Visualization of the Primo Vascular System Afloat in a Lymph Duct

Because of the potential roles of the primo vascular system (PVS) in cancer metastasis, immune function, and regeneration, understanding the molecular biology of the PVS is desirable. The current state of PVS research is comparable to that of lymph research prior to the advent of Lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1). There is very little knowledge of the molecular biology of the PVS due to difficulties in identifying and isolating primo endothelial cells. Present investigations rely on the morphology and the use of differential staining procedures to identify the PVS within tissues, making detailed molecular studies all but impossible. To overcome such difficulties, one may emulate the explosive development of lymph molecular biology. For this purpose, there is a need for a reliable method to obtain PVS specimens to initiate the molecular investigation. One of the most reliable methods is to detect the primo vessels and primo nodes afloat in the lymph flow. The protocols for observation of the PVS in the large lymph ducts in the abdominal cavity and the thoracic cavity were reported earlier. These methods require a laparectomy and skillful techniques. In this work, we present a protocol to identify and harvest PVS specimens from the lymph ducts connecting the inguinal and the axillary nodes, which are located entirely in the skin. Thus, the PVS specimen is more easily obtainable. This method is a stepping-stone toward development of a system to monitor migration of cancer cells in metastasis from a breast tumor to the axillary nodes, where cancer cells use the PVS as a survival rope in hostile lymph flow.     

Isolation and Morphological Features of Primo Vessels in Rabbit Lymph Vessels

Until now, even though intensive research has been dedicated to the primo vascular system (PVS) during these years, no statistical data on primo vessels and primo vessels in lymph flow have been available. Recently, the general morphological features of primo vessels in lymph vessels around the abdominal aorta were identified from microdissections of tissues from New Zealand White rabbits, and with Alcian blue staining, primo vessels in lymphatic vessels could be definitely identified under a digital microscope. The micro-dissected specimens in situ reveal rod-shaped nuclei stained by Acridine orange. The blue-stained nuclei, which were distributed in a broken-lined stripe, formed a tube structure of about 20 μm in diameter. The distance between the nuclei of two cells on neighboring aligned stripes, which is also the diameter of the micro tube, was measured to be about 5～10μm. The average length of the primo vessels was 2.4 mm, with the longest being 5.6 mm. The average size of the primo vessel was 50 μm, and the average diameters of the primo and the lymph vessels were 26.0 μm and 258.5 μm, respectively. Occasionally, without the use of Alcian blue staining, milk-white transparent primo vessels were observed floating in lymph vessels. Thus, we suggest that the PVS might also have an important function connected with the lymph system. We also expect the traditional Korean meridian system to leave its invisible world during the last thousands of years and soon enter the visible scientific world.     

Primo Vessel as a Novel Cancer Cell Migration Path from Testis with Nanoparticle-Labeled and GFP Expressing Cancer Cells

Background/aimRecently, a novel circulatory system, the primo vascular system (PVS), was found to be a potent metastatic route of cancer cells. The aim of the current work is to demonstrate that cancer cells injected into the testis migrate through the primo vessel (PV).Materials and methodsNCI-H460 cells labeled with fluorescent nanoparticles (FNP) or green fluorescent protein (GFP) gene transfection were injected into testicular parenchyma in 24 rats. After 24 hours of injection, the abdominal cavity was investigated via a stereomicroscope, to detect the PVS, and the samples were analyzed histologically with 4′,6-diamidino-2-phenylindole (DAPI) and hematoxylin and eosin.ResultsInjected cancer cells were detected inside the PVS distributed on the abdominal organs. Some were detected inside intestinal parenchyma into which the attached primo vessels (PVs) entered.ConclusionThe results supported the fact that the PVS may be a novel migration path of cancer cells, in addition to the lymphatic and hematogenous routes.     

Putative Primo-Vascular System in Rabbit Placenta

The primo vascular system (PVS) is a very important topic of study nowadays because of their role in transport and regeneration of tissue and in cell migration and cancer metastasis. The PVS was detected in different organs of the rabbit but not in the placenta. In this work, we observe the PVS inside the blood vessels of the placenta for the first time. The main characteristic features of the primo vessels (PVs) from the rabbit placenta were in agreement with the PVS in different organs of animals, including the rod-shaped nuclei and their arrangement.     

