褶纹冠蚌光珠与骨珠珍珠囊差异的研究

运用多种组织化学方法和电镜技术研究了褶纹冠蚌光珠和骨珠珍珠囊表皮细胞的形态结构、分泌物性质和功能等方面的差异。结果表明：骨珠珍珠囊表皮细胞合成和分泌珍珠前体物质的能力较光珠的强,故骨珠的形成速度比光珠快;光珠和骨珠珍珠囊表皮细胞合成和分泌的蛋白质的差异决定了光珠和骨珠的形成;光珠和骨珠珍珠囊表皮细胞的形态结构特征差异可作为检验和预测人工培育珍珠质量的细胞学标准。     

马氏珠母贝外套膜组织细胞体外培养技术的改进

建立一套有效的马氏珠母贝外套膜组织细胞体外分离及培养的技术和方法,同时探讨体外形成具有完整结构和分泌功能的珍珠囊并最终生成优质珍珠的最佳方法。使用0．25％胰蛋白酶消化马氏珠母贝外套膜组织,收获的细胞使用M199（含10％胎牛血清）培养基置入经多聚赖氨酸预先处理的35mm×100mm培养皿中进行培养,培养体系中添加10ng／ml的角质细胞生长因子和10％自制的、     

外周血细胞重编程为诱导性多潜能干细胞的研究进展

诱导性多潜能干细胞(iPSCs)是指分化细胞中导入特定转录因子后逆转恢复到类似胚胎干细胞的具有自我更新、多向分化等潜能的一类细胞。诱导疾病特异性iPSCs是疾病机理、再生医学等领域的研究热点。目前,人iPSCs供体细胞主要来源于皮肤成纤维细胞,需要组织活检、体外增殖等繁琐过程。利用外周血细胞(peripheral blood cells)成功诱导iPSCs,具有取材方便、诱导快速等优点,将极大地促进iPSCs研究。该文在介绍iPSCs诱导方法的基础上,重点阐述了从小鼠B细胞、T细胞,人脐带血细胞,到人外周血细胞重编程为iPSCs的研究进展,分析了该技术的特点和可能存在的问题,并进行了前景展望。     

背角无齿蚌珍珠囊形成过程中钙代谢的初步研究

本文采用同位素活体标记、人工育珠、普通石蜡切片和放射自显影的方法,对一种淡水育珠河蚌──背角无齿蚌珍珠囊形成过程中的钙代谢途径进行了研究,结果表明,由植入小片带入的钙在珍珠囊形成的过程中,主要代谢途径是：（１）随小片细胞脱落而进入游走细胞;（２）从小片进入初生珍珠囊,初生珍珠囊脱落后进入游走细胞;（３）从小片进入育珠蚌结缔组织,其中一部分再进入育珠蚌表皮,随粘液和壳质分泌;另一部分则进入次生珍珠囊表皮,分泌成为珍珠质的组成部分。实验结果说明了小片中的钙要参与育珠蚌组织的钙代谢,小片的质量对珍珠形成有重要的作用。     

背角无齿蚌珍蛛囊形成过程中钙代谢的初步研究

本文采用同位素活体标记、人工育珠、普通石蜡切片和放射自显影的方法,对一种淡水育珠河蚌－背角无齿蚌珍珠囊形成过程中的钙代谢途径进行了研究,结果表明,由植入小片带入的钙在珍珠囊形成的过程中,主要代谢途径是：（１）随小片细胞脱落而进入游走细胞;（２）从小片进入初生珍珠囊,初生珍珠囊脱落后进入游走细胞;（３）从小片进入育珠蚌结缔组织,其中一部分再进入育珠蚌表皮,随粘液和壳质分泌;另一部分则进入次生珍珠囊表     

膝关节内干细胞/祖细胞在软骨再生中作用研究进展

关节软骨(AC)由于缺乏血管、神经和淋巴,一旦损伤无法自我修复。虽然以外源性细胞为基础的治疗策略在一定程度上能够再生关节软骨,但仍然存在手术间隔长、供体有限、细胞体外培养易去分化和病原体传播等风险。成人膝关节存在许多类型干细胞/祖细胞(SCPCs),当软骨损伤时,就会被动员,迁移到损伤部位,参与再生修复。因此,基于趋化内源性SCPCs到损伤部位的AC原位再生修复策略,充分利用机体自我修复潜力,同时避免了外源性细胞策略的缺点,已经成为研究的热点。本综述主要介绍膝关节内SCPCs类型、迁移路径以及其在软骨损伤修复中的作用。重点介绍内源性AC再生修复策略的研究现状,以期吸引更多的研究人员参与这一有前景的研究领域。     

