been proved to have pharmacological effects on improving semen quality. BET also belongs to endogenous physiological active substances in the testis. However, the physiological function of BET in rat testis and its pharmacological mechanism against oligoasthenozoospermia remain unclear. PURPOSE: This research aims to prove the therapeutic effect and potential mechanism of BET on oligoasthenozoospermia rat model induced by Tripterygium wilfordii glycosides (TWGs). METHODS: The oligoasthenozoospermia rat model was established by a continuous gavage of TWGs (60 mg/kg) for 28 days. Negative control group, oligoasthenozoospermia group, positive drug group (levocarnitine, 300 mg/kg), and 200 mg/kg, 400 mg/kg, and 800 mg/kg BET groups were created for exploring the therapeutic effect of BET on the oligoasthenozoospermia rat model. The therapeutic effect was evaluated by HE and TUNEL staining. Immunofluorescence assay of DNMT3A, PIWIL1, PRMT5, SETDB1, BHMT2, and METTL3, methylation capture sequencing, Pi-RNA sequencing, and molecular docking were used to elucidate potential pharmacological mechanisms. RESULTS: It is proved that BET can significantly restore testicular pathological damage induced by TWGs, which also can significantly reverse the apoptosis of spermatogenic cells. The spermatogenic cell protein expression levels of DNMT3A, PIWIL1, PRMT5, SETDB1, BHMT2, and METTL3 significantly decreased in oligoasthenozoospermia group. 400 mg/kg and 800 mg/kg BET groups can significantly increase expression level of the above-mentioned proteins. Methylation capture sequencing showed that BET can significantly increase the 5mC methylation level of Spata, Spag, and Specc spermatogenesis-related genes. Pi-RNA sequencing proved that the above-mentioned genes produce a large number of Pi-RNA under BET intervention. Pi-RNA can form complexes with PIWI proteins to participate in DNA methylation of target genes. Molecular docking indicated that BET may not directly act as substrate for methyltransferase and instead participates in DNA methylation by promoting the methionine cycle and increasing S-adenosylmethionine synthesis. CONCLUSION: BET has a significant therapeutic effect on oligoasthenozoospermia rat model induced by TWPs. The mechanism mainly involves that BET can increase the methylation level of Spata, Specc, and Spag target genes through the PIWI/Pi-RNA pathway and up-regulation of methyltransferases (including DNA methyltransferases and histone methyltransferases).


Subject(s)
Apoptosis , Betaine , DNA Methylation , Disease Models, Animal , Oligospermia , Rats, Sprague-Dawley , Tripterygium , Male , Animals , Apoptosis/drug effects , DNA Methylation/drug effects , Betaine/pharmacology , Rats , Oligospermia/drug therapy , Tripterygium/chemistry , Asthenozoospermia/drug therapy , Up-Regulation/drug effects , Testis/drug effects , Molecular Docking Simulation , Spermatogenesis/drug effects , Methyltransferases/metabolism , Spermatozoa/drug effects
3.
Dysphagia ; 2024 Feb 06.
Article in English | MEDLINE | ID: mdl-38319366

ABSTRACT

Airway invasion is common in patients with Parkinson's disease (PD) and can cause serious complications. However, a PD-related dysphagic pattern has not been clearly elucidated. In this study, 53 patients with early to moderate PD were enrolled to undergo a videofluoroscopic study of swallowing evaluation (VFSS) and a battery of neuropsychological assessments. A set of VFSS variables (three visuoperceptual, nine temporal, and six spatial) were measured. The main effects of bolus viscosity and volume on airway invasion were calculated. Statistical analyses were performed to determine key kinematic factors of airway invasion for swallowing each bolus type. Airway invasion frequency was significantly higher for liquid boluses (liquid vs. pudding P < 0.001; liquid vs. honey P = 0.006). Laryngeal vestibule closure reaction time (LVCrt) was the key kinematic factor of airway invasion for 3 ml liquid swallow (P = 0.040), anterior displacement of hyoid bone was the key kinematic factor for both 5 ml and 10 ml liquid swallows (P = 0.010, 0.034, respectively). Male sex and advanced Hoehn and Yahr stage were significantly related to reduced anterior displacement of hyoid bone. These results reveal the dysphagic pattern related to PD, demonstrating that prolonged LVCrt and reduced anterior displacement of hyoid bone are two crucial kinematic factors contributing to airway invasion during the liquid swallow. In addition, hyoid bone dysfunction was correlated with disease severity and male sex. Our findings warrant further investigation of the pathophysiological mechanism of dysphagia in PD and would guide clinical intervention.

