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Semaphorin 3A-hypoxia inducible factor 1 subunit alpha co-overexpression enhances the osteogenic differentiation of Induced pluripotent stem cells-derived mesenchymal stem cells in vitro

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Chinese Medical Journal 2020年第3期133卷 301-309页

作者： Jing-Yi Li Ting-Ting Wang Chong Li Zhi-Fang Wang Shan Li Li Ma Li-Li Zheng Department of Plastic Surgery Capital Medical University Affiliated Beijing Tiantan HospitalBeijing 100050China Department of Endocrinology and Metabolism The First Affiliated Hospital of Zhengzhou UniversityZhengzhouHenan 450000China Department of Plastic Surgery China-Japan Friendship HospitalBeijing 100029China.

Background:Mesenchymal stem or stromal cells(MSCs)derived from the Induced pluripotent stem cells(iPSCs)have uniform biological activity,which makes the clinical application of MSCs in bone repair possible.Culturing the iPSC-MSCs onto osteoconductive materials is a promising tissue engineering-based strategy in bone regeneration.The aim of this work was to evaluate the effects of semaphorin 3A(Sema3A)and hypoxia inducible factor 1 subunit alpha(HIF1α)co-overexpression on the survival and osteogenic differentiation of iPSC-MSCs.Methods:Sema3A and HIF1αwere linked together with the three(GGGGS;G,glycine;S,serine)peptide fragment,and their co-expression in iPSC-MSCs was mediated by a lentiviral vector.The fusion protein retained the immune reactivity for both Sema3A and HIF1αas determined with Western blotting.iPSC-MSCs were infected with overexpression lentivirus(oeLenti)as negative control,oeLenti-Sema3A,oeLenti-HIF1αor oeLenti-Sema3A-HIF1αlentiviruses.Results:Sema3A overexpression alone promoted the osteogenic differentiation of iPSC-MSCs(the activity and/or expression of osteoblast markers,such as alkaline phosphatase,osteopontin,and osteocalcin,were upregulated),and suppressed cell survival.The Sema3A-HIF1αfusion protein showed a comparable osteoconductive effect to that of Sema3A without reducing cell survival.We further seeded iPSC-MSCs modified by SemaA-HIF1αoverexpression onto hydroxyapatite(HA)scaffolds,and evaluated their growth and differentiation on this three-dimensional material.Additional data indicated that,as compared to iPSC-MSCs cultured in ordinary two-dimensional dishes,cells cultured in HA scaffolds grew(blank vs.HA scaffolds:0.83 vs.1.39 for survival)and differentiated better(blank vs.HA scaffolds:11.29 vs.16.62 for alkaline phosphatase activity).Conclusion:Modifying iPSC-MSCs with pro-osteogenic(Sema3A)and pro-survival(HIF1α)factors may represent a promising strategy to optimize tissue engineering-based strategy in bone repair.

关键词： Semaphorin 3A Hypoxia inducible factor 1 subunit alpha Induced pluripotent stem cells Mesenchymal stems Cell survival Osteogenic differentiation Hydroxyapatite scaffolds

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Transfer of mitochondria from mesenchymal stem cells derived from Induced pluripotent stem cells attenuates hypoxia-ischemia-Induced mitochondrial dysfunction in PC12 cells

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Neural Regeneration Research 2020年第3期15卷 464-472页

作者： Yan Yang Gen Ye Yue-Lin Zhang Hai-Wei He Bao-Qi Yu Yi-Mei Hong Wei You Xin Li Department of Emergency Medicine Department of Emergency and Critical Care MedicineGuangdong Provincial People’s HospitalGuangdong Academy of Medical SciencesGuangzhouGuangdong ProvinceChina Department of Emergency the First Affiliated HospitalSun Yat-sen UniversityGuangzhouGuangdong ProvinceChina Shantou University Medical College ShantouGuangdong ProvinceChina Department of Physiology and Pathophysiology School of Basic Medical SciencesCapital Medical UniversityKey Laboratory of Remodelling-related Cardiovascular DiseasesMinistry of EducationBeijingChina