褶纹冠蚌珍珠囊发育的研究

以褶纹冠蚌(Cristaria plicata Leach)为实验对象,应用光学显微技术和扫描电子显微技术研究珍珠囊的发育,结果表明在水温16℃左右时约需30d形成具有单层上皮细胞的珍珠囊,6个月后稳定分泌珍珠质。构成珍珠囊的上皮细胞从高柱状逐渐变成扁平状或立方形,细胞的碳酸酐酶污性也日益增强。大部分移植细胞小片的结缔组织与母蚌的结缔组织共同成层排列在珍珠囊腔外围。游走细胞在珍珠囊的早期发育阶段十分活跃。本文还阐明了珍珠囊液是存在于上皮细胞与珍珠表面之间的一薄层流体状物质。碳酸钙结晶的核化(nucleation)和初期生长都发生在珍珠囊液中。     

背角无齿蚌珍珠囊形成过程的组织学和酶组织化学研究

本文采用Ｈ．Ｅ染色法和酶组织化学方法,对背角无齿蚌珍珠囊形成过程的组织学和酶组织化学的变化情况进行了研究,证实了“初生珍珠囊”的形成和溶解及“次生珍珠囊”的形成,并推没“次生珍珠囊”表皮细胞是由育珠蚌结缔组织转化而来的;观察了育珠手术后九种酶在珍珠囊形成的各个阶段和在珍珠囊和育珠蚌不同部位的变化情况,说明了珍珠囊珙成过程是与复杂的能量代谢,物质代谢及物质转运等有关的生理生化过程。     

生命的“河流”——血液

血液是存在于生物体内的重要组织,由血浆和多种血细胞组成,在物质运输、免疫防御、内环境稳态的调控等方面具有非常重要的作用。随着对血液学研究的不断深入,尤其是对于体内造血发生和血细胞发育过程的深度解析,促进了体外造血及再生医学的蓬勃发展。着重介绍血细胞发育、血液的组成和功能,并分析讨论现阶段以体外诱导造血干细胞为核心的血液疾病治疗等方面的研究进展及应用前景。     

VEGF-α诱导对卵巢癌血管生成拟态形成及侵袭、迁移能力的影响

目的探讨细胞因子VEGF-α诱导对卵巢癌SKOV-3和OVCAR-3细胞系血管生成拟态形成及侵袭、迁移能力的影响。方法采用不同细胞因子诱导卵巢癌SKOV-3和OVCAR-3细胞系,通过三维培养来观察卵巢癌细胞形成血管生成拟态的能力;以划痕实验和侵袭实验来判断细胞因子诱导对卵巢癌细胞侵袭、迁移能力的影响。结果三维培养结果表明在VEGF-α组SKOV-3和OVCAR-3细胞可观察到大量、明显的血管样网状结构;细胞划痕试验结果表明卵巢癌细胞系经细胞因子诱导后,其迁移能力明显增强;体外侵袭实验表明VEGF-α能够明显增强卵巢癌SKOV-3和OVCAR-3细胞的体外侵袭能力。结论 VEGF-α能促进卵巢癌细胞系血管生成拟态的形成,并明显增强卵巢癌细胞侵袭和迁移的能力。     

Wound healing after excision of mantle tissue from the Akoya pearl oyster, Pinctada fucata

Pearl oysters are usually sacrificed to donate mantle tissue for pearl production. However, if oysters are anaesthetized, they are able to survive mantle excision and regenerate this tissue. Mantle excision causes a large wound and severs the pallial artery that necessitates rapid wound repair to avoid death by bleeding. This study was undertaken to assess the wound healing process in the mantle of the Akoya pearl oyster, Pinctada fucata, following mantle excision. Forty-seven P. fucata were relaxed with 2.5 mL L(-1) propylene phenoxetol before mantle tissue was excised. Oysters were relaxed and sacrificed 1, 3, 6, 12, 25, 36, 48, 66, 80 and 105 h after excision to assess mantle healing using histological techniques. Muscular contraction that effectively reduced the size of the wound was observed within 1 h after mantle excision. Accumulation of haemocytes and connective tissue occurred 3-6 h after excision and wound plugging was achieved within 6 h of excision. Proliferation of epithelial cells to cover the wound site was observed within the first 25 h after mantle excision and growth of connective tissue and formation of the pallial artery were observed within 105 h after mantle excision.     

