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Glutamatergic CYLD deletion leads to aberrant excitatory activity in the basolateral amygdala:association with enhanced cued fear expression

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Neural Regeneration Research 2025年第11期20卷 3259-3272页

作者： Huidong Li Faqin Li Zhaoyi Chen Erwen Wu Xiaoxi Dai Danni Li Haojie An Shiyi Zeng Chunyan Wang Li Yang Cheng Long School of Life Sciences South China Normal UniversityGuangzhouGuangdong ProvinceChina School of Life Sciences Guangzhou UniversityGuangzhouGuangdong ProvinceChina Guangdong Provincial key laboratory of Major obstetric Diseases The Third Affiliated Hospital of Guangzhou Medical UniversityGuangzhouGuangdong ProvinceChina South China Normal University-Panyu Central Hospital Joint laboratory of Translational Medical Research Panyu Central HospitalGuangzhouGuangdong ProvinceChina

Neuronal activity,synaptic transmission,and molecular changes in the basolateral amygdala play critical roles in fear ***(CYLD)is a deubiquitinase that negatively regulates the nuclear factor kappa-B *** is well studied in non-neuronal cells,yet underinvestigated in the brain,where it is highly *** studies have shown involvement of CYLD in the remodeling of glutamatergic synapses,neuroinflammation,fear memory,and anxiety-and autism-like ***,the precise role of CYLD in glutamatergic neurons is largely ***,we first proposed involvement of CYLD in cued fear *** next constructed transgenic model mice with specific deletion of Cyld from glutamatergic *** results show that glutamatergic CYLD deficiency exaggerated the expression of cued fear in only male ***,loss of CYLD in glutamatergic neurons resulted in enhanced neuronal activation,impaired excitatory synaptic transmission,and altered levels of glutamate receptors accompanied by over-activation of microglia in the basolateral amygdala of male ***,our study suggests a critical role of glutamatergic CYLD in maintaining normal neuronal,synaptic,and microglial *** may contribute,at least in part,to cued fear expression.

关键词： basolateral amygdala cued fear expression cylindromatosis deubiquitinase glutamate receptor 1 glutamatergic neuron microglial activation N-methyl-D-aspartate receptor 1 neuronal activation synaptic transmission

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A conserved region in the 3' untranslated region of the human LIMK1 gene is critical for proper expression of LIMK1 at the post-transcriptional level

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Neuroscience Bulletin 2013年第3期29卷 348-354页

作者： Guang-Fei Deng Shu-Jing Liu Xun-Sha Sun Wei-Wen Sun Qi-Hua Zhao Wei-Ping Liao Yong-Hong Yi Yue-Sheng Long key laboratory of Neurogenetics and Channelopathies of Guangdong Province and the Ministry of Education of China Institute of Neuroscience and the Second Affiliated Hospital of Guangzhou Medical University key laboratory for Major obstetric Diseases of Guangdong Province and key laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes Guangzhou Medical University

LIM kinase 1 （LIMK1）, a cytosolic serine/threonine kinase, regulates actin filament dynamics and reorganization and is involved in neuronal development and brain function. Abnormal expression of LIMK1 is associated with several neurological disorders. In this study, we performed a conservation analysis using Vector NTI （8.0） software. The dualluciferase reporter assay and real-time quantitative RT-PCR were used to assess the protein and mRNA levels of the reporter gene, respectively. We found that a region ranging from nt ＋884 to ＋966 in the human LIMK1 3＇ untranslated region （UTR） was highly conserved in the mouse Limkl 3＇ UTR and formed a structure containing several loops and stems. Luciferase assay showed that the relative luciferase activity of the mutated construct with the conserved region deleted, pGL4-hLIMK1-3U-M, in SH-SY5Y and HEK-293 cells was only -60% of that of the wild-type construct pGL4-hLIMK1-3U, indicating that the conserved region is critical for the reporter gene expression. Real-time quantitative RT-PCR analysis demonstrated that the relative Luc2 mRNA levels in SH-SY5Y and HEK293 cells transfected with pGL4-hLIMK1-3U-M decreased to50% of that in cells transfected with pGL4-hLIMK1- 3U, suggesting an important role of the conserved region in maintaining Luc2 mRNA stability. Our study suggests that the conserved region in the LIMK1 3＇ UTR is involved in regulating LIMK1 expression at the post-transcriptional level, which may help reveal the mechanism underlying the regulation of LIMK1 expression in the central nervous system and explore the relationship between the 3＇-UTR mutant and neuroloqical disorders.

