Cone-rod homeobox transcriptionally activates TCF7 to promote the proliferation of retinal pigment epithelial and retinoblastoma cells in vitro

AIM:To investigate the proliferation regulatory effect of cone-rod homeobox(CRX)in retinal pigment epithelium(RPE)and retinoblastoma(RB)cells to explore the potential application and side effect(oncogenic potential)of CRXbased gene therapy in RPE-based retinopathies.METHODS:Adult human retinal pigment epithelial(ARPE)-19 and human retinal pigment epithelial(RPE)-1 cells and Y79 RB cell were used in the study.Genetic manipulation was performed by lentivirus-based technology.The cell proliferation was determined by a CellTiter-Glo Reagent.The mRNA and protein levels were determined by quantitative real-time polymerase chain reaction(qPCR)and Western blot assay.The transcriptional activity of the promoter was determined by luciferase reporter gene assay.The bindings between CRX and transcription factor 7(TCF7)promoter as well as TCF7 and the promoters of TCF7 target genes were examined by chromatin immunoprecipitation(ChIP)assay.The transcription of the TCF7 was determined by a modified nuclear run-on assay.RESULTS:CRX overexpression and knockdown significantly increased(n=3,P<0.05 in all the cells)and decreased(n=3,P<0.01 in all the cells)the proliferation of RPE and RB cells.CRX overexpression and knockdown significantly increased and deceased the mRNA levels of Wnt signaling target genes[including MYC proto-oncogene(MYC),JUN,FOS like 1(FOSL1),CCND1,cyclin D2(CCND2),cyclin D3(CCND3),cellular communication network factor 4(CCN4),peroxisome proliferator activated receptor delta(PPARD),and matrix metallopeptidase 7(MMP7)]and the luciferase activity driven by the Wnt signaling transcription factor(TCF7).TCF7 overexpression and knockdown significantly increased and decreased the proliferation of RPE and RB cells and depletion of TCF7 significantly abolished the stimulatory effect of CRX on the proliferation of RPE and RB cells.CRX overexpression and knockdown significantly increased and decreased the mRNA level of TCF7 and the promoter of TCF7 was significantly immunoprecipitated by CRX antibody.CONCLUSION:CRX transcriptionally activates TCF7 to promote the proliferation of RPE and RB cells in vitro.CRX is a potential target for RPE-based regenerative medicine.The potential risk of this strategy,tumorigenic potential,should be considered.

Mesenchymal stem cells: A promising therapeutic avenue for nonalcoholic fatty liver disease

Non-alcoholic fatty liver disease(NAFLD)is a pressing global health concern that is associated with metabolic syndrome and obesity.On the basis of the insights provided by Jiang et al,this editorial presents an exploration of the potential of mesenchymal stem cells(MSCs)for NAFLD treatment.MSCs have numerous desirable characteristics,including immunomodulation,anti-inflammatory pro-perties,and tissue regeneration promotion,rendering them attractive candidates for NAFLD treatment.Recent preclinical and early clinical studies have high-lighted the efficacy of MSCs in improving liver function and reducing disease severity in NAFLD models.However,MSC heterogeneity,long-term safety concerns,and unoptimized therapeutic protocols remain substantial challenges.Addressing these challenges through standardized protocols and rigorous clinical trials is essential to the safe and successful application of MSCs in NAFLD mana-gement.Continued research into MSC mechanisms and therapeutic optimization is required to improve treatments for NAFLD and related liver diseases.

Identification and characterization of cell cultures with various embryogenic/regenerative potential in cotton based on morphological,cytochemical,and cytogenetical assessment

Somatic embryogenesis(SE) plays a vital role in genetic transformation and massive propagation of important agronomical and economical crops.Here,we conducted a systematic assessment of the morphological,cytochemical,and cytogenetical characteristics of six culture strains with various embryogenic/regenerative potential during SE process in cotton.Results indicated that the six cell culture strains had stable ploidy levels,and did not reveal any relationship between the cytogenetic state and their morphogenetic potential.Moreover,the six culture strains were compared via double staining with Evans blue and Acetocarmine to efficiently distinguish embryogenic and non-embryogenic cells and determine the embryogenic nature of the calli.In addition,the kind of auxins added in medium affected not only growth property,color,size of cell clumps but also ploidy level and regeneration ability.By combining analysis of morphological,cytochemical,and cytogenetical characteristics of the cell cultures,we are able to obtain and maintain homogeneous cell population with high morphogenic and regeneration ability and establish efficient somatic embryogenesis and regeneration system from short-term cell cultures in upland cotton,which highlight the application of biotechnological approaches in crop breeding,and above all,to better understand totipotency of cells in higher plants.

