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In vitro rumen fermentation kinetics, metabolite production, methane and substrate degradability of polyphenol rich plant leaves and their component complete feed blocks

  • Aderao, Ganesh N.;Sahoo, A.;Bhatt, R.S.;Kumawat, P.K.;Soni, Lalit
    • Journal of Animal Science and Technology
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    • v.60 no.11
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    • pp.26.1-26.9
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    • 2018
  • Background: This experiment aimed at assessing polyphenol-rich plant biomass to use in complete feed making for the feeding of ruminants. Methods: An in vitro ruminal evaluation of complete blocks (CFB) with (Acacia nilotica, Ziziphus nummularia leaves) and without (Vigna sinensis hay) polyphenol rich plant leaves was conducted by applying Menke's in vitro gas production (IVGP) technique. A total of six substrates, viz. three forages and three CFBs were subjected to in vitro ruminal fermentation in glass syringes to assess gas and methane production, substrate degradability, and rumen fermentation metabolites. Results: Total polyphenol content (g/Kg) was 163 in A. nilotica compared to 52.5 in Z. nummularia with a contrasting difference in tannin fractions, higher hydrolysable tannins (HT) in the former (140.1 vs 2.8) and higher condensed (CT) tannins in the later (28.3 vs 7.9). The potential gas production was lower with a higher lag phase (L) in CT containing Z. nummularia and the component feed block. A. nilotica alone and as a constituent of CFB produced higher total gas but with lower methane while the partitioning factor (PF) was higher in Z. nummularia and its CFB. Substrate digestibility (both DM and OM) was lower (P < 0.001) in Z. nummularia compared to other forages and CFBs. The fermentation metabolites showed a different pattern for forages and their CFBs. The forages showed higher TCA precipitable N and lower acetate: propionate ratio in Z. nummularia while the related trend was found in CFB with V. sinensis. Total volatile fatty acid concentration was higher (P < 0.001) in A. nilotica leaves than V. sinensis hay and Z. nummularia leaves. It has implication on widening the forage resources and providing opportunity to use forage biomass rich in polyphenolic constituents in judicious proportion for reducing methane and enhancing green livestock production. Conclusion: Above all, higher substrate degradability, propionate production, lower methanogenesis in CFB with A. nilotica leaves may be considered useful. Nevertheless, CFB with Z. nummularia also proved its usefulness with higher TCA precipitable N and PF. It has implication on widening the forage resources and providing opportunity to use polyphenol-rich forage biomass for reducing methane and enhancing green livestock production.

Effect of Yijin-tang on Gastric Motility between Normal Intact and Partial Pyloric Obstructed Rats (이진탕(二陳湯)이 정상 및 위 유문부가 부분폐색된 흰 쥐의 위 운동성에 미치는 효과)

