CN109276576A - The purposes of plain boiled pork ganoderma lucidum polysaccharide in the preparation of antitumor drugs - Google Patents

The purposes of plain boiled pork ganoderma lucidum polysaccharide in the preparation of antitumor drugs Download PDF

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CN109276576A
CN109276576A CN201811387888.3A CN201811387888A CN109276576A CN 109276576 A CN109276576 A CN 109276576A CN 201811387888 A CN201811387888 A CN 201811387888A CN 109276576 A CN109276576 A CN 109276576A
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ganoderma lucidum
plain boiled
boiled pork
pork ganoderma
preparation
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CN109276576B (en
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李向敏
吴清平
谢意珍
胡惠萍
刘远超
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Guangdong Detection Center of Microbiology of Guangdong Institute of Microbiology
Guangdong Yuewei Edible Mushroom Technology Co Ltd
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Guangdong Detection Center of Microbiology of Guangdong Institute of Microbiology
Guangdong Yuewei Edible Mushroom Technology Co Ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • Nutrition Science (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention discloses the purposes of plain boiled pork ganoderma lucidum polysaccharide in the preparation of antitumor drugs, the preparation method of the plain boiled pork ganoderma lucidum polysaccharide is the following steps are included: (1) is extracted;(2) alcohol precipitation;(3) it is lyophilized;(4) it isolates and purifies.It the beneficial effects of the present invention are: providing a kind of new application of plain boiled pork ganoderma lucidum polysaccharide, can effectively inhibit tumour growth, can be used for preparing anti-tumor drug, can be used as the synergist of anti-tumor drug such as taxol etc., there is good medical value.

Description

The purposes of plain boiled pork ganoderma lucidum polysaccharide in the preparation of antitumor drugs
Technical field
The present invention relates to biomedicine technical fields, and in particular to the use of plain boiled pork ganoderma lucidum polysaccharide in the preparation of antitumor drugs On the way.
Background technique
Cancer is the world's second largest cause of the death, causes within 2015 8800000 people dead, is had become global serious public strong Kang Wenti.The important channel that independent or adjunct antineoplastic compound is still anti-tumor drug discovery is screened from natural products. Polysaccharide is the high molecular polymer connected with ketone group by glycosidic bond by aldehyde radical, serves not only as energy source and support in vivo Tissue, but prior participation biological phenomena and physiology course such as cell recognition, neurodevelopment, metastases etc., and Many natural polysaccharide, which have, adjusts the functional activities such as immune, antitumor, hypoglycemic.Ganoderma lucidum is as a kind of food medicine dual-purpose Macro fungi has long medicinal history in East Asia various countries.Ganoderma lucidum polysaccharide by modern pharmacology prove have protect liver, it is antitumor, Lowering blood pressure and blood fat adjusts a variety of effects such as immunity of organisms.
Plain boiled pork ganoderma lucidum (Ganoderma leucocontextum) is to acquire in 2011 in Linzhi Bowo County Zha Xi Gang Cun To morphological feature be different from the ganoderma lucidum of known Ganoderma kind, it is raw according to morphology and molecule by research in 4 years Object is as a result, be accredited as Ganoderma novel species (Li T et al.2014).The polysaccharide of plain boiled pork ganoderma lucidum fruitbody is detected, The content of the main component polysaccharide of individual bacterial strains is 6-8 times higher than sold ganoderma lucidum on the market as the result is shown.
The current research in relation to plain boiled pork ganoderma lucidum is only limitted to the extracting method and use of the cultivation of plain boiled pork ganoderma lucidum, identification and extract Way etc., but seldom it is related to research of the plain boiled pork ganoderma lucidum in terms of inhibition tumour growth and as anti-tumor drug synergist, because This, which carries out in-depth study in this aspect to it, has significant meaning.
Summary of the invention
In order to solve the above technical problems, the present invention provides a kind of new application of plain boiled pork ganoderma lucidum polysaccharide, that is, it is used as active constituent Purposes in the preparation of antitumor drugs has the function of inhibiting tumour growth, while can be combined with antitumor drug paclitaxel, Enhancing inhibits the effect of tumour growth.
