Optimal antigen localization in human tissues using aldehyde-fixed plastic-embedded sections

Although the utility of antigen labeling techniques in frozen tissues is well known, it is generally acknowledged that an improvement in morphologic preservation is desirable. Conventionally processed paraffin-embedded tissues are limited in the range of antigens that can be detected and newer plastic embedding techniques have been even more restricted. By using cold (4 degrees C) processing and limited fixation a wide range of antigens (including T and B markers) has been demonstrated in 2 mu plastic sections. The morphologic preservation and antigen localization are superior to other techniques. The combination of precise antigen localization and excellent morphologic preservation should expand the diagnostic and investigative uses of immunohistology.     

MicroRNAs对缺血再灌注损伤的调控作用

MicroRNA（miRNA）是一类小的（～20个核苷酸）、非编码的单链RNA分子,能够负调控基因表达,涉及多种信号途径和病理生理过程。缺血再灌注损伤(ischemia reperfusion injury,IRI)指组织器官缺血后重新获得血液的再灌注过程,灌注后组织、器官功能不能恢复,造成功能障碍和结构损伤的现象。IRI是影响多个组织与器官复杂的、系统的生理病理过程,并能够产生很多不可逆的损伤,导致级联的多器官功能障碍。现已发现多种miRNA在组织器官IRI中发生明显的变化,表明miRNA能够直接或者间接影响组织器官IRI。本文综述了miRNA的靶基因以及在心、脑、肝和肾IRI中的调控作用。miRNA 不仅参与了器官IR 损伤的病理生理过程,而且作为IR损伤的特定标志物在临床诊断和干预治疗中具有广阔的前景。     

角质细胞生长因子研究进展

角质细胞生长因子(KGF)从属于成纤维细胞生长因子家族。KGF基因表达受多种细胞因子调控。KGF与受体KGFR特异性的结合发挥其多种生物学功能：参与组织、器官的发育;参与皮肤、胃、肠、肾、膀胱、肺等上皮的损伤修复;减少放、化疗所带来的副作用,具有损伤防护功能;KGF与肿瘤密不可分。     

Animal models for radiation injury, protection and therapy

Current events throughout the world underscore the growing threat of different forms of terrorism, including radiological or nuclear attack. Pharmaceutical products and other approaches are needed to protect the civilian population from radiation and to treat those with radiation-induced injuries. In the event of an attack, radiation exposures will be heterogeneous in terms of both dose and quality, depending on the type of device used and each victim's location relative to the radiation source. Therefore, methods are needed to protect against and treat a wide range of early and slowly developing radiation-induced injuries. Equally important is the development of rapid and accurate biodosimetry methods for estimating radiation doses to individuals and guiding clinical treatment decisions. Acute effects of high-dose radiation include hematopoietic cell loss, immune suppression, mucosal damage (gastrointestinal and oral), and potential injury to other sites such as the lung, kidney and central nervous system (CNS). Long-term effects, as a result of both high- and low-dose radiation, include dysfunction or fibrosis in a wide range of organs and tissues and cancer. The availability of appropriate types of animal models, as well as adequate numbers of animals, is likely to be a major bottleneck in the development of new or improved radioprotectors, mitigators and therapeutic agents to prevent or treat radiation injuries and of biodosimetry methods to measure radiation doses to individuals.     

Epithelial stem cells of the lung: privileged few or opportunities for many?

Most reviews of adult stem cells focus on the relatively undifferentiated cells dedicated to the renewal of rapidly proliferating tissues, such as the skin, gut and blood. By contrast, there is mounting evidence that organs and tissues such as the liver and pancreatic islets, which turn over more slowly, use alternative strategies, including the self-renewal of differentiated cells. The response of these organs to injury may also reveal the potential of differentiated cells to act as stem cells. The lung shows both slow turnover and rapid repair. New experimental approaches, including those based on studies of embryonic development, are needed to identify putative lung stem cells and strategies of lung homeostasis and repair.     