  • Han, Soak-Young;Yoon, Sang-Hyub
    • The Journal of Internal Korean Medicine
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    • v.30 no.1
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    • pp.107-118
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    • 2009
  • Objectives : This study was aimed to investigated the effect of Yijin-tang on gastric motility and its mechanism of action in normal intact and partial pyloric obstructed rats. Methods : Gastric emptying was measured by the number of glass beads expelled from the stomach (containing one hundred of glass beads. ${\phi}1mm$) in 1 hour or 2 hours after glass beads and test drugs (normal saline. Yijin-tang 90mg/kg. Yijin-tang 270mg/kg) administration in normal intact and partial pyloric obstructed rats. In another series of experiments to evaluate the mechanism of Yijin-tang 270mg/kg under delayed conditions, normal intact rats were treated with atropine sulfate (1mg/kg,s.c.), cisplatin (10mg/kg,i.p.), quinpirole HCI (0.3mg/kg,i.p.) and NAME (NG-nitro-L-arginine methyl ester. 75mg/ kg,s.c.), respectively. Partial pyloric obstructed rats were modified by wrapping the nonabsorbable rubber ring (D :6mm, W:4mm, T: 1mm) around the 1st portion of the duodenum for 8 weeks. The myoelectrical activity of the gastric smooth muscle was recorded by a bipolar electrode placed at the abdominal surface in normal intact and partial pyloric obstructed rats. The gastric myoelectrical activity was measured for 30 minutes before and after orogastric administration of each solution (normal saline, Yijin-tang 270mg/kg) and expressed as dominant frequency, percent of normogastria and power ratio. Results : Yijin-tang improved gastric emptying more than normal saline in normal intact(p<0.001) and partial pyloric obstructed rats(p=0.002). Under the delayed gastric emptying induced by atropine sulfate, cisplatin, quinpirole HCI and NAME. Yijin-tang enhanced gastric emptying significantly in the cisplatin treated group(p<0.001). but didn't in other treated groups. Administration of Yijin-tang 270mg/kg has no significant effect on the myoelectrical activity of the gastric smooth muscle in both normal intact rats and partial pyloric obstructed rats. Conclusions : Yijin-tang seems to stimulate the gastric motility through suppressing the 5HT3 receptor and promoting the antroduodenal flow. We expect that Yijin-tang would be effective especially in dysmotility-like functional dyspepsia with partial pyloric obstruction or the side effects of cisplatin such as nausea, vomiting, abdominal discomfort, and delay of gastric emptying.

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Preparation of Black Ginseng and its Antitumor Activity (흑삼의 제조 및 항암효과)

  • Lee, Jee-Hyun;Shen, Gui-Nan;Kim, Eui-Keom;Shin, Hyun-Joong;Myung, Chang-Seon;Oh, Han-Jin;Kim, Dong-Hee;Roh, Seong-Soo;Cho, Won;Seo, Young-Bae;Park, Yong-Jin;Kang, Cheol-Woo;Song, Gyu-Yong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.4
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    • pp.951-956
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    • 2006
  • The purpose of this study is to prepare black ginseng and evaluate its antitumor activity. In order to achieve such aim, 5 year fresh ginsenges were steamed at 95'E for 3 hr in pottery apparatus and dried at $60^{\circ}C$ for 12-36 hr. This process was repeated again nine times in same condition. Among the ginseng saponins in black ginseng, the amount of Ginsenoside $Rg_3$ was examined by HPLC. 10.05 mE of Ginsenoside $Rg_3$ was obtained from 1 g of dried black ginseng prepared. The extract of black ginseng exhibited stronger cytotoxic activity against MCF-1, HT-1080 and Hepa 1C1C7 tumor cell lines in vitro than the extract of red ginseng. Also, the extract of black ginseng exhibited stronger antitumor activity(33%) in BDFl mice bearing Lewis lung carcinoma cells(LLC) than the extract of red ginseng(23%). From these results, it was concluded that Black ginseng had antitumor activity suggesting its application for the prevention and treatment of cancer.

A Review of Experimental study on Dementia in Oriental medicine;within Oriental medicine journal since 2000 (치매에 대한 최신 실험적 연구 동향;2000년 이후 한의학 학술지를 중심으로)