关键词： LIM kinase 1 region post-transcriptsecondary structurethree-prime untranslatedonal regulation RNA

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Regulations of m^(6)A and other RNA modifications and their roles in cancer

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Frontiers of Medicine 2024年第4期18卷 622-648页

作者： Xin-Hui Chen Kun-Xiong Guo Jing Li Shu-Hui Xu Huifang Zhu Guang-Rong Yan Biomedicine Research Center Guangdong Provincial Key Laboratory of Major Obstetric DiseaseGuangdong Provincial Clinical Research Center for Obstetrics and GynecologyState Key Laboratory of Respiratory Diseasethe Third Affiliated Hospital of Guangzhou Medical UniversityGuangzhou510150China

RNA modification is an essential component of the epitranscriptome,regulating RNA metabolism and cellular *** types of RNA modifications have been identified to date;they include N^(6)-methyladenosine(m^(6)A),N^(1)-methyladenosine(m1A),5-methylcytosine(m5C),N^(7)-methylguanosine(m^(7)G),N^(6),2′-O-dimethyladenosine(m^(6)Am),N4-acetylcytidine(ac^(4)C),*** modifications,mediated by regulators including writers,erasers,and readers,are associated with carcinogenesis,tumor microenvironment,metabolic reprogramming,immunosuppression,immunotherapy,chemotherapy,etc.A novel perspective indicates that regulatory subunits and post-translational modifications(PTMs)are involved in the regulation of writer,eraser,and reader functions in mediating RNA modifications,tumorigenesis,and anticancer *** this review,we summarize the advances made in the knowledge of different RNA modifications(especially m^(6)A)and focus on RNA modification regulators with functions modulated by a series of factors in cancer,including regulatory subunits(proteins,noncoding RNA or peptides encoded by long noncoding RNA)and PTMs(acetylation,SUMOylation,lactylation,phosphorylation,etc.).We also delineate the relationship between RNA modification regulator functions and carcinogenesis or cancer ***,inhibitors that target RNA modification regulators for anticancer therapy and their synergistic effect combined with immunotherapy or chemotherapy are discussed.

关键词： RNA modification writers erasers readers regulatory subunits PTMs cancer

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Journal of Reproduction and Contraception 2013年第2期24卷 95-100页

作者： Yu-ling HUANG Hong-zhi DU Xiang-jin KANG Yong FAN Assisted Reproductive Center of the Third Affiliated Hospital of Guangzhou Medical University Key Laboratory for Major Obstetric Diseases of Guangdong Province

Objective To find the possible factors predicting fertilization failure of in vitro fertilization-embryo transfer （1VF-ET）. Methods The IVF-ET patients with complete fertilization failure （experimental group, n =32） were analyzed retrospectively. The patients whose oocytes retrieved at the same day and cultured on the same incubators with ≥ 50% fertilization rates were matched as the control （n=56）. Results The infertility duration, superovulation days, the rates of primary case, progesterone （P） level 〉3.12 nmol/L rate and rate of severe abnormal sperm （abnormal sperm rate 〉95%） in experimental group were significantly higher than those in the control （6.4 ±3.1 years, 12.6 ±2.2 d, 56%, 43%, 43% vs 4.6±2.9years, 11.6 ±% 1.3 d, 33%, 23%, 23%, respectively, P〈0.05）. Conclusion We should pay attention to these patients with primary infertility, longer infertility duration and superovulation days （〉6.4 years and 〉12.6 d） and having increased level of P on hCG injection day （〉3.12 nmol/L）, abnormal sperm rate 〉95% at the same time. They should be included in such patients at high risk of fertilization failure.