4.
J Pharm Sci ; 113(6): 1572-1579, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38237668

ABSTRACT

Poor stability and difficult uptake of natural polysaccharides have been the main problems in their application. The purpose of this study was to optimize the preparation conditions of Polygonatum cyrtonema Hua polysaccharides liposomes (PCPL) and to investigate the immune enhancement activity of PCPL in vitro and in vivo, with a view to discovering new ways of natural polysaccharide application. The optimal preparation conditions of PCPL were as follows: the adding amount of Tween 80 of 0.5 %, the ultrasound time of 2 min and the ultrasound times of once. Under these conditions, the entrapment efficiency, drug loading rate and particle size of PCPL were 38.033 %±0.050, 2.172 %±0.003 and 146 nm, which indicated that PCPL with small particle size could be prepared by the reverse-phase evaporation method. Furthermore, PCPL promoted proliferation, phagocytosis, and secretion of nitric oxide and related cytokines in RAW264.7 cells. Moreover, PCPL improved spleen and thymus indices, increased the number or proportion of red blood cells, platelets, and lymphocytes in the blood, and ameliorated spleen and thymus atrophy in immunosuppressed mice. This study provides a new idea for applying Polygonatum cyrtonema Hua polysaccharides (PCP) and references for studying other polysaccharides.


Subject(s)
Liposomes , Phagocytosis , Polygonatum , Polysaccharides , Animals , Mice , Polysaccharides/chemistry , Polysaccharides/pharmacology , Polygonatum/chemistry , RAW 264.7 Cells , Phagocytosis/drug effects , Particle Size , Spleen/drug effects , Spleen/immunology , Nitric Oxide/metabolism , Thymus Gland/drug effects , Thymus Gland/immunology , Cell Proliferation/drug effects , Cytokines/metabolism , Male
5.
Medicine (Baltimore) ; 103(2): e35929, 2024 Jan 12.
Article in English | MEDLINE | ID: mdl-38215108

ABSTRACT

TACE has become one of the main methods for the treatment of liver cancer. The study aimed to investigate the effects of preoperative interview and prospective nursing in patients with hepatic carcinoma undergoing transcatheter chemoembolization (TACE). Eighty-six patients with hepatocellular carcinoma who underwent TACE intervention treatment at our hospital between 2020 and 2023 were selected and randomly assigned to 2 groups using computerized randomization. The control group (n = 43) received routine nursing care, while the study group (n = 43) received preoperative interviews in combination with prospective nursing during the procedure. The patients' heart rate, mean arterial pressure, and blood pressure variations were recorded, along with their mood changes after intervention. The postoperative pain and satisfaction levels were compared between the 2 groups of patients, and the incidence of postoperative complications was observed. The heart rate, systolic blood pressure, and diastolic blood pressure of the 2 groups of patients were compared 1 day before the operation (P > .05). Compared to 1 day before the operation, there was no significant change for the study group at 10 minutes after entering the room. However, the control group showed an increase. Both groups showed an increase in heart rate, systolic blood pressure, and diastolic blood pressure after the operation, with the study group having lower values than the control group (P < .05). The levels of tension, fatigue, anxiety, energy, anger, depression, self-esteem, and POMS index were compared between the 2 groups before intervention (P > .05). After intervention, there were significant differences between the 2 groups(P < .05). Immediately after the operation, the NRS scores of the 2 groups of patients were compared (P > .05). Compared to the control group, the study group showed a decrease in NRS scores at 12, 24, and 48 hours after the operation (P < .05). The nursing satisfaction rate of the study group patients was 97.67% (42/43), which was higher than the nursing satisfaction rate of the control group of 76.74% (33/43) (P < .05). Compared to routine nursing, preoperative visits and prospective nursing interventions can effectively alleviate patients' psychological stress reactions, relieve pain, reduce the incidence of complications, and improve patients' satisfaction with nursing care.


