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MGMT is down-regulated independently of promoter DNA methylation in rats with all-trans retinoic acidinduced spina bifida aperta

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Neural Regeneration Research 2019年第2期14卷 361-368页

作者： He-Nan Zhang Yi Guo Wei Ma Jia Xue Wei-Lin Wang Zheng-Wei Yuan key laboratory of health ministry for congenital malformation Shengjing Hospital China Medical University Department of Pediatric Surgery Shengjing Hospital China Medical University

O6-methylguanine DNA methyltransferase(MGMT), a DNA repair enzyme, has been reported in some congenital malformations, but it is less frequently reported in neural tube defects. This study investigated MGMT mRNA expression and methylation levels in the early embryo and in different embryonic stages, as well as the relationship between MGMT and neural tube defects. Spina bifida aperta was induced in rats by a single intragastric administration of all-trans retinoic acid on embryonic day(E) 10, whereas normal control rats received the same amount of olive oil on the same embryonic day. DNA damage was assessed by detecting γ-H2 A.X in spina bifida aperta rats. Real time-polymerase chain reaction was used to examine mRNA expression of MGMT in normal control and spina bifida aperta rats. In normal controls, the MGMT mRNA expression decreased with increasing embryonic days, and was remarkably reduced from E11 to E14, reaching a minimum at E18. In the spina bifida aperta model, γ-H2 A.X protein expression was increased, and mRNA expression of MGMT was markedly decreased on E14, E16, and E18. Bisulfite sequencing polymerase chain reaction for MGMT promoter methylation demonstrated that almost all CpG sites in the MGMT promoter remained unmethylated in both spina bifida aperta rats and normal controls, and there was no significant difference in methylation level between the two groups on either E14 or E18. Our results show that DNA damage occurs in spina bifida aperta rats. The mRNA expression of MGMT is downregulated, and this downregulation is independent of promoter DNA methylation.

关键词： nerve regeneration neural tube defects spina bifida aperta spinal cord all-trans retinoic acid O6-methylguanine DNA methyltransferase gene expression DNA methylation promoter bisulfite sequencing polymerase chain reaction neural regeneration

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Spatiotemporal expression of leukemia inhibitory factor receptor protein during neural tube development in embryos with neural tube defects

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Neural Regeneration Research 2020年第4期15卷 705-711页

作者： Dong An Xiao-Wei Wei He-Nan Zhang Dan Liu Wei Ma Zheng-Wei Yuan key laboratory of health ministry for congenital malformation Shengjing HospitalChina Medical UniversityShenyangLiaoning ProvinceChina Department of Pediatrics The First Hospital of China Medical UniversityShenyangLiaoning ProvinceChina

Leukemia inhibitory factor receptor(LIFR),as a neuroregulatory cytokine receptor,generally shows a neuroprotective effect in central nervous system *** this study,to understand the effect of LIFR on pathogenesis of neural tube defects,we explored spatiotemporal expression of LIFR at different stages of fetal development in normal and neural tube defect *** bifida aperta was induced with all-trans retinoic acid on embryonic day 10 in rats,and the spatiotemporal expression of LIFR was investigated in spina bifida aperta rats and healthy rats from embryonic day 11 to *** time-polymerase chain reaction and western blot assay were used to examine mRNA and protein expression of LIFR in healthy control and neural tube defect *** of the animal experiment demonstrated that expression of LIFR protein and mRNA in the spinal cords of normal rat embryos increased with embryonic *** was significantly downregulated in the spinal cords of spina bifida aperta rats compared with healthy rats from embryonic days 11 to *** staining showed that the expression of LIFR in placenta and spinal cord in spina bifida aperta rat embryos was decreased compared with that in control embryos at embryonic day *** from human embryo specimens showed that LIFR mRNA expression was significantly down-regulated in spinal cords of human fetuses with neural tube defects compared with normal controls at a gestational age of 24 to 33 *** results were consistent with the down-regulation of LIFR in the animal *** study revealed spatiotemporal changes in expression of LIFR during embryonic ***,LIFR might play a specific role in neural tube *** animal and human experimental procedures were approved by the Medical Ethics Committee of Shengjing Hospital of China Medical University,China(approval No.2016PS106K)on February 25,2016.