  • Choi, Sung-Youl;Kim, Dae-Hyun;Kim, Sang-Tae;Kim, Tae-Heon;Kang, Hyung-Won;Lyu, Yeong-Su
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.1
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    • pp.125-146
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    • 2008
  • Objectives : The purpose of this study is to suggest for the following experimental study of dementia by reviewing recent oriental medicine journals that have been published since 2000. Methods: We have investigated various types of studies in relation to dementia through 90 articles that have been published from 2000 to 2007 in recent oriental medicine journals were registered Korea research foundation. Results and Conclusions : 1. Since 2000, 88 articles in relation to dementia have been published and almost of them are herbal medicine-centered studies. Also they show a tendency to increase every year. The journal of oriental neuropsychiatry carries the highest number of studies in relation to dementia. 2. According to the experimental paper, there are 30 cases of using herb simplexes, 48 cases of herb-combined prescription, and 10 cases of other ways. Especially 7 cases of using herb-combined prescription relation to Sasang constitution are all for the Taeumin. 3. There are 85 cases of Animal and cellular experimental, 60 cases of using pathologic model induced cytotoxic activity, a case of using L-NAME, 3 cases of 192 saporin, 4 cases of ibotenic acid, 10 cases of focal cerebral ischemia, 3 cases of alcohol-administered, and one case of natural degradation. 4. Moms water maze, Radial arm maze Passive avoidance learning model were using for examining learning and memory of model animal 5. We propose that following studies of dementia are to he investigated of the applied method of using siRNA with tranceduced gene, sample preparation by water-soaking, oriental medical diagnosis, standardization of differentiating symptom and herb simplexes, building the database by classified prescriptions, and experiment model which are based on precise examining mechanism with cell line as like mouse H19-7 hippocampus, rat HT22 hippocampus, astrocyte, microglia, using the model of animals at APP, PS1, BACE, CT99/PS1, APOE4, Tau, APP/PSI/Tau

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A Literature Study on the Korean Acupuncture for Oral, Glottal, Labial and Dental diseases (구설순치(口舌脣齒)질환에 대한 한국 침구서적의 치료법 비교 연구)

  • Han, Chang-Hyun;Ahn, Sang-Young;Kwon, Oh-Min;Park, Sang-Young;Lee, Jeong-Hyeon;Ahn, Sang-Woo
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.1
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    • pp.182-198
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    • 2010
  • Background : The varieties of manifestations referred to oral, glottal, labial and dental diseases has been long complicated physicians in the efficient diagnosis and treatment of these diseases. Acupuncture has been widely used in Korea throughout the history and provides an efficient method in the treatment of them Objectives : Establish a distinctive and efficient acupuncture method for the treatment of oral, glottal, labial and dental diseases based in literature research Method : We reviewed four Korean medical literature, "Guide to Swollen Sore Treatment", "Treasured Mirror of Eastern Medicine", "Experiential Prescriptions of Acupuncture and Moxibustion", and "Essential Rhymes on Acupuncture and Moxibustion by Master Sa-am", and analyzed the therapeutic characteristics in the treatment of oral, glottal, labial and dental diseases Result : 1. According to "Guide to Swollen Sore Treatment", we could noted frequent application of blood letting methods in the affected area, then rinse ones mouth with salty water and further application of taro plaster. Also found acupuncture methods utilizing heated mole cricket or silkworm in the sublingual region. Regarding herbal method, Realgar was rubbed in the affected area. Mainly used acupuncture points in the treatment of oral, glottal, labial and dental diseases were GV20, GB20, LU5, and auricular anterior hairline 2. In "Treasured Mirror of Eastern Medicine", blood letting method in the sublingual region and burning needle searing method were the most frequently applied in the treatment of oral, glottal, labial and dental diseases. Moxibustion was also applied in the treatment of labial and dental diseases. Particularly, said to apply 3 moxa cones in ear zones of both sides, when no medicine is effective. This demonstrates how emphasized the importance of moxibustion in this kind of disease. Mainly used acupuncture points were GV16, CV24, LI4, EX-HN12, and EX-HN13 3. In "Experiential Prescriptions of Acupuncture and Moxibustion", no other methods beside wrist. Superstitious methods like applying moxibustion on the tooth picture drawn on the roofing tile need further confirmation. Mainly used acupuncture points were LI4, ST36, and HT7. 4. "Essential Rhymes on Acupuncture and Moxibustion by Master Sa-am", identified the cause of diseases according to visceral pattern identification. Stomach and Spleen in charge of vocal, lingual, and labial disease, Kidney for dental disease, and further scrutinizing identification according to Liver, Heart, Stomach, Lung, and Kidney manifestations. Used supplementation and draining needling methods of self meridians and other correlating meridians. Conclusions : After previous study on stroke and eye diseases, we could also find various efficient methods according to oral, glottal, labial and dental diseases, through literature research of korean medical classics. This study will concurrently result in establishing distinctive therapeutic method characteristic of Korea.

