CN102885726A - Eucommia extract product, its preparation method and application thereof - Google Patents
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Abstract
The invention discloses a preparation method of an eucommia extract product, comprising the following steps of: (1) extraction of a solvent, wherein the solvent is selected from water, methanol, ethanol, isopropanol, 1-butanol, glycol, 1,2-propylene glycol, 1,3-propylene glycol, 1,3-butylene glycol, acetone, ethyl acetate or their combination; (2) separation of an extract and residue so as to obtain the extract; (3) recovery of the optional solvent; and (4) optional dissolution or dilution. The eucommia extract product obtained by the method has antioxidation and UVA and UVB irradiation resistant effects, and can be used for ageing resistance of skin, especially light ageing resistance. The eucommia extract product can be used as a cosmetic additive or medicinal active component. The invention also discloses an application of the eucommia extract product in preparation of cosmetics or medicines for skin ageing resistance, especially for light ageing resistance.
Description
Technical field
The present invention relates to Natural Medicine Chemistry field and cosmetic field, be specifically related in cosmetics or the medicine a kind of for aging resistance, the Chinese herbal medicine Cortex Eucommiae of preferred anti-light aging.
Background technology
Studies show that, free radical and ultraviolet all are one of factors of skin aging, and photoaging mainly by the ultraviolet B radiation in the daylight (UVB, 290～320nm) and long wave ultraviolet (UVA, 320～400nm) cause jointly.Ultraviolet has obvious facilitation to application on human skin aging, and high exposed population group's skin aging is dangerous to double than low exposed population group, and the time of origin that wears out shifts to an earlier date 10 years.Long term exposure causes skin photoage in the daylight ultraviolet, and can cause serious cosmetic problem, thereby in recent years extremely industry concern, the product of aging resistance and anti-light aging comes out one after another, correlational study is like a raging fire, but natural Chinese medicinal herb anti-light aging additive is still vacancy.
Summary of the invention
The object of the invention is to provide a kind of preparation method of Cortex Eucommiae extract, and described method comprises:
(1) solvent extraction, described solvent are selected from water, methanol, ethanol, isopropyl alcohol, n-butyl alcohol, ethylene glycol, 1,2-PD, 1,3-PD, 1,3 butylene glycol, acetone, ethyl acetate or its combination;
(2) separation and Extraction liquid and residue obtain extracting solution;
(3) optional solvent recovery;
(4) optionally dissolving or dilution.
Preferably, wherein the extraction of step (1) is selected from immersion, decoction, backflow, percolation or its combination.
Preferably, wherein in the step (4) used solvent be selected from water, ethanol, isopropyl alcohol, n-butyl alcohol, ethylene glycol, 1,2-PD, 1,3-PD, 1,3 butylene glycol, acetone, ethyl acetate or its combination.
Another purpose of the present invention is to provide the preparation method of another kind of Cortex Eucommiae extract, and described method comprises:
(1) solvent extraction, described solvent are selected from methanol, ethanol, isopropyl alcohol, n-butyl alcohol, ethylene glycol, 1,2-PD, 1,3-PD, 1,3 butylene glycol, acetone, ethyl acetate or its combination;
(2) separation and Extraction liquid and residue obtain residue;
(3) residue of water extraction step (2).
(4) Separation of Water extracting solution and residue obtain aqueous extract;
(5) concentrated, the ethanol precipitate with ethanol separates obtaining precipitation;
(6) alternatively, the resolution of precipitate with step (5) is mixed with solution.
Preferably, wherein the extraction of step (1) and (3) is independently selected from immersion, decoction, backflow, percolation or its combination.
Preferably, wherein in the step (6) used solvent be selected from ethanol, isopropyl alcohol, n-butyl alcohol,, ethylene glycol, 1,2-PD, 1,3-PD, 1,3 butylene glycol, acetone, ethyl acetate or its combination.
Another purpose of the present invention provides the Cortex Eucommiae extract according to the preparation method acquisition of the invention described above.
Preferably, the form of described Cortex Eucommiae extract is selected from Emulsion, solution, powder, ointment, membrane or cream frost.
The Cortex Eucommiae extract that the method according to this invention obtains can be directly used in the prevention skin aging, and preferred light is aging, also can be used as functional additive or active component and joins in cosmetics or the medicine and use.