Subject(s)
Carcinoma, Hepatocellular , Chemoembolization, Therapeutic , Liver Neoplasms , Humans , Carcinoma, Hepatocellular/surgery , Carcinoma, Hepatocellular/pathology , Chemoembolization, Therapeutic/methods , Liver Neoplasms/surgery , Liver Neoplasms/pathology , Pain, Postoperative/etiology , Pain, Postoperative/therapy , Prospective Studies
6.
Zygote ; 32(1): 21-27, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38047349

ABSTRACT

Our previous studies have suggested that spastin, which aggregates on spindle microtubules in oocytes, may promote the assembly of mouse oocyte spindles by cutting microtubules. This action may be related to CRMP5, as knocking down CRMP5 results in reduced spindle microtubule density and maturation defects in oocytes. In this study, we found that, after knocking down CRMP5 in oocytes, spastin distribution shifted from the spindle to the spindle poles and errors in microtubule-kinetochore attachment appeared in oocyte spindles. However, CRMP5 did not interact with the other two microtubule-severing proteins, katanin-like-1 (KATNAL1) and fidgetin-like-1 (FIGNL1), which aggregate at the spindle poles. We speculate that, in oocytes, due to the reduction of spastin distribution on chromosomes after knocking down CRMP5, microtubule-kinetochore errors cannot be corrected through severing, resulting in meiotic division abnormalities and maturation defects in oocytes. This finding provides new insights into the regulatory mechanisms of spastin in oocytes and important opportunities for the study of meiotic division mechanisms.


Subject(s)
Kinetochores , Spindle Apparatus , Mice , Animals , Kinetochores/metabolism , Spastin/genetics , Spastin/metabolism , Spindle Apparatus/physiology , Microtubules/metabolism , Meiosis , Oocytes/physiology
7.
J Appl Genet ; 65(1): 31-46, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38133708

ABSTRACT

Justicia procumbens L. is a traditional medicinal plant that is widely distributed in China. However, little is known about the genetic diversity and evolution of this genus, and no genomic studies have been carried out on J. procumbens previously. In this study, we aimed to assemble and annotate the first complete chloroplast genome (cpDNA) of J. procumbens and compare it with all previously published cpDNAs within the tribe Justicieae. Genome structure and comparative and phylogenetic analyses were performed. The 150,454 bp-long J. procumbens cpDNA has a circular and quadripartite structure consisting of a large single copy, a small single copy, and two inverted repeat regions. It contains 133 genes, of which 88 are protein-coding genes, 37 are tRNA genes, and eight are rRNA genes. Twenty-four simple sequence repeats (SSRs) and 81 repeat sequences were identified. Comparative analyses with other Justicieae species revealed that the non-coding regions of J. procumbens cpDNA showed greater variation than did the coding regions. Moreover, phylogenetic analysis based on 14 cpDNA sequences from Justicieae species showed that J. procumbens and J. flava were most closely related. This study provides valuable genetic information to support further research on the genetic diversity and evolutionary development of the tribe Justicieae.