Bioelectric signaling in regeneration: Mechanisms of ionic controls of growth and form

The ability to control pattern formation is critical for the both the embryonic development of complex structures as well as for the regeneration/repair of damaged or missing tissues and organs. In addition to chemical gradients and gene regulatory networks, endogenous ion flows are key regulators of cell behavior. Not only do bioelectric cues provide information needed for the initial development of structures, they also enable the robust restoration of normal pattern after injury. In order to expand our basic understanding of morphogenetic processes responsible for the repair of complex anatomy, we need to identify the roles of endogenous voltage gradients, ion flows, and electric fields. In complement to the current focus on molecular genetics, decoding the information transduced by bioelectric cues enhances our knowledge of the dynamic control of growth and pattern formation. Recent advances in science and technology place us in an exciting time to elucidate the interplay between molecular-genetic inputs and important biophysical cues that direct the creation of tissues and organs. Moving forward, these new insights enable additional approaches to direct cell behavior and may result in profound advances in augmentation of regenerative capacity.     

The biological effects in animals related to the accident at the Chernobyl Atomic Electric Power Station. 1. The experimental model. The radiation loads on animals kept continuously under external and internal radiation exposure in the area of the Chernobyl Atomic Electric Power Station]

Irradiation conditions in which laboratory animals were kept in experimental laboratories of Chernobyl and Kiev after the accident at the Chernobyl A.P.S. are described. The data are presented on the spectral structural and activity of radionuclides in the diet as well as in the organs and tissues of the animals. The radiation loads have been estimated with regard to an external gamma component and the internal one contributed by the incorporated radionuclides. It has been shown that radiation doses received by the animals during their lifetime due to these contributions do not exceed units of cGy.     

Developmental engineering the kidney: Leveraging principles of morphogenesis for renal regeneration

Multiple methodological approaches are currently under active development for application in tissue engineering and regenerative medicine of tubular and solid organs. Most recently, developmental engineering (TE/RM), or the leveraging of embryonic and morphological paradigms to recapitulate aspects of organ development, has been proposed as a strategy for the sequential, iterative de novo assembly of tissues and organs as discrete developmental modules ex vivo, prior to implantation in vivo. In this article, we focus on the kidney to highlight in detail how principles of developmental biology are impacting approaches to TE of this complex solid organ. Ultimately, such methodologies may facilitate the establishment of clinically relevant therapeutic strategies for regeneration of renal structure and function, greatly impacting treatment regimens for chronic kidney disease. Birth Defects Research (Part C) 96:30–38, 2012. © 2012 Wiley Periodicals, Inc.     

Radiation survival of vascular smooth muscle cells as a function of age

Late damage to normal tissues is an important consideration in determining the dose of radiation which can be delivered to a given target volume in clinical radiation therapy. The response of large blood vessels to radiation injury is undoubtedly complex and is influenced by (1) the cellular composition of the vessel wall, (2) the slow turnover of vascular cells, and (3) vascular repair mechanisms. As a first order model for radiation effects in large vessels, we have studied the radiobiologic properties of cultured vascular smooth muscle cells. We have measured survival curves and repair of sublethal radiation damage in exponentially growing cultures of rat aortic smooth muscle cells as a function of animal age and site of origin (thoracic versus abdominal aorta). Radiation survival parameters (utilizing two different mathematical models for the survival curve) and repair of sublethal damage did not appear to vary significantly as a function of animal age (3-23 months) or site or origin.     

Which place for stem cell therapy in the treatment of acute radiation syndrome?

Radiation-induced (RI) tissue injuries can be caused by radiation therapy, nuclear accidents or radiological terrorism. Notwithstanding the complexity of RI pathophysiology, there are some effective approaches to treatment of both acute and chronic radiation damages. Cytokine therapy is the main strategy capable of preventing or reducing the acute radiation syndrome (ARS), and hematopoietic growth factors (GF) are particularly effective in mitigating bone marrow (BM) aplasia and stimulating hematopoietic recovery. However, first, as a consequence of RI stem and progenitor cell death, use of cytokines should be restricted to a range of intermediate radiation doses (3 to 7 Gy total body irradiation). Second, ARS is a global illness that requires treatment of damages to other tissues (epithelial, endothelial, glial, etc.), which could be achieved using pleiotropic or tissue-specific cytokines. Stem cell therapy (SCT) is a promising approach developed in the laboratory that could expand the ability to treat severe radiation injuries. Allogeneic hematopoietic stem cell transplantation (BM, mobilized peripheral blood and cord blood) transplantation has been used in radiation casualties with variable success due to limiting toxicity related to the degree of graft histocompatibility and combined injuries. Ex vivo expansion should be used to augment cord blood graft size and/or promote very immature stem cells. Autologous SCT might also be applied to radiation casualties from residual hematopoietic stem and progenitor cells (HSPC). Stem cell plasticity of different tissues such as liver or skeletal muscle, may also be used as a source of hematopoietic stem cells. Finally, other types of stem cells such as mesenchymal, endothelial stem cells or other tissue committed stem cells (TCSC), could be used for treating damages to nonhematopoietic organs.     