Further object of the present invention provides Cortex Eucommiae extract in preparation aging resistance, the cosmetics of preferred anti-light aging or the purposes in the medicine.
Preferably, described Cortex Eucommiae extract is the Cortex Eucommiae extract of above-mentioned prepared in accordance with the method for the present invention.
Further object of the present invention provides a kind of aging resistance, and cosmetics or the medicine of preferred anti-light aging wherein contain Cortex Eucommiae extract.
Preferably, described Cortex Eucommiae extract is the Cortex Eucommiae extract of above-mentioned prepared in accordance with the method for the present invention.
In the present invention, the Cortex Eucommiae refers to the bark of Eucommiaceae plant Cortex Eucommiae Eucommia ulmoides oliv.Cortex Eucommiae main product in Sichuan, Shaanxi, Hubei, Henan, Guizhou, Yunnan.Between Clear and Bright to the Summer Solstice, choose the above plant of growth 15-20, press medical material specification size, peel bark, the rough bark of shooting off dries, and puts aeration-drying place.
The Cortex Eucommiae, Gan Weixin, temperature enters the Liver and kidney warp, invigorating the liver and kidney, bone and muscle strengthening is prolonged life and is preserved youthful looks.The Cortex Eucommiae contains organic compound and the inorganic elementss such as calcium, ferrum such as lignanoids, cyclenes ether mushrooms, organic acid, mushrooms, polysaccharide, aminoacid and gutta-percha." not Lu " " ache in the main foot, do not want to trample ground ", Shennong's Herbal " make light of one's life by commiting suicide for a long time for invigorating middle warmer benefit vital essence, hard muscles and bones by clothes." modern pharmacological research, the Cortex Eucommiae has delaying senility function, can anti peroxidation of lipid, and by the effect of two-ways regulation immunologic function.
The Cortex Eucommiae extract that method of the present invention makes has the effect of the free radical of removing antioxidative; simultaneously can help skin to resist the ultraviolet radiation of UVB and UVA; protection skin; reduce cell injury; prevent the minimizing of dermal collagen; effectively prevention skin aging, especially photoaging, and to the skin nonirritant.
The specific embodiment
Embodiment 1
Get Cortex Eucommiae Eucommia ulmoides 100g, adding 8 times of amounts (doubly measures: the ratio that refers to solvent volume and Cortex Eucommiae weight, wherein the unit of solvent volume is ml, the unit of Cortex Eucommiae weight is g, lower with) 75% ethanol (v/v) merceration 48 hours, filter, filtrate recycling ethanol, the extract obtained water dissolution of using is made and is contained the solution that the crude drug amount is 1000mg/ml, obtains extract A.
Embodiment 2
Get Cortex Eucommiae Eucommia ulmoides 100g, add 8 times of amount methanol eddies and extracted 2 hours, filter, filtrate is reclaimed methanol, and the extract obtained water dissolution of using is made and contained the solution that the crude drug amount is 500mg/ml, obtains extract B.
Embodiment 3
Get Cortex Eucommiae Eucommia ulmoides 100g, decoct with 10 times of water gagings and extract approximately 2 hours, concentrated, filter, supernatant adds water to be made and contains the solution that the crude drug amount is 1000mg/ml, obtains extract C.
Embodiment 4
Get Cortex Eucommiae Eucommia ulmoides 100g, add 6 times of amount alcohol reflux, filter, residue decocts with 10 times of water gagings and extracts approximately 2 hours, concentrated, add 2 times of amount ethanol precipitations, filter, precipitation is used water dissolution, makes to contain the solution that the crude drug amount is 500mg/ml, obtains extract D.
Embodiment 5:DPPH experiment
Free radical is the chemical substance that a class character is active, have extremely strong oxidability.The irradiation of ultraviolet (UV) A/UVB can produce a large amount of free radicals, to the body injury, accelerates old and feeble.Adopt DPPH free radical scavenging method to measure the antioxidation activity in vitro of extract among the embodiment at this.
