Primo vascular system accompanying a blood vessel from tumor tissue and a method to distinguish it from the blood or the lymph system.

A primo vessel was observed in the abdominal cavity in the lung cancer mouse model, and its function as an extra metastatic path was observed. In this work, we found a primo vessel accompanying a blood vessel emanating from a tumor in the skin. We also presented simple and efficient criteria to distinguish a primo vessel from a blood or a lymph vessel and from a nerve. The criteria for using DAPI and Phalloidin will be useful in clinical situations to find and identify the primo vessels among the blood vessels, lymph vessels, or nerves in the tissue surrounding a tumor such as a melanoma or breast cancer.

Putative primo-vascular system in mesentery of rats.

Primo-vessels have been observed in the rat abdominal cavity as floating thread like structures on and not adhering to fascia-wrapped internal organs. To date their presence, locations, and lengths have been irregular and unpredictable, and their identification not regularly repeatable, thus they have remained a nagging enigma in primo-vascular system research for several years. In this work, locations were found where primo-vessels were regularly present and observed repeatedly. These vessels were not floating or freely movable but lay in a regular position in the mesentery in the abdominal cavity of the rat, being observed between the cecum and small intestine and between the colon and mesentery root. The difference between a lymph vessel and a primo-vessel is described in anatomical and histological aspects. In addition, trypan blue was found to enter primo-vessels through the surrounding membranes and filled spaces between fibers comprising the primo-vessels. It is conjectured that the previously observed floating primo-vessels had anomalously and irregularly emerged, for some unknown physiological reasons, from primo-vessels normally located in the fascia-like mesentery.

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.

Use of magnetic nanoparticles to visualize threadlike structures inside lymphatic vessels of rats.

A novel application of fluorescent magnetic nanoparticles was made to visualize a new tissue which had not been detectable by using simple stereomicroscopes. This unfamiliar threadlike structure inside the lymphatic vessels of rats was demonstrated in vivo by injecting nanoparticles into lymph nodes and applying magnetic fields on the collecting lymph vessels so that the nanoparticles were taken up by the threadlike structures. Confocal laser scanning microscope images of cryosectioned specimens exhibited that the nanoparticles were absorbed more strongly by the threadlike structure than by the lymphatic vessels. Further examination using a transmission electron microscope revealed that the nanoparticles had been captured between the reticular fibers in the extracellular matrix of the threadlike structures. The emerging technology of nanoparticles not only allows the extremely elusive threadlike structures to be visualized but also is expected to provide a magnetically controllable means to investigate their physiological functions.

Immunohistochemical and electron microscopic study of the meridian-like system on the surface of internal organs of rats.

Threadlike structures on the surfaces of mammalian internal organs were recently observed, and they can be the anatomical basis of the meridian-like network of internal organs as detected by the Bi-Digital O-Ring test imaging method. Morphological and histological studies were performed to reveal their novel features by using confocal and electron microscopy, and there is evidence that reveals that they have a ductal structure in which some liquid flows. They form a part of the network of acupuncture meridians that is thought to be a circulatory system distributed throughout an animal body. In order to confirm further the circulatory function it was necessary to provide evidences for the existence of endothelial cells at the inner boundaries of the microchannels that form the threadlike ducts. In this work we performed immunohistochemical studies for the identification of endothelial cells. In addition, we identified the extracellular matrices between the microchannels in the threadlike structure by using antifibronectin, elastin, laminin, and collagen type IV. Since immunohistochemical data are not sufficient for definite proof we supplemented the analysis with ultra structural information by using transmission electron microscopy (TEM). Yet TEM images show only ultrathin sections (50 nm) of a specimen, we needed to take 3-dimentional images to provide a visual demonstration of the microchannels in the threadlike ducts. We conducted high voltage electron microscopic studies for this purpose and obtained corroborating tomograms that exhibit the microchannels. In conclusion, the meridian-like tissues are ducts with microchannels, which implies the circulatory function of the acupuncture meridian network.

Electron microscopic study of novel threadlike structures on the surfaces of mammalian organs.

The ultrastructures of novel threadlike structures (NTSs) and corpuscles on the surfaces of internal organs of rats were investigated using electron microscopy. The samples were studied in situ by using a stereomicroscope and were taken for further morphological analysis. Scanning electron microscope (SEM) images revealed a bundle structure of threadlike tissue, which was composed of several 10-micro m-thick subducts. The surfaces of the corpuscles were rather coarse and fenestrated. The corpuscles had cucumber-like shapes with an average length of about 2 mm and a thickness of about 400 micro m. Transmission electron microscope (TEM) images disclosed disordered collagen fibers, which formed the extracellular matrix of the threadlike tissue, and immune-function cells, like macrophages, mast cells, and eosinophils. Sinuses of various diameters, which were thought to be cross-sections of the lumens of the subducts, were observed in the TEM, cryo-SEM and focused-ion-beam SEM images. These SEM images were obtained for the first time to reveal the detailed structure of the NTSs that were only recently discovered.

Feulgen reaction study of novel threadlike structures (Bonghan ducts) on the surfaces of mammalian organs.

Threadlike structures on the surfaces of internal organs, which are thought to be part of the Bonghan duct system, were first reported about 40 years ago, but have been largely ignored since then. Recently, they were rediscovered, and in this study we discuss the Feulgen reaction that specifically stains DNA in order to identify these structures on the surface of rabbit livers as part of the Bonghan system. The distribution, shapes, and sizes of their nuclei are found to be similar to those of intravascular threadlike structures. The endothelial nuclei are rod-shaped, 10-20 mum long, and aligned in a broken-line striped fashion. The threadlike structure consists of a bundle of several subducts, which is a characteristic feature of Bonghan ducts and distinguishes them morphologically from lymphatic vessels. In addition, the Feulgen reaction clearly demonstrates that the subducts pass through a corpuscle, which is usually irregular or oval-shaped and is connected to two or several threadlike structures that form a web on the surfaces of organs. Furthermore, spherical granules of about 1 mum in diameter are detected in the subducts. These granules were well stained by using the Feulgen reaction, which implies that they contain DNA. According to previous reports, a granule is a type of microcell and plays an essential role in the physiology and therapeutic effect of the Bonghan system and acupuncture. This role has yet to be elucidated.