CN103520279A - Application of cinnamon extract to preparation of medicines or healthcare products for treating or preventing tumour - Google Patents
Application of cinnamon extract to preparation of medicines or healthcare products for treating or preventing tumour Download PDFInfo
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Abstract
The invention discloses novel application of a cinnamon extract, which relates to application of cinnamon (including cinnamon, chartophyllum, cinnamomum mollifolium, howood, cinnamomum burmannii and sassafras) extract to preparation of medicines or healthcare products for treating or preventing tumour. The invention also provides a preparation method of the cinnamon extract. Researches show that the cinnamon extract is capable of activating an Nrf2 signal path, promoting anti-oxidation and expression of II phase detoxifying enzymes, increasing the intracellular glutathione level, enhancing the intracellular reduction capability and inhibiting the cytotoxicity caused by cancerogen arsenic and reactive oxygen H2O2. Consequently, the cinnamon extract can be prepared into corresponding oral preparations and injections and used for preventing the tumour.
Description
Technical field
The present invention relates to the new purposes of cinnamon extract, be specifically related to cinnamon extract in preparation treatment or the medicine of prophylaxis of tumours or the application in health product.
Background technology
Along with social development, the improvement of human living standard and medical environment, human longevity has obtained significant prolongation, but in the time of rapid economic development, environmental pollution is serious, cause the sickness rate of malignant tumor also improving, serious threat human health, brings white elephant to country and society.Realize malignant tumor effectively under the prerequisite for the treatment of there is no active drug, research and development have medicine and the health product of preventive effect, prevent the early origin of disease and become effective coping strategy.
Nuclear factor Nrf2(Nuclear factor-erythroid 2-related factor2) be the important transcription factor that regulates cellular oxidation reduction balance, by reacting original paper (ARE) in conjunction with regulating cell redox equilibrium with antioxidase and II phase detoxication enzyme promoter region antioxidation, its target gene comprises the II phase detoxication enzyme of coding GST, UGT, γ GCS, NQO1, and the endogenous antioxidation albumen such as GCL, HO-1.When body is exposed under the material that electrophilicity foreign body, chemical carcinogen, endogenous oxidant etc. have damage body; Nrf2 just can be activated and pass through to raise antioxidase and II phase detoxication enzyme is removed nuisance; protection body is avoided damage (the Lau et al. of poisonous substance; Pharmacol.Res.; 2008; 58,262-270).Therefore, by activating Nrf2 signal path, enhancing body self is removed poisonous substance and carcinogenic ability, generation that can prevention of various diseases, for example malignant tumor.
Under oxidative stress status, in the face of endogenous and the infringement of exogenous poisonous substance to body, utilize exogenous Nrf2 agonist to raise body Nrf2 level, can enhancing body self-defense ability, the damages of poisonous substance to body such as prevention endogenous active oxygen, exogenous carcinogen, significant (the Magesh et al. of generation for prevention malignant tumor, Med.Res.Rev., 2012,32,4,687-726).Activate Nrf2 signal path, can raise II phase detoxication enzyme level, remove carcinogen or reduce its toxicity, suppress DNA damage, the gene mutation of carcinogen induction, by raising Nrf2, express, can increase glutathion inside cell level, strengthen activities of antioxidant enzymes, active oxygen in scavenger cell, suppresses cytolipin peroxidating, thus the generation of prevention malignant tumor.Therefore monomeric compound, plant extract that, has Nrf2 agonism etc. can effectively prevent the incidence rate of malignant tumor.
Cinnamomum (Cinnamomum), is a genus of Lauraceae, approximately 250 kinds, is distributed in Tropical Asia, Australia to Pacific Islands and tropical America.China approximately has 46 kinds, each provinces and regions, main product south, and planting number is at most Yunnan, is secondly Guangdong and Sichuan, wherein Cortex Cinnamomi, bavin osmanthus, Gui,Chuan osmanthus, south China are important medical materials.In cinnamon, mainly contain the chemical compositions such as volatile oil, flavone, Phenylpropanoid Glycosides.
