J Acupunct Meridian Stud 2015;8(6):329e332

Available online at www.sciencedirect.com

Journal of Acupuncture and Meridian Studies

journal homepage: www.jams-kpi.com

IMPACTSHOT

Neurovascular Primo Bundles at the Kidney

Meridian Revealed Using Hemacolor Staining
Kyoung-Hee Bae 1, Hyun-Ji Gil 1, Yeong-Yung Yoo 1, Joo Ho Tai 1,
Hee-Min Kwon 2, Kwang-Sup Soh 1,*

1
Advanced Institutes of Convergence Technology, Seoul National University, Suwon,
South Korea
2
Department of Physics, Seoul National University, Seoul, South Korea

Received: Sep 1, 2015

Revised: Oct 6, 2015
Accepted: Oct 13, 2015

KEYWORDS
acupuncture;
hemacolor;
kidney meridian;
neurovascular primo
bundle;
primo vascular system;
primo vessel

Neurovascular bundles were observed earlier in connection
with the K-meridians of a rat [1]. At that time, the exis-
tence of the primo vessel (PV) in the bundle was not
noticed despite expectations in accordance with the
description of Kim [2]. Applying the hemacolor staining
technique, which was recently developed to detect the
subcutaneous primo vascular system (PVS) [3], the ex-
pected PV was, indeed, revealed. This finding of a neuro-
vascular primo (NVP) bundle is the first example of a PVS
found in the peripheral nerves. Until now, the PVS was not
regularly observable at specifically defined positions [4];
thus, the current finding is a breakthrough in PVS research
in the sense that NVP bundles are repeatedly observed in
anatomically well-defined positions. The connection of the
PVS with acupuncture was not shown until the recent report
on detection of PVS using hemacolor [3]. The NVPs exist at
acupuncture meridians such as the putative K and ST me-
ridians. Furthermore, they represent paths from the

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://
creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any
medium, provided the original work is properly cited.
* Corresponding author. 145 Gwanggyo-ro, Youngtong-Gu, Advanced Institute of Convergence Technology, Seoul National University,
Suwon 443-270, South Korea.
E-mail: kssoh1@gmail.com (K.S. Soh).
pISSN 2005-2901 eISSN 2093-8152
http://dx.doi.org/10.1016/j.jams.2015.10.002
Copyright ª 2015, Medical Association of Pharmacopuncture Institute.
330 K.H. Bae et al.

Figure 1 Neurovascular primo bundles. (A) Schematic illustration showing the distribution of the NVP bundles along the putative
kidney meridian. The NVP bundles are found near the acupuncture points (K17eK21). The midline shows the CV. (B) The NVP
bundles (arrows) in the hypodermis along the kidney meridians of a rat are displayed as a mosaic of stereomicroscope images. The
left and the right groups of five NVP bundles (those on one side are indicated by arrows) on the kidney meridians formed two lines
that were parallel to the white middle line (linea alba). The left and the right NVP bundles were located about 3e4 mm away from
the middle line. (C) Stereomicroscope image of an NVP bundle located in the hypodermis. It is stretched in order to show the whole
shape. The upper part entered the dermis layer, and the lower part entered the abdominal muscle layer. The NVP bundle was
studied earlier in order to search for the PV along the blood vessels, but no PV was seen. Only a nerve was seen to run along the
blood vessels [1]. (D) After staining with hemacolor, the PV (arrow) in the nerve of the NVP bundle emerged as a blue, threadlike
structure running along the blood vessels (arrow heads). The existence of the PV was expected from Kim’s [2] description, but was
not noticed until the recently developed hemacolor staining method [3] was applied. (E) A piece of a harvested NVP sample. The
hemacolor staining showed the parallel curves of the PVs (arrows) in the nerve. The stained parts can be clearly discerned at a
higher magnification with a phase contrast microscope in vitro, as shown in (F) and (G). (F) Magnified (400
) views of an NVP
Neurovascular Primo Bundles at the Kidney Meridian 331

Figure 1 (continued).

