Bonghan Circulatory System as an Extension of Acupuncture Meridians

Phase-contrast microscopic image of a BHD from an artery of a rat. Total length ∼4 cm [42].

BHD inside a rabbit lymphatic vessel stained with Janus Green B [43].

BHD in brain ventricles of rabbits. Stereomicroscopic images at bottom of the fourth ventricle beneath the cerebellum of same rabbit before, (A), and after, (B), hematoxylin application. No BHD visible in panel A but, after hematoxylin staining and washing, BHD (arrows) emerged near sulcus, panel B. (C) Stereomicroscopic image of BHD (arrow) in an aqueduct and third ventricle of rabbit brain after hematoxylin and washing, lifted using a needle to show it was a floating tissue in cerebrospinal fluid. Inset: wound state of threadlike structure specimen, showing its elastic nature; overlapped regions show its optical transparency; two nodes present (arrowheads); scale bar, 60 μm. (D) Stereomicroscopic image of BHD (arrow) with corpuscle (thick arrow) and node (arrowhead); one end of BHD cut at front part of third ventricle [50].

Weblike network of BHDs revealed by using trypan blue. (A) Web of BHDs on visceral peritoneum around stomach near rat spleen; several small BHCs at crossing points (arrows); blood capillaries not stained. (B) Network of BHDs on omentum below stomach and over small intestine; three small corpuscles at crossing points of BHDs (arrows). (C) Inset: another part of same omentum as (A); floating BHD (open arrow) connected to BHDs (arrows) in omentum, showing BHDs on omentum as part of larger network of freely movable BHDs on internal organ surfaces [43].

Left figure shows BHD (arrow) on rat small intestine. Right three panels show nanoparticle flow after injection at point indicated by top broken line. For first 4 minutes, nanoparticles in first region, then moved to middle after 12 minutes, and finally moved to third region in 18 minutes; speed 0.6 mm/min in only one direction [74]. Extremely slow speed due to conditions, such as temperature and humidity, and to time lapse from opening abdomen to injection of nanoparticles, about 60 minutes; peristaltic motion of BHD nearly stopped. With improved methods, including better viability conditions and shorter time lapse, we were better able to measure flow speed (0.3 mm/s) [44].

(A) Resting potential and spontaneous electrical activity of a cell in a BHC. (B) At the moment of microcapillary insertion into cell membrane of BHC, potential decreased abruptly by about 38 ± 15.5 mV (n = 11) from reference potential of bath; V_d is potential drop; potential increased slowly to resting potential of 10.5 ± 8.4 mV (n = 11); V_e is small increase (dotted line); and T_e is time of increase (18.1 ± 14.0 sec (n = 11)). Resting potential remained stable with fine background fluctuations; irregular activity of spontaneous spikes in resting potential arose for durations (D_s) of about 16.6 ± 14.9 sec (n = 11). (C) Spontaneous activity recorded in a period expanded in time and in voltage; average amplitude (V_s) was 1.2 ± 0.6 mV (n = 11) and average period (T_s) 0.8 ± 0.6 sec (n = 11); spikes had an average half-width (F_s) of 0.27 ± 0.19 sec (n = 11) [57].

Trypan blue staining of BHD and BHC inside adipose tissues. (A) BHC and connected BHD inside adipose tissue around rat small intestine. (B) BHC and two BHDs near same rat small intestine; blood vessels and adipose tissues not stained [71].

Visualization of a Bonghan system on fascia surrounding tumor tissue in mouse skin. (A) Images of tumor tissue (arrows); left, image of mouse with two tumor tissues grown for 2 weeks after subcutaneous inoculation with human lung cancer cells; right, part of tumor tissue surface after skin resection; presence of Bonghan system hardly noticeable. (B) In-situ trypan blue staining revealed Bonghan ducts (dotted arrows) and corpuscles (arrow heads) on tumor tissue fascia; right panel, magnified view of left panel; trypan blue did not stain blood vessels. (C) Sample showing multiple Bonghan ducts (dotted arrows) on tumor tissue fascia surface; right panel, magnified view of left panel, showing branching of Bonghan duct which notably, enters nearby fat layer (double arrow). (D) Trypan blue technique revealed Bonghan ducts (dotted arrows) along bundle of blood vessels and nerves; right panel, magnified view clearly showing duct along blood vessel; bundle of blood vessels and nerves connect tumor tissue (arrow) at lower left corner to outside skin. Samples A, B, C, and D from different mice [94].

(A) Threadlike structure with enshrouding fibrin observed on a slide by differential interference contrast microscopy. The fibrin and the BHD were hardly distinguishable and red blood cells scattered around; scale bar, 50 μm. (B) Threadlike structure with enshrouding fibrin observed using Acridine orange fluorescence method; scattered dotted points, white blood cells; long rod-shaped nuclei from threadlike structure; clearly distinguishes fibrin from BHD; scale bar, 50 μm [65].

Confocal laser scanning microscopic images of BHDs showing rod-shaped nuclei (arrows) distributed in broken-lined striped fashion. (A) BHD stained by YoYo-1, a DNA-specific dye, after removal from Alcian-blue-injected rabbit lymphatic vessel [46]. (B) BHD stained by Acridine orange, a DNA-specific dye, after removal from a Janus-Green-B-injected rabbit lymphatic vessel [67]. (C) BHD stained using a Feulgen reaction, a DNA-specific dye, after removal from the rabbit organ surfaces [42]. (D) BHD was stained by Acridine orange after removal from rabbit caudal vena cava [61]. Shapes, lengths, and distributions of rod-shaped nuclei similar to each other in all four cases, suggesting that BHDs in lymph vessels, in blood vessels, and on organ surfaces belong to same system.