关键词： amniotic fluid development embryogenesis leukemia inhibitory factor receptor nerve regeneration neural tube defect placenta spatiotemporal expression spina bifida aperta spinal cord serum

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Genome-Wide Screening of Aberrant Methylation Loci for Nonsyndromic Cleft Lip

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Chinese Medical Journal 2018年第17期131卷 2055-2062页

作者： Xiao-Yan Xu Xiao-Wei Wei Wei Ma Hui Gu Dan Liu Zheng-Wei Yuan key laboratory of health ministry for congenital malformation Shengjing Hospital China Medical University Shenyang Liaoning 110004 China

Background： The pathogenicity of cleft lip （CL） is pretty complicated since it is influenced by the interaction of environment and genetic factors. The purpose of this study was to conduct a genome-wide screening of aberrant methylation loci in partial lesion tissues of patients with nonsyndromic CL （NSCL） and preliminarily validate candidate dysmethylated genes associated with ***： Fifteen healthy and sixteen NSCL fetal lip tissue samples were collected. The Infinium HumanMethylation450 BeadChip was used to screen aberrant methylation loci in three NSCL and three healthy lip tissues. The differential methylation sites and functions of the annotated genes between NSCL and healthy lip tissues were analyzed using minfi package of R software, cluster analysis, Gene Ontology （GO） annotation, and metabolic pathway annotation. Gene expression was assessed in nine differentially methylated genes by real-time polymerase chain reaction （PCR）. The transcriptions mRNA levels of three out of nine candidate genes were downregulated remarkably in NSCL lip tissues, and these three genes＇ abnormal methylation loci were validated by pyrosequencing in 16 NSCL cases and 15 healthy ***： In total, 4879 sites in the genes of NSCL odinopoeia fetuses showed aberrant methylation when compared with normal lip tissue genome. Among these, 3661 sites were hypermethylated and 1218 sites were hypomethylated as compared to methylation levels in healthy specimens. These aberrant methylation sites involved 2849 genes and were widely distributed among the chromosomes. Most differentially methylated sites were located in cytosine-phosphoric acid-guanine islands. Based on GO analysis, aberrantly methylated genes were involved in 11 cellular components, 13 molecular functions, and a variety of biological processes. Notably, the transcription of DAB1, REELIN, and FYN was significantly downregulated in lesion tissues of NSCL fetus （P 〈 0.05）. Pyrosequencing results validated that there

关键词： DAB1 Methylation Nonsyndromic Cleft Lip REELIN Signaling Pathway

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Intestinal dendritic cells change in number in fulminant hepatic failure

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World Journal of Gastroenterology 2015年第16期21卷 4883-4893页

作者： Xu Cao Mei Liu Peng Wang Dong-Yan Liu Research Center Shengjing Hospital of China Medical University and Key Laboratory of Congenital Malformation ResearchMinistry of Health

AIM:To investigate the change in intestinal dendritic cell(DC)number in fulminant hepatic failure(FHF).METHODS:An animal model of FHF was *** CD11b/c was detected by immunohistochemistry and Western *** real-time polymerase chain reaction(PCR)was used to detect intestinal integrin-αm RNA *** CD83,CD86,CD74,CD3 and AKT were detected by immunohistochemistry,Western blot and ***-AKT(p-AKT)was detected by immunohistochemistry and Western ***:In the FHF group[D-galactosamine(D-Galn)+lipopolysaccharide(LPS)group],the mice began to die after 6 h;conversely,in the D-Galn and LPS groups,the activity of mice was poor,but there were no *** results showed that in FHF,the expression of CD11b/c(7988400±385941vs 1102400±132273,P0.05)

关键词： Fulminant hepatic failure Intestinal dendriticcells MHC Ⅱ CD3 AKT/Phosphorylated-AKT

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Safety of menstrual blood-derived stromal cell transplantation in treatment of intrauterine adhesion

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World Journal of Stem Cells 2020年第5期12卷 368-380页