Mitochondrial dysfunction in neurons has been implicated in hypoxia-ischemia-Induced brain injury.Although mesenchymal stem cell therapy has emerged as a novel treatment for this pathology,the mechanisms are not fully understood.To address this issue,we first co-cultured 1.5×10^5 PC12 cells with mesenchymal stem cells that were derived from Induced pluripotent stem cells at a ratio of 1:1,and then intervened with cobalt chloride(CoCl2)for 24 hours.Reactive oxygen species in PC12 cells was measured by Mito-sox.Mitochondrial membrane potential(ΔΨm)in PC12 cells was determined by JC-1 staining.Apoptosis of PC12 cells was detected by terminal deoxynucleotidal transferase-mediated dUTP nick end-labeling staining.Mitochondrial morphology in PC12 cells was examined by transmission electron microscopy.Transfer of mitochondria from the mesenchymal stem cells derived from Induced pluripotent stem cells to damaged PC12 cells was measured by flow cytometry.Mesenchymal stem cells were Induced from pluripotent stem cells by lentivirus infection containing green fluorescent protein in mitochondria.Then they were co-cultured with PC12 cells in Transwell chambers and treated with CoCl2 for 24 hours to detect adenosine triphosphate level in PC12 cells.CoCl2-Induced PC12 cell damage was dose-dependent.Co-culture with mesenchymal stem cells significantly reduced apoptosis and restoredΔΨm in the injured PC12 cells under CoCl2 challenge.Co-culture with mesenchymal stem cells ameliorated mitochondrial swelling,the disappearance of cristae,and chromatin margination in the injured PC12 cells.After direct co-culture,mitochondrial transfer from the mesenchymal stem cells stem cells to PC12 cells was detected via formed tunneling nanotubes between these two types of cells.The transfer efficiency was greatly enhanced in the presence of CoCl2.More importantly,inhibition of tunneling nanotubes partially abrogated the beneficial effects of mesenchymal stem cells on CoCl2-Induced PC12 cell injury.Mesenchymal stem cells reduced CoCl2-Induced PC12

关键词： apoptosis brain injury hypoxia-ischemia Induced pluripotent stem cells mesenchymal stem cells mitochondrial membrane potential mitochondrial transfer PC12 cells tunneling nanotubes

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Current methods for the maturation of Induced pluripotent stem cellderived cardiomyocytes

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World Journal of Stem Cells 2019年第1期11卷 33-43页

作者： Pranav Machiraju Steven C Greenway Departments of Pediatrics and Cardiac Sciences Alberta Children’s Hospital Research Institute Libin Cardiovascular Institute of Alberta Cumming School of MedicineUniversity of Calgary Departments of Pediatrics Cardiac Sciences Biochemistry & Molecular Biology Alberta Children’s Hospital Research Institute Libin Cardiovascular Institute of Alberta Cumming School of Medicine University of Calgary

Induced pluripotent stem cells(iPSCs) were first generated by Yamanaka and colleagues over a decade ago. Since then, iPSCs have been successfully differentiated into many distinct cell types, enabling tissue-, disease-, and patientspecific in vitro modelling. Cardiovascular disease is the greatest cause of mortality worldwide but encompasses rarer disorders of conduction and myocardial function for which a cellular model of study is ideal. Although methods to differentiate iPSCs into beating cardiomyocytes(iPSC-CMs) have recently been adequately optimized and commercialized, the resulting cells remain largely immature with regards to their structure and function,demonstrating fetal gene expression, disorganized morphology, reliance on predominantly glycolytic metabolism and contractile characteristics that differ from those of adult cardiomyocytes. As such, disease modelling using iPSC-CMs may be inaccurate and of limited utility. However, this limitation is widely recognized, and numerous groups have made substantial progress in addressing this problem. This review highlights successful methods that have been developed for the maturation of human iPSC-CMs using small molecules,environmental manipulation and 3-dimensional(3 D) growth approaches.