Leukocyte recruitment in inflammation: basic concepts and new mechanistic insights based on new models and microscopic imaging technologies

The immune cell system is a critical component of host defense. Recruitment of immune cells to sites of infection, immune reaction, or injury is complex and involves coordinated adhesive interactions between the leukocyte and the endothelial cell monolayer that lines blood vessels. This article reviews basic mechanisms in the recruitment of leukocytes to tissues and then selectively reviews new concepts that are emerging based on advances in live cell imaging microscopy and mouse strains. These emerging concepts are altering the conventional paradigms of inflammatory leukocyte recruitment established in the early 1990s. Indeed, recent publications have identified previously unrecognized contributions from pericytes and interstitial leukocytes and their secreted products that guide leukocytes to their targets. Investigators have also begun to design organs on a chip. Recent reports indicate that this avenue of research holds much promise.     

Regenerative biology and medicine

The replacement of damaged tissues and organs with tissue and organ transplants or bionic implants has serious drawbacks. There is now emerging a new approach to tissue and organ replacement, regenerative biology and medicine. Regenerative biology seeks to understand the cellular and molecular differences between regenerating and non-regenerating tissues. Regenerative medicine seeks to apply this understanding to restore tissue structure and function in damaged, non-regenerating tissues. Regeneration is accomplished by three mechanisms, each of which uses or produces a different kind of regeneration-competent cell. Compensatory hyperplasia is regeneration by the proliferation of cells which maintain all or most of their differentiated functions (e.g., liver). The urodele amphibians regenerate a variety of tissues by the dedifferentiation of mature cells to produce progenitor cells capable of division. Many tissues contain reserve stem or progenitor cells that are activated by injury to restore the tissue while simultaneously renewing themselves. All regeneration-competent cells have two features in common. First, they are not terminally differentiated and can re-enter the cell cycle in response to signals in the injury environment. Second, their activation is invariably accompanied by the dissolution of the extracellular matrix (ECM) surrounding the cells, suggesting that the ECM is an important regulator of their state of differentiation. Regenerative medicine uses three approaches. First is the transplantation of cells into the damaged area. Second is the construction of bioartificial tissues by seeding cells into a biodegradable scaffold where they produce a normal matrix. Third is the use of a biomaterial scaffold or drug delivery system to stimulate regeneration in vivo from regeneration-competent cells. There is substantial evidence that non-regenerating mammalian tissues harbor regeneration-competent cells that are forced into a pathway of scar tissue formation. Regeneration can be induced if the factors leading to scar formation are inhibited and the appropriate signaling environment is supplied. An overview of regenerative mechanisms, approaches of regenerative medicine, research directions, and research issues will be given.     

Voxel model of a rabbit: assessment of absorbed doses in organs after CT examination performed by two different protocols

The objective of this work was to assess absorbed doses in organs and tissues of a rabbit, following computed tomography (CT) examinations, using a dedicated 3D voxel model. Absorbed doses in relevant organs were calculated using the MCNP5 Monte Carlo software. Calculations were perfomed for two standard CT protocols, using tube voltages of 110 kVp and 130 kVp. Absorbed doses were calculated in 11 organs and tissues, i.e., skin, bones, brain, muscles, heart, lungs, liver, spleen, kidney, testicles, and fat tissue. The doses ranged from 15.3 to 28.3 mGy, and from 40.2 to 74.3 mGy, in the two investigated protocols. The organs that received the highest dose were bones and kidneys. In contrast, brain and spleen were organs that received the smallest doses. Doses in organs which are stretched along the body did not change significantly with distance. On the other hand, doses in organs which are localized in the body showed maximums and minimums. Using the voxel model, it is possible to calculate the dose distribution in the rabbit’s body after CT scans, and study the potential biological effects of CT doses in certain organs. The voxel model presented in this work can be used to calculated doses in all radiation experiments in which rabbits are used as experimental animals.

