CN103784738B - A kind of vegetable composition extracting method containing Lepidinm meyenii Walp and application - Google Patents
A kind of vegetable composition extracting method containing Lepidinm meyenii Walp and application Download PDFInfo
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- CN103784738B CN103784738B CN201410084330.3A CN201410084330A CN103784738B CN 103784738 B CN103784738 B CN 103784738B CN 201410084330 A CN201410084330 A CN 201410084330A CN 103784738 B CN103784738 B CN 103784738B
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- meyenii walp
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- 239000000203 mixture Substances 0.000 title claims abstract description 29
- 235000013311 vegetables Nutrition 0.000 title claims abstract description 29
- 238000000034 method Methods 0.000 title abstract description 13
- 238000002360 preparation method Methods 0.000 claims abstract description 18
- 239000003814 drug Substances 0.000 claims abstract description 14
- 208000032612 Glial tumor Diseases 0.000 claims abstract description 13
- 206010018338 Glioma Diseases 0.000 claims abstract description 13
- 230000004663 cell proliferation Effects 0.000 claims abstract description 9
- 239000000284 extract Substances 0.000 claims abstract description 8
- 229940079593 drug Drugs 0.000 claims abstract description 7
- 230000001629 suppression Effects 0.000 claims abstract description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 45
- 238000000605 extraction Methods 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 10
- 239000002671 adjuvant Substances 0.000 claims description 7
- 230000000274 adsorptive effect Effects 0.000 claims description 7
- 230000006837 decompression Effects 0.000 claims description 7
- 239000003480 eluent Substances 0.000 claims description 7
- 238000010828 elution Methods 0.000 claims description 7
- 238000000874 microwave-assisted extraction Methods 0.000 claims description 7
- 238000004064 recycling Methods 0.000 claims description 7
- 229920005989 resin Polymers 0.000 claims description 7
- 239000011347 resin Substances 0.000 claims description 7
- 239000008187 granular material Substances 0.000 claims description 6
- 239000002775 capsule Substances 0.000 claims description 5
- 239000003826 tablet Substances 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 4
- 229960004756 ethanol Drugs 0.000 description 15
- 238000002474 experimental method Methods 0.000 description 8
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 239000012531 culture fluid Substances 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
- QZKDXSOKZHLFHE-UHFFFAOYSA-N 4-(1,3-benzodioxol-5-yl)benzoic acid Chemical compound C1=CC(C(=O)O)=CC=C1C1=CC=C(OCO2)C2=C1 QZKDXSOKZHLFHE-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 240000000759 Lepidium meyenii Species 0.000 description 1
- 235000000421 Lepidium meyenii Nutrition 0.000 description 1
- 229920002334 Spandex Polymers 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000010165 autogamy Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012738 dissolution medium Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
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- 239000012535 impurity Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000004900 laundering Methods 0.000 description 1
- 235000012902 lepidium meyenii Nutrition 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
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- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 239000004759 spandex Substances 0.000 description 1
- 229960002385 streptomycin sulfate Drugs 0.000 description 1
- 238000009495 sugar coating Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Abstract
The invention provides a kind of vegetable composition extracting method containing Lepidinm meyenii Walp, by Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g makes as crude drug, adopts microwave extract method to be prepared from, content is improved a lot, consumption reduces, and present invention also offers the application of vegetable composition in preparation suppression rat glioma C6 cell proliferation containing Lepidinm meyenii Walp.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of vegetable composition extracting method containing Lepidinm meyenii Walp and application.
Background technology
Containing the vegetable composition of Lepidinm meyenii Walp by Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g makes as crude drug, and current extracting method decocts with water secondary, each 2-3 hour, collecting decoction, filters, is condensed into thick paste, granulates, drying, is pressed into 1000, sugar coating, obtains final product, can resisting fatigue.
In prior art, not yet there is the vegetable composition containing Lepidinm meyenii Walp extracting the report adopting microwave technology in preparation, and adopt the method for soak by water, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied clinically.
Summary of the invention
Goal of the invention: in order to solve the problem, the object of the present invention is to provide a kind of vegetable composition extracting method containing Lepidinm meyenii Walp.
Another object of the present invention is to provide a kind of application of vegetable composition in preparation suppression rat glioma C6 cell proliferation containing Lepidinm meyenii Walp.
Technical scheme: the object of the invention is by following scheme realize:
A kind of vegetable composition extracting method containing Lepidinm meyenii Walp, vegetable composition is by Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
Above-mentioned a kind of vegetable composition extracting method containing Lepidinm meyenii Walp, in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
A kind of vegetable composition containing Lepidinm meyenii Walp suppresses the application in rat glioma C6 cell proliferation in preparation, containing the vegetable composition of Lepidinm meyenii Walp by Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
Above-mentioned a kind of vegetable composition containing Lepidinm meyenii Walp suppresses the application in rat glioma C6 cell proliferation in preparation, the power of microwave extracting described in preparation method is 500W, each extraction 6 minutes.
