BACKGROUND This study aimed to explore the relationship between gene mutations and early embryonic development arrest and to provide more possibilities for the diagnosis and treatment of repeated implantation failure....BACKGROUND This study aimed to explore the relationship between gene mutations and early embryonic development arrest and to provide more possibilities for the diagnosis and treatment of repeated implantation failure.CASE SUMMARY Here,we collected and described the clinical data of a patient with early embryonic development stagnation after repeated in vitro fertilization attempts for primary infertility at the Department Reproductive Center of Zaozhuang Maternal and Child Healthcare Hospital.We also detected the whole-exon gene of the patient's spouse and parents,and conducted bioinformatics analysis to determine the pathogenesis of the gene.CONCLUSION A novel mutant of the TUBB8 gene[c.602G>T(p.C201F)]was identified,and this mutant provided new data on the genotype-phenotype relationships of related diseases.展开更多
Background Mitochondrial dysfunction induced by excessive mitochondrial reactive oxygen species(ROS)damages embryonic development and leads to growth arrest.Objective The purpose of this study is to elucidate whether ...Background Mitochondrial dysfunction induced by excessive mitochondrial reactive oxygen species(ROS)damages embryonic development and leads to growth arrest.Objective The purpose of this study is to elucidate whether maternal zinc(Zn)exert protective effect on oxidative stress targeting mitochondrial function using an avian model.Result In ovo injected tert-butyl hydroperoxide(BHP)increases(P<0.05)hepatic mitochondrial ROS,malondialdehyde(MDA)and 8-hydroxy-2-deoxyguanosine(8-OHdG),and decreases(P<0.05)mitochondrial membrane potential(MMP),mitochondrial DNA(mtDNA)copy number and adenosine triphosphate(ATP)content,contributing to mitochondrial dysfunction.In vivo and in vitro studies revealed that Zn addition enhances(P<0.05)ATP synthesis and metallothionein 4(MT4)content and expression as well as alleviates(P<0.05)the BHP-induced mitochondrial ROS generation,oxidative damage and dysfunction,exerting a protective effect on mitochondrial function by enhancing antioxidant capacity and upregulating the mRNA and protein expressions of Nrf2 and PGC-1α.Conclusions The present study provides a new way to protect offspring against oxidative damage by maternal Zn supplementation through the process of targeting mitochondria involving the activation of Nrf2/PGC-1αsignaling.展开更多
Objective Vascular smooth muscle cell(VSMC)differentiation from stem cells is one source of the increasing number of VSMCs that are involved in vascular remodeling-related diseases such as hypertension,atherosclerosis...Objective Vascular smooth muscle cell(VSMC)differentiation from stem cells is one source of the increasing number of VSMCs that are involved in vascular remodeling-related diseases such as hypertension,atherosclerosis,and restenosis.MicroRNA-146a(miR-146a)has been proven to be involved in cell proliferation,migration,and tumor metabolism.However,little is known about the functional role of miR-146a in VSMC differentiation from embryonic stem cells(ESCs).This study aimed to determine the role of miR-146a in VSMC differentiation from ESCs.Methods Mouse ESCs were differentiated into VSMCs,and the cell extracts were analyzed by Western blotting and RT-qPCR.In addition,luciferase reporter assays using ESCs transfected with miR-146a/mimic and plasmids were performed.Finally,C57BL/6J female mice were injected with mimic or miR-146a-overexpressing ESCs,and immunohistochemistry,Western blotting,and RT-qPCR assays were carried out on tissue samples from these mice.Results miR-146a was significantly upregulated during VSMC differentiation,accompanied with the VSMC-specific marker genes smooth muscle-alpha-actin(SMαA),smooth muscle 22(SM22),smooth muscle myosin heavy chain(SMMHC),and h1-calponin.Furthermore,overexpression of miR-146a enhanced the differentiation process in vitro and in vivo.Concurrently,the expression of Kruppel-like factor 4(KLF4),predicted as one of the top targets of miR-146a,was sharply decreased in miR-146a-overexpressing ESCs.Importantly,inhibiting KLF4 expression enhanced the VSMC-specific gene expression induced by miR-146a overexpression in differentiating ESCs.In addition,miR-146a upregulated the mRNA expression levels and transcriptional activity of VSMC differentiation-related transcription factors,including serum response factor(SRF)and myocyte enhancer factor 2c(MEF-2c).Conclusion Our data support that miR-146a promotes ESC-VSMC differentiation through regulating KLF4 and modulating the transcription factor activity of VSMCs.展开更多
Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embr...Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.展开更多
Background Fertility declines in high-parity sows.This study investigated whether parity-dependent declines in embryonic survival and reproductive performance could be restored by dietary coenzyme Q10(CoQ10)supplement...Background Fertility declines in high-parity sows.This study investigated whether parity-dependent declines in embryonic survival and reproductive performance could be restored by dietary coenzyme Q10(CoQ10)supplementation.Methods Two experiments were performed.In Exp.1,30 young sows that had completed their 2nd parity and 30 high-parity sows that had completed their 10^(th)parity,were fed either a control diet(CON)or a CON diet supple-mented with 1 g/kg CoQ10(+CoQ10)from mating until slaughter at day 28 of gestation.In Exp.2,a total of 314 post-weaning sows with two to nine parities were fed the CON or+CoQ10 diets from mating throughout gestation.Results In Exp.1,both young and high-parity sows had a similar number of corpora lutea,but high-parity sows had lower plasma CoQ10 concentrations,down-regulated genes involved with de novo CoQ10 synthesis in the endome-trium tissues,and greater levels of oxidative stress markers in plasma and endometrium tissues.High-parity sows had fewer total embryos and alive embryos,lower embryonic survival,and greater embryo mortality than young sows.Dietary CoQ10 supplementation increased the number of live embryos and the embryonic survival rate to levels simi-lar to those of young sows,as well as lowering the levels of oxidative stress markers.In Exp.2,sows showed a parity-dependent decline in plasma CoQ10 levels,and sows with more than four parities showed a progressive decline in the number of total births,live births,and piglets born effective.Dietary supplementation with CoQ10 increased the number of total births,live births,and born effective,and decreased the intra-litter covariation coefficients and the percentage of sows requiring farrowing assistance during parturition.Conclusions Dietary CoQ10 supplementation can improve the embryonic survival and reproductive performance of gestating sows with high parity,probably by improving the development of uterine function.展开更多
Background During mammalian pre-implantation embryonic development(PED),the process of maternal-to-zygote transition(MZT)is well orchestrated by epigenetic modification and gene sequential expression,and it is related...Background During mammalian pre-implantation embryonic development(PED),the process of maternal-to-zygote transition(MZT)is well orchestrated by epigenetic modification and gene sequential expression,and it is related to the embryonic genome activation(EGA).During MZT,the embryos are sensitive to the environment and easy to arrest at this stage in vitro.However,the timing and regulation mechanism of EGA in buffaloes remain obscure.Results Buffalo pre-implantation embryos were subjected to trace cell based RNA-seq and whole-genome bisulfite sequencing(WGBS)to draw landscapes of transcription and DNA-methylation.Four typical developmental steps were classified during buffalo PED.Buffalo major EGA was identified at the 16-cell stage by the comprehensive analy-sis of gene expression and DNA methylation dynamics.By weighted gene co-expression network analysis,stage-spe-cific modules were identified during buffalo maternal-to-zygotic transition,and key signaling pathways and biological process events were further revealed.Programmed and continuous activation of these pathways was necessary for success of buffalo EGA.In addition,the hub gene,CDK1,was identified to play a critical role in buffalo EGA.Conclusions Our study provides a landscape of transcription and DNA methylation in buffalo PED and reveals deeply the molecular mechanism of the buffalo EGA and genetic programming during buffalo MZT.It will lay a foundation for improving the in vitro development of buffalo embryos.展开更多