关键词： Induced pluripotent stem cells Induced pluripotent stem cell-derived cardiomyocytes Regenerative medicine Stem cell biology Translational research

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Lineage reprogramming of fibroblasts into Induced cardiac progenitor cells by CRISPR/Cas9-based transcriptional activators

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Acta Pharmaceutica Sinica B 2020年第2期10卷 313-326页

作者： Jianglin Wang Xueyan Jiang Lixin Zhao Shengjia Zuo Xiantong Chen Lingmin Zhang Zhongxiao Lin Xiaoya Zhao Yuyan Qin Xinke Zhou Xi-Yong Yu Key Laboratory of Molecular Target&Clinical Pharmacology and the State Key Laboratory of Respiratory Disease School of Pharmaceutical Sciences&the Fifth Affiliated HospitalGuangzhou Medical UniversityGuangzhou 511436China

Overexpression of exogenous lineage-determining factors succeeds in directly reprogramming fibroblasts to various cell types.Several studies have reported reprogramming of fibroblasts into Induced cardiac progenitor cells(iCPCs).CRISPR/Cas9-mediated gene activation is a potential approach for cellular reprogramming due to its high precision and multiplexing capacity.Here we show lineage reprogramming to iCPCs through a dead Cas9(dCas9)-based transcription activation system.Targeted and robust activation of endogenous cardiac factors,including GATA4,HAND2,MEF2 C and TBX5(G,H,M and T;GHMT),can reprogram human fibroblasts toward iCPCs.The iCPCs show potentials to differentiate into cardiomyocytes,smooth muscle cells and endothelial cells in vitro.Addition of MEIS1 to GHMT induces cell cycle arrest in G2/M and facilitates cardiac reprogramming.Lineage reprogramming of human fibroblasts into iCPCs provides a promising cellular resource for disease modeling,drug discovery and individualized cardiac cell therapy.

关键词： Lineage reprogramming Human foreskin fibroblasts Induced cardiac progenitor cells CRISPR/Cas9 SAM Cardiac transcription factors

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Decreasing intraocular pressure significantly improves retinal vessel density,cytoarchitecture and visual function in rodent oxygen Induced retinopathy

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Science China(Life Sciences) 2020年第2期63卷 290-300页

作者： Rui Du Xu Wang Ke Shen Shigang He School of Biomedical Engineering Shanghai Jiao Tong UniversityShanghai 200240China Institute of Natural Sciences Shanghai Jiao Tong UniversityShanghai 200240China Bio-X Institute Shanghai Jiao Tong UniversityShanghai 200240China

We attempted to explore a noninvasive,easily applicable and economically affordable therapy for retinopathy of prematurity(ROP).Rat pups were raised in 80%oxygen from postnatal day 7 to P12,and returned to room air.Travoprost eye drops were administered twice a day for 7 days,to reduce intraocular pressure(IOP)by about 20%.Immunohistochemical staining was performed to visualize vessel endothelial cells,to analyze retinal neurons and cytoarchitecture.Behavioral experiments were carried out to test visual acuity and contrast sensitivity.At the end of the 7-day treatment,the number of vessels extending to the vitreous body was significantly reduced and retinal vessel density increased.This improvement was maintained to the end of the12 th week.In the central retina of the model group,the horizontal cells were completely wiped out,the outer plexiform layer was undetectable,and the rod bipolar cell dendrites sprouted into the outer nuclear layer.The treatment partially reverted these architectural changes.Most importantly,behavioral experiments revealed significantly improved visual acuity and contrast sensitivity in the treated group.Therefore,reducing IOP could potentially serve as a safe and economical measure to treat ROP.

关键词： oxygen Induced retinopathy vessel plasticity visual acuity contrast sensitivity rat

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Pseudohalide Induced tunable electronic and excitonic properties in two-dimensional single-layer perovskite for photovoltaics and photoelectronic applications

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Journal of Energy Chemistry 2019年第9期28卷 106-113页

