Volume 3, Issue 1 , Pages 10-15, March 2010
Anatomical Discrimination of the Differences Between Torn Mesentery Tissue and Internal Organ-surface Primo-vessels
Article Outline
- Abstract
- 1.. Introduction
- 2.. Materials and Methods
- 3.. Results
- 4.. Discussion
- Acknowledgments
- References
- Copyright
Abstract
The most difficult step in the morphological study of the internal organ-surface primo-vessel (Bong-Han duct) system is the correct identification of samples, due to similarities between torn mesentery and primo-vessel tissue. Without proper sample discernment, the subsequent parts of morphological studies cannot be trusted. Here, we present differences between torn mesentery and primo-vessel tissues as determined by minimal operation, using stereoscopic and microscopic observation. Stereoscopic observation revealed that torn mesentery is tightly connected to the organ surface and does not branch; the detached margin has a fan-shaped membrane without any swollen portions. Primo-vessels are slightly connected to the organ surface and branched, while detached margins lack a membrane and possess a swelling termed the primo-node (Bong-Han corpuscle). Microscopic observation shows bundle patterns in primo-vessel tissue, but irregular arrangement in torn mesentery tissue. These characteristics can be used to distinguish torn mesentery from primo-vessels.
Key Words: Bong-Han theory , morphological study , primo-vessel , torn mesentery
1. Introduction
The Bong-Han theory was first introduced by BongHan Kim in 1965 [1]. This theory has been developed and is now more sophisticated since an early study relating to acupuncture and meridian was carried out [2]. Kim reported that primo-vessels (Bong-Han ducts) are the anatomical structures underlying acupunctural meridians, with superficial primo-nodes (Bong-Han corpuscles) being the targets of acupoints. He claimed that primo-vessels are present throughout the whole body, i.e., on the surface of internal organs, inside blood and lymph vessels and under the skin [1]. In 1965, Kim reported that liquid with DNA granules, termed primo-fluid (Bong-Han liquid), flows through the primo-vascular system, a new circulatory system entirely different from the vascular, nervous, and lymphatic systems [3].
However, due to difficulties in reproducing Kim's results, his work has been neglected for the past 40 years [4]. A few research groups, such as Fujiwara and Yu [5], Lee [6], and Cho et al [7] were able to partially confirm Kim's results, but their work has received limited attention.
In 2003, Soh and members of the Biomedical Phy sics Laboratory of Seoul National University (SNU) published “Acridine orange staining method to reveal characteristic features of an intravascular threadlike structure”, a crucial paper in establishing the existence of primo-vessels [8]. They also discovered methods utilizing Janus Green B [9] and Alcian Blue [10] for staining primo-vessels inside lymphatic vessels and used the Feulgen Reaction to observe the canaliculus bundle structure of the primo-vessel on the surface of internal organs under an optical microscope [11]. In addition, the SNU team injected fluorescent nanoparticles into primo-vessels and found that primo-fluid flows at a velocity of 2 mm per minute through the vessels [12].
However, few papers concerning the primo-vessel system have been published without collaboration with SNU, as it is difficult to observe primo-vessels and corpuscles unaided, and for untrained researchers to discriminate between primo-vessels and other connective tissues. Even if primo-vessels are found, the system used for verification often lacks sufficient detail for confirmation. A method for finding primo-vessels and primo-nodes was reported by Lee et al in 2006, but did not give a detailed explanation of how to discriminate between primo-vessels and other tissues. The purpose of the present article is to present a new method allowing researchers to distinguish between internal organ-surface primovessels and torn mesentery, which is a similar and potentially confusing tissue that can be encountered during the gathering process.
2. Materials and Methods
2.1. Experimental animals
Sprague-Dawley rats (males, 250–320g weight) were housed in a constant temperature-controlled environment (22±1°C) with 55±10% relative humidity under a 12-hour light/dark cycle for approximately 5 days prior to the start of the experiment. All animals had ad libitum access to food and water but were fasted for 24–48 hours before the experiment. The experimental protocol was in full compliance with current international laws and policies [13].
2.2. Anesthesia
Rats were anesthetized with 1/5 diluted urethane (1.5 mg/kg) administered intraperitoneally, and all surgical procedures were performed under general anesthesia.
2.3. Surgical methods
The abdominal skin of the rat was incised along the linea alba, making sure that it did not bleed. The abdominal wall was lifted and incised carefully so as to avoid damage to primo-vessels from surgical instruments.