Primo vascular system of murine melanoma and heterogeneity of tissue oxygenation of the melanoma

Murine melanoma requires the complex development of lymphatic, vascular, and non-vascular structures. A possible relationship between the primo vascular system (PVS) and the melanoma metastasis has been proposed. In particular, the PVS may be involved in oxygen transport. Vasculogenic-like networks, similar to the PVS, have been found within melanoma tumors, but their functional relationship with the PVS and meridian structures are unclear. Herein, we report on the use of an electrochemical O(2) sensor to study oxygenation levels of melanoma tumors in mice. We consistently found higher tissue oxygenation in specific sites of tumors (n=5). These sites were strongly associated with vascular structures or the PVS. Furthermore, the PVS on the tumor surface was associated with adipose tissue. Our findings suggest that the PVS is involved in the regulation of metastasis.     

Primo Vessel Inside a Lymph Vessel Emerging From a Cancer Tissue

Primo vessels were observed inside the lymph vessels near the caudal vena cava of a rabbit and a rat and in the thoracic lymph duct of a mouse. In the current work we found a primo vessel inside the lymph vessel that came out from the tumor tissue of a mouse. A cancer model of a nude mouse was made with human lung cancer cell line NCI-H460. We injected fluorescent nanoparticles into the xenografted tumor tissue and studied their flow in blood, lymph, and primo vessels. Fluorescent nanoparticles flowed through the blood vessels quickly in few minutes, and but slowly in the lymph vessels. The bright fluorescent signals of nanoparticles disappeared within one hour in the blood vessels but remained much longer up to several hours in the case of lymph vessels. We found an exceptional case of lymph vessels that remained bright with fluorescence up to 24 hours. After detailed examination we found that the bright fluorescence was due to a putative primo vessel inside the lymph vessel. This rare observation is consistent with Bong-Han Kim’s claim on the presence of a primo vascular system in lymph vessels. It provides a significant suggestion on the cancer metastasis through primo vessels and lymph vessels.     

Primo Vascular System in Human Umbilical Cord and Placenta

The primo vascular system (PVS) has been observed in various animals such as mice, rats, rabbits, dogs, swine, and cow, but not in humans. In this work, we report on the observation of a human PVS on both the epithelial fascia and inside the blood vessels of the umbilical cord (UC). The main morphological characteristics of the primo vessels (PVs) and primo nodes (PNs) from the human UC were in agreement with those of the PVS in various animal organs, including the thicknesses and the transparency of the PVs, the sizes of the PNs, the broken-line arrangement of the rod-shaped nuclei, the sparse distribution of nuclei, and the presence of hollow lumens in the central inner parts of the PNs. It was rather surprising that the human PV was not thicker than the PVs from small animals. The difference between the PVS and blood/lymph vessels was confirmed using immunofluorescence staining of von Willebrand factor, CD31, LYVE-1, and D2-40. The positive expression of the PVS to proliferating cell nuclear antigen, a cell-proliferation marker, was consistent with the recent finding of very small embryonic-like stem cells in the PVS of mice.     

VEGF-C在非小细胞肺癌血清及淋巴液中的表达及意义

目的通过对非小细胞肺癌(NSCLC)患者血清中、淋巴液中血管内皮生长因子C(VEGF-C)水平表达的测定来探讨VEGF-C和VEGFR-3在NSCLC肿瘤淋巴转移中的意义。方法采取肺部良性病变手术患者术前血清及术中采取胸导管内淋巴液作为对照组,肺癌患者术前血清及术中行肺癌根治术前,采胸导管内淋巴液为肺癌组,应用酶联免疫(ELISA)法检测VEGF-C浓度。结果55例NSCLC患者淋巴液中VEGF-C水平较血清中VEGF-C水平显著升高,而二者较肺部良性病变对照组淋巴液血清中VEGF-C浓度显著升高(P均<0.01)。肿瘤淋巴结转移组中淋巴液血清中VEGF-C浓度显著高于未有淋巴结转移患者(P均<0.01)。淋巴结转移组中,淋巴液VEGF-C浓度较血清VEGF-C浓度明显增高(P<0.01)。结论NSCLC患者淋巴液及血清中VEGF-C水平高低与肺癌淋巴结转移有关,对其检测可以作为肺癌侵袭转移的量化指标,特别是对淋巴液VEGF-C的研究,可为抑制肿瘤淋巴管生成的靶向治疗提供理论依据。     