In prior art, the every sheet 0.25g of vegetable composition containing Lepidinm meyenii Walp, each 4,3 times on the one, adopt the every sheet 0.25g of vegetable composition containing Lepidinm meyenii Walp that the present invention is prepared into, but the medical material amount contained is original 2 times, therefore only needs 2 at every turn, within 1st, take 3 times, under the condition with more active component, greatly reduce dose.
Detailed description of the invention
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, every sheet 0.25g.
Embodiment 2
Get Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collects 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into capsule.
Embodiment 3
Get Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extracts 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collects 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into granule.
In the present invention, Lepidinm meyenii Walp is the under ground portion root of crucifer Lepidium meyenii, and all the other are pharmacopeia kind.
Embodiment 4: the vegetable composition containing Lepidinm meyenii Walp suppresses the experimentation data of rat glioma C6 cell proliferation
1 experiment material
1.1 experiment cell strains
Rat glioma C6 cell, Nanjing Medical University's Experimental Animal Center laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: the present invention is containing the vegetable composition of Lepidinm meyenii Walp: prepare by embodiment 1 method.
Medicinal liquid liquid storage: take 100mg containing the vegetable composition of Lepidinm meyenii Walp, be dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ ldoff pipe subpackages ,-20 DEG C of storage, 0.2 μm of frit dehydrated alcohol prepares against the use of matched group simultaneously.
1.3 experiment reagent
DMEM (GIBCO company Cat.No.12100-061Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3 (Shanghai hundred million chemical reagent company limited Cat.No.11810-033Lot.No.1088387 of a specified duration); Trypsin (AMRESCO company lot number: 2010/04); EDTA (AMRESCO company lot number: 2009/10); PenicillinGSodiumSalt (AMRESCO company lot number: 2010242); StreptomycinSulfate (AMRESCO company lot number: 2010382); Dehydrated alcohol (Nanjing Chemistry Reagent Co., Ltd.'s lot number: 080310182); MTT (Biosharp lot number: 0793); PBS (laboratory autogamy);
1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRAMAX190); CO2 incubator (FORMA model: 3111); Super-clean bench (safe and sound Inc. of Su Jing group moulding number: SW-CJ-ZFD); Pure water instrument (Spring company of U.S. model: S/N020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (the accurate experimental facilities company in Shanghai model: DHG9123A); Refrigerator (Siemens Company's model: KG18V21TI); Liquid nitrogen container (CBS model: 2001); Low speed centrifuge (Anting Scientific Instrument Factory, Shanghai's model: KA-1000); 0.2 μm of filter (MILLIPORE model: SLGP033RB); 10cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2 experimental techniques
1) rat glioma C6 cell DMEM+10%FBS in 37 DEG C, 5%CO2 carries out cellar culture (10cm culture dish), when Growth of Cells is to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml0.25% trypsin-0.04%EDTA, after 37 DEG C of digestion 2min, add 5ml complete medium neutralization reaction wherein, proceeded in centrifuge tube after piping and druming cell, the centrifugal 5min of 1000rpm, adjustment concentration of cell suspension 3 × 10
4individual/ml.
2) enter in 96 well culture plates by cell kind, every hole adds cell suspension 180 μ l, and culture plate puts into cell culture incubator (37 DEG C, 5%CO2) cellar culture.
3) according to cell growth status, generally grow to 50%-70%, add the vegetable composition solution containing Lepidinm meyenii Walp, continue to cultivate 24h.
4) add 20 μ lMTT solution (5mg/ml, i.e. 0.5%MTT) after 24h, continue to cultivate 4h.
5) after 4h, buckle method removes supernatant, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, crystal is fully dissolved.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm place.
6) background (do not add cell, only add culture fluid) is set, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide) simultaneously, often organizes the multiple hole of setting 6.
7) result represents with the suppression ratio of medicine to cell:
Cell proliferation suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%.Experiment repetition 3 times.
3 statistical dispositions
Adopt the correlation analysis in MicrosoftExcel2003 software and Studentt inspection, data represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to rat glioma C6 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has pole significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 contains the vegetable composition of Lepidinm meyenii Walp to rat glioma C6 cell inhibitory effect influence research
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
Vegetable composition containing Lepidinm meyenii Walp can suppress rat glioma C6 cell proliferation, and reduce the Growth of Cells number of rat glioma C6 cell, this effect is dose dependent.
Claims (4)
1. the vegetable composition preparation method containing Lepidinm meyenii Walp, it is characterized in that vegetable composition is by Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
2. a kind of vegetable composition preparation method containing Lepidinm meyenii Walp according to claim 1, is characterized in that in preparation method, microwave extracting power is 500W, each extraction 6 minutes.