Summary of the invention
For above-mentioned prior art, the invention discloses the new purposes of cinnamon extract---cinnamon (comprising Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree) extract, in preparation treatment or the medicine of prophylaxis of tumours or the application in health product.The present invention also provides the preparation method of cinnamon extract.
The present invention is achieved by the following technical solutions:
The present inventor finds by research, cinnamon (comprises Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy fragrant, yellow Camphor tree) extract, can activate Nrf2 signal path, and can resist the cytotoxicity of carcinogen arsenic and the generation of active oxygen hydrogen peroxide by Cell protection, therefore, the above-mentioned cinnamon extract of can take is prepared the health product of prophylaxis of tumours as raw material, also can prepare treat tumor medicine (through By consulting literatures, there are no cinnamon extract, activate up to now the report of Nrf2 signal path, also have no cinnamon for the report of tumor prevention and treatment).
During concrete application, cinnamon (comprising Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), the yellow Camphor tree) extract of can take is raw material, add medically acceptable adjuvant, make the dosage forms such as capsule (comprising soft capsule), tablet, powder, granule, oral liquid, medicated wine, pill, pellet, mixture, tincture, preferred particulates agent, capsule, tablet; Or cinnamon extract is further purified and makes ejection preparation.Also cinnamon extract and other can be had to the medicament mixed for the treatment of or preventive effect to tumor, add again or do not add medically acceptable adjuvant, making the dosage forms such as capsule (comprising soft capsule), tablet, powder, granule, oral liquid, medicated wine, pill, pellet, mixture, tincture, injection.
Described cinnamon extract, prepares by the following method:
Cinnamon (comprising Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree) aerial parts is pulverized, by 0～95% ethanol (percentage by volume) reflux, extract, 1～8 time, each solvent load is 1～20 times of medical material weight, and each extraction time is 0.5～10 hour; Merge extractive liquid,, filters, and reclaims ethanol, concentrates, is dried, and obtains cinnamon extract.
Preferably, with 30～90% alcohol reflux 2～4 times, the consumption of each solvent is 3～7 times of medical material weight, and each extraction time is 2～4 hours.
Described any one cinnamon extract, all can be for the preparation of medicine or the health product for the treatment of or prophylaxis of tumours.
The present inventor has completed cinnamon (comprising Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree) extract to the effect of Nrf2 signal activation, and the research of the Cytotoxic protective effect that carcinogen arsenic and active oxygen hydrogen peroxide are caused.Result shows, above-mentioned cinnamon extract can activate Nrf2 at protein level, increases the expression of Nrf2 and downstream gene NQO1 and GCS, by increasing the level of glutathion inside cell, strengthens cell reducing power.Inventor also finds that they can suppress the cytotoxicity of carcinogen arsenic and the induction of active oxygen hydrogen peroxide, show the ability that suppresses cell carcinogenesis.With cinnamon, comprise Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree below, introduce it to Nrf2 signal path and Cytotoxic effect that carcinogen arsenic and active oxygen hydrogen peroxide are caused:
The MDA-MB-231-Luc cell line of the luciferase plasmids that employing can stably express relies on containing ARE, has evaluated the effect of cinnamon extract to Nrf2, and result shows that cinnamon extract has the effect (Fig. 1) that activates Nrf2 signal path.
Select human breast carcinoma MDA-MB-231 cell as detecting cell, evaluate the effect of cinnamon extract to Nrf2 signal path.Western blot analysis result shows, cinnamon extract is to raising the expression of Nrf2 albumen, and can promote the expression (Fig. 2) of downstream antioxidation and II phase detoxication enzyme albumen NQO1 and γ GCS.
Select people's normal lung epithelium HBE cell, detected the impact of cinnamon extract on glutathion inside cell level.Result demonstration, cinnamon extract, to increasing glutathion inside cell level (Fig. 3), strengthens reducing power and Scavenger of ROS ability in cell.