作者： Zhuo Xu Ming Chen Shengzhong(Frank)Liu Key Laboratory of Applied Surface and Colloid Chemistry National Ministry of Education Shaanxi Normal UniversityXi'an 710119 Shaanxi China Shaanxi Key Laboratory for Advanced Energy Devices Shaanxi Normal UniversityXi'an 710119 Shaanxi China Shaanxi Engineering Lab for Advanced Energy Technology Institute for Advanced Energy Materials Shaanxi Normal UniversityXi'an 710119 Shaanxi China School of Materials Science and Engineering Shaanxi Normal UniversityXi'an 710119 Shaanxi China iChEM Dalian National Laboratory for Clean Energy Dalian Institute of Chemical Physics Chinese Academy of Sciences Dalian 116023 Liaoning China University of the Chinese Academy of Sciences Beijing 100039 China

Two-dimensional(2D) layered organic-inorganic hybrid perovskites have attracted much more attention for some applications than their three-dimensional(3D) perovskite counterparts due to their promising thermal and moisture stabilities.In particular, the 2D perovskite devices have shown better promise for optoelectronic applications.However, tunability of optoelectronic properties is often demanded to improve the device performance.Herein, we adopt a newly method to tune the electronic properties of 2D perovskite by introducing pseudohalide into the structure.In this work, we designed a pseudohalidesubstituted 2D perovskite by substituting the out-of-plane halide with pseudohalide and studied the electronic and excitonic properties of 2D-BA2MX4 and 2D-BA2MX2Ps2(M=Ge^(2+), Sn^(2+), and Pb^(2+);X=I;Ps=NCO, NCS, OCN, SCN, Se CN).We revealed the dependence of electronic properties including band gaps, composition of band edges, bonding characteristics, work functions, effective masses, and exciton binding energies on different pseudohalides substituted in 2D perovskite.Our results indicate that the substitution of pseudohalide in 2D perovskites is energetically favorable and can significantly affect the bonding characteristics as well as the CBM and VBM that often play major role in determining their performance in optoelectronic devices.It is expected that the pseudohalide substitution will be helpful in developing more advanced optoelectronic device based on 2D perovskite by optimizing band alignment and promoting charge extraction.

关键词： Pseudohalide Induced tunable electronic and excitonic properties in two-dimensional single-layer perovskite for photovoltaics and photoelectronic applications

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Using Induced pluripotent stem cells for modeling Parkinson’s disease

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World Journal of Stem Cells 2019年第9期11卷 634-649页

作者： Minjing Ke Cheong-Meng Chong Huanxing Su State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical Sciences University of Macao

Parkinson’s disease(PD)is an age-related neurodegenerative disease caused by the progressive loss of dopaminergic(DA)neurons in the substantia nigra.As DA neurons degenerate,PD patients gradually lose their ability of movement.To date no effective therapies are available for the treatment of PD and its pathogenesis remains unknown.Experimental models that appropriately mimic the development of PD are certainly needed for gaining mechanistic insights into PD pathogenesis and identifying new therapeutic targets.Human Induced pluripotent stem cells(iPSCs)could provide a promising model for fundamental research and drug screening.In this review,we summarize various iPSCs-based PD models either derived from PD patients through reprogramming technology or established by gene-editing technology,and the promising application of iPSC-based PD models for mechanistic studies and drug testing.

关键词： Parkinson’s disease Dopaminergic neurons Induced pluripotent stem cells Somatic cell reprogramming Aging

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Differentiation of retinal ganglion cells from Induced pluripotent stem cells: a review

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International Journal of Ophthalmology(English edition) 2019年第1期12卷 152-160页

作者： Shang-Li Ji Shi-Bo Tang Aier Eye Institute Aier School of Ophthalmology Central South University

Glaucoma is a common optic neuropathy that is characterized by the progressive degeneration of axons and the loss of retinal ganglion cells(RGCs). Glaucoma is one of the leading causes of irreversible blindness worldwide. Current glaucoma treatments only slow the progression of RGCs loss. Induced pluripotent stem cells(iPSCs) are capable of differentiating into all three germ layer cell lineages. iPSCs can be patient-specific,making iPSC-derived RGCs a promising candidate for cell replacement. In this review, we focus on discussing the detailed approaches used to differentiate iPSCs into RGCs.