作者： Qi-Yuan Chang Si-Wen Zhang Ping-Ping Li Zheng-Wei Yuan Ji-Chun Tan Department of Obstetrics and Gynecology Shengjing Hospital of China Medical UniversityShenyang 110022Liaoning ProvinceChina key laboratory of Reproductive Dysfunction Diseases and Fertility Remodeling of Liaoning Province Shenyang 110004Liaoning ProvinceChina key laboratory of health ministry for congenital malformation Shengjing Hospital of China Medical UniversityBenxi 117004Liaoning ProvinceChina

BACKGROUND Intrauterine adhesion(IUA)can cause serious damage to women’s reproductive health,yet current treatment methods are difficult to achieve satisfactory *** our previous studies,we demonstrated that menstrual-derived stromal stem cells(MenSCs),with high proliferative capacity and self-renewal ability,have a powerful therapeutic effect in patients with severe ***,safety assessment of MenSCs transplantation is essential for its further *** To evaluate the short-,medium-,and long-term biosafety of MenSCs via intrauterine transplantation in a rat model of IUA,with a focus on toxicity and *** MenSCs were injected into the sub-serosal layer of the uterus in an IUA rat model,for 3 d,3 mo,and 6 mo separately,to monitor the corresponding acute,sub-chronic,and chronic *** rats of the same age served as negative *** effects were evaluated by body weight,organ weight,histopathology,hematology,and biochemistry *** of MenSCs was investigated in Balb/c-nu mice in vivo and by colony formation assays in *** Compared with the same week-old control group,all of the IUA rats receiving MenSC transplantation demonstrated no obvious changes in body weight,mainorgan weight,or blood cell composition during the acute,sub-chronic,and chronic observation *** the same time,serum biochemical tests showed no adverse effects on metabolism or liver and kidney *** 4 wk of subcutaneous injection of Men SCs in Balb/c-nu nude mice,no tumor formation or cell metastasis was ***,there was no tumor colony formation of Men SCs during soft agar culture in *** There is no acute,sub-chronic,or chronic poisoning,infection,tumorigenesis,or endometriosis in rats with IUA after Men SC *** above results suggest that intrauterine transplantation of Men SCs is safe for endometrial treatment.

关键词： Menstrual blood-derived stromal cells Endometrial treatment Intrauterine adhesion Stem cell transplantation Biosafety Toxicity

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Inhibition of renin activity by aliskiren ameliorates diabetic nephropathy in type 1 diabetes mouse model

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Journal of Diabetes Mellitus 2012年第3期2卷 353-360页

作者： Yan Zhang Youli Wang Yunzi Chen Dilip K. Deb Tao Sun Qun Zhao Yan Chun Li Department of Medicine Division of Biological Sciences The University of Chicago Chicago USA ministry of health key laboratory of congenital malformation Shengjing Hospital China Medical University Shenyang China

Renin is the rate-limiting enzyme of the reninangiotensin system (RAS). In addition to its enzymatic activity to generate angiotensin I, renin also signals through the (pro)renin receptor to exert angiotensin II-independent effects. In this study we examined the effect of renin inhibition on the development of diabetic nephropathy. Male DBA/2J mice were induced to diabetes with streptozotocin, and the diabetic mice were treated for 16 weeks with saline or aliskiren, a renin enzymatic inhibitor. Aliskiren treatment had little effects on blood glucose and blood pressure in diabetic mice. Saline-treated mice developed progressive albuminuria and glome-rulosclerosis, and aliskiren treatment effectively alleviated albumiuria and glomerulosclerosis. Morphologically aliskiren treatment prevented the thickening of the glomerular basement membrane and reduced podocyte loss. At the molecular levels, aliskiren prevented the decline of slit diaphragm proteins and blocked the synthesis of extracellular matrix and pro-fibrotic factors in the diabetic kidney. Aliskiren treatment results in compensatory renin increase in the glomeruli due to blockade of the negative feedback loop, and also partially suppressed the intracellular signaling mediated by the (pro)renin receptor activated in hyperglycemia. These observations suggest that the therapeutic activity of aliskiren to prevent diabetic renal injury is contributed by inhibition of both the angiotensin II-dependent and -independent pathways. Taken together, it is concluded that inhibition of renin enzymatic activity ameliorates diabetic renal injury in type 1 diabetes, and support the use of aliskiren in diabetes kidney disease.