Select the cytotoxicity model of carcinogen arsenic induction, evaluate cinnamon extract to the protective effect to people's normal lung epithelium HBE cell.Through cinnamon extract, to after processing, cells survival rate is significantly higher than not dosing processed group, proves that cinnamon extract, to suppressing the cytotoxicity (Fig. 4) of trivalent arsenic induction, shows that cinnamon extract is to can be used for prevention and the treatment of tumor.
Select active oxygen H
2o
2the cytotoxicity model of induction, evaluates the protective effect of cinnamon extract to people's normal lung epithelium HBE cell.Result shows that plant extract can suppress H
2o
2the toxicity producing, improves cells survival rate (Fig. 5).The generation toxicity many and that active oxygen produces of tumor is closely related, and this result proof cinnamon extract can be used for prevention and the treatment of tumor.Accompanying drawing explanation
The luciferase reporting test that Fig. 1: ARE relies on shows that cinnamon extract is to activating Nrf2 signal path, cinnamon extract concentrations unit in figure is μ g/mL, the positive contrast of sulforaphen 2.5 μ M, wherein A is Cortex Cinnamomi, B is hard leaf Camphor tree, and C is Mao Yezhang, and D is Lignum cinnamomi camphorae, E is cloudy fragrant, and F is yellow Camphor tree.
Fig. 2: western blot analysis shows that cinnamon extract can activate its downstream gene of Nrf2 at protein level, cinnamon extract concentrations unit in figure is μ g/mL, the positive contrast of sulforaphen 2.5 μ M, wherein A is Cortex Cinnamomi, B is hard leaf Camphor tree, and C is Mao Yezhang, and D is Lignum cinnamomi camphorae, E is cloudy fragrant, and F is yellow Camphor tree.
Fig. 3: cinnamon extract can increase glutathion inside cell level, the positive contrast of sulforaphen 2.5 μ M in figure, cinnamon concentration is respectively: Cortex Cinnamomi (60 μ g/mL), hard leaf Camphor tree (60 μ g/mL), Mao Yezhang (90 μ g/mL), Lignum cinnamomi camphorae (60 μ g/mL), cloudy fragrant (90 μ g/mL), yellow Camphor tree (400 μ g/mL).
Fig. 4: cinnamon extract can reduce the cytotoxicity (* p < 0.05) of arsenic (As) induction, wherein A is Cortex Cinnamomi (60 μ g/mL), B is hard leaf Camphor tree (60 μ g/mL), C is Mao Yezhang (90 μ g/mL), D is Lignum cinnamomi camphorae (60 μ g/mL), E is cloudy fragrant (90 μ g/mL), and F is yellow Camphor tree (400 μ g/mL).
Fig. 5: cinnamon extract can reduce hydrogen peroxide (H<sub TranNum="95">2</sub>o<sub TranNum="96">2</sub>) induction cytotoxicity (* p<0.05), wherein A is Cortex Cinnamomi (60 μ g/mL), B is hard leaf Camphor tree (60 μ g/mL), C is Mao Yezhang (90 μ g/mL), D is Lignum cinnamomi camphorae (60 μ g/mL), and E is cloudy fragrant (90 μ g/mL), and F is yellow Camphor tree (400 μ g/mL).
The specific embodiment
Below in conjunction with embodiment, the present invention is further illustrated, and following embodiment is only the object of explanation, not for limiting the present invention.
Embodiment 1: the activation of cinnamon extract to Nrf2 signal path
Method: the preparation of (1) cinnamon extract
Cinnamon (is respectively Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree, carry out altogether the preparation of 6 kinds of Lignum cinnamomi camphorae plant extracts) aerial parts 200g, pulverize, 75% ethanol 800mL, extract 2 times, each 1 hour, merge extractive liquid,, concentrate drying obtains 6 kinds of Lignum cinnamomi camphorae plant extracts, for activity rating.