2.4. Observations
Phosphate-buffered saline (PBS) was maintained at 40°C and administered periodically to keep the organ surface moist. The physical characteristics of primo-vessels and torn mesentery were observed and recorded using a stereoscopic microscope and digital camera (Olympus SZ30 and Olympus E-330; Olympus Optical, Tokyo, Japan).
A sample was detached from the organ surface. Each of the detached samples were placed on a slide treated with 0.1 M PBS solution without any other processes. Characteristic features were observed with an optical microscope (BS 41 TF; Olympus Optical) and images were archived using a screen capture program (Focus Lite, version 2.88; NET GmbH Lerchenberg, Finning, Germany).
3. Results
3.1. Observation and comparison by stereoscopic microscopy
3.1.1. Discovery of a milk-white internal organ-surface primo-vessel on the surface of the small intestine, not attached to the internal organAfter the abdominal incision was made, a thread like structure was exposed, passing from the surface of the small intestine to the large intestine (Figures 1A and 1B). Appearing semi-transparent and milk-white in color, the threadlike structure was easy to lift as it had either no attachment or a very slight connection to the organ surface.
Figure 1.
(A) Sample of a primo-vessel on the surface of the small intestine after incision and exposure of the abdominal cavity. (B) Magnified view of the rectangular area highlighted in Figure 1A (the primo-vessel is visibly held by forceps). (C) A primo-node inside the folds of the colon, found by following the primo-vessel on the surface of the small intestine. (D) A primo-vessel on the surface of the small intestine and branches to the large intestine.
The primo-vessel was traced to a milk-white-colored lump, presumed to be a primo-node, located inside the folds of the colon (Figure 1C). Another primo-vessel was found on the surface of small intestine and it branched to the large intestine. The primo-vessel was connected slightly on the surface of the small intestine, and after detachment with forceps, the features of the structure were revealed (Figure 1D). These features are in agreement with those reported by Fujiwara and Yu [5] and Soh [8], confirming that the sampled tissue is the same as reported previously.
3.1.2. Observation and comparison of sampled mesenteric tissueA membranous tissue similar to primo-vessel tissue was found on part of the mesentery covering the small and large intestine. The membrane was connected to the surface of the organ, which distinguished from the tissue observed in Figures 1B and 1D. A fan-shaped membrane was exposed by pulling up the surface of the membrane with forceps (Figure 2A). Even applying a force sufficient to lift the entire organ, the membrane did not separate from the intestine surface. Moreover, when the fan-shaped membrane was intentionally removed, the connective membrane proved to be strong and stretched without snapping when placed under tension (Figure 2B). When the mesentery connected with the intestines was lifted, a fan-shaped membrane was seen (Figure 2C). The structure was not observed during the detachment of the primo-vessels from organ surfaces. The fan shape appears due to separation of the membrane structure. As this does not appear with primovessels, the presence of the fan shape allows the two tissues to be distinguished. In addition, the primo-vessels tended to branch and were connected to the primo-node, while torn mesentery did not connect to any tissues resembling the primonode. Therefore, torn mesentery does not resemble primo-vessels except in the shape of the torn section.
Figure 2.
(A) Mesentery under tension from forceps (arrows). The mesentery appears similar to a primovessel, but is connected tightly to the internal organ. The transparent membrane is connected within the dashed triangle. (B) Tearing of mesenteric tissue causes it to appear similar to a primo-vessel, but it is strong enough to withstand tension. (C) The membrane structure (outlined in the dashed triangle) remains at the end of the torn mesentery (arrows). Unlike a primo-vessel, torn mesentery does not branch or reach a primo-node, but connects to the mesentery.
3.2. Observation and comparison by optical microscopy
Primo-vessel tissue has been reported to have a canaliculus bundle structure containing DNA granules and rod-shaped nuclei [3]. However, it takes time to verify that samples contain DNA granules and rod-shaped nuclei using fluorescein staining, making it difficult to identify samples as primo-vessels or other organ tissues. In this study, the sampled specimens were instantly observed after applying only 0.1 M PBS solution and were identified as primovessels or other similar tissues.
3.2.1. The morphological structure of the internal organ-surface primo-vessel observed by optical microscopyThe primo-vessel specimen from the small intestine was placed on a slide with 0.1 M PBS solution and observed with an optical microscope. Bundle patterns were observed (Figure 3). This feature is in agreement with Kim [2] and Shin et al [11], confirming that the sampled tissue was the same as previously reported.