The preparation method of above-mentioned plain boiled pork ganoderma lucidum polysaccharide specifically comprises the following steps:
(1) it extracts: plain boiled pork ganoderma lucidum fruitbody being smashed, is soaked in water, and is heated, refluxing extraction is multiple, merges filter Filtrate is concentrated liquid, obtains concentrate;
(2) alcohol precipitation: concentrate is subjected to alcohol precipitation, stands overnight, is centrifuged later, precipitated;
(3) it is lyophilized: being dissolved and precipitated with pure water, freeze-drying obtains Thick many candies, and kept dry is spare;
(4) it isolates and purifies: the plain boiled pork ganoderma lucidum crude polysaccharide of acquisition is dissolved in water, stepwise elution is carried out by ion exchange column, Plain boiled pork ganoderma lucidum polysaccharide is obtained after being freeze-dried again.
Preferably, the preparation method of above-mentioned plain boiled pork ganoderma lucidum polysaccharide may further include following steps:
(1) it extracts: plain boiled pork ganoderma lucidum fruitbody being smashed, is soaked in water half an hour according to the ratio of solid-liquid ratio 1:10~20, And heated at 80~100 DEG C, three times, merging filtrate concentrates the filtrate to 10th to ten/7th of stoste to refluxing extraction / mono-, obtain concentrate;
(2) alcohol precipitation: being sink to final concentration of the 40~80% of alcoholic solvent for concentrate progress alcohol, stand overnight, be centrifuged later, It is precipitated;
(3) it is lyophilized: being dissolved and precipitated with pure water, freeze-drying obtains Thick many candies, and kept dry is spare;
(4) it isolates and purifies: the plain boiled pork ganoderma lucidum crude polysaccharide of acquisition is dissolved in water, stepwise elution is carried out by ion exchange column, Plain boiled pork ganoderma lucidum polysaccharide is obtained after being freeze-dried again.
Preferably, the time of refluxing extraction is respectively 2h, 2h, 1h three times in above-mentioned steps (1).
Preferably, the concentrate in above-mentioned steps (2) after alcohol precipitation is stood overnight at a temperature of 4 DEG C, and/or with 4000~ The revolving speed of 8000rpm is centrifuged 10~30min.
It is further preferred that the concentrate in above-mentioned steps (2) after alcohol precipitation is centrifuged 10min with the revolving speed of 6000rpm.
Preferably, above-mentioned steps (2) alcohol is sink to final concentration of the 75% of alcoholic solvent.
Preferably, the plain boiled pork ganoderma lucidum is plain boiled pork ganoderma lucidum Ganoderma leucocontextum HMGIM- Z110122, deposit number are CCTCC NO:M2016087.
The above-mentioned plain boiled pork ganoderma lucidum of the present invention is picked up from the broad-leaf forest of Linzhi Area of Tibet Bowo County Zha Xi Gang Xiang, is identified as Plain boiled pork Ganoderma Varieties By Uv Induced, and separation is organized to obtain original strain, it is named as plain boiled pork ganoderma lucidum Ganoderma Leucocontextum HMGIM-Z110122 has been preserved in China typical culture collection center (letter on March 7th, 2016 Claim CCTCC, address is Wuhan City, Hubei Province Wuchang District Bayi Road Luo Jia Shan), deposit number is CCTCC NO:M2016087.
Further, the present invention also provides a kind of plain boiled pork ganoderma lucidum polysaccharide, and the plain boiled pork ganoderma lucidum polysaccharide is by above-mentioned preparation method It obtains.
Secondly, the present invention also provides a kind of antineoplastic pharmaceutical compositions, wherein the antineoplastic pharmaceutical compositions contain Effective quantity plain boiled pork ganoderma lucidum polysaccharide as described above.
Preferably, the antineoplastic pharmaceutical compositions further comprise one or more pharmaceutically acceptable auxiliary materials, The auxiliary material may include acid, alkali, salt, hydrate or ester and other auxiliary materials.
Compared with prior art, the invention has the following beneficial effects:
The present invention provides a kind of new applications of plain boiled pork ganoderma lucidum polysaccharide, can effectively inhibit tumour growth, can be used for preparing anti- Tumour medicine, anti-cancer food or health care product can be used as the synergist of anti-tumor drug, have good medical value.
Detailed description of the invention
Fig. 1 is that the tumor weight of each experimental group in experimental example 1 compares figure;
Fig. 2 is that the tumor weight of each experimental group in experimental example 2 compares figure.