(2) cultivation of human breast cancer cell MDA-MB-231 cell line
Human breast cancer cell MDA-MB-231, purchased from US mode culture collection warehousing (ATCC), adopts the MEM culture medium containing 10% hyclone (FBS), 2mMHEPES and 6ng/mL insulin, is placed in 37 ℃, 5%CO
2in incubator, cultivate.
(3) the luciferase reporting test that ARE relies on
MDA-MB-231-Luc cell is inoculated on 96 orifice plates, the testing compound that adds variable concentrations after 24 hours, process 16 hours, adopt cell pyrolysis liquid [0.1M potassium phosphate and 1mM dithiothreitol, DTT] cell lysis, with detecting liquid (25mM glycylglycine, 15mM magnesium sulfate, 500 μ MATP, 250 μ M fluoresceins and 250 μ M coenzyme As), measure fluorescence intensity.
Result: as shown in Figure 1, cinnamon extract can significantly strengthen the uciferase activity that ARE relies on, under arbitrary extract optium concentration, intensity is 4 times of blank group, and positive control medicine sulforaphen (2.5 μ M) is 2 times of blank group.The above results shows, six kinds of cinnamons (comprising Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree) extract can be induced Nrf2 signal path, and body is had to better protective effect.
Embodiment 2: cinnamon extract can raise Nrf2 and with the expression of downstream gene protein level
Method: western blot analysis (Westernblot) detects the variation of protein level in cell
MDA-MB-231 cell is inoculated in diameter 35mm culture dish, after being cultured to density and reaching 70%～80%, add the cinnamon extract (cinnamon extract is embodiment 1 preparation) of variable concentrations to process 16h, PBS washing 2 times, adds cell pyrolysis liquid (50 μ g/ml aprotiniies, 0.5mM Phenylmethanesulfonyl fluoride, 1mM sodium vanadate, 10mM sodium fluoride, 10mM β-phosphoglycerol), collect albumen and adopt Bradford method to measure protein concentration.Each sample thief albumen (100 μ g) loading, SDS-PAGE protein isolate component also adopts electrotransfer method that protein band is transferred on cellulose nitrate film.Thin film is after the defatted milk powder solution room temperature sealing 1h of TBS preparation 5%, respectively with 4 ℃ of overnight incubation of each testing protein antibody.After TBS washing, add respectively two anti-hatching after 1h of horseradish peroxidase, with enhancement mode ECL chemiluminescence, carry out analysis of protein.
Result: as shown in Figure 2, cell is after cinnamon extract-treated 16h, Nrf2 and downstream antioxidation thereof and II phase detoxication enzyme albumen NQO1 and γ GCS protein level present dose dependent to be increased, and confirms that six kinds of cinnamons (comprising Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree) extract can plan Nrf2 signal path on protein level.
Embodiment 3: cinnamon extract increases glutathion inside cell level
Method: the cultivation of (1) people's normal lung epithelial cell HBE cell
People's normal lung epithelial cell HBE, purchased from US mode culture collection warehousing (ATCC), adopts MEM culture medium, and adds wherein 10% hyclone (FBS), and 5% glutamine, is placed in 37 ℃, 5%CO
2in incubator, cultivate.
(2) mensuration of glutathion inside cell content
HBE cell is inoculated in diameter 35mm culture dish, after being cultured to density and reaching 70%～80%, add the cinnamon extract (cinnamon extract is embodiment 1 preparation) of variable concentrations to process 24 hours, PBS washing 2 times, adds 0.5mL50mM sodium phosphate and 1mMEDTA buffer to receive cell, ultrasonic 1min, centrifugal 15 minutes of 10000g, get supernatant, according to glutathion, measure the operation of test kit description, 412nm measures absorbance and calculates glutathione content.
Result: as shown in Figure 3, six kinds of cinnamons (comprising Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree) extract can significantly increase glutathion inside cell level, strengthens reducing power in cell.