Objective:To compare embryonic development,ploidy status and clinical outcomes between fresh and frozen-thawed oocytes.Methods:This retrospective cohort study evaluated 83 fertilization cycles including both fresh and...Objective:To compare embryonic development,ploidy status and clinical outcomes between fresh and frozen-thawed oocytes.Methods:This retrospective cohort study evaluated 83 fertilization cycles including both fresh and frozen oocytes from 79 patients at the HP Fertility Center of Hai Phong International Hospital of Obstetrics and Pediatrics in Vietnam.The patient underwent several ovarian stimulation cycles to accumulate a certain number of oocytes that would be vitrified.In the last oocyte retrieval,all patient’s oocytes including both frozen and fresh would be fertilized.The outcomes included the rates of oocyte survival,cleavage embryo,blastocyst,ploidy status,pregnancy,biochemical pregnancy and clinical pregnancy.Results:The oocyte survival rate after thawing was 96.5%.No statistically significant difference was found when comparing fresh and frozen oocytes regarding fertilization rate(78.1%vs.75.5%,P=0.461),usable cleavage embryo rate(86.9%vs.87.2%,P=0.916)but usable blastocyst rate was found higher statistically in the frozen oocyte group(44.4%vs.54.0%,P=0.049).The percentages of euploid,aneuploid and mosaic embryos between the fresh group and the vitrified group had no significant differences(33.8%vs.31.6%,P=0.682;51.0%vs.54.2%,P=0.569;15.2%vs.12.4%,P=0.787;respectively).The rates of pregnancy,biochemical pregnancy and clinical pregnancy had no statistical difference(68.8%vs.64.8%,P=0.764;12.5%vs.3.6%,P=0.258;37.5%vs.46.4%,P=0.565).17 Mature oocytes are the minimum to have at least one euploid embryo.Conclusions:Oocyte vitrification does not affect embryonic,genetic and clinical results.The number of mature oocytes should be considered for fertilization in some cases.展开更多
This paper describes a new species of the snout moth Berastagia tainanica sp. nov. (Lepidoptera: Pyralidae) from Taiwan. From 2009 to 2016, a biology study was conducted on population dynamics and embryonic developmen...This paper describes a new species of the snout moth Berastagia tainanica sp. nov. (Lepidoptera: Pyralidae) from Taiwan. From 2009 to 2016, a biology study was conducted on population dynamics and embryonic development. Spring season is the peak of the eclosion of overwintering larvae or pupae. The average longevity of adult was 14.8 ± 6.2 days (N = 174), the average number of eggs laid was 259 ± 3 eggs/moth (N = 2), the hatching rate of eggs was 95.4% (N = 262), and the average hatching time of eggs was 99.6 ± 18.6 hours (N = 68). The average body length of males was 5.64 mm ± 0.91 mm (N = 30), and the average body length of females was 6.28 mm ± 0.84 mm (N = 30). This finding indicates that female snout moths are larger than males (Global R = 0.058, P = 0.012). The snout moth eclosion rate was 16.9 moths/100 pods in the first year (2010/2011, N = 2,224 pods) and 10.9 moths/100 pods in the second year (2014/2015, N = 6,382 pods). The pod borer rate was 31.8% (N = 707) and the seed borer rate was 41.2% (N = 3,628) in the first year, whereas the pod borer rate was 76.2% (N = 6,382) in the second year.展开更多
During in vitro fertilization, human embryos are incubated without light, and these conditions do not ensure embryo survival. This study explored whether environmental conditions can influence the embryo viability rat...During in vitro fertilization, human embryos are incubated without light, and these conditions do not ensure embryo survival. This study explored whether environmental conditions can influence the embryo viability rates of the house cricket, Acheta domesticus. In particular, the experiment tested what colors of visible light provide the best incubation conditions to ensure cricket embryo viability. The concept was to use house cricket embryos to represent human embryos. Cricket embryos were chosen as their eggs have soft outer membrane casings and resemble human embryos during the first few days after fertilization. During the experiment, the adult crickets laid their eggs into one of six soil-filled boxes called substrates. Each substrate was placed into one of six storage containers filled with adult crickets and lit with a different colored visible light (red, yellow, green, blue, white, or no light). After two days of breeding, the egg-filled substrates were removed from the adult crickets and placed in another storage container of the same color light. After incubation under heat-emitting lamps and under one of six light colors, nymphs were counted after hatching to determine embryo viability. After three trials, the red light provided the significantly highest viability rate, with yellow and no light being comparable seconds. The green, blue, and white lights showed significantly lower viability rates than no visible light. My results raise the speculation that exposing fertilized mammal eggs to visible light colors might have the same effects during the in vitro fertilization process.展开更多
Objective:To understand the clinical characteristics of patients with embryonic posterior cerebral artery and its correlation with abnormal vascular development.Methods:The clinical data of 396 patients with embryonic...Objective:To understand the clinical characteristics of patients with embryonic posterior cerebral artery and its correlation with abnormal vascular development.Methods:The clinical data of 396 patients with embryonic posterior cerebral artery confirmed by magnetic resonance angiography(MRA)and computed tomography angiography(CTA)were analyzed.Results:Two-hundred patients had clinical manifestations of posterior circulation ischemia,including recurrent dizziness,vertigo,and tinnitus;45 had headaches,97 had limb weakness,and 16 patients had syncope or impaired consciousness.Seventy-six patients with circulatory infarction were admitted to the hospital.There were 251 patients with history of hypertension,74 with diabetes,113 with hyperlipidemia,13 with dominant vertebral artery,10 with intracranial aneurysm,and 19 with absence of A1 segment of the anterior cerebral artery(considering developmental variation).Conclusion:Embryonic posterior cerebral artery develops abnormally during the embryonic period,often accompanied by abnormal vascular access.Due to abnormal hemodynamics,the incidence of posterior circulation ischemia,aneurysm,and infarction increases in such patients.展开更多
Background: The endometrium is the tissue that lines the inside of the uterus. It undergoes multiple changes during the menstrual cycle and prepares to welcome the fertilized egg. Endovaginal ultrasound is a first-lin...Background: The endometrium is the tissue that lines the inside of the uterus. It undergoes multiple changes during the menstrual cycle and prepares to welcome the fertilized egg. Endovaginal ultrasound is a first-line examination in the assessment of female infertility. Objective: We aimed to determine the impact of endometrial status by endovaginal ultrasound on the outcome of embryo transfers. Subjects and Methods: This was a prospective cross-sectional study of 150 women collected between January and October 2020 at the “Le Diafounou” medically assisted procreation laboratory. Were included in the study, women presenting for desire of pregnancy and having continued the treatment until the transfer of fresh or frozen embryos. An uterine score was used. This score took into account the thickness of endometrium, it is coffee bean aspect, the presence of the notch, the echogenicity of the endometrium, the pulsatility index, the presence of the flow at the end of diastole and sub-endometrial flow. Ultrasounds were performed using a General Electric logic 500, logic 9 and Voluson E8 device. The data was entered into Excel and then analyzed using SPSS version 21. Results: 150 women were selected for the study. 83 women or 55.3% were between 17 and 35 years old and 44.7% were over 35 years old with an upper limit of 50 years old. The number of women who became pregnant was 69% or 46%. The average thickness of the endometrium was 11.40 mm with extremes of 4.65 mm and 18.6 mm. There was a correlation between the thickness of the endometrium and obtaining a pregnancy (p i.e. 88.7%. The pulsatility index was relatively low for those who started a pregnancy, i.e. 90%. The high pulsatility index is one of the reasons for the failure of embryo implantation. The correlation was significant (p = 0.009). 141 women had no notch in the uterine arteries, i.e. 94%. 94 women or 62.7% had a triple line endometrium or typical coffee bean appearance. End-diastolic flow was observed in 127 women, i.e. 84.7%. The sub-endometrial flow was found in 128 women or 85.3%. Conclusion: The Knowledge of the status of the endometrium is essential and has an impact on the outcome of embryo transfers. The thinner (7 and <15). Doppler factors also play an important role.展开更多