Specific embodiment
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited In this.
Unless otherwise indicated, reagent and instrument involved in following embodiment can be bought from conventional on the market.
Embodiment 1
The preparation method of plain boiled pork ganoderma lucidum polysaccharide, comprising the following steps:
1) it extracts: plain boiled pork ganoderma lucidum fruitbody being smashed, is soaked in water half an hour according to the ratio of solid-liquid ratio 1:10~20, and It is heated at 80~100 DEG C, three times, each reflux extracting time is respectively 2h, 2h, 1h to refluxing extraction, and merging filtrate will Filtrate is concentrated into 1/10th of stoste, obtains concentrate;
2) alcohol precipitation: being sink to final concentration of the 75% of alcoholic solvent for concentrate progress alcohol, stand overnight at a temperature of 4 DEG C, it 10min is centrifuged with the revolving speed of 6000rpm afterwards, is precipitated;
3) it is lyophilized: being dissolved and precipitated with pure water, freeze-drying obtains Thick many candies, and kept dry is spare;
4) it isolates and purifies: the plain boiled pork ganoderma lucidum crude polysaccharide of acquisition is dissolved in water, stepwise elution is carried out by ion exchange column, then Plain boiled pork ganoderma lucidum polysaccharide is obtained after secondary freeze-drying.
Experimental example 1
1. reagent and instrument
Cell strain: breast cancer mouse cell strain (4T1);
Mouse: BALB/c mouse.
2. cell culture
Mouse breast cancer is carried out with the DMEM culture solution containing 1% penicillin, 1% streptomysin and 10% fetal calf serum (FBS) 4T1 culture is placed in the middle culture of Thermo Forma3110 incubator for tissue culture (originating from German Thermo company) of 37 DEG C, 5%CO2 To logarithmic growth phase.
3. tumor model zoopery
3.1 is subcutaneous to BABL/C mouse by logarithmic growth phase tumor cell inoculation, and is grouped into model group at random, taxol Group, plain boiled pork ganoderma lucidum polysaccharide group, taxol+plain boiled pork ganoderma lucidum polysaccharide group;
3.2 second days, start that each group is administered respectively, administration mode is intraperitoneal injection, dosage 50mg/kg/day, mould Type group be injecting normal saline, successive administration 4 weeks;
Second day after 3.3 last time administrations, mouse was sacrificed in a manner of carbon dioxide, takes tumour, measures tumour weight Amount and size;
It is antitumor to inhibition mice tumors grew and auxiliary synergy taxol that the plain boiled pork ganoderma lucidum polysaccharide is analyzed in 3.4 calculating It influences, determines optimal dose.
4. as the result is shown
Table 1
Group Tumor weight (g) Inhibiting rate (%)
Model group 0.255±0.13 0
Taxol group 0.178±0.08 30.2±3.0
Polysaccharide group 0.195±0.1 23.5±4.0
Note: inhibition rate of tumor growth calculates compared with model group.
Experimental example 2
1. reagent and instrument
Cell strain: breast cancer mouse cell strain (4T1);
Mouse: BALB/c mouse.
2. cell culture
Mouse breast cancer is carried out with the DMEM culture solution containing 1% penicillin, 1% streptomysin and 10% fetal calf serum (FBS) 4T1 culture, is placed in 37 DEG C, 5%CO2Thermo Forma3110 incubator for tissue culture (originating from German Thermo company) in culture To logarithmic growth phase.
3. tumor model zoopery
3.1 is subcutaneous to BABL/C mouse by logarithmic growth phase tumor cell inoculation, and is grouped into model group at random, taxol Group, plain boiled pork ganoderma lucidum polysaccharide group, taxol+plain boiled pork ganoderma lucidum polysaccharide group;
3.2 second days, start that each group is administered respectively, administration mode is intraperitoneal injection, dosage 100mg/kg/day, mould Type group be injecting normal saline, successive administration 4 weeks;
Second day after 3.3 last time administrations, mouse was sacrificed in a manner of carbon dioxide, takes tumour, measures tumour weight Amount and size;
It is antitumor to inhibition mice tumors grew and auxiliary synergy taxol that the plain boiled pork ganoderma lucidum polysaccharide is analyzed in 3.4 calculating It influences, determines optimal dose.