     

Stem cell-based composite tissue constructs for regenerative medicine

A major task of contemporary medicine and dentistry is restoration of human tissues and organs lost to diseases and trauma. A decade-long intense effort in tissue engineering has provided the proof of concept for cell-based replacement of a number of individual tissues such as the skin, cartilage, and bone. Recent work in stem cell-based in vivo restoration of multiple tissue phenotypes by composite tissue constructs such as osteochondral and fibro-osseous grafts has demonstrated probable clues for bioengineered replacement of complex anatomical structures consisting of multiple cell lineages such as the synovial joint condyle, tendon-bone complex, bone-ligament junction, and the periodontium. Of greater significance is a tangible contribution by current attempts to restore the structure and function of multitissue structures using cell-based composite tissue constructs to the understanding of ultimate biological restoration of complex organs such as the kidney or liver. The present review focuses on recent advances in stem cell-based composite tissue constructs and attempts to outline challenges for the manipulation of stem cells in tailored biomaterials in alignment with approaches potentially utilizable in regenerative medicine of human tissues and organs.     

Glucosamine protects against radiation‐induced lung injury via inhibition of epithelial‐mesenchymal transition

Radiotherapy is one of the most important treatments for chest tumours. Although there are plenty of strategies to prevent damage to normal lung tissues, it cannot be avoided with the emergence of radiation‐induced lung injury. The purpose of this study was to investigate the potential radioprotective effects of glucosamine, which exerted anti‐inflammatory activity in joint inflammation. In this study, we found glucosamine relieved inflammatory response and structural damages in lung tissues after radiation via HE staining. Then, we detected the level of epithelial‐mesenchymal transition marker in vitro and in vivo, which we could clearly observe that glucosamine treatment inhibited epithelial‐mesenchymal transition. Besides, we found glucosamine could inhibit apoptosis and promote proliferation of normal lung epithelial cells in vitro caused by radiation. In conclusion, our data showed that glucosamine alleviated radiation‐induced lung injury via inhibiting epithelial‐mesenchymal transition, which indicated glucosamine could be a novel potential radioprotector for radiation‐induced lung injury.     

Morphological types of radiation-induced hematopoietic tissue cell death,its biological essence and significance at various stages of developing acute radiation injury

The results of morphologic investigation of radiation-induced cell death types in human and animal (dogs) hematopoietic tissue within an acute radiation injury are presented. It has been shown that early and late necrobiosis of myelokaryocytes occurs via apoptosis. An attempt to designate the pathogenic role of apoptosis in hematologic syndrome of acute radiation sickness was performed.     

Lymphoid tissue- and inflammation-specific endothelial cell differentiation defined by monoclonal antibodies

Endothelial cells play an essential role in immune responses by regulating the entry of leukocytes into lymphoid tissues and sites of inflammation. As an initial approach to analyzing endothelial cell specialization in relation to such immune function, we have produced monoclonal antibodies (MAB) against mouse lymph node endothelium. Three antibodies were selected: MECA-20, recognizing the endothelium of all blood vessels in lymphoid as well as non-lymphoid organs; MECA-217, which stains the endothelium lining large elastic arteries, but among small vessels is specific for post-capillary venules within lymphoid organs and tissues exposed to exogenous antigen, such as skin and uterus; and MECA-325, an antibody that demonstrates specificity for the specialized high endothelial venules (HEV) that control lymphocyte homing into lymph nodes and Peyer's patches. MECA-325 failed to stain vessels in any non-lymphoid organs tested. Immunoperoxidase studies of HEV in lymph node frozen sections, and of isolated high endothelial cells in suspensions, demonstrated that the antigens recognized by all three antibodies are expressed at the cell surface; those defined by MECA-20 and MECA-325 are also present in the cytoplasm. To study the regulation of the antigens defined by these MAB in relation to extra-lymphoid immune reactions, we assessed their expression in induced s.c. granulomas as a model for chronic inflammation. Small vessels in the granulomas were already stained by MECA-217 in the first days of development. In contrast MECA-325 detected postcapillary venules (which frequently displayed the morphologic characteristics of HEV) only from approximately 1 wk, in parallel with the development of a persistent mononuclear cell infiltrate including numerous lymphocytes. The selective appearance of the MECA-325 antigen on vascular endothelium supporting lymphocyte traffic in both lymphoid and extra-lymphoid sites suggests that this antigen may play an important role in the process of lymphocyte extravasation. The demonstration of lymphoid organ- and inflammation-specific microvascular antigens offers direct evidence for a complex specialization of endothelium in relation to immune stimuli, and supports the concept that microvascular differentiation may play an important role in local immune responses.     