With distilled water the extract among the embodiment half-and-half is diluted to 0.625%v/v since 5%) totally 4 concentration.Then get embodiment extract 2ml, concentration is DPPH alcoholic solution 0.5ml and the 1.5ml water of 500 μ mol/L, successively add in the same test tube, shake up, leave standstill, measure its absorbance (OD) behind the 30min at the 517nm place, and with corresponding mixed liquor as blank.The embodiment extract calculates according to following formula the clearance rate of DPPH:
Wherein:
The absorbance of T-DPPH+ embodiment extract,
The absorbance of T0-embodiment extract+solvent,
The absorbance of C-DPPH+ solvent.
Experimental result sees Table 1.
Table 1, the assessment of Cortex Eucommiae extract oxidation resistance
Experimental result shows, embodiment 1 extract A, embodiment 2 extract B, embodiment 23 get thing C, embodiment 4 extract D all have better removing free radical under external four concentration oxidation resistance, and be wherein best with embodiment 4 extract D effects.
Embodiment 6: the fibroblast proliferation test
Fibroblast in the skin is the main cell of synthetic collagen protein, and the enhancing of collagen protein ability is synthesized and secreted to fibroblastic a large amount of propagation or fibroblast, and the collagen protein total amount is increased, thus the aging that slows down skin.Therefore, to facilitate fibrocellular proliferation function be one of important method of assess sample activity of fighting against senium in research.The mechanism of the above skin aging of this experimental evidence adopts the fibroblastic method of vitro culture of human, comes the senile-resistant efficacy of the Cortex Eucommiae extract among preliminary assessment the present invention.
(being mixed with concentration is 5*10 with the cell suspension for preparing
3Individual/as ml) to be inoculated in 96 orifice plates, behind cell attachment, except the blank group, other every holes add the Chinese herbal medicine extract of the embodiment of 20ul respective concentration, establish 4 concentration groups, every group of 6 holes, put incubator (37 ℃, 5%CO
2) in cultivate 24h, then add people MTT liquid 20ul, continue incubator (37 ℃, 5%CO
2) the middle 4h that cultivates.After taking out ELISA Plate, carefully suck the culture fluid in every hole, add DMSO, mix homogeneously.Measure optical density (570nm) with enzyme-linked immunosorbent assay instrument, by with the relative rate of increase that relatively obtains sample sets of matched group.
Cortex Eucommiae extract is as shown in table 2 to fibroblastic proliferation activity result.
Table 2, Cortex Eucommiae extract are to fibroblastic proliferation activity
Experimental result shows, embodiment 1 extract A, embodiment 2 extract B, embodiment 3 extract C, embodiment 4 extract D all can promote fibroblastic propagation, effect with embodiment 3 extract C is best, and its 0.001% concentration facilitates fibrocyte propagation to reach 11.91%.
Embodiment 7:UV irradiation is to the protective effect of fibroblast damage
Skin photoage mainly by the ultraviolet B radiation in the daylight (UVB, 290～320nm) and long wave ultraviolet (UVA, 320～400nm) cause jointly, ultraviolet has obvious facilitation to application on human skin aging.The UVA/UVB irradiation can cause that fibroblastic damage is dead, and skin is damaged, and causes photoaging.Investigate the Chinese herbal medicine extract of embodiment to the photoaging effect of resisting of the cytoprotective of UV irradiation at this by UV irradiation experiment.
Get respectively extract 0.5ml among the embodiment, add 4.5ml PBS and dilute, be made into 1/10 concentration (v/v) and carry out sucking filtration, then get 0.1ml and add the aseptic PBS of 9.9ml and be diluted at last 1/1000 concentration (v/v).The skin flbroblast (Fb) of cultivating take normal DMEM liquid is as matched group, usefulness UVA (5-20J/cm after cultivating with normal DMEM liquid
2)/UVB UVB (20-80J/cm
2) the Fb cell of irradiation is model group, take the embodiment extract that contains 1/1000 concentration and DMEM liquid is cultivated altogether and through the Fb cell of UVA/UVB irradiation as the dosing group.24h after the UV treatment measures the absorbance (OD value) in each hole with microplate reader by CCK-8, cell proliferation rate accounts for the percentage rate of negative control group OD value for each processed group OD value, and each tests triplicate.
Experimental result sees Table 3 and table 4.