Gene Expression Patterns in the Outer Mantle Epithelial Cells Associated with Pearl Sac Formation

For pearl culture, nucleus and mantle grafts are implanted into the gonad of the host oyster. The epithelial cells of the implanted mantle graft elongate and surround the nucleus, and a pearl sac is formed. Shell matrix proteins secreted by the pearl sac play an important role in pearl formation. We studied the gene expression patterns of six shell matrix proteins (msi60, n16, nacrein, msi31, prismalin-14, and aspein) in the epithelial cells associated with pearl sac formation. There were differences in the expression patterns of the six genes in the epithelial cells, and the relative expression levels for msi60 and aspein differed between the mantle graft and pearl sac (48 days after implantation). Therefore, the gene expression patterns of the epithelial cells were genetically undetermined, and changed between before and after pearl sac formation. The gene expression patterns of the epithelial cells of the pearl sac may be regulated by the host oysters.     

培养条件对三角帆蚌外套膜离体上皮组织珍珠质分泌能力的影响（简报）

珍珠是由珍珠贝外套膜的上皮细胞受到外源刺激物刺激形成珍珠囊（pearl—sac）,并由珍珠囊产生的钙质分泌物．其分泌物逐渐包围刺激原．使之体积急剧增长而形成的,珍珠质（nacre）是由大于95％的碳酸钙晶体与约5％的角壳蛋白组成的生物矿化产物。因此珍珠贝的外套膜在珍珠形成中起着重要的作用。珍珠贝外套膜的体外培养已开展了一些初步的研究工作。     

Comparison of proliferation and motile activity between human keratinocytes isolated from skin and oral mucosa

OBJECTIVE: Epithelial wound repair assures the recovery of the epithelial barrier after wounding. During wound healing epithelial cells migrate to cover the wound surface. For healing of skin wounds the skin keratinocytes can be replaced by oral mucosa epithelial cells grown in vitro. The presented experiments were carried out in order to compare the proliferation, morphology, and migration between human keratinocytes isolated from human skin and oral mucosa. MATERIALS AND METHODS: Human epidermal and oral mucosa keratinocytes from primary culture were used in all experiments. Cell motility and shape were determined using computer-aided methods. RESULTS AND CONCLUSIONS: It was demonstrated that although both cell types exhibit the same typical epithelial morphology, oral mucosa keratinocytes locomote significantly faster than skin keratinocytes. They also differ in proliferation activity. Oral mucosa keratinocytes exhibited faster growth and different actin cytoskeleton organisation than skin keratinocytes under in vitro conditions. Autologous oral mucosa keratinocytes may be expanded in vitro and used for skin wound healing in vivo.     

The role of electrical signals in murine corneal wound re-epithelialization

Ion flow from intact tissue into epithelial wound sites results in lateral electric currents that may represent a major driver of wound healing cell migration. Use of applied electric fields (EF) to promote wound healing is the basis of Medicare-approved electric stimulation therapy. This study investigated the roles for EFs in wound re-epithelialization, using the Pax6(+/-) mouse model of the human ocular surface abnormality aniridic keratopathy (in which wound healing and corneal epithelial cell migration are disrupted). Both wild-type (WT) and Pax6(+/-) corneal epithelial cells showed increased migration speeds in response to applied EFs in vitro. However, only Pax6(+/+) cells demonstrated consistent directional galvanotaxis towards the cathode, with activation of pSrc signaling, polarized to the leading edges of cells. In vivo, the epithelial wound site normally represents a cathode, but 43% of Pax6(+/-) corneas exhibited reversed endogenous wound-induced currents (the wound was an anode). These corneas healed at the same rate as WT. Surprisingly, epithelial migration did not correlate with direction or magnitude of endogenous currents for WT or mutant corneas. Furthermore, during healing in vivo, no polarization of pSrc was observed. We found little evidence that Src-dependent mechanisms of cell migration, observed in response to applied EFs in vitro, normally exist in vivo. It is concluded that endogenous EFs do not drive long-term directionality of sustained healing migration in this mouse corneal epithelial model. Ion flow from wounds may nevertheless represent an important component of wound signaling initiation.     