中医药治疗胃癌淋巴转移的研究进展

胃癌的发病率和死亡率在所有肿瘤中均较高,我国因为胃癌死亡的人数每年高达49. 8万人。由于早期胃癌的发病特征不明显,多数到医院治疗的患者均已是中晚期胃癌,给家庭和社会都带来了沉重的负担。中晚期胃癌的术后生存期显著低于早期胃癌,其中最重要的原因在于胃癌的淋巴转移。淋巴转移是胃癌的主要转移途径,进展期胃癌的淋巴转移率高达70%左右,早期胃癌也可有淋巴转移。胃癌的淋巴结转移率和癌灶的浸润深度呈正相关。目前的治疗手段除手术切除、淋巴化疗及分子靶向治疗外,中医药因其中毒性低、治疗效果好、价格低的优势也发挥了重要的作用。中医认为胃癌的发生多因饮食不洁、情志失调、素体亏虚而致痰凝、气阻、血瘀与胃发为本病,病机总属本虚标实,临床中医治疗多以化痰解毒、健脾益气、温中散寒为主。近年来,多项研究证明中医药治疗胃癌淋巴转移在临床中疗效显著、作用广泛,其可能通过抑制基质金属蛋白酶对基底膜和细胞外基质(ECM)的水解,激发肿瘤失巢凋亡,抑制淋巴管新生,调控与肿瘤相关基因的表达等途径来治疗胃癌淋巴转移。文章从近年来中医药在治疗胃癌淋巴转移中的理论研究、实验研究以及临床研究的进展进行综述,揭示中药作用的机制及其作用靶点,以期为中医药治疗胃癌淋巴转移的深入研究以及临床上的应用提供科学依据。     

右旋柠烯对裸鼠人乳腺癌模型淋巴转移的抑制作用

目的:观察右旋柠烯对人乳腺癌淋巴管生成和淋巴结转移的影响,探讨其抗肿瘤转移机制。方法:人乳癌细胞株MDA-MB-435裸鼠乳腺原位种植建立乳癌模型,免疫组化法检测右旋柠烯对乳癌组织微淋巴管密度(LMVD)和VEGF-C的影响。结果:右旋柠烯组乳癌生长和淋巴结转移受到抑制,LMVD降低,VEGF-C表达下调。结论:右旋柠烯通过影响VEGF-C诱导的淋巴管生成抑制乳癌淋巴结转移。     

Bonghan Ducts as Possible Pathways for Cancer Metastasis

ObjectiveThe 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 MethodsTumor 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.ResultsWe demonstrate that the trypan blue staining technique allows the first visualization of BHDs which are connected to tumor tissues.ConclusionSince BHDs are known to make up a circulatory system corresponding to acu puncture 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.     

Primo Vascular System Floating in Lymph Ducts of Rats

An epoch-making development in the gross anatomy of the lymph system has emerged: the observation of the primo vascular system (PVS), which is a threadlike structure floating in lymph ducts. The PVS, which was proposed as the conduit for the acupuncture Qi, is a complex network distributed throughout an animal's body. The lymph-PVS, which is a subsystem of the PVS, is one of the most convincing visual demonstrations of the PVS. Because its existence is not easily demonstrated, even with a microscope, due to its transparency, in current anatomy its existence is largely unknown despite its potential significance in physiology and medicine. The lymph-PVS has been observed in rabbits, rats, and mice by several independent teams. Because the involved techniques are rather complicated, we provide detailed protocols for surgery, for injection of the staining dye, and for detection, extraction, and identification of the PVS in a rat.     