Controversies regarding transplantation of mesenchymal stem cells

Mesenchymal stem cells(MSCs)have tantalized regenerative medicine with their therapeutic potential,yet a cloud of controversies looms over their clinical tran-splantation.This comprehensive review navigates the intricate landscape of MSC controversies,drawing upon 15 years of clinical experience and research.We delve into the fundamental properties of MSCs,exploring their unique immuno-modulatory capabilities and surface markers.The heart of our inquiry lies in the controversial applications of MSC transplantation,including the perennial debate between autologous and allogeneic sources,concerns about efficacy,and lingering safety apprehensions.Moreover,we unravel the enigmatic mechanisms surro-unding MSC transplantation,such as homing,integration,and the delicate balance between differentiation and paracrine effects.We also assess the current status of clinical trials and the ever-evolving regulatory landscape.As we peer into the future,we examine emerging trends,envisioning personalized medicine and innovative delivery methods.Our review provides a balanced and informed perspective on the controversies,offering readers a clear understanding of the complexities,challenges,and potential solutions in MSC transplantation.

Dental mesenchymal stromal/stem cells in different microenvironments— implications in regenerative therapy

Current research data reveal microenvironment as a significant modifier of physical functions,pathologic changes,as well as the therapeutic effects of stem cells.When comparing regeneration potential of various stem cell types used for cytotherapy and tissue engineering,mesenchymal stem cells(MSCs)are currently the most attractive cell source for bone and tooth regeneration due to their differentiation and immunomodulatory potential and lack of ethical issues associated with their use.The microenvironment of donors and recipients selected in cytotherapy plays a crucial role in regenerative potential of transplanted MSCs,indicating interactions of cells with their microenvironment indispensable in MSC-mediated bone and dental regeneration.Since a variety of MSC populations have been procured from different parts of the tooth and tooth-supporting tissues,MSCs of dental origin and their achievements in capacity to reconstitute various dental tissues have gained attention of many research groups over the years.This review discusses recent advances in comparative analyses of dental MSC regeneration potential with regards to their tissue origin and specific microenvironmental conditions,giving additional insight into the current clinical application of these cells.

启动人体PRC的再生生命

尊敬的IEF经济俱乐部执行主席兼行政长官Dr.Peter Kasalovsky及各位大使、世界商坐联合会主席Peter Mihok先生、斯洛伐克科学院副主席Dagmar Podmakova女士、IEF经济俱乐部的各位成员、世界各国的在线网友们、女士们、先生们：今年我讨论的题目是“启动人PRC的再生生命”。

Insight into generation of induced mesenchymal stem cells from induced pluripotent cells

Mesenchymal stem cells(MSCs)have the potential for use in cell-based regenerative therapies.Currently,hundreds of clinical trials are using MSCs for the treatment of various diseases.However,MSCs are low in number in adult tissues;they show heterogeneity depending upon the cell source and exhibit limited proliferative potential and early senescence in in vitro cultures.These factors negatively impact the regenerative potential of MSCs and therefore restrict their use for clinical applications.As a result,novel methods to generate induced MSCs(iMSCs)from induced pluripotent stem cells have been explored.The development and optimization of protocols for generation of iMSCs from induced pluripotent stem cells is necessary to evaluate their regenerative potential in vivo and in vitro.In addition,it is important to compare iMSCs with primary MSCs(isolated from adult tissues)in terms of their safety and efficacy.Careful investigation of the properties of iMSCs in vitro and their long term behavior in animals is important for their translation from bench to bedside.

Disease modifying treatment of spinal cord injury with directly reprogrammed neural precursor cells in non-human primates

