Vyacheslav Ogay

Comparison of the Characteristic Features of Bonghan Ducts, Blood and Lymphatic Capillaries

OBJECTIVE To show that the characteristic morphological and ultrastructural features of a Bonghan corpuscle and duct presented here are consistent with the description given in the early reports of Bonghan Kim. MATERIALS AND METHODS We compared the morphological aspects of Bonghan ducts with those of blood and lymphatic capillaries on the ultrastructural level to display the manifestly distinctive nature of the Bonghan system. RESULTS The walls of the ductules were observed to be composed of a single layer of endothelial cells with characteristic rod-shaped nuclei and were not surrounded by a basal lamina or by accessory cells, such as pericytes or smooth muscle cells. The abluminal cell membranes of Bonghan ductules were not attached by anchoring filaments to the fibers of extracellular matrices as observed in lymphatic capillaries. The cytoplasmic processes of ductule endothelial cells appear to form overlapping and interdigitated interconnections which completely lack junctional elements. Although the cytoplasm of ductule endothelial cells contained a well-developed rough endoplasmic reticulum and many free ribosomes and polysomes, there was a relatively small number of pinocytotic vesicles and lacks specific organelles, such as Weibel-Palade bodies. CONCLUSIONS The Bonghan corpuscles are specialized structures consisting of different types of immune cells randomly scattered as single cells in the matrix or clustered in follicle-like formations. Moreover, the Bonghan ductules in the corpuscle contain flowing immune cells and occasionally basophilic bodies.

Bonghan Ducts as Possible Pathways for Cancer Metastasis

OBJECTIVE The present study has been designed to find a possible new route for the metastasis of cancer cells on the fascia surrounding tumor tissue using a novel technique of trypan blue staining. MATERIALS AND METHODS Tumor tissues were grown in the skin of nude mice after sub-cutaneous inoculation with human lung cancer cells. Trypan blue was recently identified as a dye with specificity for Bonghan ducts (BHDs) and not other tissues, such as blood or lymph vessels or nerves. RESULTS We demonstrate that the trypan blue staining technique allows the first visualization of BHDs which are connected to tumor tissues. CONCLUSION Since BHDs are known to make up a circulatory system corresponding to acupuncture meridians or collaterals, we propose that, in addition to the currently known blood or lymph vessels, BHDs on tumor tissue fascia may be a novel pathway for metastasis.

Visualization of Bonghan Microcells by Electron and Atomic Force Microscopy

OBJECTIVES The origin of adult stem cells remains an open question. If they derive from embryos, it is difficult to determine the mechanism which interrupts their differentiation during tissue formation. In the 1960s, the Bonghan microcell was suggested as one possible, yet to be described, route of stem cell production, such that they have the potential to proliferate to produce normal cells. MATERIALS AND METHODS In this study, Bonghan microcells were isolated from Bonghan tissues on rat organ surfaces, and their detailed morphology examined by electron and atomic force microscopy. RESULTS The ultrastructure observed distinguished them from apoptotic bodies and other microorganisms, and their unique, possible proliferation feature, as protruding threads, was imaged by atomic force microscopy. CONCLUSIONS The unique threadlike structure of the Bonghan microcell is consistent with Prof. Kims observation in the first step of making a cell. Understanding of the functions of this threadlike structure may give a clue to understand the origin or the differentiation cue of adult stem cells.

The Flow Path of Alcian Blue From the Acupoint BL23 to the Surface of Abdominal Organs

Two hours after Alcian Blue (AB) dye was injected at the rat acupoint BL23, the abdominal cavity was examined and AB-stained threadlike structures were observed on the right abdominal cavity. Those threadlike structures were mainly distributed on the surfaces of the duodenum, colon and cecum. These threadlike structures were thin (about 50 microm) and moved freely, and were connected to corpuscles that were about 500 x 200 microm wide and also stained with AB. On analyzing the histology of the threadlike structures, rod-shaped nuclei, bundles of collagen fibers, reticulofibers, and squamous-like epithelial cells were observed. Immune cells and some sinuses were inside the threadlike structures. These characteristics describe those of Bonghan ducts. The flow paths from the acupoint to internal organs can possibly be used as paths for drug delivery.

Identification and Characterization of Small Stem-Like Cells in the Primo Vascular System of Adult Animals

Pluripotent stem cells have been recently identified in many tissues and intensively studied for their cell-therapeutic potential to renew and replace lost cells in any damaged tissue or organ. However, the true nature and origin of these cells is still unclear. Based on data obtained by Dr. Bonghan Kim (1965), we put forward a hypothesis that a rare population of small stem-like (SSL) cells found in the primo-vascular system (Bonghan system) possesses a broad tissue differentiation potential and that these cells are capable of differentiation into all specialized cells in the adult body. To examine the hypothesis in this study, we isolated SSL cells from primo-vascular system of rats and characterized their morphological and phenotypic features. Our microscopic examinations indicated that SSL cells are very small, approximately 3–4 μm in diameter. Hematoxyline-eosin staining revealed that SSL cells have an extremely dense eccentrically located round nucleus surrounded by basophilic cytoplasm. Immunofluorescence analysis showed that SSL cells expressed embryonic stem cell markers: CD133, Oct 4, and Nanog. Thus, based on our preliminary data, we suppose that SSL cells in the primo-vascular system might be variants of embryonic stem-like cells which can differentiate into tissue-specific cells and regenerate damaged tissue or organ.