Subject(s)
Genome, Chloroplast , Justicia , Phylogeny , Justicia/genetics , Genomics , Repetitive Sequences, Nucleic Acid
8.
Chem Biodivers ; 20(11): e202300980, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37831331

ABSTRACT

Dendrobium huoshanense is an important Traditional Chinese medicine that thickens the stomach and intestines. Its active ingredient Dendrobium huoshanense polysaccharide (DHP), was revealed to relieve the symptoms of liver injury. However, its mechanism of action remains poorly understood. This study aimed to investigate the mechanism of DHP in protecting the liver. The effects of DHP on lipid levels, liver function, and intestinal barrier function were investigated in mice with high-fat diet-induced liver damage. Changes in the gut flora and their metabolites were analyzed using 16S rRNA sequencing and metabolomics. The results showed that DHP reduced lipid levels, liver injury, and intestinal permeability. DHP altered the intestinal flora structure and increased the relative abundance of Bifidobacterium animalis and Clostridium disporicum. Furthermore, fecal metabolomics revealed that DHP altered fecal metabolites and significantly increased levels of gut-derived metabolites, spermidine, and indole, which have been reported to inhibit liver injury and improve lipid metabolism and the intestinal barrier. Correlation analysis showed that spermidine and indole levels were significantly negatively correlated with liver injury-related parameters and positively correlated with the intestinal species B. animalis enriched by DHP. Overall, this study confirmed that DHP prevented liver injury by regulating intestinal microbiota dysbiosis and fecal metabolites.


Subject(s)
Chemical and Drug Induced Liver Injury, Chronic , Dendrobium , Animals , Mice , Dendrobium/chemistry , Diet, High-Fat/adverse effects , RNA, Ribosomal, 16S , Spermidine , Polysaccharides/pharmacology , Polysaccharides/chemistry , Indoles , Lipids
9.
Article in Chinese | MEDLINE | ID: mdl-37138397

ABSTRACT

Objective:To analyze the risk factors of recurrence and canceration for premalignant vocal fold lesions after surgery, and to provide a reasonable basis for preoperative evaluation and postoperative follow-up. Methods:This study retrospective analyzed the relationship between clinicopathological factors and clinical outcome(recurrence, canceration, recurrence-free survival, and canceration-free survival) in 148 patients undergoing surgical treatment in Chongqing General Hospital from 2014 to 2017. Results:The five-year overall recurrence rate was 14.86% and the overall recurrence rate was 8.78%. Univariate analysis showed that smoking index, laryngopharyngeal reflux and lesion range were significantly associated with recurrence(P<0.05), and smoking index and lesion range were significantly associated with canceration(P<0.05). Multivariate logistic regression analysis showed that smoking index ≥600 and laryngopharyngeal reflux were independent risk factors for recurrence(P<0.05), and smoking index ≥600 and lesion range ≥1/2 vocal cord were independent risk factors for canceration(P<0.05). The mean carcinogenesis interval for the postoperative smoking cessation group was significantly longer(P<0.05). Conclusion:Excessive smoking, laryngopharyngeal reflux and a wide range of lesions may be related to postoperative recurrence or malignant progression of precancerous lesions in the vocal cord, and further large-scale multi-center prospective randomized controlled studies are needed to clarify the effects of the above factors on recurrence and malignant changes in the future.


Subject(s)
Laryngopharyngeal Reflux , Precancerous Conditions , Humans , Vocal Cords/pathology , Retrospective Studies , Laryngopharyngeal Reflux/complications , Prospective Studies , Precancerous Conditions/complications , Precancerous Conditions/pathology , Risk Factors
10.
Zygote ; 31(2): 140-148, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36533678