关键词： in vitro fertilization-embryo transfer (IVF-ET) complete fertilization failure proportion of sperm abnormality

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Addressing challenges in the clinical applications associated with CRISPR/Cas9 technology and ethical questions to prevent its misuse

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Protein & Cell 2017年第11期8卷 791-795页

作者： Xiang Jin Kang Chiong Isabella Noelle Caparas Boon Seng Soh Yong Fan key laboratory for Major obstetric Diseases of Guangdong Province Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes Center for Reproductive Medicine The Third Affiliated Hospital of Guangzhou Medical University Guangzhou 510150 China Disease Modeling and Therapeutics laboratory ASTAR Institute of Molecular and Cell Biology 61 Biopolis Drive Proteos Singapore 138673 Singapore

The recently developed RNA-guided clustered regularly interspaced short palindromic repeat （CRISPR）/CRISPR-associated 9 （Cas9） nuclease system has progressed to be an invaluable technology for genome manipulation in somatic cell types and germline model organisms. While the unprecedented advance in human embryo gene editing research has great potential in next-generation therapeutics, it raises various ethical concerns that need to be addressed before being translated for clinical use. Here, we discuss the current and potential applications of CRISPR/Cas9 technology and its limitations in clinical applications, as well as ethical and legal considerations in the treatment, disease prevention or disability in somatic cells or human embryo via gene editing.

关键词： Addressing challenges clinical applications associated CRISPR/Cas9 technology ethical questions prevent its misuse

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Activation of AMPK/MnSOD signaling mediates anti-apoptotic effect of hepatitis B virus in hepatoma cells

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World Journal of Gastroenterology 2016年第17期22卷 4345-4353页

作者： Lei Li Hong-Hai Hong Shi-Ping Chen Cai-Qi Ma Han-Yan Liu Ya-Chao Yao key laboratory for Major obstetric Diseases of Guangdong Province Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes The Third Affiliated Hospital of Guangzhou Medical University Department of Biochemistry Zhongshan School of Medicine Sun Yat-sen University Department of Clinical laboratory Medicine the Second People’s Hospital of Guangdong Province

AIM: To investigate the anti-apoptotic capability of the hepatitis B virus(HBV) in the HepG2 hepatoma cell line and the underlying ***: Cell viability and apoptosis were measured by MTT assay and flow cytometry, respectively. Targeted knockdown of manganese superoxide dismutase(Mn SOD), AMP-activated protein kinase(AMPK) and hepatitis B virus X protein(HBx) genes as well as AMPK agonist AICAR and antagonist compound C were employed to determine the correlations of expression of these ***: HBV markedly protected the hepatoma cells from growth suppression and cell death in the condition of serum deprivation. A decrease of superoxide anion production accompanied with an increase of Mn SOD expression and activity was found in Hep G2.215 cells. Moreover, AMPK activation contributed to the up-regulation of Mn SOD. HBx protein was identified to induce the expression of AMPK and Mn SOD. CONCLUSION: Our results suggest that HBV suppresses mitochondrial superoxide level and exerts an antiapoptotic effect by activating AMPK/Mn SOD signaling pathway, which may provide a novel pharmacological strategy to prevent HCC.

关键词： Hepatitis B virus Reactive oxygen species Apoptosis Manganese superoxide dismutase AMP-activated protein kinase

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Impact of Blood Clot in Oocyte-Corona-Cumulus Complex during in vitro Fertilization and Embryo Transfer

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Journal of Reproduction and Contraception 2012年第1期23卷 57-63页

作者： Yu-ling HUANG Xiao-ling LONG Yong FAN Wen-hong ZHANG Hong-zi DU Assisted Reproductive Center the Third Affiliated Hospital of Guangzhou Medical CollegeKey Laboratory for the Major Obstetric Disease in Guangdong ProvinceGuangzhou 510150China