关键词： Renin Inhibitor Renin-Angiotensin System Diabetic Nephropathy Aliskiren

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IRF7 and CTSS are pivotal for cutaneous wound healing and may serve as therapeutic targets

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Signal Transduction and Targeted Therapy 2023年第9期8卷 3927-3930页

作者： Jiali Yin Dongxin Shi Yan Sun Peiyao Zhu Yiping Zhao Xuegang Xu Hongduo Chen Yan Wu Zhengwei Yuan Xing-Hua Gao Department of Dermatology The First Hospital of China Medical UniversityShenyangChina key laboratory of Immunodermatology Ministry of Education and NHCNational joint Engineering Research Center for Theranostics of Immunological Skin DiseasesShenyangChina Department of Otolaryngology Head and Neck SurgeryYantai Yuhuangding HospitalQingdao UniversityYantaiChina key laboratory of health ministry for congenital malformation Shengjing HospitalChina Medical UniversityShenyangChina College of Medicine and Biological Information Engineering Northeastern UniversityNO.3-11Wenhua RoadHeping DistrictShenyangChina

Dear Editor,Adult cutaneous wounds typically result in deficient regenera-tion due to abnormal fibro-proliferative responses.1 The scarring,while conducive to preventing extended infection and loss of nutrients at the injury site,can be troublesome.2 In contrast,wound healing in embryos showes lower levels of inflammation and a remarkable capacity for remodeling.3 Interestingly,the capability of fetal scarless healing is *** contrasted with the near-normal healing after the injury that occurs in the early gestational period(embryonic day 16,E16),wound healing in late gestation(embryonic day 18,E18)has prominent *** mechanisms responsible for this transition are largely unknown.

关键词： healing wound inflammation

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Predictive factors for pituitary response to pulsatile GnRH therapy in patients with congenital hypogonadotropic hypogonadism

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Asian Journal of Andrology 2018年第4期20卷 319-323页

作者： Jiang-Feng Mao Xi Wang Jun-Jie Zheng Zhao-Xiang Liu Hong-Li Xu Bing-Kun Huang Min Nie Xue-Yan Wu Department of Endocrinology Peking Union Medical College Hospital Key Laboratory of Endocrinology Ministry of Health Beijing 100730 China

Pulsatile gonadotropin-releasing hormone （GnRH） may induce spermatogenesis in most patients with congenital hypogonadotropic hypogonadism （CHH） by stimulating gonadotropin production, while the predictors for a pituitary response to pulsatile GnRH therapy were rarely investigated. Therefore, the aim of our study is to investigate predictors of the pituitary response to pulsatile GnRH therapy. This retrospective cohort study included 82 CHH patients who received subcutaneous pulsatile GnRH therapy for at least 1 month. Patients were categorized into poor or normal luteinizing hormone （LH） response subgroups according to their LH level （LH 〈2 IU 1-1 or LH P_2 IU 1-1） 1 month into pulsatile GnRH therapy. Gonadotropin and testosterone levels, testicular size, and sperm count were compared between the two subgroups before and after GnRH therapy. Among all patients, LH increased from 0.4±0.5 IUI^-1 to 7.5±4.4 IUI^-1 and follicle-stimulating hormone （FSH） increased from 1.1±0.9 IUI^-1 to 8.8±5.3 IU 1-1. A Cox regression analysis showed that basal testosterone level （6 = 0.252, P = 0.029） and triptorelin-stimulated FSH60min（13 = 0.518, P = 0.01） were two favorable predictors for pituitary response to GnRH therapy. Nine patients （9/82, 11.0%） with low LH response to GnRH therapy were classified into the poor LH response subgroup. After pulsatile GnRH therapy, total serum testosterone level was 39± 28 ng dl^-1 versus 248±158 ng d1^-1 （P=0.001）, and testicular size was 4.0±3.1 ml versus 7.9±4.5 ml （P= 0.005） in the poor and normal LH response subgroups, respectively. It is concluded that higher levels of triptorelin-stimulated FSH60min. and basal total serum testosterone are favorable predictors of pituitary LH response to GnRH therapy.