4. as the result is shown
Table 2
Group Tumor weight (g) Inhibiting rate (%) Increasing Efficiency (%)
Model group 0.143±0.06 0
Taxol group 0.103±0.07 27.8±7
Polysaccharide group 0.067±0.05 52.9±15
Polysaccharide+taxol group 0.034±0.01** 75.6±21 66±14*
Note: inhibition rate of tumor growth calculates compared with model group;
Increasing Efficiency is polysaccharide+taxol group compared with taxol group.
According to Fig. 1,2 and table 1,2, it can be found that: plain boiled pork ganoderma lucidum polysaccharide has the function of inhibition mouse interior tumor growth, And the effect that antitumor drug paclitaxel inhibits tumour growth can be significantly increased, is the ganoderma lucidum product with potentiality to be exploited Kind.
The above, preferable specific embodiment only of the invention, but scope of protection of the present invention is not limited thereto, Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its Design is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.

Claims (10)

1. purposes of the plain boiled pork ganoderma lucidum polysaccharide as active constituent in the preparation of antitumor drugs.
2. the preparation method of plain boiled pork ganoderma lucidum polysaccharide, which comprises the steps of:
(1) it extracts: plain boiled pork ganoderma lucidum fruitbody being smashed, is soaked in water, and is heated, refluxing extraction is multiple, merging filtrate, will Filtrate concentration, obtains concentrate;
(2) alcohol precipitation: concentrate is subjected to alcohol precipitation, stands overnight, is centrifuged later, precipitated;
(3) it is lyophilized: being dissolved and precipitated with pure water, freeze-drying obtains Thick many candies, and kept dry is spare;
(4) it isolates and purifies: the plain boiled pork ganoderma lucidum crude polysaccharide of acquisition is dissolved in water, stepwise elution is carried out by ion exchange column, again Plain boiled pork ganoderma lucidum polysaccharide is obtained after freeze-drying.
3. the preparation method of plain boiled pork ganoderma lucidum polysaccharide according to claim 2, which is characterized in that the preparation method is further Include the following steps:
(1) it extracts: plain boiled pork ganoderma lucidum fruitbody being smashed, is soaked in water half an hour according to the ratio of solid-liquid ratio 1:10~20, and Heated at 80~100 DEG C, refluxing extraction three times, merging filtrate, concentrate the filtrate to stoste 10 to ten/7th/ One, obtain concentrate;
(2) alcohol precipitation: concentrate progress alcohol is sink to final concentration of the 40~80% of alcoholic solvent, stands overnight, is centrifuged later, obtain Precipitating;
(3) it is lyophilized: being dissolved and precipitated with pure water, freeze-drying obtains Thick many candies, and kept dry is spare;
(4) it isolates and purifies: the plain boiled pork ganoderma lucidum crude polysaccharide of acquisition is dissolved in water, stepwise elution is carried out by ion exchange column, again Plain boiled pork ganoderma lucidum polysaccharide is obtained after freeze-drying.
4. the preparation method of plain boiled pork ganoderma lucidum polysaccharide according to claim 3, which is characterized in that step flows back three times in (1) The time of extraction is respectively 2h, 2h, 1h.
5. the preparation method of plain boiled pork ganoderma lucidum polysaccharide according to claim 3, which is characterized in that in step (2) after alcohol precipitation Concentrate is stood overnight at a temperature of 4 DEG C, and/or is centrifuged 10~30min with the revolving speed of 4000~8000rpm;And/or step (2) alcohol is sink to final concentration of the 75% of alcoholic solvent.
6. the preparation method of plain boiled pork ganoderma lucidum polysaccharide according to claim 5, which is characterized in that in step (2) after alcohol precipitation Concentrate is centrifuged 10min with the revolving speed of 6000rpm.
7. the preparation method of plain boiled pork ganoderma lucidum polysaccharide according to claim 3, which is characterized in that the plain boiled pork ganoderma lucidum is plain boiled pork Ganoderma lucidum Ganoderma leucocontextum HMGIM-Z110122, deposit number are CCTCC NO:M2016087.
8. a kind of plain boiled pork ganoderma lucidum polysaccharide, wherein plain boiled pork ganoderma lucidum polysaccharide method according to claim 2 to 7 any one It is prepared.