Table 3, UVA irradiation
| Cell proliferation rate | The P value | |
| Embodiment 1 extract A | (90.13±4.38)％ | ＜0.01 |
| Embodiment 2 extract B | (90.25±4.20)％ | ＜0.01 |
| Embodiment 3 extract C | (90.46±3.64)％ | ＜0.01 |
| Embodiment 4 extract D | (99.77±6.59)％ | ＜0.01 |
| Matched group | (100±2.66)％ | |
| Model group | (78.25±3.98)％ | ＜0.01 |
Table 4, UVB irradiation
| Cell proliferation rate | The P value | |
| Embodiment 1 extract A | (97.88±3.53)％ | ＜0.01 |
| Embodiment 2 extract B | (98.06±2.55)％ | ＜0.01 |
| Embodiment 3 extract C | (98.19±5.58)％ | ＜0.01 |
| Embodiment 4 extract D | (99.33±4.60)％ | ＜0.01 |
| Matched group | (100±5.68)％ | |
| Model group | (83.63±5.15)％ | ＜0.01 |
Experimental result shows, embodiment 1 extract A, embodiment 2 extract B, embodiment 3 extract C, embodiment 4 extract D all can reduce the UV irradiation to fibroblastic damage, the cell appreciation rate is all returned to more than 90%, and best with the effect of embodiment 4 extract D.
Embodiment 8:UV irradiation is to dermal fibroblast MMP-1, the impact that MMP-3mRNA expresses and the protective effect of the Cortex Eucommiae
MMP-1 (interstitial collagenase) and MMP-3 (stromlysin) are two kinds of main enzymes of degraded hypodermal cell epimatrix, unusually the hypodermal cell epimatrix that will cause of their expressions is synthesized with catabolic unbalance, is the major reason that dermis is destroyed in the photoaging.
Dermal fibroblast is inoculated in the 60mm culture dish, and to contain the embodiment extract incubated cell of 1/1000 concentration, 24h after cell is accepted the UV irradiation carries out the extracting of cell total rna, and the reverse transcription of RNA according to the design primer, is carried out Real-time PCR.
Experimental result sees Table 5 and table 6.
Table 5
Compare with not irradiation group:
#P＜0.01, compare with the UVA group:
*P＜0.05,
*P＜0.01
Table 6
Compare with not irradiation group:
#P＜0.01, compare with the UVB group:
*P＜0.05,
*P＜0.01
Experimental result shows, embodiment 1 extract A, embodiment 2 extract B, embodiment 3 extract C, embodiment 4 extract D all can weaken the dermal fibroblast MMP-1 that the UV irradiation causes, the unconventionality expression of MMP-3mRNA, and best with the effect of embodiment 4 extract D.
Embodiment 9:UV shines dermal fibroblast I, the impact of III procollagen type protein expression level and the protective effect of the Cortex Eucommiae
The main pathological change of photoaging is at skin corium, and dermal fibroblast is the main cell of keeping corium normal configuration and function, and I, III Collagen Type VI are the main components that consists of the hypodermal cell epimatrix.Studies show that, photoaging skin collagen produces and reduces, and precollagen can significantly reduce behind the UV irradiation 24h.
Skin flbroblast is inoculated in the 35mm culture dish.With containing the embodiment extract incubated cell of 1/1000 concentration, 24h after the UV irradiation, the collecting cell culture supernatant adopts double-antibody sandwich ABC-ELISA method, with microplate reader at 450nm place survey light absorption value (OD value).On standard curve, read sample I and III procollagen type protein content (ng/ml) according to the OD value.The while collecting cell, in-20 ℃ of multigelation cells three times, 4 ℃ of centrifugal 15min of 14000g, getting supernatant, to measure cell protein quantitative, add standard substance or testing sample 20ul in 96 orifice plates, each hole adds the 200ulBCA working solution, places 20-30min for 37 ℃, survey light absorption value (OD value) at the 562nm place with microplate reader, calculate the protein content (mg/ml) of testing sample according to standard curve.I and III procollagen type protein expression level are corrected by cell protein content in the cell culture supernatant, represent with ng/mg.
Experimental result sees Table 7 and table 8.
Table 7
Compare with not irradiation group:
#P＜0.01, compare with the UVA1 group:
*P＜0.05,
*P＜0.01
Table 8
Compare with not irradiation group:
#P＜0.01, compare with the UVA1 group:
*P＜0.05,
*P＜0.01
Experimental result shows, the UV irradiation has significantly suppressed fibroblast and expressed I, III procollagen type albumen (P＜0.01).Irradiation adds each embodiment extract A, B, C and the D contain 1/1000 concentration simultaneously, and the content that can make I type and III procollagen type albumen in the cell culture supernatant increasing in various degree is best with the effect of embodiment 4 extract D.