中医药对淋巴管新生与回流功能的作用研究进展＊

淋巴管作为循环系统的重要组成部分之一，具有调节机体体液稳态，协助免疫监视和肠道脂质吸收等重要作用。淋巴管新生是机体生理和病理过程中维持脉管系统结构和功能正常的重要手段，淋巴管新生调控对于防治肿瘤、心血管等诸多疾病有着潜在的临床转化意义；淋巴回流功能则与关节炎症等疾病发病机制关系密切。在循环系统中，相较于中医药调控血管相关疾病的发病机制已取得很大进展，近年来对于淋巴管的研究则明显相对滞后。本文从中医药作用于淋巴管新生及回流功能角度对这一领域的研究进展作一综述，以期为临床上中医药治疗相关疾病提供新的思路与方法。     

Expression of keratin 10 in rat organ surface primo-vascular tissues

The primo-vascular system is described as the anatomical structure corresponding to acupuncture meridians and has been identified in several tissues in the body, but its detailed anatomy and physiology are not well understood. Recently, the presence of keratin 10 (Krt10) in primo-vascular tissue was reported, but this finding has not yet been confirmed. In this study, we compared Krt10 expression in primo-vascular tissues located on the surface of rat abdominal organs with Krt10 expression on blood and lymphatic vessels. Krt10 protein (approximately 56.5 kDa) was evaluated by western blot analysis and immunohistochemistry. Krt10 (IR) in the primo-node was visualized as patchy spots around each cell or as a follicle-like structure containing a group of cells. Krt10 IR was also identified in vascular and lymphatic tissues, but its distribution was diffuse over the extracellular matrix of the vessels. Thus Krt10 protein was expressed in all three tissues tested, but the expression pattern of Krt10 in primo-vascular tissue differed from those of blood and lymphatic vascular tissues, suggesting that structural and the regulatory roles of Krt10 in primo-vascular system are different from those in blood and lymphatic vessels.     

Effects of the Chinese herbal medicines Bupleuri radix, Ginseng radix, and Zingiberis rhizoma on lymphatic vessel activity in rats

The aim of the present study was to determine the effects of the Chinese herbal medicines Bupleuri radix, Ginseng radix and Zingiberis rhizoma on spontaneous lymphatic vessel activity. The effect of each herbal medicine on in vivo lymphatic flow was examined by injection of dye into the femoral regions of rats after feeding with the herbal medicines. In an in vitro study, spontaneous changes in diameter of the rat thoracic duct were monitored, and each segment was exposed to each herbal medicine. In the in vivo study, 100% of the right iliac lymphatic node were positively stained in the herbal medicine group, whereas only 40% of the node were positively stained in the control group. In the in vitro study, Bupleuri radix and Ginseng radix increased the amplitude of spontaneous activity of lymphatic vessels in a concentration-dependent manner with or without L-NAME, an NO synthase inhibitor. The results indicated that the herbal medicines Bupleuri radix and Ginseng radix activated spontaneous lymphatic vasomotion and lymph flow, and the mechanisms of this effect seem to be independent of endothelial cells.     

Primo Bundles Identified by Microcomputed Tomography in Primo Vascular Tissue on the Surface of Rat Abdominal Organs