ABSTRACT

The low maturation rate of oocytes is an important reason for female infertility and failure of assisted pregnancy. The germinal vesicle breakdown (GVBD) is a landmark event of oocyte maturation. In our previous studies, we found that zona pellucida 3 (ZP3) was strongly concentrated in the nuclear region of germinal vesicle (GV) oocytes and interacted with aryl hydrocarbon receptor-interacting protein-like 1 (AIPL1) and lamin A to promote GVBD. In the current study, we found that lamin A is mainly concentrated in the nuclear membrane. When ZP3 is knocked down, lamin A will be partially transferred to the nucleus of oocytes. The prelamin A is increased in both the nuclear membrane and nucleus, while phosphorylated lamin A (p-lamin A) is significantly reduced. AIPL1 was also proved to accumulate in the GV region of oocytes, and ZP3 deletion can significantly inhibit the aggregation of AIPL1 in the nuclear region. Similar to ZP3 knockdown, the absence of AIPL1 resulted in a decrease in the occurrence of GVBD, an increase in the amount of prelamin A, and a significant decrease in p-lamin A in oocytes developed in vitro. Finally, we propose the hypothesis that ZP3 can stabilize farnesylated prelamin A on the nuclear membrane of AIPL1, and promote its further processing into mature lamin A, therefore promoting the occurrence of GVBD. This study may be an important supplement for the mechanism of oocyte meiotic resumption and provide new diagnostic targets and treatment clues for infertility patients with oocyte maturation disorder.


Subject(s)
Lamin Type A , Zona Pellucida , Pregnancy , Female , Mice , Animals , Zona Pellucida/metabolism , Lamin Type A/genetics , Lamin Type A/metabolism , Nuclear Envelope/metabolism , Oocytes/metabolism , Meiosis , Adaptor Proteins, Signal Transducing , Zona Pellucida Glycoproteins/genetics , Zona Pellucida Glycoproteins/metabolism
11.
Theriogenology ; 192: 89-96, 2022 Oct 15.
Article in English | MEDLINE | ID: mdl-36084388

ABSTRACT

An optimal lipid droplet (LD) content is essential for successful mammalian embryonic development. Salidroside (SAL) is a traditional Chinese medicine and one of the important active components of the Rhodiola plant. SAL possesses antioxidative, anti-aging, and cardiovascular properties. Here, we studied the effects of SAL on in vitro maturation (IVM) of porcine oocytes and the subsequent embryonic development after parthenogenetic activation (PA). We found that 100 µM of SAL had no effect on the extrusion rate of the first polar body of porcine oocytes but significantly improved the subsequent blastocyst formation rate and embryo quality. Our study further revealed that SAL treatment altered the morphology, increased the lipid content in oocytes, increased mitochondrial number. Further analysis revealed that SAL upregulated the expression of genes related to lipid metabolism (FASN, FADS1, HSL, and CPT1a) and the mitochondria function-related genes (PGC-1α). These results suggest that SAL supplementation enhances oocyte maturation and subsequent embryonic development by promoting lipid metabolism, providing the necessary energy for the aforementioned processes.


Subject(s)
In Vitro Oocyte Maturation Techniques , Lipid Metabolism , Animals , Blastocyst/physiology , Embryonic Development , Glucosides , In Vitro Oocyte Maturation Techniques/methods , In Vitro Oocyte Maturation Techniques/veterinary , Lipids/pharmacology , Mammals , Oocytes/physiology , Phenols , Reactive Oxygen Species/metabolism , Swine
12.
Zygote ; 30(6): 872-881, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36148793

ABSTRACT

Microtubule-severing proteins (MTSPs) play important roles in mitosis and interphase. However, to the best of our knowledge, no previous studies have evaluated the role of MTSPs in female meiosis in mammals. It was found that FIGNL1, a member of MTSPs, was predominantly expressed in mouse oocytes and distributed at the spindle poles during meiosis in the present study. FIGNL1 was co-localized and interacted with γ-tubulin, an important component of the microtubule tissue centre (MTOC). Fignl1 knockdown by specific small interfering RNA caused spindle defects characterized by an abnormal length:width ratio and decreased microtubule density, which consequently led to aberrant chromosome arrangement, oocyte maturation and fertilization obstacles. In conclusion, the present results suggested that FIGNL1 may be an essential factor in oocyte maturation by influencing the meiosis process via the formation of spindles.