9. a kind of antineoplastic pharmaceutical compositions, wherein the antineoplastic pharmaceutical compositions contain described in a effective amount of claim 8 Plain boiled pork ganoderma lucidum polysaccharide.
10. antineoplastic pharmaceutical compositions according to claim 9, which is characterized in that the antineoplastic pharmaceutical compositions into One step includes one or more pharmaceutically acceptable auxiliary materials.
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CN112094358A (en) * 2020-09-22 2020-12-18 广东省微生物研究所(广东省微生物分析检测中心) Tibetan ganoderma lucidum polysaccharide GLP-1 with antioxidant effect, and preparation method and application thereof
CN112569336A (en) * 2020-12-16 2021-03-30 广东省微生物研究所(广东省微生物分析检测中心) Application of grifola frondosa glycoprotein in preparation of drugs or health foods for enhancing body immunity and preventing and/or treating tumor diseases
CN113122458A (en) * 2020-06-11 2021-07-16 广东省科学院微生物研究所(广东省微生物分析检测中心) Ganoderma leucocontextum Z160097 and cultivation method and application thereof
CN113278535A (en) * 2021-06-30 2021-08-20 四川省中医药科学院 New ganoderma lucidum strain ZL167 and new application thereof
CN114766285A (en) * 2022-04-24 2022-07-22 云南省农业科学院生物技术与种质资源研究所 Ganoderma leucocontextum strain L4495 and cultivation method and application thereof
WO2023040033A1 (en) * 2021-09-18 2023-03-23 广东粤微食用菌技术有限公司 Anti-oxidant repair ganoderma lucidum extract and preparation method therefor
CN115886242A (en) * 2022-11-23 2023-04-04 河北岁神本草肉灵芝生物科技有限公司 Ganoderma lucidum, high-concentration oral liquid containing ganoderma lucidum and preparation method of oral liquid

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CN113122458A (en) * 2020-06-11 2021-07-16 广东省科学院微生物研究所(广东省微生物分析检测中心) Ganoderma leucocontextum Z160097 and cultivation method and application thereof
CN112094358A (en) * 2020-09-22 2020-12-18 广东省微生物研究所(广东省微生物分析检测中心) Tibetan ganoderma lucidum polysaccharide GLP-1 with antioxidant effect, and preparation method and application thereof
WO2022062380A1 (en) * 2020-09-22 2022-03-31 广东省微生物研究所(广东省微生物分析检测中心) Tibetan ganoderma lucidum polysaccharide glp-1 with antioxidant effect, preparation method and use
CN112094358B (en) * 2020-09-22 2022-05-27 广东省微生物研究所(广东省微生物分析检测中心) Tibetan ganoderma lucidum polysaccharide GLP-1 with antioxidant effect, and preparation method and application thereof
CN112569336A (en) * 2020-12-16 2021-03-30 广东省微生物研究所(广东省微生物分析检测中心) Application of grifola frondosa glycoprotein in preparation of drugs or health foods for enhancing body immunity and preventing and/or treating tumor diseases
CN112569336B (en) * 2020-12-16 2023-08-11 广东省微生物研究所(广东省微生物分析检测中心) Application of russula cinerea glycoprotein in preparation of medicines or health foods for enhancing organism immunity and preventing and/or treating tumor diseases
CN113278535A (en) * 2021-06-30 2021-08-20 四川省中医药科学院 New ganoderma lucidum strain ZL167 and new application thereof
WO2023040033A1 (en) * 2021-09-18 2023-03-23 广东粤微食用菌技术有限公司 Anti-oxidant repair ganoderma lucidum extract and preparation method therefor
CN114766285A (en) * 2022-04-24 2022-07-22 云南省农业科学院生物技术与种质资源研究所 Ganoderma leucocontextum strain L4495 and cultivation method and application thereof
CN114766285B (en) * 2022-04-24 2023-10-27 云南省农业科学院生物技术与种质资源研究所 Ganoderma lucidum strain L4495 and cultivation method and application thereof
CN115886242A (en) * 2022-11-23 2023-04-04 河北岁神本草肉灵芝生物科技有限公司 Ganoderma lucidum, high-concentration oral liquid containing ganoderma lucidum and preparation method of oral liquid

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