| Fig1.CT Results of a herniated lumber disc patient after taking Chuna medicine for 6 months |
The left arrow indicates a herniated disc. The right arrow indicates the restored disc
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| Fig 2. MRI Results of a disc-disruption patient |
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The arrow indicates a herniated disc.
The disc has been restored by Chuna medicine treatment. |
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 Animal experimentation for the edema restriction effect of Chuna medicine |
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| Fig 3. Edema induction on feet of a rat |
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(Bottom left) After the induction, the edema in the control group degenerated in 70 days
(Bottom right) After the induction of Chuna Medicine; the edema was eliminated in 70 days
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(1) Objective : The objective of this study was to examine the anti-inflammation effect of Chuna medicine. Edema may give rise to thighbone disruption (degree of restoration), osteoporosis, and necrosis or disc disruption.
(2) Method : The control group was administered Aymogen and Adjuvent on their feet, to induce edema. In order for the edema to develop the inducers were left on for two weeks, and the rats were observed for 70 days.
The experimental group was administered Aymogena and Adjuvent on their feet as well. After two weeks of development the rats were administered Chuna Medicine and the rats were observed for 70 days. Chuna Medicine [Prescription concentration; 0.6%/ml (Chuna medicine/water)] The rats were orally administered 1kg/ml per day.
(3) Result : As [Fig. 3], the edema in the control group showed degenerative changes, but the experimental group recovered from the edema. This shows that the Chuna medicine pharmacologically may eliminate or reduce edema.
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| Animal experimentation for the effect of bone regeneration and disc treatment of Chuna medicine |
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| [Experiment. 3-1, 3-2] Induction experimentation of herniated or disrupted disc |
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| Fig 4. Tissue staining of a rat's lumbar disc |
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(L) Lumbar disc of a healthy rat
(M) Control group without any medication, its bone tissues were disrupted due to disc-herniation induction and the disc collapsed.
(R) Experimental group with medication, its disc recovered
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(1) Objective : The objective of this study was to see whether Shinbarometin [CBB-13] the substance abstracted from Chuna medicine has an effect on bone tissue regeneration and the restoration of disrupted disc or nerve paralysis.
(2) Method : Inflammation, degeneration or disruption on a disc was induced in both groups with a mixture of Aymogen and Adjuvent to L4-L5.
The rats in the experimental group were orally administered 0.25mg/ml Shinbarometin (CBB-13) from Chuna medicine for 6 months. The results were determined by observation of H/E tissue staining using a microscope and the movement of the experimental rat.
(3) Result : As [Fig 4, 5], comparing the control group (without any medication), with the experimental group (with medication), new bone growth was regenerated in the experimental group Also, the collapsed disc was restored and 85% of disc disruption was restored. It was proven that Chuna Medicine regenerates bone growth, and restores disrupted disc. The movement ability of an experimental rat gradually returned to its healthy state at the 3 month point of the medication intake and it was restored to normal in 5~6 months. |
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| Fig 5. Tissue staining of a disrupted disc |
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(L) Bone abrasion and pores developed due to the degeneration and pressure on the disc after induction.
(R) After the intake of Chuna medicine, bone density become compact and the number or size of trabecular bone was restored and the disrupted disc was recovered. |
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| [Experiment 3-3] Experiment for the effect of bone regeneration through osteoporosis induction |
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| Fig 6. Regeneration of a disrupted thighbone |
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(L) Normal thighbone
(M) Discomposed and empty thighbone due to bone clastic enzyme
(R) Following Chuna medicine treatment, the disrupted thighbone was restored |
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(1) Objective : The objective of this study was to examine the efficacy of bone regeneration and disc restoration by Shinbarometin (CBB-13), which was abstracted from Chuna Medicine.
(2) Method : In order to induce Osteoporosis we removed the ovary of white rats. Then the experimental rat was orally administered 0.25mg/ml of Shinbarometin (CBB-13) for 6 months, and we observed the thighbone tissue to check the degree of bone damage. (3) Result : The trabecular bone of both rats was disrupted due to the induced osteoporosis. [Fig 6] The condition of the control group remained the same. The 85% of the thighbone of the experimental group was restored following the administration of Chuna medicine. This shows that Chuna medicine may help to regenerate bone tissue growth and facilitate the growth of the trabecular bone. |
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| [Experiment 4-1] RT-PCR Experiment to restrict the revelation of COX-II, iNOS on raw 264.7 cells |
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(1) Objective: Since bone disorders frequently go with pain, edema and inflammation, the experiment was designed to examine whether Shinbarometin may reduce inflammation.
