Seong-Hun Ahn

A cytological observation of the fluid in the primo-nodes and vessels on the surfaces of mammalian internal organs

We report on the preliminary cytological observation of fluid in the primo-nodes and vessels on the surfaces of the internal organs of mammals. With some microsurgical procedures, we observed many cells and microcells that spread out of the nodes on the organ surfaces of rats and rabbits. These cells generally showed the following morphologies: (1) round or oval cells, 10 μm in size, with predominantly little cytoplasm; (2) cells with nuclei that exhibited a collapsed shape; (3) binucleated cells, 20 μm in size; (4) spherical granules, ranging 0.5–2.0 μm in size (primo-microcells); and (5) aggregations of such granules. These findings on the existence of cells with diverse morphologies in the fluid of primo-nodes and vessels could be evidence supporting the hypothesis that the anatomical basis of acupuncture meridians (i.e., primo-vascular system) may be a migration channel for various kind of cells.

Anatomical Discrimination of the Differences Between Torn Mesentery Tissue and Internal Organ-surface Primo-vessels

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.

Induction of oral tolerance to Japanese cedar pollen

Oral tolerance is thought to play a role in preventing allergic responses and immune-mediated diseases. An improved mouse model of the oral tolerance to Japanese cedar pollen (JCP) as antigen was developed in order to detect induction of the tolerance, and the immunological characteristics of this model were also elucidated. Oral tolerance was induced by C3H/ HeN mice given an oral administration of 10mg JCP 7 days before immunization with an i.p. injection of 0.1 mg JCP in complete Freunds adjuvant (CFA). The effects of oral JCP on systemic immunity were assessed by enzyme-linked immunosorbent assay (ELISA) of immunoglobulin (Ig) levels in serum collected on day 7 or 14 after immunization. Oral tolerance to JCP was adequately induced on day 7 after immunization and was more effective in C3H/HeN mice than in BALB/c mice. The tolerance was primarily concerned with the decreased serum levels of antigen-specific IgG. In these mice, oral administration of JCP also suppressed various immune responses to the antigen including delayed-type hypersensitivity (DTH), total IgE level and anti-JCP lgG1 level. The suppression of these immune responses by the oral antigen was associated with a significant reduction in interleukin-4 (IL-4) production. These findings therefore indicate that this C3H/HeN mice model has potential use in detecting the induction of oral tolerance by JCP, and suggest that this tolerance model may be effective in the treatment and prevention of allergic responses caused by the antigen.

Primo Microcell in a Primo Node as a Possible Origin of Adult Stem Cells

Adult stem cells have been intensively studied for their cell-therapeutic potential to renew damaged tissues or organs because their use eliminates the ethical, legal, and political concerns with using embryonic stem cells. On the other hand, a microcell contains a micronucleus that has one or a few chromosomes with a small amount of cytoplasm. Recently, microcells have become a possible origin of adult stem cells because of their ability to induce undifferentiated cells. In the early 1960s, after its first observation by Kim, the primo vascular system (PVS) was thought to be an anatomical basis for traditional acupuncture systems (termed kyungrak theory or meridians). Kim also claimed that about 1.0-μm-sized DNA-containing granules, named primo microcells or sanals, flowed through the PVS consisting of primo vessels and primo nodes to internal organs to proliferate and differentiate into specific cells in the organs in a manner similar to that of adult stem cells. In this study, we harvested primo microcells from Sprague Dawley rats about 250–300 g. The primo microcells were observed to grow into cell-like body entities. These interesting results suggest that Kim’s manual pictures from his article 1965 and Sanal theory may be true based on experimental studies similar to ours. Also, primo microcells in primo nodes might be one possible origin of adult stem cells. Understanding the primo microcell is thought to be one of the most fundamental studies for cell therapy.

Biological Activities of Scolopendrid Pharmacopuncture

Reactive Oxygen Species(ROS) are continuously produced at a high rate as a by-product of aerobic metabolism. Since tissue damage by free radical increases with age, the reactive oxygen species(ROS) such as hydrogen peroxide(), nitric oxide(NO). Several lines of evidence provided that ROS appears to cause to develop aging-related various diseases such as cancer, arthritis, cardiovascular disease. Our reserch objective was to examine the in vitro biological activity of Scolopendrid Pharmacopuncture, including the total poly-phenol content, DPPH radical scavenging, ABTS radical scavenging, Superoxide dismutase(SOD)-like activity, Nitrite scavenging ability. The total poly-phenol contents of Scolopendrid Pharmacopuncture was 35.859mg/L. Elctron donation ability on DPPH was 36.82%. The 2,2`-azinobis-3-ehtlbezothiazoline-6-sulfonic acid radical decolorization (ABTS) was 84.7%. The superoxide dismutase (SOD)-like activities of Scolopendrid Pharmacopuncture was 44.33%. The nitrite scavenging effects were pH dependent, and were highest at pH 1.5(45.2%) and lowest at pH 6.0(11.3%). We conclude that Scolopendrid Pharmacopuncture may be useful as potential sources of antioxidant.

Study on a Single-Dose Toxicity Test of D-Amino Acid Oxidase (DAAO) Extracts Injected into the Tail Vein of Rats.

Objective: This study was performed to analyze the single-dose toxicity of D-amino acid oxidase (DAAO) extracts. Methods: All experiments were conducted at the Korea Testing & Research Institute (KTR), an institution authorized to perform non-clinical studies, under the regulations of Good Laboratory Practice (GLP). Sprague-Dawley rats were chosen for the pilot study. Doses of DAAO extracts, 0.1 to 0.3 cc, were administered to the experimental group, and the same doses of normal saline solution were administered to the control group. This study was conducted under the approval of the Institutional Animal Ethics Committee. Results: In all 4 groups, no deaths occurred, and the LD50 of DAAO extracts administered by IV was over 0.3 ml/kg. No significant changes in the weight between the control group and the experimental group were observed. To check for abnormalities in organs and tissues, we used microscopy to examine representative histological sections of each specified organ, the results showed no significant differences in any organs or tissues. Conclusion: The above findings suggest that treatment with D-amino acid oxidase extracts is relatively safe. Further studies on this subject should be conducted to yield more concrete evidence.