Comparison of Flavonoid Content and Antioxidant Effect of Extracts from Stachys sieboldii Miq. and Lycopus lucidus Turcz (초석잠 및 택란 추출물의 플라보노이드 함량 및 항산화 활성 비교)

  • Lee, Jung Woo;Lim, Sun Young
    • Journal of Life Science
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    • v.28 no.7
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    • pp.841-848
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    • 2018
  • The flavonoid content and antioxidant effects of extracts from Stachys sieboldii Miq. and Lycopus lucidus Turcz were compared. The flavonoid content of the acetone + methylene chloride (A+M) extract of L. lucidus Turcz was 233.2 mg/g, suggesting that the extract was greater than that of S. sieboldii Miq. In the DPPH assay and the A+M and methanol (MeOH) extracts from L. lucidus Turcz had greater scavenging effects than those of S. sieboldii Miq. (p<0.05). The A+M extract from L. lucidus Turcz (0.5 mg/ml concentration) had an 82% scavenging effect in the DPPH assay. In the ABTS assay, A+M extracts from both S. sieboldii Miq. and L. lucidus Turcz (0.5 mg/ml concentration) had scavenging effects of 90% and 88%, respectively (p<0.05), suggesting that both A+M extracts had greater scavenging effects than those of both MeOH extracts. In a 120 min ROS production assay, all tested extracts dose-dependently decreased the cellular ROS production that was induced by $H_2O_2$, as compared to those produced by exposure to the extract-free control. The A+M extracts from both S. sieboldii Miq. and L. lucidus Turcz had greater inhibitory effects on cellular ROS production than those of both MeOH extracts at all concentrations tested. Treatment with the A+M extracts from S. sieboldii Miq. and L. lucidus Turcz (0.25 mg/ml concentration) inhibited the cellular ROS production by 60% and 86%, respectively. These results suggest that the A+M extracts of Stachys sieboldii Miq. and L. lucidus Turcz inhibit cellular oxidation and may contain valuable bioactive compounds, such as flavonoids.

Thermoradiotherapy in the Treatment of Advanced Stomach Cancer - To compare the difference between the results of Pre- and post-radiotherapy hyperthermia - (진행성 위암 환자에서 방사선 치료 전후에 실시한 국소온열치료)

  • Kay Chul Seung;Choi Ihl Bohng;Jang Ji Young;Kim In Ah
    • Radiation Oncology Journal
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    • v.16 no.1
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    • pp.27-33
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    • 1998
  • Purpose : To improve the therapeutic results of postoperative recurrent disease and inoperable disease of stomach cancer, we used the thermoradiotherapy. We conducted a retrospective analysis of the results and compared the results of hyperthermia before radiotherapy and those of hyperthermia after radiotherapy Materials and Methods : From July 1994 to November 1996, we treated twenty patients with locally advanced stomach cancer and recurrent stomach cancer with thermoradiotherapy. We divided those patients into two groups : hyperthermia before radiotherapy group (PreRT group : 13 Patients) and hyperthermia after radiotherapy group (PostRT group : 7 patients). We performed radiation therapy with the total tumor dose of 3000-5040cGy in a traction of 180-300cGy and 5 fractions per week. Hyperthermia was performed with 8 MHz radiofrequency apparatus. PreRT group patients were treated daily for 30 minutes before the radiation therapy within the interval of ten minutes. And PostRT group patients were treated with 1-2 sessions Per week for 40-60 minutes after the radiation therapy within the interval of 10 minutes. Results : Overall response rate was $33.3\%$. This response rate appeared the same in both groups. Mean survival and 1 rear survival rate were 10.3 months and $16.5\%$. In PreRT group, mean survival and 1 year survival rate were 6.8 months and $9.0\%$, and in PostRT group, mean survival and 1 year survival rate were 7.7 months and $34\%$. There were no statistically significant difference between the prognostic factors and therapeutic results. Conclusion : The thermoradiotherapy was a safe treatment method in advanced and recurrent gastric cancer when compared with other treatment. Because the number of patients we treated was small and the follow up period was short. we were not able to draw any conclusions about the therapeutic efficacy of the sequence of radiation therapy and hyperthermia. Therefore, further clinical trials of thermoradiotherauy for stomach cancer appear to be warranted.

