mountain ginseng pharmacopuncture ( mgp ) is the first intravenous injection pharmacopuncture using an extract distilled from mountain cultivated ginseng or mountain wild ginseng in korean traditional medicine .
mountain wild ginseng refers to ginseng ( panax ginseng c. a. meyer ) of the araliaceae family that is naturally grown in the mountain .
based on the growth environment and the cultivation , commercial ginseng can be classified into three grades ; cultivated ginseng , mountain cultivated wild ginseng and mountain wild ginseng .
cultivated ginseng is grown artificially on farms and account for the major quantity of ginseng in the current market .
mountain wild ginseng is slower in growth and more sensitive to environmental changes than cultivated ginseng , showing a preference for are - as with fluctuating daily temperatures and less exposure to direct sunlight .
these differences may result in a variation of the active compounds between cultivated ginseng and mountain wild ginseng .
mountain cultivated wild ginseng can be considered as mimicry mountain wild ginseng ; it is seeded and grown in forests and mountains .
it is widely accepted in both korea and china that mountain wild ginseng produces more potent medicinal activity than mountain cultivated wild ginseng or cultivated ginseng . however
, only a few studies have been conducted to compare the food components and the pharmacological activities between mountain wild ginseng and both cultivated and mountain cultivated ginseng [ 3 , 4 ] . because authenticity tests of mountain wild ginseng are very difficult and confusing , scientific criteria of authenticity are needed .
thus , an objective diagnostic method that used gene analysis to identify mountain ginseng ( mg ) was developed . comparing gene activation between numerous mg samples and regular ginseng ,
genes only existing in mg , such as p - nrt2 , p - rpoc1 , p - psbb , and p - gapdh , were found [ 3 - 6 ] . because of
the high - cost and authenticity issues associated with mountain wild ginseng , mountain cultivated ginseng which is similar to mountain wild ginseng according to the gene analysis , is used for pharmacopuncture .
although continuous clinical studies have been performed to obtain more objective data on the efficacies of mgp , safety tests are still necessary .
the aims of this study were to evaluate the safety of mgp , and to conduct single- dose toxicity tests of mgp .
18 sprague - dawley ( sd ) rats of each gender were obtained from a specific pathogen - free facility ( biotoxtech , oh chang , korea ) at 5 weeks of age and were used after a week of quarantine and acclimatization .
the animals were housed in a room maintained at 19.0  23.2 under a relative humidity of 33.0%  59.5% .
the room was illuminated with artificial lighting for 07:00 to 19:00 hours and had 10  15 air changes per hour .
three animals were housed in suspended stainless - steel wire - mesh cages and were allowed sterilized tap water and commercial rodent chow ( teklad certified irradiated global 18% protein rodent diet 2918c , harlan laboratories , inc .
this study protocol was approved by the institutional animal care of biotoxtech co. ( oh chang , korea ) .
the process for the manufacturing mgp was performed in a pathogen - free facility ( korean pharmacopuncture institute , seoul , korea ) , as described previously .
mg of over 10 years which was cultivated from the seeds of mountain wild ginseng , was selected for the mgp .
first , we washed mg with running water and dried it in the air - conditioned room .
second , dried mg was powdered in to fine particles ( size are below 10  ) and decocted for 2 hours in distilled water .
remnants were then removed , and the decoction was distilled before yielding the desired herbal acupuncture . then the pharmacopuncture was filtered using 0.1  filter paper and was then kept in the container . finally , the pharmacopuncture was sterilized before being used .
healthy male and female rats , 5 of each sex were selected and assigned to 1 of 3 groups : control group ( normal saline ) , low - dose group ( 10 mg / kg ) , and high - dose group ( 20 mg / kg ) .
all animals were observed for clinical signs at 30 minutes , 1 hour and 2 hours after the first injection and at 30 minutes , 1 hour , 2 hours , 4 hours and 6 hours after the second injection .
clinical signs were observed daily from the injection day to 14 day after the first injection .
the body weight of each rat was measured at the initiation of treatment , and at 3 days , 7 days and 14 days after the injection .
the scheduled day of study termination , all surviving animals were anesthetized by co2 inhalation .
tissues were obtained from the following organs of all surviving animals and fixed with 10% neutral buffered formalin solution : brain , heart , liver , spleen , kidney , lung , spinal nerves .
data on body weights were tested by using sas ( version 9.2 , sas institute inc , usa ) .
if the variance was homogeneous , the data were subjected to one - way analysis of variance ( anova ) .