Subject(s)
Meiosis , Spindle Apparatus , Female , Mice , Animals , Spindle Apparatus/metabolism , Oocytes/metabolism , Microtubules/metabolism , Tubulin/genetics , Tubulin/metabolism , Mammals
13.
J Biomed Res ; 36(4): 269-279, 2022 Jul 10.
Article in English | MEDLINE | ID: mdl-35965436

ABSTRACT

Microtubule-severing proteins (MTSPs), are a family of proteins which use adenosine triphosphate to sever microtubules. MTSPs have been shown to play an important role in multiple microtubule-involved cellular processes. One member of this family, fidgetin ( FIGN), is also involved in male fertility; however, no studies have explored its roles in female fertility. In this study, we found mouse fidgetin is rich within oocyte zona pellucida (ZP) and is the only MTSP member to do so. Fidgetin also appears to interact with all three ZP proteins. These findings prompted us to propose that fidgetin might prevent polyspermy. Results from in vitro maturation oocytes analysis showed that fidgetin knockdown did cause polyspermy. We then deleted all three fidgetin isoforms with CRISPR/Cas9 technologies; however, female mice remained healthy and with normal fertility. Of all mouse MTSPs, only the mRNA level of fidgetin-like 1 ( FIGNL1) significantly increased. Therefore, we assert that fidgetin-like 1 compensates fidgetin's roles in fidgetin knockout female mice.

14.
J Exp Psychol Gen ; 151(11): 2756-2770, 2022 Nov.
Article in English | MEDLINE | ID: mdl-35951375

ABSTRACT

Many resource allocation tasks involve the assignment of multiple units of a resource (e.g., money, time, labor) to multiple targets (e.g., stocks, expenditure categories, projects to be carried out). The decision-maker can either focus on the individual targets and decide how many resource units each target should receive (allocation-by-target), or focus on the individual resource units and decide which target each unit should be assigned to (allocation-by-unit). We suggest that the two allocation procedures might result in different outcomes. Specifically, we predict that the allocation-by-unit procedure leads to more variety-seeking than the allocation-by-target procedure. Nine experiments (N = 4,152) provided evidence consistent with this procedure dependence hypothesis. We further demonstrate that the effect occurs because the allocation-by-target procedure encourages people to consider the differences between targets whereas the allocation-by-unit procedure induces people to focus more on each undifferentiated unit. As a result, allocation-by-target, relative to allocation-by-unit, leads to a lower variety-seeking, which manifests as a more concentrated distribution of the resource units across the targets. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Subject(s)
Resource Allocation , Humans , Resource Allocation/methods
15.
Front Pharmacol ; 13: 920823, 2022.
Article in English | MEDLINE | ID: mdl-35903345

ABSTRACT

Dendrobium huoshanense, a traditional medicinal and food homologous plant, belongs to the family Orchidaceae and has a long history of medicinal use. It is reported that the stem of D. huoshanense has a variety of bioactive ingredients such as polysaccharides, flavonoids, sesquiterpenes, phenols, etc. These bioactive ingredients make D. huoshanense remarkable for its pharmacological effects on anti-tumor, immunomodulation, hepatoprotective, antioxidant, and anticataract activities. In recent years, its rich pharmacological activities have attracted extensive attention. However, there is no systematic review focusing on the chemical compositions and pharmacological effects of D. huoshanense. Therefore, the present review aims to summarize current research on the chemical compositions and pharmacological activities of D. huoshanense. This study provides valuable references and promising ideas for further investigations of D. huoshanense.

16.
Mol Reprod Dev ; 89(8): 325-336, 2022 08.
Article in English | MEDLINE | ID: mdl-35734898

ABSTRACT

Mammalian sperm and oocytes are haploid cells that carry parental genetic and epigenetic information for their progeny. The cytoplasm of oocytes is also capable of reprograming somatic cells to establish totipotency through somatic cell nuclear transfer (SCNT). However, epigenetic barriers seriously counteract SCNT reprogramming. Here, we found that sperm-derived RNAs elevated chromatin accessibility of nuclear donor cells concurrent with the appearance of increased RNA amount and decreased cell proliferation, instead of activating DNA damage response. Additionally, tri-methylation of lysine 9 on histone H3 (H3K9me3) and the H3K9 methyltransferase SUV39H2 were significantly downregulated by the sperm-derived RNA treatment. Our findings thus raise a fascinating possibility that sperm RNA-induced R-loops may activate gene expression and chromatin structure, thereby promoting SCNT reprogramming.