Embodiment 4: cinnamon extract can suppress the toxicity of trivalent arsenic to people's pulmonary branches tracheal epithelium HBE cell
Method: mtt assay is measured the Cytotoxic protective effect of compound to arsenic (As) induction
HBE cell is inoculated on 96 orifice plates, add variable concentrations cinnamon extract-treated to be measured 16 hours, then add arsenic and the concentration cinnamon extract to be measured (cinnamon extract is embodiment 1 preparation) of variable concentrations to process 24 hours, add after MTT3 hour, 590nm measures absorbance and calculates cells survival rate.
Result: as shown in Figure 4; adopt cinnamon extract pretreatment cell 16 hours; then add the arsenic of 20 μ M, 40 μ M and cinnamon extract-treated cell 24 hours, result shows that cinnamon extract has protective effect to HBE cell, can suppress the cytotoxicity of arsenic induction.The cells survival rate of cinnamon extract-treated groups is apparently higher than not adding medicine processed group.Result proves, cinnamon extract (comprising Cortex Cinnamomi, hard leaf Camphor tree, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree) can suppress the toxicity of carcinogen arsenic induction, can be used in prevention and the treatment of tumor.
Embodiment 5: cinnamon extract can suppress active oxygen hydrogen peroxide (H
2o
2) toxicity to people's pulmonary branches tracheal epithelium HBE cell
Method is with embodiment 4.
Result: shown in Fig. 5, adopt respective concentration cinnamon extract pretreatment cell 16 hours, then add the H of 100 μ M, 200 μ M
2o
2with respective concentration cinnamon extract-treated cell 24 hours, result showed that cinnamon extract has protective effect to HBE cell, can suppress H
2o
2the cytotoxicity of induction.The cells survival rate of cinnamon extract-treated groups is apparently higher than not adding medicine processed group.Result proves, cinnamon extract (comprising Cortex Cinnamomi, hard leaf Camphor tree, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree) can suppress active oxygen H
2o
2the neurotoxicity producing, can be used in prevention and the treatment of tumor.
Embodiment 6: the preparation of Cortex Cinnamomi extract
Cortex Cinnamomi aerial parts 5kg, pulverizes, and with 90% alcohol reflux of 3 times of medical material weight 2 times, each extraction time is 4 hours, and merge extractive liquid, filters, and reclaims ethanol, and drying under reduced pressure, obtains cinnamon extract 315g, and yield is 6.3%.
Embodiment 7: the preparation of Lignum cinnamomi camphorae extract
Lignum cinnamomi camphorae aerial parts 5kg, pulverizes, and with 75% alcohol reflux of 5 times of medical material weight 3 times, each extraction time is 3 hours, and merge extractive liquid, filters, and reclaims ethanol, and spraying is dry, obtains cinnamon extract 350g, and yield is 7.0%.
Embodiment 8: the preparation of yellow Cinnomomum Sps. Extract
Yellow Camphor tree aerial parts 5kg, pulverizes, and with 35% alcohol reflux of 7 times of medical material weight 4 times, each extraction time is 2.5 hours, and merge extractive liquid, filters, and reclaims ethanol, and lyophilization, obtains cinnamon extract 412g, and yield is 8.24%.
Embodiment 9: the preparation of tablet
Yellow Cinnomomum Sps. Extract (embodiment 8 preparations) 20g, adds corn starch 30g, and dextrin 15g, fully mixes, granule processed, and dry, granulate, adds appropriate Pulvis Talci, and tabletting, obtains.
Embodiment 10: the preparation of capsule
Yellow Cinnomomum Sps. Extract (embodiment 8 preparations) 20g, adds starch 20g, dextrin 15g, and microcrystalline Cellulose 8g, fully mixes, granule processed, and dry, granulate, adds appropriate magnesium stearate, encapsulated, obtains.
Claims (8)
1. cinnamon extract is in preparation treatment or the medicine of prophylaxis of tumours or the application in health product.