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Study on the Anti-cancer, Anti-metastasis and Immune response improvement of Aqua-acupuncture with Cistanches Herba infusion solution (육종용약침의 항암작용(抗癌作用) 및 면역효과(免疫效果)에 대(對)한 실험적(實驗的) 연구(硏究))

  • Lee, Eun-Yong;Lee, Byung-Ryul
    • Journal of Acupuncture Research
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    • v.17 no.1
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    • pp.251-286
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    • 2000
  • To study the effects of anti-cancer, anti-metastasis and immune response improvement of aqua-acupuncture with Cistanches Herba infusion solution, we used Cistanches Herba infusion solution(taken by water-alcohol method) put into Chung-wan(CV12) and Chok-Samni(S36) of BALB/c or C57BL/6 which are corresponding to human body. We observed the cytotoxicity, the effect on the expression of MMP-9 gene, the ability to control cancer cell proliferation, change of body weight, surviving number, MST, ILS, changes in amount of WBC, RBC, PLT, total protein, creatinine, glucose and LDH, weight of spleen, number of pulmonary colony, histological analysis on tissue metastasis of lung and liver, splenic cell proliferation, the expression of cytokine gene, the number of $CD4^+$, $CD8^+$, $CD19^+$ and NK cell, and concluded like this. The results were obtained as follows : 1. The cytotoxicity about B16-F10 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-5}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 2. The cytotoxicity about HT1080 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-5}$, $2^{-8}$ diluent groups in Cistanches Herba infusion solution treatment was inhibited significantly, compared with control group. 3. The effect on expression of MMP-9 gene was decreased in all the sample groups, compared with control group. 4. The effect on the control-ability on the cancer cell proliferation showed cytotooicity significantly in $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-4}$, $2^{-5}$ diluent groups. 5. S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight increase in all the sample groups, compared with control group. The surviving number increased in almost sample groups, except one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group that showed same number of the control group. 6. S-180 cancer cell line transplants in BALB/c mice showed high MST and ILS significantly in almost sample groups, compared with control group. But one group put into Chok-Samni(S36) with 20% Cistanches Herba infusion solution treatment group showed low MST and ILS than control group. 7. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice showed increased ILS compared with control group significantly in anti-metastasis test. 8. The sample group injected in vein with B16-F10 cancer cell line in C57BL/6 mice were increased significantly in the number of WBC and glucose, and decreased significantly in the amount of LDH, compared with control group. However, there's no significant increase or decrease in number of RBC, PLT, total protein and creatinine. 9. We couldn't find any significant relation in spleen weight of the sample group. 10. In pulmonary colony, sample group was decreased significantly, compared with control group. 11. Histological analysis of sample group inhivited compared with that of control group in both of lung and liver. 12. In immune system, all the sample groups showed having more relevancy to the effect on splenic cell proliferation than normal group. 13. The effect on cytokine gene expression of all the sample groups were increased than control group. 14. In flow cytometry there's no significant relation in number of $CD8^+$, $CD19^+$ cell, however, the number of $CD4^+$ cell and NK cell in sample groups were increased than in control group. Above the results showed that aqua-acupuncture of Cistanches Herba infusion solution has effects of anti-cancer, anti-metastasis and immune response improvement.