Neurons derived from embryonic stem cells(ESCs) have gained great merit in both basic research and regenerative medicine. Here we review and summarize the signaling pathways that have been reported to be involved in t...Neurons derived from embryonic stem cells(ESCs) have gained great merit in both basic research and regenerative medicine. Here we review and summarize the signaling pathways that have been reported to be involved in the neuronal differentiation of ESCs,particularly those associated with in vitro differentiation. The inducers and pathways explored include retinoic acid, Wnt/b-catenin, transforming growth factor/bone morphogenetic protein, Notch, fibroblast growth factor, cytokine, Hedgehog, c-Jun N-terminal kinase/mitogen-activated protein kinase and others. Some other miscellaneous molecular factors that have been reported in the literature are also summarized and discussed. These include calcium, calcium receptor, calcineurin, estrogen receptor, Hox protein, ceramide, glycosaminioglycan, ginsenoside Rg1, opioids, two pore channel 2, nitric oxide, chemically defined medium, cellcell interactions, and physical stimuli. The interaction or crosstalk between these signaling pathways and factors will be explored. Elucidating these signals in detail should make a significant contribution to future progress in stem cell biology and allow, for example, better comparisons to be made between differentiation in vivo and in vitro. Of equal importance, a comprehensive understanding of the pathways that are involved in the development of neurons from ESCs in vitro will also accelerate their application as part of translational medicine.展开更多
BACKGROUND Premature ovarian failure(POF)affects many adult women less than 40 years of age and leads to infertility.According to previous reports,various tissue-specific stem cells can restore ovarian function and fo...BACKGROUND Premature ovarian failure(POF)affects many adult women less than 40 years of age and leads to infertility.According to previous reports,various tissue-specific stem cells can restore ovarian function and folliculogenesis in mice with chemotherapy-induced POF.Human embryonic stem cells(ES)provide an alternative source for mesenchymal stem cells(MSCs)because of their similarities in phenotype and immunomodulatory and anti-inflammatory characteristics.Embryonic stem cell-derived mesenchymal stem cells(ES-MSCs)are attractive candidates for regenerative medicine because of their high proliferation and lack of barriers for harvesting tissue-specific MSCs.However,possible therapeutic effects and underlying mechanisms of transplanted ES-MSCs on cyclophosphamide and busulfan-induced mouse ovarian damage have not been evaluated.AIM To evaluate ES-MSCs vs bone marrow-derived mesenchymal stem cells(BMMSCs)in restoring ovarian function in a mouse model of chemotherapy-induced premature ovarian failure.METHODS Female mice received intraperitoneal injections of different doses of cyclophosphamide and busulfan to induce POF.Either human ES-MSCs or BMMSCs were transplanted into these mice.Ten days after the mice were injected with cyclophosphamide and busulfan and 4 wk after transplantation of the ESMSCs and/or BM-MSCs,we evaluated body weight,estrous cyclicity,folliclestimulating hormone and estradiol hormone concentrations and follicle count were used to evaluate the POF model and cell transplantation.Moreover,terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling,real-time PCR,Western blot analysis and immunohistochemistry and mating was used to evaluate cell transplantation.Enzyme-linked immunosorbent assay was used to analyze vascular endothelial growth factor,insulin-like growth factor 2 and hepatocyte growth factor levels in ES-MSC condition medium in order to investigate the mechanisms that underlie their function.RESULTS The human ES-MSCs significantly restored hormone secretion,survival rate and reproductive function in POF mice,which was similar to the results obtained with BM-MSCs.Gene expression analysis and the terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling assay results indicated that the ES-MSCs and/or BM-MSCs reduced apoptosis in the follicles.Notably,the transplanted mice generated new offspring.The results of different analyses showed increases in antiapoptotic and trophic proteins and genes.CONCLUSION These results suggested that transplantation of human ES-MSCs were similar to BM-MSCs in that they could restore the structure of the injured ovarian tissue and its function in chemotherapy-induced damaged POF mice and rescue fertility.The possible mechanisms of human ES-MSC were related to promotion of follicular development,ovarian secretion,fertility via a paracrine effect and ovarian cell survival.展开更多
AIM To investigate the role of embryonic liver fordin(ELF) in liver fibrosis by regulating hepatic stellate cells(HSCs) glucose glycolysis.METHODS The expression of ELF and the glucose glycolysisrelated proteins were ...AIM To investigate the role of embryonic liver fordin(ELF) in liver fibrosis by regulating hepatic stellate cells(HSCs) glucose glycolysis.METHODS The expression of ELF and the glucose glycolysisrelated proteins were evaluated in activated HSCs. si RNA was used to silence ELF expression in activated HSCs in vitro and the subsequent changes in PI3K/Akt signaling and glucose glycolysis-related proteins were observed.RESULTS The expression of ELF increased remarkably in HSCs of the fibrosis mouse model and HSCs that were cultured for 3 wk in vitro. Glucose glycolysis-related proteins showed an obvious increase in the activated HSCs, such as phosphofructokinase, platelet and glucose transporter 1. ELF-si RNA, which perfectly silenced the expression of ELF in activated HSCs, led to the induction of glucose glycolysis-related proteins and extracellular matrix(ECM) components. Moreover, p Akt, which is an important downstream factor in PI3K/Akt signaling, showed a significant change in response to the ELF silencing. The expression of glucose glycolysisrelated proteins and ECM components decreased remarkably when the PI3K/Akt signaling was blocked by Ly294002 in the activated HSCs. CONCLUSION ELF is involved in HSC glucose glycolysis by regulating PI3K/Akt signaling.展开更多
Pancreatic ductal adenocarcinoma(PDAC),the most common type of pancreatic tumor,is a highly aggressive human cancer with the lowest five-year survival rate of any human maligancy primarily due to its earlymetastasis a...Pancreatic ductal adenocarcinoma(PDAC),the most common type of pancreatic tumor,is a highly aggressive human cancer with the lowest five-year survival rate of any human maligancy primarily due to its earlymetastasis and lack of response to chemotherapy and radiation.Recent research suggests that PDAC cells comprise a hierarchy of tumor cells that develop around a population of cancer stem cells(CSCs),a small and distinct population of cancer cells that mediates tumoregenesis,metastasis and resistance to standard treatments.Thus,CSCs could be a target for more effective treatment options.Interestingly,pancreatic CSCs are subject to regulation by some of key embryonic stem cell(ESC)transctiption factors abberently expressed in PDAC,such as SOX2,OCT4 and NANOG.ESC transcription factors are important DNA-binding proteins present in both embryonic and adult somatic cells.The critical role of these factors in reprogramming processes makes them essential not only for embryonic development but also tumorigenesis.Here we provide an overview of stem cell transcription factors,particularly SOX2,OCT4,and NANOG,on their expression and function in pancreatic cancer.In contrast to embryonic stem cells,in which OCT4 and SOX2 are tightly regulated and physically interact to regulate a wide spectrum of target genes,de novo SOX2 expression alone in pancreatic cancer cells is sufficient to promote self-renewal,dedifferentiation and imparting stemness characteristics via impacting specific cell cycle regulatory genes and epithelial-mesnechymal transtion driver genes.Thus,targeting ESC factors,particularly SOX2,could be a worthy strategy for pancreatic cancer therapy.展开更多