Subject(s)
R-Loop Structures , Semen , Animals , Cellular Reprogramming/genetics , Chromatin/metabolism , Embryo, Mammalian/metabolism , Male , Mammals/genetics , Nuclear Transfer Techniques , RNA/genetics , RNA/metabolism , Spermatozoa
17.
Oxid Med Cell Longev ; 2022: 7113793, 2022.
Article in English | MEDLINE | ID: mdl-35237383

ABSTRACT

α-Ketoglutarate (α-KG) is a metabolite in the tricarboxylic acid cycle. It has a strong antioxidant function and can effectively prevent oxidative damage. Previous studies have shown that α-KG exists in porcine follicles, and its content gradually increases as the follicles grow and mature. However, the potential mechanism of supplementation of α-KG on porcine oocytes during in vitro maturation (IVM) has not yet been reported. The purpose of this study was to explore the effect of α-KG on the early embryonic development of pigs and the mechanisms underlying these effects. We found that α-KG can enhance the development of early pig embryos. Adding 20 µM α-KG to the in vitro culture medium significantly increased the rate of blastocyst formation and the total cell number. Compared with to that of the control group, apoptosis in blastocysts of the supplement group was significantly reduced. α-KG reduced the production of reactive oxygen species and glutathione levels in cells. α-KG not only improved the activity of mitochondria but also inhibited the occurrence of apoptosis. After supplementation with α-KG, pig embryo pluripotency-related genes (OCT4, NANOG, and SOX2) and antiapoptotic genes (Bcl2) were upregulated. In terms of mechanism, α-KG activates the Nrf2/ARE signaling pathway to regulate the expression of antioxidant-related targets, thus combating oxidative stress during the in vitro culture of oocytes. Activated Nrf2 promotes the transcription of Bcl2 genes and inhibits cell apoptosis. These results indicate that α-KG supplements have a beneficial effect on IVM by regulating oxidative stress during the IVM of porcine oocytes and can be used as a potential antioxidant for IVM of porcine oocytes.


Subject(s)
Antioxidants/pharmacology , Embryonic Development/drug effects , Ketoglutaric Acids/pharmacology , Meiosis/drug effects , NF-E2-Related Factor 2/metabolism , Oocytes/metabolism , Oogenesis/drug effects , Oxidative Stress/drug effects , Signal Transduction/drug effects , Animals , Apoptosis/drug effects , Blastocyst/metabolism , Culture Media/chemistry , Dietary Supplements , Embryo, Mammalian/drug effects , Embryo, Mammalian/metabolism , Female , Glutathione/metabolism , In Vitro Oocyte Maturation Techniques/methods , Mitochondria/metabolism , Oocytes/drug effects , Pregnancy , Reactive Oxygen Species/metabolism , Swine
18.
Int J Mol Sci ; 23(4)2022 Feb 13.
Article in English | MEDLINE | ID: mdl-35216182

ABSTRACT

The double homeobox (Dux) gene, encoding a double homeobox transcription factor, is one of the key drivers of totipotency in mice. Recent studies showed Dux was temporally expressed at the 2-cell stage and acted as a transcriptional activator during zygotic genome activation (ZGA) in embryos. A similar activation occurs in mouse embryonic stem cells, giving rise to 2-cell-like cells (2CLCs). Though the molecular mechanism underlying this expanded 2CLC potency caused by Dux activation has been partially revealed, the regulation mechanisms controlling Dux expression remain elusive. Here, we discuss the latest advancements in the multiple levels of regulation of Dux expression, as well as Dux function in 2CLCs transition, aiming to provide a theoretical framework for understanding the mechanisms that regulate totipotency.