Objective To study the impact of blood clots in the oocyte-corona-cumulus complexes （OCCC） during in vitro fertilization and embryo transfer （IVF-ET）.Methods The OCCCs were harvested from the patients undergoing long protocol ovarian hyperstimulation. The OCCCs with blood clots removed or not, were randomly grouped into A or B. The OCCCs without blood clots were group C （the control）.Results The patient＇s age, infertility duration, the average GN consumption, the average days of superovulation and an average number of harvested oocytes showed no significant difference in the 3 groups. The fertilization rate and 2PN rate in group A were the highest, which were 85.4% and 71.1%, respectively, followed by group C, which were 77.5% and 64.9%, respectively. The lowest fertilization rate and 2PN rate were in group B, 75.8% and 62.2%, respectively. Those in group A were significantly higher than those in groups B and C （P〈0.01）, while there was no significant difference between group B and group C. The implantation rates and pregnancy rates showed no significant difference in the 3 groups after transplantation, even if group A got the highest rate among the 3 groups. Conclusion Removing the blood clots in OCCC can improve the outcome of IVF-ET without increasing the cost and complexity of the operation.

关键词： in vitro fertilization and embryo transfer (IVF-ET) blood clot fertilization rate pregnancy

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Genome wide abnormal DNA methylome of human blastocyst in assisted reproductive technology

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Journal of Genetics and Genomics 2017年第10期44卷 475-481页

作者： Guoqiang Li Yang Yu Yong Fan Congru Li Xiaocui Xu Jialei Duan Rong Li Xiangjin Kang Xin Ma Xuepeng Chen Yuwen Ke Jie Yan Ying Lian Ping Liu Yue Zhao Hongcui Zhao Yaoyong Chen Xiaofang Sun Jianqiao Liu Jie Qiao Jiang Liu Ministry of Education key laboratory of Assisted Reproduction Beijing Key Laboratory of Reproductive Endocrinology and Assisted ReproductiveTechnology Center of Reproductive Medicine Peking University Third Hospital Beijing 100191 China GAS key laboratory of Genome Sciences and Information Collaborative Innovation Center of Genetics and Development Beijing Institute of GenomicsGAS Beijing 160101 China key laboratory for Major obstetric Diseases of Guangdong Province The Third Affiliated Hospital of Guangzhou Medical University Guangzhou 510150China University of Chinese Academy of Sciences Beijing 100029 China

Proper reprogramming of parental DNA methylomes is essential for mammalian embryonic development. However, it is unknown whether abnormal methylome reprogramming occurs and is associated with the failure of embryonic development. Here we analyzed the DNA methylomes of 57 blastocysts and 29 trophectoderm samples with different morphological grades during assisted reproductive technology （ART） practices. Our data reveal that the global methylation levels of high-quality blastocysts are similar （0.30 ± 0.02, mean ± SD）, while the methylation levels of low-quality blastocysts are divergent and away from those of high-quality blastocysts. The proportion of blastocysts with a methylation level falling within the range of 0.30± 0.02 in different grades correlates with the live birth rate for that grade. Moreover, abnormal methylated regions are associated with the failure of embryonic development. Furthermore, we can use the methylation data of cells biopsied from trophectoderm to predict the blastocyst methylation level as well as to detect the aneuploidy of the blastocysts. Our data indicate that global abnormal methylome reprogramming often occurs in human embryos, and suggest that DNA methylome is a potential biomarker in blastocyst selection in ART.

关键词： Human Abnormal Blastocyst Methylome ART

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Influence of gonadotropin-releasing hormone agonist on the effect of chemotherapy upon ovarian cancer and the prevention of chemotherapy-induced ovarian damage:an experimental study with nu/nu athymic mice

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2012年第11期13卷 894-903页

作者： Qiong-yan LIN Yi-feng WANG Hui-nan WENG Xiu-jie SHENG Qing-ping JIANG Zhi-ying YANG Department of obstetrics and Gynecology Key Laboratory for Major Obstetric Diseases of Guangdong Province the Third Affiliated Hospital of Guangzhou Medical University Guangzhou 510150 China Department of obstetrics and Gynecology Zhujiang Hospital Nanfang Medical University Guangzhou 510282 China