BACKGROUND: Choosing proper donor cells is one of keys in experimental and clinical studies on cell replacement therapy (CRT) for treating Parkinson disease (PD). Embryonic mesencephalic precursor cells (MPCs) can sta...BACKGROUND: Choosing proper donor cells is one of keys in experimental and clinical studies on cell replacement therapy (CRT) for treating Parkinson disease (PD). Embryonic mesencephalic precursor cells (MPCs) can stably differentiate into dopaminergic neuron after in vitro proliferated culture. As compared with embryonic stem cell and neural stem cell strains, cell composition of embryonic MPCs after primary culture is also the most close to that of embryonic mesencephalic ventral cell suspension without proliferated culture. Successful experience accumulated in the latter suggests that primary cultured embryonic MPCs might be the most potential donor cells in clinical application with CRT for treating PD so far. OBJECTIVE: To investigate the feasibility of primary cultured embryonic precursor cells cultured primarily as donor cells in CRT for treating PD in rats. DESIGN: A randomized and controlled trial taking SD rats as experimental animals. SETTING: Department of Neurosurgery, Huashan Hospital Affiliated to Fudan University. MATERIALS: This experiment was carried out at the Institute of Neuroscience, Shanghai Institute for Biological Science, Chinese Academy of Sciences from July 2003 to June 2004. Totally 26 female SD rats, with body mass of 200 to 220 g, were provided by Shanghai Experimental Animal Center of Chinese Academy of Sciences. METHODS: Stereotaxic injection of 6-hydroxydopamine into the medial forebrain bundle were perfored to develop PD model rat. Among 26 SD rats, 20 rats achieved a more than 5 turns/min in apomorphine induced rotation test, reaching the standard of PD model rats. Immunohistochemical detection was performed on 1 out of 20 model rats after execution, and the other 19 rats were randomly divided into control group (n=5), sham transplantation group (n=5)and cell grafted group (n=9). Primary cultured E12 MPC cell suspension (1.2×1011 L-1)were used as donor cells. 4 μL primary cultured E12 MPC cell suspension prepared freshly was injected into the lesioned corpus striatum of rats in cell grafted group, and 4 μL D-Hank’s solution was injected in sham transplantation group in the same way. There was no injection in control group. Apomorphine-induced rotation rate of PD rats were recorded respectively in cell grafted group and sham transplantation group pre-operation (initial value) and at postoperative 2, 4, 6 and 16 weeks. Apomorphine-induced rotation rate of PD rats was recorded in control group at postoperative 2 months (initial value) and following 2,4,6 and 16 weeks. To determine TH antigen with immunohistological ABC method (DAB developing) at 6 months post-transplantation to investigate the differentiation and survival of donor cells in the host body. MAIN OUTCOME MEASURES: Apomorphine-induced rotation behavior before and after transplantation and the survival and differentiation of implanted cells in the host body at 6 months post-transplantation. RESULTS: Among 19 model rats, one rat died after transplantation respectively in the cell grafted group and sham transplantation group; finally 17 model rats entered the stage of result analysis. Relative apomorphine-induced rotation rate was significantly decreased in the cell grafted group as compared with that before transplantation, with significant difference (P < 0.01,P < 0.05);the mean value of relative apomorphine-induced rotation rate was significantly decreased at postoperative 16 weeks in cell grafted group as compared with that of corresponding relative rotation rate in control group, also with significant difference (P < 0.05).Immunohistological results showed that donor cells could differentiate into large and multi-polar dopaminergic neurons in the host body. CONCLUSION: Primary cultured embryonic MPCs can be used as the donor cells in CRT for treating PD.展开更多
Pluripotency of embryonic stem cells(ESCs) and induced pluripotent stem cells is regulated by a well characterized gene transcription circuitry. The circuitry is assembled by ESC specific transcription factors, signal...Pluripotency of embryonic stem cells(ESCs) and induced pluripotent stem cells is regulated by a well characterized gene transcription circuitry. The circuitry is assembled by ESC specific transcription factors, signal trans-ducing molecules and epigenetic regulators. Growing understanding of stem-like cells, albeit of more complex phenotypes, present in tumors(cancer stem cells), provides a common conceptual and research frame-work for basic and applied stem cell biology. In this review, we highlight current results on biomarkers, gene signatures, signaling pathways and epigenetic regulators that are common in embryonic and cancer stem cells. We discuss their role in determining the cell phenotype and finally, their potential use to design next generation biological and pharmaceutical approaches for regenerative medicine and cancer therapies.展开更多
Acute respiratory distress syndrome(ARDS)is one of the most fatal diseases worldwide.Pulmonary fibrosis occurs early in ARDS,and its severity plays a crucial role in ARDS mortality rate.Some studies suggested that fib...Acute respiratory distress syndrome(ARDS)is one of the most fatal diseases worldwide.Pulmonary fibrosis occurs early in ARDS,and its severity plays a crucial role in ARDS mortality rate.Some studies suggested that fibroproliferation is an essential mechanism in ARDS.Mitofusion2(Mfn2)overexpression plays a role in inhibiting cell proliferation.However,the role and potential mechanism of Mfn2 on the proliferation of fibroblasts is still unknown.In this study,we aimed at exploring the effect of Mfn2 on the human embryonic lung fibroblasts(HELF)and discussed its related mechanism.The HELF were treated with the Mfn2 overexpressing lentivirus(adv-Mfn2).The cell cycle was detected by flow cytometry.MTT,PCR and Western blotting were used to investigate the effect of Mfn2 on the proliferation of the HELF,collagen expression,the RAS-RAF-1-ERK1/2 pathway and the expression of cycle-related proteins(p21,p27,Rb,Raf-1,p-Raf-1,Erk1/2 and p-Erk1/2).The co-immunoprecipitation assay was used to explore the interaction between Mfn2 and Ras.The results showed that the overexpression of Mfn2 inhibited the proliferation of the HELF and induced the cell cycle arrest at the G0/G1 phase.Meanwhile,Mfn2 also inhibited the expression of collagen I,p-Erk and p-Raf-1.In addition,an interaction between Mfn2 and Ras existed in the HELF.This study suggests that the overexpression of Mfn2 can decrease the proliferation of HELF in ARDS,which was associated with the inhibition of the RAS-RAF-1-ERK1/2 pathway.The results may offer a potential therapeutic intervention for patients with ARDS.展开更多
AIM: To investigate the epigenetic states and expres- sion of imprinted genes in five human embryonic stem cell (hESC) lines derived in Taiwan. METHODS: The heterozygous alleles of single nucleo- tide polymorphisms (S...AIM: To investigate the epigenetic states and expres- sion of imprinted genes in five human embryonic stem cell (hESC) lines derived in Taiwan. METHODS: The heterozygous alleles of single nucleo- tide polymorphisms (SNPs) at imprinted genes were analyzed by sequencing genomic DNAs of hESC lines and the monoallelic expression of the imprinted genes were confirmed by sequencing the cDNAs. The expres- sion profiles of 32 known imprinted genes of five hESC lines were determined using Affymetrix human genome U133 plus 2.0 DNA microarray. RESULTS: The heterozygous alleles of SNPs at seven imprinted genes, IPW , PEG10 , NESP55 , KCNQ1 , ATP10A ,TCEB3C and IGF2 , were identified and the monoallelic expression of these imprinted genes except IGF2 were confirmed. The IGF2 gene was found to be imprinted in hESC line T2 but partially imprinted in line T3 and not imprinted in line T4 embryoid bodies. Ten imprinted genes, namely GRB10 , PEG10 , SGCE, MEST , SDHD , SN- RPN , SNURF , NDN , IPW and NESP55 , were found to be highly expressed in the undifferentiated hESC lines and down-regulated in differentiated derivatives. The UBE3A gene abundantly expressed in undifferentiated hESC lines and further up-regulated in differentiated tissues. The expression levels of other 21 imprinted genes were relatively low in undifferentiated hESC lines and five of these genes (TP73 , COPG2 , OSBPL5 , IGF2 and ATP10A ) were found to be up-regulated in differentiated tissues. CONCLUSION: The epigenetic states and expression of imprinted genes in hESC lines should be thoroughly studied after extended culture and upon differentiation in order to understand epigenetic stability in hESC lines before their clinical applications.展开更多
The study of embryonic stem cells is in the spotlight in many laboratories that study the structure and function of chromatin and epigenetic processes. The key properties of embryonic stem cells are their capacity for...The study of embryonic stem cells is in the spotlight in many laboratories that study the structure and function of chromatin and epigenetic processes. The key properties of embryonic stem cells are their capacity for selfrenewal and their pluripotency. Pluripotent stem cells are able to differentiate into the cells of all three germ layers, and because of this property they represent a promising therapeutic tool in the treatment of diseases such as Parkinson's disease and diabetes, or in the healing of lesions after heart attack. As the basic nuclear unit, chromatin is responsible for the regulation of the functional status of cells, including pluripotency and differentiation. Therefore, in this review we discuss the functional changes in chromatin during differentiation and the correlation between epigenetics events and the differentiation potential of embryonic stem cells. In particular we focus on post-translational histone modification, DNA methylation and the heterochromatin protein HP1 and its unique function in mouse and human embryonic stem cells.展开更多
基金Supported by the Shandong Provincial Traditional Chinese Medicine Science and Technology Development Program,No.C-262the 2021 Science and Technology Innovation Research Project of Shandong Maternal and Child Health Association,No.2021-19-24.