bundle; the view was obtained using a phase contrast microscope. The PVs (arrows) were stained with hemacolor. Some mast cells
are indicated by arrowheads. (G) The yellow lines are PVs illustrated by Kim [2]. The PVs (arrows) are located at the perineurium
and the endoneuria. The artery and vein are indicated by red and blue arrows, respectively. (H) A phase contrast microscope image
of the cross-section of an NVP bundle in Optimal Cutting Temperature (OCT) before defreezing. The blue stained spots (arrows)
exhibit the positions of the PVs. Most of them are just inside the perineurium, but some are in the fascicle (yellow double-headed
arrow). Some blood vessels are indicated by double-headed arrows. (I) After tissue processing and H&E staining, the blue stained
spots that appeared in Fig. 1H, implying the positions of the PVs (arrows) just outside of the perineurium, remained clearly,
however, there appeared more stained spots either inside or outside of the nerve bundle. This was due to insufficient washing of
the hemacolor. Three-dimensional digital histopathology will be performed to show more explicitly what the destiny of the
threadlike structure of the PV would be in a three-dimensional way. Two blood vessels are indicated with double-headed arrows.
After 40 , 6-Diamidino-2-phenylindole (DAPI) staining of nuclei and washing, the hemacolor disappeared, and (J) the bright mode
shows fiber-like structures (arrows) of the nerve with the PV, whereas (K) the fluorescent mode revealed the putative endothelial
nuclei (blue rod-shaped structures indicated with arrows) of the PV. The aligned distribution of rod-shaped nuclei (length
10e20 mm) is the hallmark of a PV [2e4]. (L) The NVP is located in the HD layer under the D and penetrates the F to the M. The
peripheral nerve goes to the thoracic spinal cord through the ganglion. The insets are (F) and (G), and indicate their respective
locations in the peripheral nerve. The nervous PV from the kidney acupoint can provide a path for electric signal transmission from
the acupoint to the spine. It can also be a conduit for the herbal medicine that is injected into the acupoint. This path further
proceeds to various parts of the brain. Therefore, acupuncture can influence the brain either by electric signal transmissions or by
herbal medicine delivery. CV Z conception vessel; D Z dermis; F Z deep fascia; HD Z hypodermis; H&E Z hematoxylin and eosin;
M Z abdominal muscle; NVP Z neurovascular primo; PV Z primo vessel.
332
acupoints in the skin to the brain for either transmission of
signals, such as the electric signal of electroacupuncture,
or flow of the herbal medicine of pharmacopuncture. This
result gives rise to a hope that acupuncture may possibly be
used to treat diseases of the brain such as Alzheimer’s and
Parkinson’s disease (Fig. 1).
Disclosure statement
The authors declare that they have no conflicts of interest
and no financial interests related to the material of this
manuscript.
Acknowledgments
This research was supported by the Basic Science Research
Program through the National Research Foundation of
Korea (NRF) funded by the Ministry of Science, ICT & Future
K.H. Bae et al.
Planning (Grant numbers: 2013 R1A1A2011526 and
2013R1A1A2008343).
References
[1] Ogay V, Min F, Bae KH, Kim JS, Han SC, Soh KS. Observation of
coiled blood plexus in rat skin with diffusive light illumination.
J Acupunct Meridian Stud. 2009;2:56e65.
[2] Kim BH. Theory of Sanal. J Jo Sun Med. 1965;108:1e38 [In
Korean]. [English translation available at http://ispvs.org/
publications-archives].
[3] Lim CJ, Lee SY, Ryu PD. Identification of primo-vascular system
in abdominal subcutaneous tissue layer of rats. Evid Based
Complement Alternat Med. 2015;2015:1e13. http://dx.doi.
org/10.1155/2015/751937. Article ID 751937.
[4] Shin HS, Johng H, Lee BC, Cho S, Baik KY, Yoo JS, et al.
Feulgen reaction study of novel threadlike structures on
the surface of rabbit livers. Anat Rec B New Anat. 2005;284:
35e40.