BACKGROUND The development of regenerative therapy for human spinal cord injury(SCI)is dramatically restricted by two main challenges:the need for a safe source of functionally active and reproducible neural stem cells and the need of adequate animal models for preclinical testing.Direct reprogramming of somatic cells into neuronal and glial precursors might be a promising solution to the first challenge.The use of non-human primates for preclinical studies exploring new treatment paradigms in SCI results in data with more translational relevance to human SCI.AIM To investigate the safety and efficacy of intraspinal transplantation of directly reprogrammed neural precursor cells(drNPCs).METHODS Seven non-human primates with verified complete thoracic SCI were divided into two groups:drNPC group(n=4)was subjected to intraspinal transplantation of 5 million drNPCs rostral and caudal to the lesion site 2 wk post injury,and lesion control(n=3)was injected identically with the equivalent volume of vehicle.RESULTS Follow-up for 12 wk revealed that animals in the drNPC group demonstrated a significant recovery of the paralyzed hindlimb as well as recovery of somatosensory evoked potential and motor evoked potential of injured pathways.Magnetic resonance diffusion tensor imaging data confirmed the intraspinal transplantation of drNPCs did not adversely affect the morphology of the central nervous system or cerebrospinal fluid circulation.Subsequent immunohistochemical analysis showed that drNPCs maintained SOX2 expression characteristic of multipotency in the transplanted spinal cord for at least 12 wk,migrating to areas of axon growth cones.CONCLUSION Our data demonstrated that drNPC transplantation was safe and contributed to improvement of spinal cord function after acute SCI,based on neurological status assessment and neurophysiological recovery within 12 wk after transplantation.The functional improvement described was not associated with neuronal differentiation of the allogeneic drNPCs.Instead,directed drNPCs migration to the areas of active growth cone formation may provide exosome and paracrine trophic support,thereby further supporting the regeneration processes.

Calcium channels and their role in regenerative medicine

Stem cells hold indefinite self-renewable capability that can be differentiated into all desired cell types.Based on their plasticity potential,they are divided into totipotent(morula stage cells),pluripotent(embryonic stem cells),multipotent(hematopoietic stem cells,multipotent adult progenitor stem cells,and mesenchymal stem cells[MSCs]),and unipotent(progenitor cells that differentiate into a single lineage)cells.Though bone marrow is the primary source of multipotent stem cells in adults,other tissues such as adipose tissues,placenta,amniotic fluid,umbilical cord blood,periodontal ligament,and dental pulp also harbor stem cells that can be used for regenerative therapy.In addition,induced pluripotent stem cells also exhibit fundamental properties of self-renewal and differentiation into specialized cells,and thus could be another source for regenerative medicine.Several diseases including neurodegenerative diseases,cardiovascular diseases,autoimmune diseases,virus infection(also coronavirus disease 2019)have limited success with conventional medicine,and stem cell transplantation is assumed to be the best therapy to treat these disorders.Importantly,MSCs,are by far the best for regenerative medicine due to their limited immune modulation and adequate tissue repair.Moreover,MSCs have the potential to migrate towards the damaged area,which is regulated by various factors and signaling processes.Recent studies have shown that extracellular calcium(Ca^(2+))promotes the proliferation of MSCs,and thus can assist in transplantation therapy.Ca^(2+)signaling is a highly adaptable intracellular signal that contains several components such as cell-surface receptors,Ca^(2+)channels/pumps/exchangers,Ca^(2+)buffers,and Ca^(2+)sensors,which together are essential for the appropriate functioning of stem cells and thus modulate their proliferative and regenerative capacity,which will be discussed in this review.

The Mechanism of Burn Regenerative Therapy and Wound Healing

Objective: From molecular, cell and systemic clinical treatment three levels, this report revealed the mechanism of wound healing, the basic theory and the importance of clinical practice for the Burn Regenerative Therapy with MEBT/MEBO (BRT & MEBT/MEBO).Method: This report analyzed the importance and clinical curative effect for standardized appliance of BRT & MEBT/MEBO from two key points: 1) Liquefaction of necrotic tissue from burn wound; 2) Skin regeneration in situ . Result: There are three necessary conditions for skin physiologically repair and regeneration of burn wound: 1) The formation of moist physiological environment on burn wound; 2) The material foundation of life regenerative substances and histology for keratin-19 stem cells’ regeneration in situ ; 3) Standardized procedures and appliance of BRT & MEBT/MEBO, which is the guarantee of burn wound healing physiologically. Conclusion: The theory of “Potential Regenerative Cell" and the technique of “Stem cell in situ regeneration" are the basic theory and clinical treatment for BRT. Full-thickness burn skin can be healed physiologically and the skin tissues and organs can be regenerated ONLY by standardized procedures and timely appliance of BRT & MEBT/MEBO.