Embodiment 10: the Emulsion one, two, three and four that extracts respectively Cortex Eucommiae extract A, the B, C and the D preparation that obtain with embodiment of the invention 1-4
Preparation method: after caprylic/capric triglyceride, ethylhexyl palmitate, spermol, behenyl alcohol polyethers-25, glyceryl stearate mixing, the heated and stirred dissolving evenly; With butanediol, 1,2-pentanediol, glycerol, xanthan gum, the dissolving of deionized water heated and stirred are evenly; With two parts mix homogeneously, then add Sepigel305 under the processing condition.Be cooled to 50 degree and add Liquid Germall Plus and Cortex Eucommiae extract of the present invention, stirring gets product.
Embodiment 11: the solution one, two, three and four that extracts respectively Cortex Eucommiae extract A, the B, C and the D preparation that obtain with embodiment of the invention 1-4
Preparation method: with water, butanediol, propylene glycol, 1,2-pentanediol, EDETATE SODIUM, hyaluronate sodium mix, heating, after stirring and dissolving is even, be cooled to 55 degree, add Liquid Germall Plus and Cortex Eucommiae extract of the present invention, stirring and dissolving is even, gets product.
According to the cosmetic formulations that the prescription of above embodiment 10 and 11 makes, has the effect of certain antioxidation and anti-light aging.
Skin photoage mainly by the ultraviolet B radiation in the daylight (UVB, 290～320nm) and long wave ultraviolet (UVA, 320～400nm) cause jointly, ultraviolet has obvious facilitation to application on human skin aging.The UVA/UVB irradiation can cause that fibroblastic damage is dead, and the minimizing of dermal collagen, and skin is damaged.The skin flbroblast that the present invention cultivates with the UVA/UVB irradiation of some strength; hatch with Cortex Eucommiae extract; can effectively reduce the UV irradiation to the impact of fibroblast proliferation; can effectively reduce simultaneously the variation of dermal fibroblast collagen protein; protection skin; illustrate that Cortex Eucommiae extract has the protective effect of ultra-vioket radiation damage to skin flbroblast, can defend photoaging.
Claims (12)
1. the preparation method of a Cortex Eucommiae extract comprises:
(1) solvent extraction, described solvent are selected from water, methanol, ethanol, isopropyl alcohol, n-butyl alcohol, ethylene glycol, 1,2-PD, 1,3-PD, 1,3 butylene glycol, acetone, ethyl acetate or its combination;
(2) separation and Extraction liquid and residue obtain extracting solution;
(3) optional solvent recovery;
(4) optionally dissolving or dilution.
2. preparation method as claimed in claim 1, wherein the extraction of step (1) is selected from immersion, decoction, backflow, percolation or its combination.
3. preparation method as claimed in claim 1, wherein used solvent is selected from water, ethanol, isopropyl alcohol, n-butyl alcohol, ethylene glycol, 1,2-PD, 1,3-PD, 1,3 butylene glycol, acetone, ethyl acetate or its combination in the step (4).
4. the preparation method of a Cortex Eucommiae extract comprises:
(1) solvent extraction, described solvent are selected from methanol, ethanol, isopropyl alcohol, n-butyl alcohol, ethylene glycol, 1,2-PD, 1,3-PD, 1,3 butylene glycol, acetone, ethyl acetate or its combination;
(2) separation and Extraction liquid and residue obtain residue;
(3) residue of water extraction step (2).
(4) Separation of Water extracting solution and residue obtain aqueous extract;
(5) concentrated, the ethanol precipitate with ethanol separates obtaining precipitation;
(6) alternatively, the resolution of precipitate with step (5) is mixed with solution.
5. preparation method as claimed in claim 4, wherein the extraction of step (1) and (3) is independently selected from immersion, decoction, backflow, percolation or its combination.
6. preparation method as claimed in claim 4, wherein used solvent is selected from ethanol, isopropyl alcohol, n-butyl alcohol, ethylene glycol, 1,2-PD, 1,3-PD, 1,3 butylene glycol, acetone, ethyl acetate or its combination in the step (6).