3. the application of vegetable composition in preparation suppression rat glioma C6 cell proliferation containing Lepidinm meyenii Walp, it is characterized in that the vegetable composition containing Lepidinm meyenii Walp is by Lepidinm meyenii Walp 100g, Rhizoma Polygonati 80g, Radix Angelicae Sinensis 60g, Radix Salviae Miltiorrhizae 50g makes as crude drug, preparation method is made up of the following step: get above-mentioned raw materials medicine, pulverize, add 70% ethanol of 2L, drop in microwave extracting apparatus and carry out microwave extracting, extraction power is 400-600W, extract 2 times, each 4-8 minute, combining extraction liquid, concentrated, be added on D101 macroporous adsorptive resins, 50% ethanol elution, collect 5 times amount column volume eluents, decompression recycling ethanol, concentrated and dry, obtain microwave extraction thing, add customary adjuvant and be prepared into tablet, capsule or granule.
4. a kind of vegetable composition containing Lepidinm meyenii Walp suppresses the application in rat glioma C6 cell proliferation in preparation according to claim 3, it is characterized in that the power of microwave extracting described in preparation method is 500W, each extraction 6 minutes.
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| CN104920762A (en) * | 2015-06-30 | 2015-09-23 | 福建德广中草药有限公司 | Huangjing Maka tablet sugar and production method thereof |
| CN105193967A (en) * | 2015-09-12 | 2015-12-30 | 南京正亮医药科技有限公司 | Method for extracting plant composition containing cimicifugae foetidae and application |
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| CN105106331A (en) * | 2015-10-02 | 2015-12-02 | 陈爱华 | Extraction method and application of plant composition containing oenothera biennis |
| CN105125989A (en) * | 2015-10-02 | 2015-12-09 | 陈爱华 | Extracting method and application of plant composite containing hawthorns |
| CN105125943A (en) * | 2015-10-03 | 2015-12-09 | 陈爱华 | Extracting method for plant composition containing American ginseng and application |
| CN105106264A (en) * | 2015-10-03 | 2015-12-02 | 陈爱华 | Extraction method and application of plant composition containing shiitake |
| CN105147869A (en) * | 2015-10-03 | 2015-12-16 | 陈爱华 | Extracting method and application of plant composition containing green tea |
| CN105581333A (en) * | 2015-12-30 | 2016-05-18 | 重庆骄王天然产物股份有限公司 | Fully-nutritional Maca powder and preparation method thereof |
| CN105582132A (en) * | 2016-03-07 | 2016-05-18 | 南京多宝生物科技有限公司 | Preparation method and application of liquorice root formulation granules |
| CN106108008A (en) * | 2016-06-24 | 2016-11-16 | 丽江师范高等专科学校 | A kind of extracting method of effective component in Maka |
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| CN102885302A (en) * | 2012-09-29 | 2013-01-23 | 楚雄州百草岭药业发展有限公司 | Maca composition and preparation method thereof and preparation |
| CN103040896A (en) * | 2013-01-06 | 2013-04-17 | 云南圣草峰生物科技有限公司 | Preparation method of maca extractive |
| CN103566046A (en) * | 2013-11-25 | 2014-02-12 | 楚雄三平裕康商贸有限公司 | Traditional Chinese medicine composition containing maca, traditional Chinese medicine preparation, and preparation method of traditional Chinese medicine preparation |
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| CN102885302A (en) * | 2012-09-29 | 2013-01-23 | 楚雄州百草岭药业发展有限公司 | Maca composition and preparation method thereof and preparation |
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Address after: 100085 Beijing city Haidian District four Station Road West. Cottage 12-22 No. 17 room 310 Patentee after: BEIJING FORRIDA BIOLOGICAL HEALTH TECHNOLOGY CO.,LTD. Address before: 100085 Beijing city Haidian District four Station Road West. Cottage 12-22 No. 17 room 310 Patentee before: BEIJING FORRIDA TECHNOLOGY DEVELOPMENT CO.,LTD. |
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Denomination of invention: Extraction method and application of plant composition containing Maca Effective date of registration: 20161220 Granted publication date: 20160413 Pledgee: Haidian Beijing science and technology enterprise financing Company limited by guarantee Pledgor: BEIJING FORRIDA BIOLOGICAL HEALTH TECHNOLOGY CO.,LTD. Registration number: 2016990001136 |
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Date of cancellation: 20181026 Granted publication date: 20160413 Pledgee: Haidian Beijing science and technology enterprise financing Company limited by guarantee Pledgor: BEIJING FORRIDA BIOLOGICAL HEALTH TECHNOLOGY CO.,LTD. Registration number: 2016990001136 |
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