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Short Tenn Reactions to Acupuncture Treatment and Adverse Events Following Acupuncture in Korea a Cross-sectional Survey of Patient Reports (침치료 직 후 자가 설문지를 이용한 침반응(針感)과 부작용에 대한 단면적 연구)

  • Park, Seong-Uk;Jung, Woo-Sang;Moon, Sang-Kwan;Ko, Chang-Nam;Cho, Ki-Ho;Kim, Young-Suk;Bae, Hyung-Sup;Park, Jung-Mi
    • The Journal of Korean Medicine
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    • v.28 no.2 s.70
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    • pp.66-79
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    • 2007
  • Objectives : To explore the type and frequency of short term reactions, de Qi associated with acupuncture treatment and to determine the incidence of adverse events following acupuncture in Korea. Subjects and methods : This study is a retrospective and cross-sectional survey of patient reports. 1095 subjects, 585 of out-patients of the Oriental Medicine of Stroke & Neurological Disorders Center, East-West NEO Medical Center of Kyunghee University and 510 of out-patients of the Department of Cardiovascular & Neurologic Diseases (Stoke Center), Hospital of Oriental Medicine, Kyunghee Medical Center, from June through November of 2006, who had acupuncture, gave informed consent and completed one survey form. On this form, patients were asked to report short term acupuncture reactions, de Qi, patient satisfaction measurement (using VAS), and adverse events relating to acupuncture treatment. The acupuncturists of this study are Korean Medicine Doctors (KMD) who had worked as practitioners for 3-30 years or more. Results : The average age of the 1095 subjects was 58 years old. Positive short term acupuncture reactions after treatment were reported by 878 (80.2%), negative short term acupuncture reactions by 75 (6.8%) and no reactions were reported by 142 (13.0%). The most common positive short term acupuncture reactions were feeling 'relaxed', 472 (43.1%), followed by feeling 'less pain' 90 (8.2%), 'energized' 16 ( 1.5%), 'tingling' 16 (1.5%), 'heat feeling or Cold feeling' 10 (0.9%), and others 274 (25.0%), respectively. Negative short term acupuncture reactions were feeling 'pain' 37 (3.4%), tiredness 24 (2.2%), dizziness 9 (0.8%), and others 5 (0.2%), respectively. Traditionally described needling sensations of de Qi refer to a patient's response to distention, pulling, soreness, heaviness, numbness. 39.7% of subjects reported de Qi during needling, experiencing 'distention' 333 (30.4%), 'soreness' 52 (4.7%), 'pulling' 22 (2.0%), 'heaviness' 18 (1.6%), and 'numbness' 10 (0.9%) respectively. Positive short term acupuncture reactions and de Qi rate were the highest in the less than 40 years group (83/96 86.5%, 50/96 52.1%). No acupuncture reactions were highly seen in the over 70 years old group (31/187, 16.6%). Patient satisfaction level using VAS was a comparatively high $72.9{\pm}19.9$. Adverse events were only bleeding in 92 (8.4%) of the total subjects. High sensitive acupoints were 95 points as GV26 (54 times), LI4 (54 times), ST36 (53 times), GB20 (37 times), HT8(34 times), LV3 (29 times), SI3 (29 times), and LI11 (27 times) in order. Main impressions were stroke patients 430 (16.9%), headache 185 (16.9%), hypertension 97 (8.9%), and dizziness 85 (7.8%). Conclusions : Although 8 different Korean Medicine Doctors participated in this research, we obtained similar results from each. There were no significantly different results between the two hospitals. Short term acupuncture reactions and de Qi were most related to age. Except for bleeding there were no adverse events relating to acupuncture treatment in this study. We consider acupuncture treatment as very safe depending on practitioners. Positive short term acupuncture reactions after treatment were 12 times higher than negative short term acupuncture reactions. Subjects were comparatively satisfied with acupuncture treatment.