Acupuncture Muscle Channel in the Subcutaneous Layer of Rat Skin

Using a mixed-dye injection technique, we found a novel kind of muscle fiber with a lumen, established its precise location in the subcutaneous muscle layer along the acupuncture muscle of the bladder line, and determined its detailed ultrastructure. The channels with flowing liquid were a novel kind of muscle fibers with lumens and they were located in the subcutaneous muscle layer of rat. Their detection was realized by using chrome-hematoxylin and a mixture of fluorescent nanoparticles and commercial Pelikan ink. These acupuncture muscle channels were hidden among the neighboring skin skeletal muscle fibers and were barely distinguishable from them with light microscopes. Only with a transmission electron microscope were their characteristic features shown to be different from normal skin skeletal muscle. These features included undifferentiated muscle fibers that resembled immature myofibrils without Z-lines and reassembled telophase nuclei.

Primo-Vessels as New Flow Paths for Intratesticular Injected Dye in Rats

After intratesticular injection of a chromium hematoxylin and fluorescent nanoparticle solution, we found a novel flow path in the abdominal cavity consisting of primo-vessels and primo-nodes. This flow path formed a network that crossed over the surfaces of abdominal organs, and generally linked to the greater omentum and adjacent visceral peritoneum. Some of these structures terminated at organs such as the small intestine and the urinary bladder; occasionally, the network entered the parenchyma of organs. The semitransparent primo-vessels and nodes were wholly or partially stained dark-blue by chromium hematoxylin. Injected nanoparticles were also observed in primo-vessels and nodes as well as the parenchyma of organs which were the site of primo-vessel termination. Transmission electron microscopy showed that the primo-vessels consist of many sinuses (4-6 mum), surrounded by collagen fibers, specific granules, cellular remnants, immune cells, extracellular matrices, and hematopoietic cells. These histological features are completely different from blood and lymph vessels indicating that primo-vessels are novel structures that allow the flow of dye.

Catecholamine-storing Cells at Acupuncture Points of Rabbits

Recent studies have shown that specific sites of the skin related to the acupoints contain a high concentration of catecholamines, especially noradrenaline (NA). Considering this newly discovered property of the acupoints we assumed that heterogeneous distribution of cutaneous catecholamines could be associated with a specific location of catecholamine-storing cells in acupoint sites. In the present work we used an immunohistochemical method and confocal laser scanning microscopy to examine the presence of catecholamine-storing cells at acupoints of rabbits. Double immunofluorescence staining with antibodies against adrenaline and NA revealed only the cells storing NA in the dermal layer of rabbit skin. NA-storing cells were randomly scattered as single cells as well as existing in small clusters in a globular tissue formation surrounded by blood vessels and capillaries. Microscopic analysis of histological sections also revealed that the distribution of NA-storing cells was closely associated with the location of acupoints. Thus results from our study strongly suggest that acupoint areas of rabbit skin contain catecholamine-storing cells which can release a high level of NA during acupuncture stimulation.

Observation of Coiled Blood Plexus in Rat Skin with Diffusive Light Illumination

Blood plexuses are characteristic anatomical features of acupuncture points (APs). We developed an optical technique using diffusive light illumination to increase the brightened area of skin for observation of the blood plexuses in skin. We found that the blood plexuses were coiled blood vessels which came out of the perforations in the fascia of muscle. The coiled vessels could be straightened by stretching the skin. We observed a series of blood plexuses at the putative APs along the left and right kidney meridian lines in the abdominal skin of rats. In addition, the locations of the plexuses on the muscle fascia were just above the putative acupuncture muscle channels along the kidney meridians. Furthermore, immunohistochemical analysis of the skin specimens of the plexuses revealed its neurovascular bundle nature as expected from known anatomical features of the APs.

Novel Threadlike Structures on the Surfaces of Mammalian Abdominal Organs are Loose Bundles of Fibrous Stroma with Microchannels Embedded with Fibroblasts and Inflammatory Cells

Novel threadlike structures (NTSs) on the surfaces of mammalian abdominal organs have recently attracted interests regarding their ability to transport fluid, enable cell migration, and possibly facilitate cancer metastasis. Nevertheless, histological studies of NTSs have been sporadic and often have inconsistent interpretations of the NTS internal structure. In this article, we provide a synthetic and consistent view of the NTS internal structure: the NTS is a loose bundle of fibrous stroma that forms interstitial channels and microsinusoids infiltrated with inflammatory cells. The fibroblasts are embedded in the stroma and mostly aligned along the major axis of the NTS. The sinusoids, which are in inconsecutive cross sections, have boundaries more or less delineated by extracellular fibers, partly surrounded by endothelial-like cells, or both. We compare these morphological features to other well-known connective tissues (i.e., trabecular meshwork and lymphatic capillary) and discuss the biomechanical and biological functions of NTSs based on their structural characteristics.