Postquadrantectomy breast deformities: classification and techniques of surgical correction

The deformity which is encountered following quadrantectomy (or similar procedures such as segmentectomy or partial mastectomy) and radiation therapy is difficult to evaluate objectively, and subjective assessment of the cosmetic outcome is extremely variable. In a group of 54 patients who underwent the procedure between 1979 and 1983, the types of cosmetic changes were evaluated and classified according to morphologic criteria. Four types of deformities and their related etiopathologic factors were identified. Type I is characterized by malposition and distortion of the nipple-areola complex and is mainly due to postoperative fibrosis and scar contracture. In type II deformity, localized tissue insufficiency is observed, which may be due to skin deficiency (type IIa), subcutaneous tissue deficiency (type IIb), or both (type IIab). Type III deformity is characterized by breast retraction and shrinkage and is mainly due to the effects of radiotherapy on residual breast parenchyma. In type IV deformity, severe radiation-induced damage to the skin, nipple-areola complex, and subcutaneous and glandular tissues is present. Surgical correction of each type of deformity is discussed, and examples are reported.     

SHARPIN is a key regulator of immune and inflammatory responses

Mice with spontaneous mutations in the Sharpin gene develop chronic proliferative dermatitis that is characterized by eosinophilic inflammation of the skin and other organs with increased expression of type 2 cytokines and dysregulated development of lymphoid tissues. The mutant mice share phenotypic features with human hypereosinophilic syndromes. The biological function of SHARPIN and how its absence leads to such a complex inflammatory phenotype in mice are poorly understood. However, recent studies identified SHARPIN as a novel modulator of immune and inflammatory responses. The emerging mechanistic model suggests that SHARPIN functions as an important adaptor component of the linear ubiquitin chain assembly complex that modulates activation of NF-κB signalling pathway, thereby regulating cell survival and apoptosis, cytokine production and development of lymphoid tissues. In this review, we will summarize the current understanding of the ubiquitin-dependent regulatory mechanisms involved in NF-κB signalling, and incorporate the recently obtained molecular insights of SHARPIN into this pathway. Recent studies identified SHARPIN as an inhibitor of β1-integrin activation and signalling, and this may be another mechanism by which SHARPIN regulates inflammation. Furthermore, the disrupted lymphoid organogenesis in SHARPIN-deficient mice suggests that SHARPIN-mediated NF-κB regulation is important for de novo development of lymphoid tissues.     

Vitrification of large tissues with dielectric warming: biological problems and some approaches to their solution

If large pieces of tissue and organs are to be successfully stored at low temperatures, some means must be found to minimize the disruption of extracellular structures by the ice that develops during conventional cryopreservation methods. The use of sufficiently high concentrations of cryoprotectant (CPA) to vitrify rather than freeze the tissue is a possible solution to this problem, and the retention of function of embryos and elastic arteries after vitrification suggests that some cells and tissues at least can withstand exposure to the high concentrations of CPA necessary for this process to occur. There are, however, additional problems in applying vitrifying techniques to bulky tissues and organs. These are related to the additional time required for tissue equilibration of CPA to occur and the consequences for toxic injury, the difficulty in achieving sufficiently rapid and uniform cooling rates to produce the required glassy state, and the even more rapid and uniform warming rates that are necessary to avoid devitrification. Non-uniformity of temperature will increase the risk of mechanical stresses and fractures developing in the glass during rapid warming. This paper reviews possible strategies and the progress that has been made in overcoming these problems. This will include the permeation of CPA mixtures into whole tissues and possibilities for reducing their toxicity by the inclusion of adjuncts such as ice inhibitors and sugars. The warming of tissues by dielectric heating is currently the only practical means by which sufficiently rapid rates can be achieved in bulky tissues given that the tolerable limits of CPA concentration will most likely be insufficient to prevent the development of ice nuclei during cooling. The biological effects of microwaves are reviewed and their effectiveness in producing the required uniformity in warming of tissue models of various shapes are discussed.