Mechanical Stimulation of the HT7 Acupuncture Point to Reduce Ethanol Self-Administration in Rats.

Background Alcoholism, which is a disabling addiction disorder, is a major public health problem worldwide. The present study was designed to determine whether the application of acupuncture at the Shenmen (HT7) point suppresses voluntary alcohol consumption in addicted rats and whether this suppressive effect is potentiated by the administration of naltrexone. Methods Rats were initially trained to self-administer a sucrose solution by operating a lever. A mechanical acupuncture instrument (MAI) for objective mechanical stimulation was used on rats whose baseline response had been determined. In addition, the effect of HT7 acupuncture on beta-endorphin concentration and ethanol intake via naltrexone were investigated in different groups. Results We found that ethanol intake and beta-endorphin level in rats being treated with the MAI at the HT7 point reduced significantly. The treatment of naltrexone at high doses reduced the ethanol intake and low-dose injection of naltrexone in conjunction with the MAI also suppressed ethanol intake. Conclusions The results of the current study indicate that using the MAI at the HT7 point effectively reduces ethanol consumption in rats. Furthermore, the coadministration of the MAI and a low dose of naltrexone can produce some more potent reducing effect of ethanol intake than can acupuncture alone.

Safety recommendations for moxa use based on the concentration of noxious substances produced during commercial indirect moxibustion

Background Moxibustion therapy has been used historically for thousands of years and there are many clinical trials supporting its efficacy and effectiveness for various conditions. Moxa smoke has been a major reason for avoiding moxibustion due to its smell and potential risks to the human body. Methods 10 units of commercial indirect moxa (CIM) from six manufacturers (A–F) were burnt in a 2.5×2.5×2.5 m chamber without ventilation, and concentrations of carbon oxides (CO and CO2), nitrogen oxides (NOx), and volatile organic compounds (VOCs) from the indoor air samples were measured. Results For brands A, B, C, D, E, and F, respectively, relative to baseline values, we observed an increase in CO (from 0.002 to 0.007, 0.006, 0.005, 0.006, 0.005, and 0.006 parts per billion (ppb)), NOx (from 0.009 to 0.051, 0.025, 0.015, 0.050, 0.019, and 0.020 ppb), and total VOCs (TVOC; from 48.06 to 288.83, 227.93, 140.82, 223.22, 260.15, and 161.35 μg/m3), while the concentration of CO2 was not elevated. Each CIM brand demonstrated different VOC emission characteristics, which could be divided into three groups. On average, we estimated that 20 units of CIM or 2.41 g moxa floss would need to be combusted in order to exceed the safe levels set by Korean environmental law. This limit is likely to be greater in the case of a larger room or use of ventilation. Conclusions Despite increased CO/NOx/VOC concentrations, overall levels remained within safe limits. These findings may help clinicians to maintain safe moxibustion treatment conditions to help keep both patients and clinicians safe from the pollutants generated by moxa combustion.

Single Intramuscular-dose Toxicity of Samgihwalryeok-Pharmacopuncture in Sprague-Dawley Rats.

Objectives: This study was performed to examine the single-dose toxicity of Samgihwalryeok pharmacopuncture. Methods: Forty six-week-old Sprague-Dawley (SD) rats were divided into four groups of 10 rats each; each group was then sub-divided into two smaller groups, one of five males and the other of five females. Group 1 (G1, control) received 1.0 mL of normal saline solution, while group 2 (G2, low-dose group), group 3 (G3, mid-does group, and group 4 (G4, high-dose group) received 0.1, 0.5, and 1.0 mL of Samgihwalryeok pharmacopuncture, respectively. Results: No mortalities or clinical signs were observed in the four groups. Also, no significant changes in body weights were observed among the group, and no significant differences in hematology/biochemistry, necropsy, or histopathology results were noted. Conclusion: The above findings suggest that treatment with Samgihwalryeok pharmacopuncture is relatively safe. Further studies on this subject are needed.

Single Dose Toxicity of Chukyu (spine-healing) Pharmacopuncture Injection in the Muscle of Rats

Objectives: This study was performed to analyze the single dose toxicity of Chukyu (spine-healing) pharmacopuncture. Methods: All experiments were conducted at the Biotoxtech, an institution authorized to perform non-clinical studies under the regulations of Good Laboratory Practice (GLP) regulations. Sprague-Dawley rats were chosen for the pilot study. Doses of Chukyu (spine-healing) pharmacopuncture, 0.1, 0.5 and 1.0 mL, were administered to the experimental groups, and a dose of normal saline solution, 1.0 mL, was administered to the control group. This study was conducted under the approval of the Institutional Animal Ethic Committee. Results: No deaths or abnormalities occurred in any of the four groups. No significant changes in weight, hematological parameters or clinical chemistry between the control group and the experimental groups were observed. To check for abnormalities in organs and tissues, we used microscopy to examine representative histological sections of each specified organ; the results showed no significant differences in any of the organs or tissues except in one case, where interstitial infiltrating macrophages were found in one female rat in the 0.5-mL/animal experimental group. Conclusion: The above findings suggest that treatment with Chukyu (spine-healing) pharmacopuncture is relatively safe. Further studies on this subject are needed to yield more concrete evidence.