文摘BACKGROUND This study aimed to explore the relationship between gene mutations and early embryonic development arrest and to provide more possibilities for the diagnosis and treatment of repeated implantation failure.CASE SUMMARY Here,we collected and described the clinical data of a patient with early embryonic development stagnation after repeated in vitro fertilization attempts for primary infertility at the Department Reproductive Center of Zaozhuang Maternal and Child Healthcare Hospital.We also detected the whole-exon gene of the patient's spouse and parents,and conducted bioinformatics analysis to determine the pathogenesis of the gene.CONCLUSION A novel mutant of the TUBB8 gene[c.602G>T(p.C201F)]was identified,and this mutant provided new data on the genotype-phenotype relationships of related diseases.
基金sponsored by the National Key R&D Program of China(2022YFD1301800 and1300400)National Natural Science Foundation of China(31802080 and 3197200131)+1 种基金Key Open Laboratory of Chinese Veterinary Medicine of State Ethnic Affairs Commission&National Local Joint Engineering Research Centre for the Separation and Purification Technology of Ethnic Chinese Veterinary Medicine([2022]09)Guangdong Provincial Science and Technology Special Foundation(210723106900762 and 2021020103-2)。
文摘Background Mitochondrial dysfunction induced by excessive mitochondrial reactive oxygen species(ROS)damages embryonic development and leads to growth arrest.Objective The purpose of this study is to elucidate whether maternal zinc(Zn)exert protective effect on oxidative stress targeting mitochondrial function using an avian model.Result In ovo injected tert-butyl hydroperoxide(BHP)increases(P<0.05)hepatic mitochondrial ROS,malondialdehyde(MDA)and 8-hydroxy-2-deoxyguanosine(8-OHdG),and decreases(P<0.05)mitochondrial membrane potential(MMP),mitochondrial DNA(mtDNA)copy number and adenosine triphosphate(ATP)content,contributing to mitochondrial dysfunction.In vivo and in vitro studies revealed that Zn addition enhances(P<0.05)ATP synthesis and metallothionein 4(MT4)content and expression as well as alleviates(P<0.05)the BHP-induced mitochondrial ROS generation,oxidative damage and dysfunction,exerting a protective effect on mitochondrial function by enhancing antioxidant capacity and upregulating the mRNA and protein expressions of Nrf2 and PGC-1α.Conclusions The present study provides a new way to protect offspring against oxidative damage by maternal Zn supplementation through the process of targeting mitochondria involving the activation of Nrf2/PGC-1αsignaling.
基金funded by the National Natural Science Foundation of China(No.82070376 and No.81873491)the Natural Science Foundation of Zhejiang Province(No.LY21H020005)+1 种基金the Zhejiang Medical Science and Technology Project(No.2019KY376 and No.2018KY071)a Ningbo Science and Technology Project(No.202002N3173).
文摘Objective Vascular smooth muscle cell(VSMC)differentiation from stem cells is one source of the increasing number of VSMCs that are involved in vascular remodeling-related diseases such as hypertension,atherosclerosis,and restenosis.MicroRNA-146a(miR-146a)has been proven to be involved in cell proliferation,migration,and tumor metabolism.However,little is known about the functional role of miR-146a in VSMC differentiation from embryonic stem cells(ESCs).This study aimed to determine the role of miR-146a in VSMC differentiation from ESCs.Methods Mouse ESCs were differentiated into VSMCs,and the cell extracts were analyzed by Western blotting and RT-qPCR.In addition,luciferase reporter assays using ESCs transfected with miR-146a/mimic and plasmids were performed.Finally,C57BL/6J female mice were injected with mimic or miR-146a-overexpressing ESCs,and immunohistochemistry,Western blotting,and RT-qPCR assays were carried out on tissue samples from these mice.Results miR-146a was significantly upregulated during VSMC differentiation,accompanied with the VSMC-specific marker genes smooth muscle-alpha-actin(SMαA),smooth muscle 22(SM22),smooth muscle myosin heavy chain(SMMHC),and h1-calponin.Furthermore,overexpression of miR-146a enhanced the differentiation process in vitro and in vivo.Concurrently,the expression of Kruppel-like factor 4(KLF4),predicted as one of the top targets of miR-146a,was sharply decreased in miR-146a-overexpressing ESCs.Importantly,inhibiting KLF4 expression enhanced the VSMC-specific gene expression induced by miR-146a overexpression in differentiating ESCs.In addition,miR-146a upregulated the mRNA expression levels and transcriptional activity of VSMC differentiation-related transcription factors,including serum response factor(SRF)and myocyte enhancer factor 2c(MEF-2c).Conclusion Our data support that miR-146a promotes ESC-VSMC differentiation through regulating KLF4 and modulating the transcription factor activity of VSMCs.
基金financially supported by the Science and Technology Major Project of the Inner Mongolia Autonomous Region of China(2020ZD0007)the Major Program of the Inner Mongolia Natural Science Foundation,China(2020ZD10)+3 种基金the National Natural Science Foundation of China(32160172)the Natural Science Foundation of Inner Mongolia Autonomous Region(2020BS03003 and 2020BS03022)the National Transgenic Project of China(2016ZX0801000-002 and 2016ZX08010005-001)the Science and Technology Major Project of the Inner Mongolia Autonomous Region of China(zdzx2018065)。
文摘Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.
基金supported by the National Key Research and Development Program of China(2022YFD1301300).
文摘Background Fertility declines in high-parity sows.This study investigated whether parity-dependent declines in embryonic survival and reproductive performance could be restored by dietary coenzyme Q10(CoQ10)supplementation.Methods Two experiments were performed.In Exp.1,30 young sows that had completed their 2nd parity and 30 high-parity sows that had completed their 10^(th)parity,were fed either a control diet(CON)or a CON diet supple-mented with 1 g/kg CoQ10(+CoQ10)from mating until slaughter at day 28 of gestation.In Exp.2,a total of 314 post-weaning sows with two to nine parities were fed the CON or+CoQ10 diets from mating throughout gestation.Results In Exp.1,both young and high-parity sows had a similar number of corpora lutea,but high-parity sows had lower plasma CoQ10 concentrations,down-regulated genes involved with de novo CoQ10 synthesis in the endome-trium tissues,and greater levels of oxidative stress markers in plasma and endometrium tissues.High-parity sows had fewer total embryos and alive embryos,lower embryonic survival,and greater embryo mortality than young sows.Dietary CoQ10 supplementation increased the number of live embryos and the embryonic survival rate to levels simi-lar to those of young sows,as well as lowering the levels of oxidative stress markers.In Exp.2,sows showed a parity-dependent decline in plasma CoQ10 levels,and sows with more than four parities showed a progressive decline in the number of total births,live births,and piglets born effective.Dietary supplementation with CoQ10 increased the number of total births,live births,and born effective,and decreased the intra-litter covariation coefficients and the percentage of sows requiring farrowing assistance during parturition.Conclusions Dietary CoQ10 supplementation can improve the embryonic survival and reproductive performance of gestating sows with high parity,probably by improving the development of uterine function.