(2) Method: Checked the effect of inflammation reduction through RT-PCR analysis. Once an inflammation inducer such as LPS is applied to cells during germ culture, iNOS, which is related to |
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inflammation, is regenerated. Since it regenerates the same protein as the protein to induce inflammation, we've examined the restriction of NO (very strong inflammatory inducers within blood) regeneration by iNOS and COX- II (enzyme engaging in inflammation) after the treatment of each ingredient and checked the reduction of inflammation.
(3) Result: We could see that the ingredients, lane 5, 6, 7 and 8 are a little engaged in the regeneration of COX-II, iNOS. In the image, the white region represents the induced inflammation in both groups while the black region represents the restricted regeneration of inflammation. |
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| [Experiment 4-2] Experiment to restrict the revelation of COX-II, iNOS on raw 264.7 cells |
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(1) Objective : Since bone disorders frequently go with pain, edema and inflammation, the experiment was designed to examine whether Shinbarometin may reduce inflammation. Checked the effect of inflammation reduction through RT-PCR analysis. Once an inflammation inducer
(2) Method : such as LPS is applied to cells during germ culture, iNOS, which is related to inflammation, is regenerated. Since it regenerates the same protein as the protein to induce inflammation, we've examined the restriction of NO (very strong inflammatory inducers within blood) regeneration by iNOS and COX- II (enzyme engaging in inflammation) after the treatment of each ingredient and checked the reduction of inflammation. |
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(3) Result : In comparing the controlled group to the experimental group, the experimental group showed that Shinbarometin may restrict the generation of inflammation. In short, Shinbarometin was deeply involved in the restriction of inflammation and edema. |
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| [Experiment 5] The Effect of the restriction of NOGO gene inducing nerve paralysis |
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(1) Objective : The experimentation was designed to examine the treatment effect of Shinbarometin for blocking lumbar nerve paralysis |
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(2) Method : To induce nerve paralysis the AXON, an axis of nerve cell with NOGO-A was melted, used to block the nerve. Then we examined to see whether Shinbarometin might effectively restrict nerve paralysis (NOGO-A dissolves myelin to induce the necrosis of cells and it obstructs the regeneration of nerves). During the germ culture on SK-N-SH, the nerve cell line, which originated from the brain was administered protein NOGO-A and we examined the nerve regeneration by Shinbarometin. We then observed it with a microscope at X200 and produced images
(3) Result : The controlled group's nerve was damaged by NOGO-A but the experimental group who were treated with Shinbarometin, the nerve protrusions were actively reproduced from day 3 and AXON was regenerated to normal nerves. We could see that Shinbarometin restricted the necrosis of nerves. . |
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| Experimentations & Summarized Analyses |
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| When we clinically treated patients with herniated lumbago disc or disc disruption using Chuna medicine, it was demonstrated by MIR image (Fig.2) and C.T (Fig.1) that the treatment was effective or that the subjective symptom was recovered. Therefore, we've determined to conduct an animal experimentation to see the scientific efficacy of Chuna medicine. Through Animal experimentation for the edema restriction effect of Chuna medicine (Fig.3), Tissue staining of a rat's lumbar disc (Fig. 4), Tissue staining of a disrupted disc (Fig. 5) and Regeneration of a disrupted thighbone (Fig. 6), it was proven that Chuna medicine may restrict the disruption of soft bone neighboring arthritis and facilitate the reproduction of bone cells. With the evidence supplied from the animal experimentation to show the efficacy of Chuna medicine, we tried to find the most predominant ingredient, which restricts edema and to regenerates nerve. We finally refined three single substances, CBB-11, CBB-12 and CBB-13 during the several procedures of separation and refinement. As the result of examination of their chemical structures, CBB-11 and CBB-12 were Ornithine and Daucoterol that have already been known, while CBB-13 was a new chemical that has never been known, 'Shinbarometin'. Meanwhile, through cell experiments and molecular biological experiments, we've proven that Shinbarometin may restrict the increase of cells in bone disorders and efficiently facilitate the promotion of nerve regeneration. |