Molecular cloning and characterization of perlucin from the freshwater pearl mussel, Hyriopsis cumingii

Perlucin is an important functional protein that regulates shell and pearl formation. In this study, we cloned the perlucin gene from the freshwater pearl mussel Hyriopsis cumingii, designated as Hcperlucin. The full-length cDNA transcribed from the Hcperlucin gene was 1460 bp long, encoding a putative signal peptide of 20 amino acids and a mature protein of 141 amino acids. The mature Hcperlucin peptide contained six conserved cysteine residues and a carbohydrate recognition domain, similar to other members of the C-type lectin families. In addition, a “QPS” and an invariant “WND” motif near the C-terminal region were also found, which are extremely important for polysaccharide recognition and calcium binding of lectins. The mRNA of Hcperlucin was constitutively expressed in all tested H. cumingii tissues, with the highest expression levels observed in the mantle, adductor, gill and hemocytes. In situ hybridization was used to detect the presence of Hcperlucin mRNA in the mantle, and the result showed that the mRNA was specifically expressed in the epithelial cells of the dorsal mantle pallial, an area known to express genes involved in the biosynthesis of the nacreous layer of the shell. The significant Hcperlucin mRNA expression was detected on day 14 post shell damage and implantation, suggesting that the Hcperlucin might be an important gene in shell nacreous layer and pearl formation. The change of perlucin expression in pearl sac also confirmed that the mantle transplantation results in a new expression pattern of perlucin genes in pearl sac cells that are required for pearl biomineralization. These findings could help better understanding the function of perlucin in the shell and pearl formation.     

Roles of wound geometry, wound size, and extracellular matrix in the healing response of bovine corneal endothelial cells in culture

It has classically been accepted that the healing of narrow wounds in epithelia occurs by the formation of a contractile actin cable, while wide wounds are resurfaced by lamellipodia-dependent migration of border cells into the denuded area. To further investigate the general validity of this idea, we performed systematic experiments of the roles of wound geometry, wound size, and extracellular matrix (ECM) in wound healing in monolayers of bovine corneal endothelial cells, a system shown here to predominantly display any of the two healing mechanisms according to the experimental conditions. We found that, in this system, it is the absence or presence of the ECM on the wound surface that determines the specific healing mode. Our observations demonstrate that, independent of their size and geometry, wounds created maintaining the ECM heal by migration of cells into the wound area, while ECM removal from the wound surface determines the predominant formation of an actin cable. While the latter mechanism is slower, the actin cable permits the maintainance of the epithelial phenotype to a larger extent during the healing process, as also confirmed by our finding of a more conserved localization of cadherin and vinculin. We also introduce a model that simulates experimental findings about the dynamics of healing mechanisms, both for the maintenance or removal of the ECM on the wound surface. The findings of this study may contribute to the understanding of physiological and pathological aspects of epithelial wound healing and to the design of therapeutic strategies.     

Roles of lactoferrin on skin wound healing

Skin wound healing is a complex biological process that requires the regulation of different cell types, including immune cells, keratinocytes, fibroblasts, and endothelial cells. It consists of 5 stages: hemostasis, inflammation, granulation tissue formation, re-epithelialization, and wound remodeling. While inflammation is essential for successful wound healing, prolonged or excess inflammation can result in nonhealing chronic wounds. Lactoferrin, an iron-binding glycoprotein secreted from glandular epithelial cells into body fluids, promotes skin wound healing by enhancing the initial inflammatory phase. Lactoferrin also exhibits anti-inflammatory activity that neutralizes overabundant immune response. Accumulating evidence suggests that lactoferrin directly promotes both the formation of granulation tissue and re-epithelialization. Lactoferrin stimulates the proliferation and migration of fibroblasts and keratinocytes and enhances the synthesis of extracellular matrix components, such as collagen and hyaluronan. In an in vitro model of wound contraction, lactoferrin promoted fibroblast-mediated collagen gel contraction. These observations indicate that lactoferrin supports multiple biological processes involved in wound healing.     

Activin B regulates adipose-derived mesenchymal stem cells to promote skin wound healing via activation of the MAPK signaling pathway

Adipose-derived stem cells (ADSCs) are multipotent stromal cells that can differentiate into a variety of cell types, including skin cells, and they can provide an abundant source of cells for skin tissue engineering and skin wound healing. The purpose of this study is to explore the therapeutic effects of activin B in combination with ADSCs and the possible signaling mechanism. In this study, we found that activin B was able to promote ADSC migration by inducing actin stress fiber formation in vitro. In vivo, activin B in combination with ADSCs was capable of enhancing α-SMA expression and wound closure. This combined treatment also promoted fibroblast and keratinocyte proliferation and accelerated re-epithelialization and collagen deposition. Moreover, activin B in combination with ADSCs boosted angiogenesis in the wound area. Further study of the mechanism revealed that activation of JNK and ERK signaling, but not p38 signaling, were required for activin B-induced ADSC actin stress fiber formation and cell migration. These results showed that activin B was able to activate JNK and ERK signaling pathways to induce actin stress fiber formation and ADSC migration to promote wound healing. These results suggest that combined treatment with activin B and ADSCs is a promising therapeutic strategy for the management of serious skin wounds.