Figure 3.
(A) Primo-vessel observed by optical microscopy. Bundle patterns are clearly observable (arrows, 100×). (B) Higher magnification of Figure 3A (400×; scale bar, 50 μm).
The torn mesentery specimen from the small intestine was placed on a slide with buffer solution and observed with an optical microscope. An irregular pattern could be observed (Figure 4), rather than the bundle structure of the primo-vessel, indicating that the sample was a different tissue.
Figure 4.
(A) Torn mesentery by optical microscopy. At low (100×) magnification, the elongated structure of torn mesentery resembles a primo-vessel. (B) A higher magnification of Figure 4A. Irregular patterns are seen in place of the bundle structure of the primo-vessel tissue (400×; scale bar, 50 μm).
4. Discussion
Bong-Han theory has been controversial for some time, with researchers divided into supporters, including Fujiwara and Yu and others 5, 6, 7, 8, 14, and those who deny the existence of primo-vessels, for example Kellner [15]. This disagreement is due to the difficulty of finding primo-vessels. Even when they are found, they are easily confused with other tissues. Torn mesentery is particularly similar to primovessel tissue, causing difficulties in the study of the duct system.
Previously, a misconception existed that the Bong-Han theory was simply a study of lymphatic vessels [15]. However, due to reports regarding the presence of primo-vessels inside lymphatic and blood vessels, this issue has now been resolved. Yet, there is still potential for separated, torn mesentery tissue to be confused with primo-vessel tissue. In this study, we have analyzed and compared samples of torn mesentery and primo-vessel tissue and present methods for differentiating the two, based on anatomical characteristics. Kim's description of primo-vessel histology states that primo-vessels are bundles of several ductules, which show striped lines and consist of thin endothelial cells, with the ductule walls being about 0.1-0.2 mm in thickness. The average diameter of primo-vessels is about 10 mm and ranges from 1 mm to 50 mm. The endothelial cells in the walls of primo-vessels lack a clear boundary and have rod-shaped nuclei 12-20 mm long. Spindle-shaped adventitial cells surround primovessels, which also have rod-shaped nuclei. Basophil granulocytes, located inside primo-vessels, can be identified by histological staining 2, 3. Soh [8] and Lee et al 9, 16, 17 found that primo-vessels are separate from the surface of internal organs. They suggest that branches, rod-shaped nuclei, swollen portions (primo-node), a transparent color, weak connection, threadlike structure, and DNA granules as characteristics to be used in differentiating primovessels from other tissue types.
In this study, two differentiating characteristics are proposed, neither of which require extra effort during experimentation, such as staining. These are the distinctive features of primo-vessels in situ and the bundle patterns observed by optical microscopy. Torn mesentery can be distinguished from primo-vessels using the above criteria. Particular differences are shown in Table 1. These criteria for discriminating tissue types may assist researchers in identifying ambiguous tissue samples more accurately in the future, aiding in the study of the primovascular system. We expect further research and more detailed results to be generated using other methods for discriminating primo-vessels from torn omentum, fibrin strings, nerves, or lymphatic ducts.
Table 1. Similarities and differences between internal organ-surface primo-vessel and torn mesentery
| Internal organ-surface primo-vessel | Torn mesentery | ||
|---|---|---|---|
| Observation using a stereoscope | Similarity | Milky-white colored and string-like feathers | |
| Difference | Weakly connected to the organ surface | Tightly connected to the organ surface, with attachment strong enough to withstand force sufficient to lift the organ | |
| Branch onto other organ surfaces | |||
| Connected to corpuscles | Connected to the organ surface with a fan-shaped membrane and does not branch | ||
| Observation using an optical microscope | Similarity | Both have string-like feathers at low magnification | |
| Difference | Bundle patterns | Irregular pattern | |
Acknowledgments
This work was supported by the “Characterization Study of Electrophysiology in Primo-vessel” Project funded by the Ministry for Health, Welfare and Family Affairs, and a Korean Science and Engineering Foundation (KOSEF) grant from the Korean Government (MOST; No. R13-2008-028-01003-0).
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PII: S2005-2901(10)60002-7
doi:10.1016/S2005-2901(10)60002-7
© 2010 Korean Pharmacopuncture Institute. Published by Elsevier Inc. All rights reserved.
Volume 3, Issue 1 , Pages 10-15, March 2010