关键词： congenital hypogonadotropic hypogonadism pulsatile GnRH therapy treatment

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Progress in Biomarkers Related to Biliary Atresia

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Journal of Clinical and Translational Hepatology 2024年第3期12卷 305-315页

作者： Fanyang Kong Rui Dong Gong Chen Song Sun Yifan Yang Jingying Jiang Lingdu Meng Huifen Chen Jiajie Zhu Shan Zheng Department of Pediatric Surgery Children’s Hospital of Fudan UniversityShanghai Key Laboratory of Birth Defectand Key Laboratory of Neonatal DiseaseMinistry of HealthShanghaiChina

Biliary atresia(BA)is a congenital cholestatic disease that can seriously damage children’s liver *** is one of the main reasons for liver transplantation in *** diagnosis of BA is crucial to the prognosis of patients,but there is still a lack of reliable non-invasive diagnostic ***,as some children are in urgent need of liver transplantation,evaluating the stage of liver fibrosis and postoperative native liver survival in children with BA using a straightforward,efficient,and less traumatic method is a major focus of *** recent years,an increasing number of BA-related biomarkers have been identified and have shown great potential in the following three aspects of clinical practice:diagnosis,evaluation of the stage of liver fibrosis,and prediction of native liver *** review focuses on the pathophysiological function and clinical application of three novel BA-related biomarkers,namely MMP-7,FGF-19,and ***,progress in well-known biomarkers of BA such as gamma-glutamyltransferase,circulating cytokines,and other potential biomarkers is discussed,aiming to provide a reference for clinical practice.

关键词： Biliary atresia Biomarkers Diagnosis Liver fibrosis Native liver survival

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Different sperm sources and parameters can influence intracytoplasmic sperm injection outcomes before embryo implantation

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2012年第1期13卷 1-10页

作者： Yue-hong LU1,2, Hui-juan GAO1, Bai-jia LI1, Ying-ming ZHENG1, Ying-hui YE1, Yu-li QIAN1, Chen-ming XU1, He-feng HUANG1, Fan JIN1 (1Department of Reproductive Endocrinology, key laboratory of Reproductive Genetics, ministry of Education, and key laboratory of Women’s Reproductive health of Zhejiang Province, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou 310006, China) (2Department of Reproductive health, Shaoxing Women and Children Hospital, Shaoxing 312000, China) Department of Reproductive Endocrinology Key Laboratory of Reproductive Genetics Ministry of Education and Key Laboratory of Women's Reproductive Health of Zhejiang Province Women's Hospital School of Medicine Zhejiang University Hangzhou 310006 China

To evaluate the effects of sperm with different parameters and sources on the outcomes of intracytoplasmic sperm injection (ICSI), 1972 ICSI cycles were analyzed retrospectively. Groups 1 to 5 were composed of cycles using ejaculated sperm and were grouped according to sperm quantity, quality, and morphology into normal (288 cycles), or mild (329 cycles), moderate (522 cycles), severe (332 cycles), and extremely severe (171 cycles) oligozoospermia and/or asthenozoospermia and/or teratozoospermia (OAT) groups. Group 6 was composed of 250 cycles using testicular or epididymal sperm, and Group 7 consisted of 80 cycles using frozen-thawed sperm. We found that fertilization rates were gradually reduced from Groups 1 to 6, and reached statistical difference in Groups 5 and 6 (P0.05). A total of nine cases of malformation were observed, with a malformation rate of 1.25% (9/719). In conclusion, different sperm sources and parameters can affect ICSI outcomes before embryo implantation. A full assessment of offspring mal-formation will require further study using a larger sample size.

关键词： Intracytoplasmic sperm injection (ICSI)i Sperm Sperm source Sperm parameters malformation

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