烧伤再生医学与疗法创面治疗原则及机理阐述

目的探讨、了解烧伤再生医学与疗法的创面治疗机理,阐述坚持再生医学与疗法创面治疗原则的重要性,规范创面换药操作程序。方法通过实验室细胞学观察研究和模拟对照研究的方法 ,阐述和论证在实施湿润暴露疗法、湿润半暴露疗法和湿润包扎疗法时,坚持烧伤再生医学与疗法创面治疗原则的依据和重要性。结果 水、消毒液、抗生素等均会直接损伤人体组织细胞,不宜应用于创面;湿润烧伤膏(MEBO)药纱在半暴露疗法和包扎疗法中起到较好的稳定MEBO框架剂型、吸附、引流、保持创面立体式生理湿润环境的作用,能够有效延长和维持MEBO对创面的治疗时间,疗效基本等同于湿润暴露疗法。结论在烧伤再生医学与疗法的临床应用中,无论是采用湿润暴露疗法、湿润半暴露疗法和湿润包扎疗法治疗创面,只要严格遵守创面治疗原则和规范的换药操作程序,就能够实现较好的皮肤组织器官原位再生效果。

再生医学技术在多学科的临床应用与展望

目的:总结了解近年再生医学及技术在临床多学科的应用与进展情况。方法:对1999年以来再生医学及技术的临床研究和应用报告进行分类、总结和综述。结果:再生医学技术不仅被广泛应用于皮肤烧伤、慢性溃疡的救治,而且已被更广泛地应用于普通外科、骨科、创伤外科、手外科、肛肠科、妇产科、眼科、口腔科、皮肤科、耳鼻喉科等多学科、多领域。结论:再生医学技术能实现人体多组织器官(皮肤、黏膜、神经、肌肉、肌腱等)的原位再生修复,适合临床多学科推广应用,特别是在医治肿瘤方面,经临床实验研究发现,再生医学技术能促进骨髓细胞增长,使癌细胞停止增殖,直至死亡,使我们看到了攻克癌症的希望。

仔猪神经干细胞体外培养及鉴定

由仔猪脑部海马体齿状回分离得到神经干细胞,利用无血清培养基添加特定生长因子方式培养;采用RT-PCR鉴定神经干细胞和诱导分化后细胞生物特性;利用免疫荧光化学技术检测克隆的细胞抗原和分化后特异性成熟神经细胞抗原表达。由海马体齿状回分离的细胞群具有连续克隆能力;根据免疫荧光检测,脑神经干细胞表达巢蛋白、配对盒因子6和解整合素样金属蛋白酶10;体外培养下NSCs可被诱导分化为神经源性细胞,且表达微管相关蛋白2、胶质纤维酸性蛋白和髓鞘碱性蛋白。分离和培育NSCs细胞结果显示,其具有更新能力和分化为神经元和胶质细胞的潜能,可为干细胞移植提供基础数据。

生命科学的未来掌握在我们中国人手中 生命之美 美在细胞——在第二次世界中西医结合大会烧伤卫星会议上的即席学术演讲

作者在第二次世界中西医结合大会烧伤卫星会议上作了题为“生命之美 ,美在细胞”的学术演讲 ,演讲主要包括烧伤皮肤再生医学理论 ,国际烧伤治疗的进展以及皮肤再生医学发展规划等三大方面的内容。在第一部分内容中 ,结合烧伤临床实践 ,作者详细阐述了潜能再生细胞的理论基础和应用价值 ,以及以该理论为核心思想形成的烧伤皮肤再生医学的主要内容。第二部分则就目前国际烧伤治疗研究趋势进行了论述 ,即保持创面的生理性湿润环境是烧伤创面处理的发展方向和用药基础。在第三部分 ,作者强调了皮肤再生医学学术发展的一些具体事项 ,以及专业委员会 ,科技中心 ,全国医疗网 。

再生医学理论与实践——在第七届全国烧伤创疡学术会议上的报告

作者在本文中 ,从哲学思想入手深入浅出地论述了再生医学的八大基本理论 ,其中心内容是以生命哲学思想“三维论”为指导 ,通过“整体与个体论”的研究方法 ,揭示了“生命延续的机理”是存在组织器官中的“潜能再生细胞” ,据此以“顺应生命的营养论”为基础 ,按照“天人合一 ,扶正固本”的临床医疗保障理论 ,实现器官、组织和细胞在人体原位再生 ,最终达到保障人体健康的目的。同时作者运用成熟的再生医学实践工作系统的阐述了再生医学各种理论的形成和具体的实践过程 。

全能干细胞与潜能再生细胞的实验研究进展

全能干细胞(totipotent stem cell,TSC)和潜能再生细胞(potential regenerative cell,PRCs)在实现各组织器官的原位再生,修复或改善损伤组织和器官的结构及功能方面发挥着重要作用,是目前医学领域研究的热点。笔者根据自身研究经验,并通过查阅大量文献资料,综述了全能干细胞与潜能再生细胞的生理作用,旨在为再生医学的后续研究提供一定的方向与思路。