if significant changes were observed on the one - way anova , the data were analyzed by using the multiple comparison procedure of dunnett s test   ( p < 0.05 , p < 0.01 ) .
no treatment - related mortalities occurred in sd rats treated with any dose of the mgp .
also , no clinical signs were observed in the male and the female sd rats of the experimental groups during the observation period .
hematuria was observed in 1 of the rats of control groups , but it was not caused by mpg .
no significant changes in weight were observed in the experimental groups compared to the control group ( fig 1 , 2 ) .
no abnormal macroscopic appearances were observed on necropsy in either the control or the experimental groups .
all experimental groups showed no abnormal signs caused by intravenous injection of mgp . in the control and the high - dosage groups ,
some changes were observed in the kidney , lung , liver and spleen , but those changes seemed to have occurred naturally or accidentally ( fig 3 - 6 ) .
mgp is administered intravenously , and intravenous injection is enlisted as a legal manipulation in korean classification procedures in medicine and resource . for this reason , intravenous injection of mgp has been studied . in a randomized controlled trial of mgp , no significant changes in blood pressure , hematology , biochemistry and urine analysis were reported .
however , immune protein , which acts against microbes and pathogenic organisms , vitamin d binding protein , which protects the lung at the time of inflammatory response , ras - related protein ral - a , which controls phospholipid metabolism , cytoskeletal formation , and membrane traffic , and transferrin which balances the iron level in the body were reported to be increased significantly , after the administration of mgp , but antitrypsin , which is secreted with inflammatory responses in the lungs , were reduced [ 13 , 14 ] .
also no effects on heart rate variability   and pulse - wave factors were revealed in middle - aged women .
there are many types of flavonoid compounds , such as quercetin , hesperidin and anthocyacanidins in mgp , and these constituents were reported to have actions that improved liver functions [ 17 , 18 ] , and that produced anti - oxidant [ 19 - 23 ] , anti - cancer [ 7 , 24 - 30 ] , anti - hypertensive , anti - diabetic , anti - obesity   and anti - depressant effects .
thus , mgp has been used to treat progressive disorders such as cancer [ 35 - 40 ] and amyotropic lateral sclerosis .
the present study was conducted to evaluate the safety of mgp , and the results of this study showed no mortalities and no significant changes in sd rats caused by intravenous injection of mgp .
these results correspond with those of an earlier study which reported that mgp caused negligible toxicity .
although the safety of mgp has been demonstrated through a single - dose toxicity test , still further study , like a 4-week repeated toxicity test , is needed .
based on the results of this study , we can conclude with certainly that the lethal dose of mgp is higher than 20 ml / kg in sd rats and that intravenous injection of mgp in sd rats is a safe treatment considering that the usual clinical dose of mgp is about 0.3 ml / kg .
the field of traditional korean medicine ( tkm ) has a long waited technology that uses the vessel as a treatment medium , as the vessel provides the quickest path of drug ab - sorption .
a limitation on treating patients is the difficulty of oral administration of medicines , but pharmacopuncture , especially mgp has opened the door for immediate treatment benefits . as the result of this study , we can conclude with certainly that intravenous injection of mgp is safe .
 objectives : mountain ginseng pharmacopuncture ( mgp ) is an extract distilled from either mountain cultivated ginseng or mountain wild ginseng . 
 this is the first intravenous injection of pharmacopuncture in korea . 
 the word intravenous does not discriminate between arteries , veins , and capillaries in oriental medicine , but only the vein is used for mgp . 
 the aim of this study is to evaluate the intravenous injection toxicity of mgp through a single - dose test in sprague - dawley ( sd ) rats.methods:male and female 6-week - old sd rats were injected intravenously with mgp ( high dosage of 20 ml / kg or low dosage of 10 ml / kg ) . 
 normal saline was injected into the rats in the control group by using the same method . 
 after the rats has treated , we conducted clinical observations , body - weight measurements and histological observations.results:in this study , no mortalities were observed in any of the experimental groups . also , no significant changes by the intravenous injection of mgp were observed in the body weights , or the histological observations in any of the experimental groups compared to the control group . 
 the lethal dose for intravenous injection of mgp was found to be over 20 ml / kg in sd rats.conclusion:considering that the dosage of mgp generally used each time in clinical practice is about 0.3 ml / kg , we concluded with confidence that mgp is safe pharmacopuncture . 
1. Introduction
2. Materials and Methods
3. Results
4. Discussion
5. Conclusion