2. application according to claim 1, it is characterized in that: while specifically applying, take cinnamon extract as raw material, add medically acceptable adjuvant, make capsule, tablet, powder, granule, oral liquid, medicated wine, pill, pellet, mixture or tincture; Or cinnamon extract purification is made to ejection preparation; Or: cinnamon extract and other are had to the medicament mixed for the treatment of or preventive effect to tumor, add again or do not add medically acceptable adjuvant, making capsule, tablet, powder, granule, oral liquid, medicated wine, pill, pellet, mixture, tincture or injection; The extract that described cinnamon extract divides for this plant shoot.
3. application according to claim 1 and 2, is characterized in that: described cinnamon is Cortex Cinnamomi (Cinnamomum cassia Presl.), hard leaf Camphor tree (Cinnamomum chartophyllum H.W.Li), Mao Yezhang (Cinnamomum mollifolium H.W.Li), Lignum cinnamomi camphorae (Cinnamomum camphora var.linaloolifera Y.Fujita), cloudy fragrant (Cinnamomum burmannii (C.G.etTh.Nees) Bl.), yellow Camphor tree (Cinnamomum porrectum (Roxb.) Kosterm).
4. a cinnamon extract, it is characterized in that: prepare by the following method: cinnamon aerial parts is pulverized, with 0～95% alcohol reflux 1～8 time, each solvent load is 1～20 times of medical material weight, and each extraction time is 0.5～10 hour; Merge extractive liquid,, filters, and reclaims ethanol, concentrates, is dried, and obtains cinnamon extract.
5. cinnamon extract according to claim 4, is characterized in that: described cinnamon is selected from Cortex Cinnamomi, hard leaf Camphor tree, Mao Yezhang, Lignum cinnamomi camphorae, cloudy perfume (or spice), yellow Camphor tree.
6. according to the cinnamon extract described in claim 4 or 5, it is characterized in that: cinnamon aerial parts is pulverized, and with 30～90% alcohol reflux 2～4 times, each solvent load is 3～7 times of medical material weight, and extraction time is 2～4 hours at every turn; Merge extractive liquid,, filters, and reclaims ethanol, concentrates, is dried, and obtains cinnamon extract.
7. the cinnamon extract described in claim 4～6 is in preparation treatment or the medicine of prophylaxis of tumours or the application in health product.
8. application according to claim 7, it is characterized in that: while specifically applying, take any one cinnamon extract is raw material, adds medically acceptable adjuvant, makes capsule, tablet, powder, granule, oral liquid, medicated wine, pill, pellet, mixture or tincture; Or cinnamon extract purification is made to ejection preparation; Or: cinnamon extract and other are had to the medicament mixed for the treatment of or preventive effect to tumor, add again or do not add medically acceptable adjuvant, making capsule, tablet, powder, granule, oral liquid, medicated wine, pill, pellet, mixture, tincture or injection.
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| CN107496433A (en) * | 2017-08-24 | 2017-12-22 | 山东大学 | A kind of biphenyl compound is in the application for preparing oxidative stress and inducing an illness in medicine |
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| CN107496433A (en) * | 2017-08-24 | 2017-12-22 | 山东大学 | A kind of biphenyl compound is in the application for preparing oxidative stress and inducing an illness in medicine |
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Effective date of registration: 20190605 Address after: 276002 Bancheng Town, Lanshan District, Linyi City, Shandong Province Patentee after: Linyi Shansong Pharmaceutical Co. Ltd. Address before: 250061 culture West Road, Lishi District, Ji'nan, Shandong Province, No. 44 Patentee before: Shandong University |
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Effective date of registration: 20190710 Address after: 276002 Bancheng Town, Lanshan District, Linyi City, Shandong Province Co-patentee after: Linyi Jinluo Health Science and Technology Research Institute Patentee after: Linyi Shansong Pharmaceutical Co. Ltd. Address before: 276002 Bancheng Town, Lanshan District, Linyi City, Shandong Province Patentee before: Linyi Shansong Pharmaceutical Co. Ltd. |
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