BackgroundThe primo vascular system (PVS) is a novel network composed of primo nodes (PNs) and primo vessels (PVs). Currently, its anatomy is not fully understood.ObjectivesThe aim of this study was to elucidate the three-dimensional PN–PV structure.MethodsOrgan-surface PVS tissue was isolated from healthy and anemic rats. The tissues were analyzed by X-ray microcomputed tomography (CT), hematoxylin and eosin staining, and scanning electron microscopy.ResultsFrom CT images, we identified one or more bundles in a PV. In the PN, the bundles were enlarged and existed in isolation and/or in anastomosis. The transverse CT images revealed four areas of distinct intensities: zero, low, intermediate, and high. The first two were considered to be the sinuses and the subvessels of the PVS and were identified in the hematoxylin and eosin–stained PN sections. The enlargement of the PN from anemic rats was associated with an increase in the intermediate-intensity area. The high-intensity area demarcated the bundle and was overlapped with the mesothelial cells. In scanning electron microscopy, the PV bundles branched out, tapering down to a single bundle at some distance from the PN. Each bundle was composed of several subvessels (∼5 μm). Clustered round microcells (1–25 μm), scattered flat oval cells (∼15 μm), and amorphous extracellular matrix were observed on the surface of the PVS tissue.ConclusionsThe results newly showed that the primo bundle is a structural unit of both PVs and PNs. A bundle was demarcated by high CT intensity and mesothelial cells and consisted of multiple subvessels. The PN bundles contained also sinuses.     

Historical Observations on the Half-Century Freeze in Research between the Bonghan System and the Primo Vascular System

This article provides potential reasons for the past 45-year halt in research between the time of the Bonghan system of Bong Han Kim (B.H. Kim) and that of the primo vascular system (PVS) of Kwang-Sup Soh (K.S. Soh), briefly but more accurately in its history. Over the years, numerous questions related to the Bonghan system and the PVS have arisen, especially from researchers interested in pursuing PVS research: When and how did B.H. Kim's study results on the Bonghan system become known to public? Why did B.H. Kim and his publications disappear after 1966? Why was little study performed on the system for almost 50 years after Kim? Why and how was the research on the system reinitiated in 2002 by Kwang-Sup Soh? Why did the Bonghan system become the PVS? These questions, as well as technical difficulties in identifying the system, have discouraged many researchers from becoming involved in research on the system. The motivation for preparing this article was to remove doubts about the existence of this important organ, which might have been caused by its unusual and unclear historical background, by providing an accurate history.     

收涩类中药对小鼠肝癌血道及淋巴道转移的影响

目的观察收涩类中药对小鼠H22腹水型肝癌细胞血道和淋巴道转移的影响。方法血道转移实验,取昆明种小鼠50只,随机分为5组,每组各10只,即阴性对照组、参一胶囊组(120 mg·kg~(-1)·d~(-1))、椿皮组(62.5g·kg~(-1)·d~(-1))、乌梅组(250 g·kg~(-1)·d~(-1))和山莱萸组(75 g·kg~(-1)·d~(-1))。,将小鼠尾静脉接种H22肝癌细胞悬液,自造模之日起,连续灌胃给药14 d后,脱颈椎处死小鼠,观察小鼠的肺部转移瘤生长情况及血液流变学指标的变化;淋巴道转移实验分组与给药同血道转移实验,采用腹水型肝癌H22瘤株,在小鼠右下肢爪垫内侧皮下注射接种,连续给药21 d后,脱颈椎处死小鼠,观察小鼠实体瘤的瘤重量、抑瘤率、肺部转移率、淋巴结转移等相关指标的变化情况。结果血道转移实验中,椿皮组和山茱萸组的肺部转移结节数较模型组明显减少,椿皮组能明显改善血液流变学指标;淋巴道转移实验中,乌梅组和山茱萸组的肺部转移结节较模型组明显减少,抑瘤率明显增加;三味收涩药的淋巴结转移显著减少。结论收涩类中药对H22腹水型肝癌细胞血道及淋巴道两种转移途径均有一定的抑制作用。     

收涩中药对小鼠肝癌淋巴道转移的影响

目的:观察收涩中药对小鼠肝癌淋巴道转移的影响。方法:以腹水型肝癌H22瘤株在小鼠右下肢爪垫内侧皮下注射接种,复制淋巴道转移模型,观察小鼠实体瘤的瘤重量抑瘤率、肺部转移率、淋巴结转移情况等的指标变化。结果:乌梅组和山茱萸组的肺部转移结节较模型组明显减少,抑瘤率明显增加;三味收涩药的淋巴结转移显著减少。结论:收涩中药对H22肿瘤淋巴道转移有一定的抑制作用。