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Study on the Anti-cancer, Anti-metastasis and Immune response improvement Effects of Aqua-acupuncture with Rubi Fructus infusion solution (항암(抗癌) 및 면역효과(免疫效果)에 복분자약침(覆盆子藥鍼)이 미치는 영향(影響))

  • Lee, Seon-Hee;Lee, Byung-Ryul
    • Journal of Acupuncture Research
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    • v.17 no.1
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    • pp.189-219
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    • 2000
  • To study the effects of anti-cancer, anti-metastasis and immune response improvement effects of aqua-acupuncture with Rubi Fructus infusion solution, we used Rubi Fructus infusion solution(taken by water-alcohol method) put into Chung-wan (CV12) and Chok-Samni(ST36) of BALB/c or C57BL/6 which are corresponding to humanbody. We observed the cytotoxicity, the effect on the expression of MMP-9 gene, the ability to control cancer cell proliferation, change of body weight, surviving number, median surviving time, increase of life span, changes in amount of leukocyte, erythrocyte, platelet, total protein, creatinine, glucose and LDH, weight of spleen, number of pulmonary colony, histological analysis on tissue metastasis of lung and liver, splenic cell proliferation, the expression of cytokine gene, the number of $CD4^+$, $CD8^+$, $CD19^+$ and NK cell, and concluded like this. The results were obtained as follows : 1. Effects of Anti-cancer 1) The cytotoxicity about B16-F10 cell line of $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-5}$, $2^{-6}$, $2^{-7}$, $2^{-8}$ diluent groups in Rubi Fructus infusion solution treatment was inhibited significantly, compared with control group. 2) The cytotoxicity about HT1080 cell line of $2^0{\sim}2^{-8}$ diluent groups in Rubi Fructus infusion solution treatment was inhibited significantly, compared with control group. 3) The effect on expression of MMP-9 gene was inhibited significantly in all the sample groups, compared with control group. 4) The effect on the control-ability on the cancer cell proliferation showed cytotoxicity significantly in $2^0$, $2^{-1}$, $2^{-2}$, $2^{-3}$, $2^{-4}$, $2^{-5}$, $2^{-6}$, $2^{-7}$, diluent groups. 2. Effects of Anti-metastasis 1) S-180 cancer cell line transplants in BALB/c mice were inhibited significantly in weight increase in all the sample groups, compared with control group. The surviving number increased in almost sample groups, except one group put into Chok-Samni(ST36) with 20% Rubi Fructus infusion solution treatment group that showed same number of the control group. 2) S-180 cancer cell line transplants in BALB/c mice showed high MST significantly in almost sample groups, compared with control group. But one group put into Chok-Samni(ST36) with 20% Rubi Fructus infusion solution showed low MST than control group. 3) The group injected in vein with B16-F10 cancer cell line in C57BL/6 mice showed increased ILS than control group significantly in anti-metastasis test. 3. Effects of Immune response improvement 1) The group injected in vein with B16-F10 cancer cell line in C57BL/6 mice were increased significantly in the number of leukocyte and glucose, and decreased significantly in the amount of platelet and LDH, compared with control group. However, there's no significant increase or decrease in number of erythrocyte, total protein and creatinine. 2) We couldn't find any significant relation in spleen weight of the sample group. 3) In pulmonary colony, sample group was decreased significantly, compared with control group. 4) Histological analysis of sample group inhivited compared with that of control group in both of lung and liver. 5) In immune system, all the sample groups showed having more relevancy to the effect on splenic cell proliferation than normal group. 6) Cytokine gene increased in almost sample groups, except one group treated with $50{\mu}g/m{\ell}$ Rubi Fructus infusion solution on IL-12. 7) In flow cytometry there's no significant relation in number of $CD8^+$ cell, however, the number of $CD4^+$, $CD19^+$ cell and NK cell in sample group had more relation than in control group. Above the results showed that aqua-acupuncture of Rubi Fructus solution has effects of anti-cancer, and-metastasis and immune response improvement.

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