从再生医学角度探讨深度烧伤界定与坏死组织层处理方法

目的:加深对再生医学的理解,探讨皮肤再生医学对“深度烧伤”的界定与相应的治疗方法。方法:通过查阅传统疗法关于“深度烧伤”概念的认识和针对焦痂所采取的处理方法文献复习,结合“皮肤再生医学”的现代理论与实践,从病理组织学与临床特点方面对徐荣祥提出的三度六分法中六个烧伤深度界面综合分析,“深度烧伤”以传统方法治疗是否瘢痕愈合为基准,施加于创面上的一切疗法是否利于创面愈合为原则。结果:①传统疗法对“深度烧伤”概念的认识并不一致,对焦痂采用的处理方法有碍于皮肤再生修复;②从“皮肤再生医学”角度拟定,三度六分法中的深Ⅱ度深型是“深度烧伤”的最低界定深度;③“激活潜能再生细胞疗法”是治疗一切烧伤创面的核心技术,Ⅲ度烧伤辅以“皮肤耕耘减张术”或“坏死层薄化技术”处理,能充分发挥“激活潜能再生细胞疗法”的作用。结论:深Ⅱ度深型和浅Ⅲ度与深Ⅲ度合称为“深度烧伤”,“激活潜能再生细胞疗法”是皮肤再生的核心技术,“皮肤耕耘减张术”的最佳适应证为浅Ⅲ度创面,“坏死层薄化技术”有利于深Ⅲ度创面再生修复。

干法转湿法治愈21例大面积深度烧伤临床体会

目的 :总结在大面积深度烧伤病人传统疗法治疗无效后改用烧伤湿性医疗技术 (MEBT/MEBO)的临床经验。方法 :总结我科 1 999年 9月至 2 0 0 4年 3月收治的院外采用传统疗法治疗无效改用烧伤皮肤再生疗法 (MEBT/MEBO)治疗的 2 1例特重度烧伤病人临床资料 ,其创面处理全程采用MEBT/MEBO ,结合全身综合治疗。结果 :本组 2 1例全部治愈 ,未植皮自行愈合者 1 8例 ,占 80 95 %,深Ⅲ度创面自愿要求植皮 3例 ,占 1 4 2 0 %。经随访无增生性瘢痕 ,无功能障碍。结论 :烧伤湿性医疗技术对传统疗法治疗无效的大面积深度烧伤病人临床疗效显著。

再生医学技术在烧伤传统治疗无效病人中的应用(附38例报告)

目的:总结重度烧伤传统疗法无效改为再生医学(MEBT/MEBO)技术治疗变化规律和临床经验。方法:将我科1995年5月至2002年5月收治的院外采用传统疗法无效改用MEBT/MEBO技术治疗的38例重度烧伤病人临床资料进行回顾性总结,病人一旦入院创面均改用MEBT/MEBO治疗,初始予以彻底清创,规范用药、规范操作,规范认识;全身实行系统综合治疗措施,以临床观察和病人感觉评价治疗效果。结果:本组38例全部治愈,未植皮自行愈合者21例,占55.26％,深Ⅲ度创面自愿要求植皮者17例,占44.74％。经随访多数无增生性瘢痕,部分愈后有局限性瘢痕,质软无残废。结论:重度烧伤经传统疗法久治不愈或疗效欠佳病人,病情复杂,并发症较多治疗难度也较大,再生医学可有效改善以上缺陷和病症,是重度烧伤病人传统治疗无效时的理想疗法。

皮肤原位再生复原技术对深Ⅱ度烧伤治疗的临床疗效总结

目的论证皮肤原位再生复原技术对深Ⅱ度烧伤治疗中的临床机理,总结"体细胞转化为干细胞、再原位再生皮肤组织器官"的治疗效果。方法对应用皮肤原位再生复原技术治疗深Ⅱ度烧伤创面过程中的组织学观察情况和物理检查体征进行描述,并对规范治疗后的全身治疗和创面各期组织学变化与临床表现进行分析总结。结果规范应用皮肤原位再生复原技术对深Ⅱ度烧伤局部创面和全身器官都有明显的原位再生复原的治疗作用。结论皮肤原位再生复原技术为深Ⅱ度烧伤患者提供了全新的治疗方法,临床结果显示其具有显著的疗效,较强的实用性和先进性。