基金funded by the National Natural Science Foundation of China (31972996 and 32160790)Guangxi Bagui Scholar ProgramGuangxi Innovation-Driven Development Project (AA17204051)
文摘Background During mammalian pre-implantation embryonic development(PED),the process of maternal-to-zygote transition(MZT)is well orchestrated by epigenetic modification and gene sequential expression,and it is related to the embryonic genome activation(EGA).During MZT,the embryos are sensitive to the environment and easy to arrest at this stage in vitro.However,the timing and regulation mechanism of EGA in buffaloes remain obscure.Results Buffalo pre-implantation embryos were subjected to trace cell based RNA-seq and whole-genome bisulfite sequencing(WGBS)to draw landscapes of transcription and DNA-methylation.Four typical developmental steps were classified during buffalo PED.Buffalo major EGA was identified at the 16-cell stage by the comprehensive analy-sis of gene expression and DNA methylation dynamics.By weighted gene co-expression network analysis,stage-spe-cific modules were identified during buffalo maternal-to-zygotic transition,and key signaling pathways and biological process events were further revealed.Programmed and continuous activation of these pathways was necessary for success of buffalo EGA.In addition,the hub gene,CDK1,was identified to play a critical role in buffalo EGA.Conclusions Our study provides a landscape of transcription and DNA methylation in buffalo PED and reveals deeply the molecular mechanism of the buffalo EGA and genetic programming during buffalo MZT.It will lay a foundation for improving the in vitro development of buffalo embryos.
文摘Objective:To compare embryonic development,ploidy status and clinical outcomes between fresh and frozen-thawed oocytes.Methods:This retrospective cohort study evaluated 83 fertilization cycles including both fresh and frozen oocytes from 79 patients at the HP Fertility Center of Hai Phong International Hospital of Obstetrics and Pediatrics in Vietnam.The patient underwent several ovarian stimulation cycles to accumulate a certain number of oocytes that would be vitrified.In the last oocyte retrieval,all patient’s oocytes including both frozen and fresh would be fertilized.The outcomes included the rates of oocyte survival,cleavage embryo,blastocyst,ploidy status,pregnancy,biochemical pregnancy and clinical pregnancy.Results:The oocyte survival rate after thawing was 96.5%.No statistically significant difference was found when comparing fresh and frozen oocytes regarding fertilization rate(78.1%vs.75.5%,P=0.461),usable cleavage embryo rate(86.9%vs.87.2%,P=0.916)but usable blastocyst rate was found higher statistically in the frozen oocyte group(44.4%vs.54.0%,P=0.049).The percentages of euploid,aneuploid and mosaic embryos between the fresh group and the vitrified group had no significant differences(33.8%vs.31.6%,P=0.682;51.0%vs.54.2%,P=0.569;15.2%vs.12.4%,P=0.787;respectively).The rates of pregnancy,biochemical pregnancy and clinical pregnancy had no statistical difference(68.8%vs.64.8%,P=0.764;12.5%vs.3.6%,P=0.258;37.5%vs.46.4%,P=0.565).17 Mature oocytes are the minimum to have at least one euploid embryo.Conclusions:Oocyte vitrification does not affect embryonic,genetic and clinical results.The number of mature oocytes should be considered for fertilization in some cases.
文摘This paper describes a new species of the snout moth Berastagia tainanica sp. nov. (Lepidoptera: Pyralidae) from Taiwan. From 2009 to 2016, a biology study was conducted on population dynamics and embryonic development. Spring season is the peak of the eclosion of overwintering larvae or pupae. The average longevity of adult was 14.8 ± 6.2 days (N = 174), the average number of eggs laid was 259 ± 3 eggs/moth (N = 2), the hatching rate of eggs was 95.4% (N = 262), and the average hatching time of eggs was 99.6 ± 18.6 hours (N = 68). The average body length of males was 5.64 mm ± 0.91 mm (N = 30), and the average body length of females was 6.28 mm ± 0.84 mm (N = 30). This finding indicates that female snout moths are larger than males (Global R = 0.058, P = 0.012). The snout moth eclosion rate was 16.9 moths/100 pods in the first year (2010/2011, N = 2,224 pods) and 10.9 moths/100 pods in the second year (2014/2015, N = 6,382 pods). The pod borer rate was 31.8% (N = 707) and the seed borer rate was 41.2% (N = 3,628) in the first year, whereas the pod borer rate was 76.2% (N = 6,382) in the second year.
文摘During in vitro fertilization, human embryos are incubated without light, and these conditions do not ensure embryo survival. This study explored whether environmental conditions can influence the embryo viability rates of the house cricket, Acheta domesticus. In particular, the experiment tested what colors of visible light provide the best incubation conditions to ensure cricket embryo viability. The concept was to use house cricket embryos to represent human embryos. Cricket embryos were chosen as their eggs have soft outer membrane casings and resemble human embryos during the first few days after fertilization. During the experiment, the adult crickets laid their eggs into one of six soil-filled boxes called substrates. Each substrate was placed into one of six storage containers filled with adult crickets and lit with a different colored visible light (red, yellow, green, blue, white, or no light). After two days of breeding, the egg-filled substrates were removed from the adult crickets and placed in another storage container of the same color light. After incubation under heat-emitting lamps and under one of six light colors, nymphs were counted after hatching to determine embryo viability. After three trials, the red light provided the significantly highest viability rate, with yellow and no light being comparable seconds. The green, blue, and white lights showed significantly lower viability rates than no visible light. My results raise the speculation that exposing fertilized mammal eggs to visible light colors might have the same effects during the in vitro fertilization process.
文摘Objective:To understand the clinical characteristics of patients with embryonic posterior cerebral artery and its correlation with abnormal vascular development.Methods:The clinical data of 396 patients with embryonic posterior cerebral artery confirmed by magnetic resonance angiography(MRA)and computed tomography angiography(CTA)were analyzed.Results:Two-hundred patients had clinical manifestations of posterior circulation ischemia,including recurrent dizziness,vertigo,and tinnitus;45 had headaches,97 had limb weakness,and 16 patients had syncope or impaired consciousness.Seventy-six patients with circulatory infarction were admitted to the hospital.There were 251 patients with history of hypertension,74 with diabetes,113 with hyperlipidemia,13 with dominant vertebral artery,10 with intracranial aneurysm,and 19 with absence of A1 segment of the anterior cerebral artery(considering developmental variation).Conclusion:Embryonic posterior cerebral artery develops abnormally during the embryonic period,often accompanied by abnormal vascular access.Due to abnormal hemodynamics,the incidence of posterior circulation ischemia,aneurysm,and infarction increases in such patients.
文摘Background: The endometrium is the tissue that lines the inside of the uterus. It undergoes multiple changes during the menstrual cycle and prepares to welcome the fertilized egg. Endovaginal ultrasound is a first-line examination in the assessment of female infertility. Objective: We aimed to determine the impact of endometrial status by endovaginal ultrasound on the outcome of embryo transfers. Subjects and Methods: This was a prospective cross-sectional study of 150 women collected between January and October 2020 at the “Le Diafounou” medically assisted procreation laboratory. Were included in the study, women presenting for desire of pregnancy and having continued the treatment until the transfer of fresh or frozen embryos. An uterine score was used. This score took into account the thickness of endometrium, it is coffee bean aspect, the presence of the notch, the echogenicity of the endometrium, the pulsatility index, the presence of the flow at the end of diastole and sub-endometrial flow. Ultrasounds were performed using a General Electric logic 500, logic 9 and Voluson E8 device. The data was entered into Excel and then analyzed using SPSS version 21. Results: 150 women were selected for the study. 83 women or 55.3% were between 17 and 35 years old and 44.7% were over 35 years old with an upper limit of 50 years old. The number of women who became pregnant was 69% or 46%. The average thickness of the endometrium was 11.40 mm with extremes of 4.65 mm and 18.6 mm. There was a correlation between the thickness of the endometrium and obtaining a pregnancy (p i.e. 88.7%. The pulsatility index was relatively low for those who started a pregnancy, i.e. 90%. The high pulsatility index is one of the reasons for the failure of embryo implantation. The correlation was significant (p = 0.009). 141 women had no notch in the uterine arteries, i.e. 94%. 94 women or 62.7% had a triple line endometrium or typical coffee bean appearance. End-diastolic flow was observed in 127 women, i.e. 84.7%. The sub-endometrial flow was found in 128 women or 85.3%. Conclusion: The Knowledge of the status of the endometrium is essential and has an impact on the outcome of embryo transfers. The thinner (7 and <15). Doppler factors also play an important role.