关键词： glaucoma retinal ganglion cells Induced pluripotent stem cells differentiation

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Thermoresponsive Fluorescent Semicrystalline Polymers Decorated with Aggregation Induced Emission Luminogens

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Chinese Journal of Polymer Science 2019年第4期37卷 394-400,I0007页

作者： Jia-Long Wu Chi Zhang Wei Qin Da-Ping Quan Ming-Liang Ge Guo-Dong Liang PCFM and GDHPPC labs School of Materials Science and EngineeringSun Yat-sen University School of Chemistry Sun Yat-sen University Key Laboratory of Polymer Processing Engineering South China University of TechnologyMinistry of Education

Thermoresponsive fluorescent polymers(TFPs) with unique temperature-dependent luminescent properties are of great importance for the development of new functional devices in recent years. Herein, we facilely synthesized an efficient blue-emissive polymer, abbreviated as PCB-TPE, using tetraphenylethene(TPE) as the main building block. PCB-TPE is thermally stable with a novel property of aggregation Induced emission(AIE). The thermoresponsive property and mechanism of PCB-TPE were investigated. Its emission shows temperature-dependent features and reveals fine details in the thermal transitions from-10 °C to 60 °C. The polymer offers a platform for the development of efficient luminescent materials for further biological and optoelectronic applications.

关键词： Aggregation Induced emission Thermoresponsive Fluorescence Crystallization Polymer

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Diagnostic value of gamma-glutamyltransferase/aspartate aminotransferase ratio, protein Induced by vitamin K absence or antagonist II, and alpha-fetoprotein in hepatitis B virus-related hepatocellular carcinoma

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World Journal of Gastroenterology 2019年第36期25卷 5515-5529页

作者： Qiang Wang Qi Chen Xia Zhang Xiao-Lan Lu Qin Du Tao Zhu Guo-Yuan Zhang Dong-Sheng Wang Qu-Ming Fan Department of Laboratory Medicine Affiliated Hospital of North Sichuan Medical College Nanchong 637000 Sichuan Province China Faculty of Laboratory Medicine Center for Translational Medicine North Sichuan Medical College Nanchong 637000 Sichuan Province China Department of Laboratory Medicine Affiliated Sichuan Ba-Yi Rehabilitation Center of Chengdu University of TCM Chengdu 610000 Sichuan Province China Department of Laboratory Medicine of the Ninth People’s Hospital of Chongqing Chongqing 400700 China Department of Preventive Medicine North Sichuan Medical College Nanchong 637000 Sichuan Province China

BACKGROUND Researchers have investigated the diagnostic value of protein Induced by vitamin K absence or antagonist II (PIVKA-II) and alpha-fetoprotein (AFP) in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), and obtained abundant clinical diagnostic data. However, PIVKA-II and AFP have unsatisfactory specificity and sensitivity in the diagnosis of early-stage HBV-related HCC. Gamma-glutamyltransferase (γ-GT) and aspartate aminotransferase (AST) are common biomarkers for evaluating liver function, and we hypothesized that the γ-GT/AST ratio in combination with PIVKA-II and AFP would improve the diagnosis of early-stage HBV-related HCC. AIM To evaluate the diagnostic value of γ-GT/AST ratio alone or in combination with PIVKA-II and AFP in HBV-related HCC. METHODS Serum levels of γ-GT, AST, PIVKA-II, and AFP were detected and analysed in 176 patients with HBV-related HCC and in 359 patients with chronic hepatitis B. According to tumour size and serum level of HBV DNA, HBV-related HCC patients were divided into the following categories: Early-stage HCC patients, HCC patients, HBV DNA positive (HBV DNA+) HCC patients, and HBV DNA negative (HBV DNA-) HCC patients. Receiver-operating characteristic (ROC) curves were used to analyse and compare the diagnostic value of the single and combined detection of various biomarkers in different types of HBV-related HCC. RESULTS Tumour size was positively correlated with serum levels of PIVKA-II and AFP in HCC patients (r = 0.529, aP 0.05). When PIVKA-II was combined with the γ-GT/AST ratio in the diagnosis of earlystage HCC, HCC, and H

关键词： Gamma-glutamyltransferase Aspartate aminotransferase Protein Induced by vitamin K absence or antagonist II Alpha-fetoprotein Hepatitis B virus Hepatocellular carcinoma

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