Subject(s)
Genes, Homeobox/genetics , Transcription Factors/genetics , Animals , Embryonic Development/genetics , Gene Expression Regulation, Developmental/genetics , Genome/genetics , Homeodomain Proteins , Humans , Zygote/metabolism
19.
Asia Pac J Clin Oncol ; 18(1): 70-75, 2022 Feb.
Article in English | MEDLINE | ID: mdl-33629514

ABSTRACT

AIM: Metabolic syndrome (MS) is tightly associated with the oncogenesis and prognosis of endometrioid adenocarcinoma, but the underlying mechanism is unclear. Here, we studied the relation between the expression status of WW domain-containing oxidoreductase (WWOX) and the clinicopathological features of endometrioid adenocarcinoma patients with MS. METHODS: Fifty-seven samples of endometrial adenocarcinoma were chosen for detection of expression level of WWOX. Overall survival (OS) time of these patients was analyzed by univariate and multivariate analysis. Survival analysis of patients with different WWOX expression levels from the Cancer Genome Atlas (TCGA) database was also performed. RESULTS: The WWOX expression is significantly higher in MS group than that in non-MS group (36.4% vs 65.7%, P = .03). WWOX was closely related to MS (P = .03) and muscle invasion of tumor cells (P = .04), but age, tumor grade, status of lymphatic metastasis, and FIGO (International Federation of Gynecology and Obstetrics) stage were not significantly different between the two WWOX expression status. Univariate analysis revealed that lymphatic metastasis (P = .023) and lower stage (P = .006) are significantly associated with OS. Multivariate analysis demonstrated that stage was an independent prognostic factor for OS (hazard ratio = 0.197; 95% CI, 0.043-0.896). Downregulation of WWOX was statistically associated with OS in patients from TCGA database (P = .04). CONCLUSION: WWOX may play an important role in the progression of endometrial cancer with MS.


Subject(s)
Carcinoma, Endometrioid , Endometrial Neoplasms , Metabolic Syndrome , Tumor Suppressor Proteins , WW Domain-Containing Oxidoreductase , Female , Humans , Metabolic Syndrome/genetics , Prognosis
20.
Zygote ; 30(1): 80-91, 2022 Feb.
Article in English | MEDLINE | ID: mdl-34034836

ABSTRACT

Microtubule-severing protein (MTSP) is critical for the survival of both mitotic and postmitotic cells. However, the study of MTSP during meiosis of mammalian oocytes has not been reported. We found that spastin, a member of the MTSP family, was highly expressed in oocytes and aggregated in spindle microtubules. After knocking down spastin by specific siRNA, the spindle microtubule density of meiotic oocytes decreased significantly. When the oocytes were cultured in vitro, the oocytes lacking spastin showed an obvious maturation disorder. Considering the microtubule-severing activity of spastin, we speculate that spastin on spindles may increase the number of microtubule broken ends by severing the microtubules, therefore playing a nucleating role, promoting spindle assembly and ensuring normal meiosis. In addition, we found the colocalization and interaction of collapsin response mediator protein 5 (CRMP5) and spastin in oocytes. CRMP5 can provide structural support and promote microtubule aggregation, creating transportation routes, and can interact with spastin in the microtubule activity of nerve cells (30). Knocking down CRMP5 may lead to spindle abnormalities and developmental disorders in oocytes. Overexpression of spastin may reverse the abnormal phenotype caused by the deletion of CRMP5. In summary, our data support a model in which the interaction between spastin and CRMP5 promotes the assembly of spindle microtubules in oocytes by controlling microtubule dynamics, therefore ensuring normal meiosis.


Subject(s)
Hydrolases/metabolism , Microtubule-Associated Proteins/metabolism , Microtubules , Oocytes , Spastin , Animals , Meiosis , Mice , Microtubules/metabolism , Oocytes/metabolism , RNA, Small Interfering/genetics , Spastin/metabolism , Spindle Apparatus/metabolism
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