基金Supported by National Science Council,No.NSC101-2311-B-003-005 and NSC102-2311-B-003-003National Taiwan Normal University,No.103T3040B06,103T3040C06 and 104T3040C06
文摘Neurons derived from embryonic stem cells(ESCs) have gained great merit in both basic research and regenerative medicine. Here we review and summarize the signaling pathways that have been reported to be involved in the neuronal differentiation of ESCs,particularly those associated with in vitro differentiation. The inducers and pathways explored include retinoic acid, Wnt/b-catenin, transforming growth factor/bone morphogenetic protein, Notch, fibroblast growth factor, cytokine, Hedgehog, c-Jun N-terminal kinase/mitogen-activated protein kinase and others. Some other miscellaneous molecular factors that have been reported in the literature are also summarized and discussed. These include calcium, calcium receptor, calcineurin, estrogen receptor, Hox protein, ceramide, glycosaminioglycan, ginsenoside Rg1, opioids, two pore channel 2, nitric oxide, chemically defined medium, cellcell interactions, and physical stimuli. The interaction or crosstalk between these signaling pathways and factors will be explored. Elucidating these signals in detail should make a significant contribution to future progress in stem cell biology and allow, for example, better comparisons to be made between differentiation in vivo and in vitro. Of equal importance, a comprehensive understanding of the pathways that are involved in the development of neurons from ESCs in vitro will also accelerate their application as part of translational medicine.
文摘BACKGROUND Premature ovarian failure(POF)affects many adult women less than 40 years of age and leads to infertility.According to previous reports,various tissue-specific stem cells can restore ovarian function and folliculogenesis in mice with chemotherapy-induced POF.Human embryonic stem cells(ES)provide an alternative source for mesenchymal stem cells(MSCs)because of their similarities in phenotype and immunomodulatory and anti-inflammatory characteristics.Embryonic stem cell-derived mesenchymal stem cells(ES-MSCs)are attractive candidates for regenerative medicine because of their high proliferation and lack of barriers for harvesting tissue-specific MSCs.However,possible therapeutic effects and underlying mechanisms of transplanted ES-MSCs on cyclophosphamide and busulfan-induced mouse ovarian damage have not been evaluated.AIM To evaluate ES-MSCs vs bone marrow-derived mesenchymal stem cells(BMMSCs)in restoring ovarian function in a mouse model of chemotherapy-induced premature ovarian failure.METHODS Female mice received intraperitoneal injections of different doses of cyclophosphamide and busulfan to induce POF.Either human ES-MSCs or BMMSCs were transplanted into these mice.Ten days after the mice were injected with cyclophosphamide and busulfan and 4 wk after transplantation of the ESMSCs and/or BM-MSCs,we evaluated body weight,estrous cyclicity,folliclestimulating hormone and estradiol hormone concentrations and follicle count were used to evaluate the POF model and cell transplantation.Moreover,terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling,real-time PCR,Western blot analysis and immunohistochemistry and mating was used to evaluate cell transplantation.Enzyme-linked immunosorbent assay was used to analyze vascular endothelial growth factor,insulin-like growth factor 2 and hepatocyte growth factor levels in ES-MSC condition medium in order to investigate the mechanisms that underlie their function.RESULTS The human ES-MSCs significantly restored hormone secretion,survival rate and reproductive function in POF mice,which was similar to the results obtained with BM-MSCs.Gene expression analysis and the terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling assay results indicated that the ES-MSCs and/or BM-MSCs reduced apoptosis in the follicles.Notably,the transplanted mice generated new offspring.The results of different analyses showed increases in antiapoptotic and trophic proteins and genes.CONCLUSION These results suggested that transplantation of human ES-MSCs were similar to BM-MSCs in that they could restore the structure of the injured ovarian tissue and its function in chemotherapy-induced damaged POF mice and rescue fertility.The possible mechanisms of human ES-MSC were related to promotion of follicular development,ovarian secretion,fertility via a paracrine effect and ovarian cell survival.
基金Supported by National Natural Science Foundation of China,No.81300329 and No.81401992
文摘AIM To investigate the role of embryonic liver fordin(ELF) in liver fibrosis by regulating hepatic stellate cells(HSCs) glucose glycolysis.METHODS The expression of ELF and the glucose glycolysisrelated proteins were evaluated in activated HSCs. si RNA was used to silence ELF expression in activated HSCs in vitro and the subsequent changes in PI3K/Akt signaling and glucose glycolysis-related proteins were observed.RESULTS The expression of ELF increased remarkably in HSCs of the fibrosis mouse model and HSCs that were cultured for 3 wk in vitro. Glucose glycolysis-related proteins showed an obvious increase in the activated HSCs, such as phosphofructokinase, platelet and glucose transporter 1. ELF-si RNA, which perfectly silenced the expression of ELF in activated HSCs, led to the induction of glucose glycolysis-related proteins and extracellular matrix(ECM) components. Moreover, p Akt, which is an important downstream factor in PI3K/Akt signaling, showed a significant change in response to the ELF silencing. The expression of glucose glycolysisrelated proteins and ECM components decreased remarkably when the PI3K/Akt signaling was blocked by Ly294002 in the activated HSCs. CONCLUSION ELF is involved in HSC glucose glycolysis by regulating PI3K/Akt signaling.
基金Supported by Universidad del Pais Vasco,Instituto Biodonostia,San Sebastian,and CIBERehd(red de enfermedades hepaticas y digestivas)American Cancer Society institutional awardMayo Clinic Pancreatic Cancer SPORE,No.CA102701
文摘Pancreatic ductal adenocarcinoma(PDAC),the most common type of pancreatic tumor,is a highly aggressive human cancer with the lowest five-year survival rate of any human maligancy primarily due to its earlymetastasis and lack of response to chemotherapy and radiation.Recent research suggests that PDAC cells comprise a hierarchy of tumor cells that develop around a population of cancer stem cells(CSCs),a small and distinct population of cancer cells that mediates tumoregenesis,metastasis and resistance to standard treatments.Thus,CSCs could be a target for more effective treatment options.Interestingly,pancreatic CSCs are subject to regulation by some of key embryonic stem cell(ESC)transctiption factors abberently expressed in PDAC,such as SOX2,OCT4 and NANOG.ESC transcription factors are important DNA-binding proteins present in both embryonic and adult somatic cells.The critical role of these factors in reprogramming processes makes them essential not only for embryonic development but also tumorigenesis.Here we provide an overview of stem cell transcription factors,particularly SOX2,OCT4,and NANOG,on their expression and function in pancreatic cancer.In contrast to embryonic stem cells,in which OCT4 and SOX2 are tightly regulated and physically interact to regulate a wide spectrum of target genes,de novo SOX2 expression alone in pancreatic cancer cells is sufficient to promote self-renewal,dedifferentiation and imparting stemness characteristics via impacting specific cell cycle regulatory genes and epithelial-mesnechymal transtion driver genes.Thus,targeting ESC factors,particularly SOX2,could be a worthy strategy for pancreatic cancer therapy.