Background and objective:Gonadotropin-releasing hormone (GnRH) plays an important role in the regulation of ovarian function and ovarian cancer cell growth. In this study, we determined whether administration of the GnRH agonist (GnRHa), triporelin, prior to cisplatin treatment affects cisplatin and/or prevents cisplatin-induced ovarian damage. Methods:nu/nu mice were injected with ovarian cancer OVCAR-3 cells intraperitoneally. After two weeks, the mice were treated with saline (control), cisplatin, GnRHa, or cisplatin plus GnRHa for four weeks. At the end of the experimental protocol, blood, tumor, ovary, and uterine tissues were resected for hematoxylin and eosin (H&E) staining, immunohistochemical analyses of Ki67, nuclear factor-κB (NF-κB), and caspase-3, transmission electron microscopy of apoptosis, or enzyme-linked immunosorbent assay (ELISA) analyses of anti-Mullerian hormone (AMH). Results:Cisplatin treatment effectively inhibited tumor growth in mice treated with human ovarian cancer cells; however the treatment also induced considerable toxicity. Immunohistochemical analyses showed that Ki67 expression was reduced in cisplatin-treated mice compared to control (P0.05), while expressions of NF-κB and caspase-3 were reduced and induced, respectively, in cisplatin-treated mice and cisplatin plus GnRHa-treated mice. Apoptosis occurred in the GnRHa, cisplatin, and cisplatin plus GnRHa-treated mice, but not in control mice. Ovaries exposed to GnRHa in both GnRHa mice and cisplatin-treated mice (combination group) had significantly more primordial and growth follicles and serum levels of AMH than those in the control mice and cisplatin-treated mice (P<0.05). Conclusions:Administration of GnRHa to mice significantly decreased the extent of ovarian damage induced by cisplatin, but did not affect the anti-tumor activity of cisplatin

关键词： Gonadotrophin-releasing hormone Cisplatin Ovarian cancer Animal experiment

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Selective Short-term Fertilization Combined with Early Rescue ICSI: An Optimal Strategy for Patients at High Risk for Fertilization Failure

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Journal of Reproduction and Contraception 2014年第4期25卷 207-217页

作者： Yu-ling HUANG Ai-hua WU Jian-qiao LIU Center for Reproductive Medicine the Third Affiliated Hospital of Guangzhou Medical University key laboratory for Reproductive Medicine of Guangdong Province key laboratory for Major obstetric Diseases of Guangdong Province obstetrics and Gynecology Department the Second Affiliated Hospital of Guangzhou Medical University

Objective To investigate clinical outcomes in patients who were at more precise criteria risks for fertilization failure and were treated with selective, short-term fertilization （oocytes and sperm co-incubated for 4 h） and early rescue intracytoplasmic sperm injection （ICSI）. Methods A retrospective analysis was performed on 2023 women undergoing assisted reproductive technology （ART）. They were assigned to 4 groups： short-term in vitro fertilization （short-term IVF,, group A, n=217）, regular IVF （oocytes and sperm coincubated overnight, group B, n=1475）, short-term IVF and early rescue ICSI （shortterm ICSI, group C, n=94）, and regular ICSI （group D, n=237）. Results In group A, 69.8% （217/311） achieved normal fertilization rates, and the complete fertilization failure rate （fertilization rate was 0%） was 12.9% （40/311）. But all of the fertilization failure oocytes got rescue ICSI. In group B, the complete fertilization failure rate was 1.1% （19/1 692）. The fertilization rate, 2 PN （pronucleus） rate, and i PN rate were significantly lower in group A than those in group B （70.9% vs 80.8%, 57.8% vs 66.3%, and 3.5% vs 6.2%, respectively）. No significant differences were observed in clinical pregnancy rates and birth defect rates between groups A and B. The fertilization rates in groups C and D did not significantly differ （77.9% vs 76.2%）, which was also true for birth defect rates. The clinical pregnancy rate of group C was higher than that of group D （51.2% vs 42.3%）, but this difference was not significant （P〉0. 05）.Conclusion These results suggested that selective, short-term fertilization can result in effective outcomes for patients who were at high risk for fertilization failure.

关键词： in vitro fertilization and embryo transfer (IVF-ET) short-term fertilization early rescue intracytoplasmic sperm injections (ICSI) pregnancy

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