文摘BACKGROUND: Choosing proper donor cells is one of keys in experimental and clinical studies on cell replacement therapy (CRT) for treating Parkinson disease (PD). Embryonic mesencephalic precursor cells (MPCs) can stably differentiate into dopaminergic neuron after in vitro proliferated culture. As compared with embryonic stem cell and neural stem cell strains, cell composition of embryonic MPCs after primary culture is also the most close to that of embryonic mesencephalic ventral cell suspension without proliferated culture. Successful experience accumulated in the latter suggests that primary cultured embryonic MPCs might be the most potential donor cells in clinical application with CRT for treating PD so far. OBJECTIVE: To investigate the feasibility of primary cultured embryonic precursor cells cultured primarily as donor cells in CRT for treating PD in rats. DESIGN: A randomized and controlled trial taking SD rats as experimental animals. SETTING: Department of Neurosurgery, Huashan Hospital Affiliated to Fudan University. MATERIALS: This experiment was carried out at the Institute of Neuroscience, Shanghai Institute for Biological Science, Chinese Academy of Sciences from July 2003 to June 2004. Totally 26 female SD rats, with body mass of 200 to 220 g, were provided by Shanghai Experimental Animal Center of Chinese Academy of Sciences. METHODS: Stereotaxic injection of 6-hydroxydopamine into the medial forebrain bundle were perfored to develop PD model rat. Among 26 SD rats, 20 rats achieved a more than 5 turns/min in apomorphine induced rotation test, reaching the standard of PD model rats. Immunohistochemical detection was performed on 1 out of 20 model rats after execution, and the other 19 rats were randomly divided into control group (n=5), sham transplantation group (n=5)and cell grafted group (n=9). Primary cultured E12 MPC cell suspension (1.2×1011 L-1)were used as donor cells. 4 μL primary cultured E12 MPC cell suspension prepared freshly was injected into the lesioned corpus striatum of rats in cell grafted group, and 4 μL D-Hank’s solution was injected in sham transplantation group in the same way. There was no injection in control group. Apomorphine-induced rotation rate of PD rats were recorded respectively in cell grafted group and sham transplantation group pre-operation (initial value) and at postoperative 2, 4, 6 and 16 weeks. Apomorphine-induced rotation rate of PD rats was recorded in control group at postoperative 2 months (initial value) and following 2,4,6 and 16 weeks. To determine TH antigen with immunohistological ABC method (DAB developing) at 6 months post-transplantation to investigate the differentiation and survival of donor cells in the host body. MAIN OUTCOME MEASURES: Apomorphine-induced rotation behavior before and after transplantation and the survival and differentiation of implanted cells in the host body at 6 months post-transplantation. RESULTS: Among 19 model rats, one rat died after transplantation respectively in the cell grafted group and sham transplantation group; finally 17 model rats entered the stage of result analysis. Relative apomorphine-induced rotation rate was significantly decreased in the cell grafted group as compared with that before transplantation, with significant difference (P < 0.01,P < 0.05);the mean value of relative apomorphine-induced rotation rate was significantly decreased at postoperative 16 weeks in cell grafted group as compared with that of corresponding relative rotation rate in control group, also with significant difference (P < 0.05).Immunohistological results showed that donor cells could differentiate into large and multi-polar dopaminergic neurons in the host body. CONCLUSION: Primary cultured embryonic MPCs can be used as the donor cells in CRT for treating PD.
文摘Pluripotency of embryonic stem cells(ESCs) and induced pluripotent stem cells is regulated by a well characterized gene transcription circuitry. The circuitry is assembled by ESC specific transcription factors, signal trans-ducing molecules and epigenetic regulators. Growing understanding of stem-like cells, albeit of more complex phenotypes, present in tumors(cancer stem cells), provides a common conceptual and research frame-work for basic and applied stem cell biology. In this review, we highlight current results on biomarkers, gene signatures, signaling pathways and epigenetic regulators that are common in embryonic and cancer stem cells. We discuss their role in determining the cell phenotype and finally, their potential use to design next generation biological and pharmaceutical approaches for regenerative medicine and cancer therapies.
基金This project was supported by Wuhan Medical Science Foundation of China(No.WX17B07,No.WX19A09,and No.WJ2019H324).
文摘Acute respiratory distress syndrome(ARDS)is one of the most fatal diseases worldwide.Pulmonary fibrosis occurs early in ARDS,and its severity plays a crucial role in ARDS mortality rate.Some studies suggested that fibroproliferation is an essential mechanism in ARDS.Mitofusion2(Mfn2)overexpression plays a role in inhibiting cell proliferation.However,the role and potential mechanism of Mfn2 on the proliferation of fibroblasts is still unknown.In this study,we aimed at exploring the effect of Mfn2 on the human embryonic lung fibroblasts(HELF)and discussed its related mechanism.The HELF were treated with the Mfn2 overexpressing lentivirus(adv-Mfn2).The cell cycle was detected by flow cytometry.MTT,PCR and Western blotting were used to investigate the effect of Mfn2 on the proliferation of the HELF,collagen expression,the RAS-RAF-1-ERK1/2 pathway and the expression of cycle-related proteins(p21,p27,Rb,Raf-1,p-Raf-1,Erk1/2 and p-Erk1/2).The co-immunoprecipitation assay was used to explore the interaction between Mfn2 and Ras.The results showed that the overexpression of Mfn2 inhibited the proliferation of the HELF and induced the cell cycle arrest at the G0/G1 phase.Meanwhile,Mfn2 also inhibited the expression of collagen I,p-Erk and p-Raf-1.In addition,an interaction between Mfn2 and Ras existed in the HELF.This study suggests that the overexpression of Mfn2 can decrease the proliferation of HELF in ARDS,which was associated with the inhibition of the RAS-RAF-1-ERK1/2 pathway.The results may offer a potential therapeutic intervention for patients with ARDS.
基金Supported by National Program for Genomic Medicine GrantsNSC95/96/97-3112-B-037-002 of National Science Council inTaiwan (to Li SS)a Chair Professorship of The Medical Educationand Development Foundation of Kaohsiung Medical University (to Li SS)
文摘AIM: To investigate the epigenetic states and expres- sion of imprinted genes in five human embryonic stem cell (hESC) lines derived in Taiwan. METHODS: The heterozygous alleles of single nucleo- tide polymorphisms (SNPs) at imprinted genes were analyzed by sequencing genomic DNAs of hESC lines and the monoallelic expression of the imprinted genes were confirmed by sequencing the cDNAs. The expres- sion profiles of 32 known imprinted genes of five hESC lines were determined using Affymetrix human genome U133 plus 2.0 DNA microarray. RESULTS: The heterozygous alleles of SNPs at seven imprinted genes, IPW , PEG10 , NESP55 , KCNQ1 , ATP10A ,TCEB3C and IGF2 , were identified and the monoallelic expression of these imprinted genes except IGF2 were confirmed. The IGF2 gene was found to be imprinted in hESC line T2 but partially imprinted in line T3 and not imprinted in line T4 embryoid bodies. Ten imprinted genes, namely GRB10 , PEG10 , SGCE, MEST , SDHD , SN- RPN , SNURF , NDN , IPW and NESP55 , were found to be highly expressed in the undifferentiated hESC lines and down-regulated in differentiated derivatives. The UBE3A gene abundantly expressed in undifferentiated hESC lines and further up-regulated in differentiated tissues. The expression levels of other 21 imprinted genes were relatively low in undifferentiated hESC lines and five of these genes (TP73 , COPG2 , OSBPL5 , IGF2 and ATP10A ) were found to be up-regulated in differentiated tissues. CONCLUSION: The epigenetic states and expression of imprinted genes in hESC lines should be thoroughly studied after extended culture and upon differentiation in order to understand epigenetic stability in hESC lines before their clinical applications.
基金Supported by Grants P302/12/G157 and 13-07822S from the Grant Agency of the Czech Republicby COST-CZ project LD11020 of the Ministry of Education Youth and Sport of the Czech RepublicBártová E is a coordinator of the EU Marie Curie Project PIRSES-GA-2010-269156-LCS
文摘The study of embryonic stem cells is in the spotlight in many laboratories that study the structure and function of chromatin and epigenetic processes. The key properties of embryonic stem cells are their capacity for selfrenewal and their pluripotency. Pluripotent stem cells are able to differentiate into the cells of all three germ layers, and because of this property they represent a promising therapeutic tool in the treatment of diseases such as Parkinson's disease and diabetes, or in the healing of lesions after heart attack. As the basic nuclear unit, chromatin is responsible for the regulation of the functional status of cells, including pluripotency and differentiation. Therefore, in this review we discuss the functional changes in chromatin during differentiation and the correlation between epigenetics events and the differentiation potential of embryonic stem cells. In particular we focus on post-translational histone modification, DNA methylation and the heterochromatin protein HP1 and its unique function in mouse and human embryonic stem cells.