7. Cortex Eucommiae extract, it makes according to any one described preparation method of claim 1-6.
8. Cortex Eucommiae extract as claimed in claim 7, wherein, the form of described Cortex Eucommiae extract is selected from Emulsion, solution, powder, ointment, membrane or cream frost.
9. Cortex Eucommiae extract is in preparation aging resistance, the cosmetics of preferred anti-light aging or the purposes in the medicine.
10. purposes as claimed in claim 9, wherein said Cortex Eucommiae extract is claim 7 or 8 described Cortex Eucommiae extract.
11. an aging resistance, cosmetics or the medicine of preferred anti-light aging wherein contain Cortex Eucommiae extract.
12. cosmetics as claimed in claim 11 or medicine, wherein said Cortex Eucommiae extract are claim 7 or 8 described Cortex Eucommiae extract.
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| CN201110202162XA CN102885726A (en) | 2011-07-19 | 2011-07-19 | Eucommia extract product, its preparation method and application thereof |
| PCT/CN2012/078865 WO2013010489A1 (en) | 2011-07-19 | 2012-07-19 | Eucommia extract, preparation method therefor and use thereof |
| AU2012286357A AU2012286357A1 (en) | 2011-07-19 | 2012-07-19 | Eucommia extract, preparation method therefor and use thereof |
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| CN105616467A (en) * | 2015-12-23 | 2016-06-01 | 庄信修 | Application of cortex parabarii and cortex parabarii extract both serving as whitening effective ingredients |
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| CN112933018A (en) * | 2021-04-02 | 2021-06-11 | 科丝美诗(中国)化妆品有限公司 | Composition with barrier repairing and anti-photoaging effects and preparation method and application thereof |
| CN113116796A (en) * | 2021-03-30 | 2021-07-16 | 中国科学院合肥物质科学研究院 | Preparation method of eucommia anti-aging composition, eucommia anti-aging composition prepared by preparation method and application of eucommia anti-aging composition |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105456078A (en) * | 2015-06-26 | 2016-04-06 | 普正药业股份有限公司 | Cortex eucommiae exfoliating abrasive particles and preparation method and application thereof |
| CN105456078B (en) * | 2015-06-26 | 2018-01-23 | 普正药业股份有限公司 | A kind of bark of eucommia exfoliating abrasive particle and preparation method and application |
| CN104938285A (en) * | 2015-07-02 | 2015-09-30 | 安徽恩赐方生物科技有限公司 | Magnesium-rich eucommia plantation method |
| CN105616467A (en) * | 2015-12-23 | 2016-06-01 | 庄信修 | Application of cortex parabarii and cortex parabarii extract both serving as whitening effective ingredients |
| CN110547975A (en) * | 2018-06-04 | 2019-12-10 | 科丝美诗株式会社 | Cosmetic material composition containing epipinoresinol |
| CN110547975B (en) * | 2018-06-04 | 2022-07-05 | 科丝美诗株式会社 | Cosmetic material composition containing epipinoresinol |
| CN110934787A (en) * | 2019-11-27 | 2020-03-31 | 陈少威 | Anti-aging composition and preparation method thereof |
| CN110934787B (en) * | 2019-11-27 | 2022-04-19 | 陈少威 | Anti-aging composition and preparation method thereof |
| CN110755334A (en) * | 2019-12-05 | 2020-02-07 | 上海中医药大学 | Eucommia bark emulsion and preparation process thereof |
| CN113116796A (en) * | 2021-03-30 | 2021-07-16 | 中国科学院合肥物质科学研究院 | Preparation method of eucommia anti-aging composition, eucommia anti-aging composition prepared by preparation method and application of eucommia anti-aging composition |
| CN112933018A (en) * | 2021-04-02 | 2021-06-11 | 科丝美诗(中国)化妆品有限公司 | Composition with barrier repairing and anti-photoaging effects and preparation method and application thereof |
| CN119139190A (en) * | 2024-09-29 | 2024-12-17 | 诺德溯源(广州)生物科技有限公司 | Plant sun-proof repair composition applicable to sensitive muscles and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2013010489A1 (en) | 2013-01-24 |
| AU2012286357A1 (en) | 2014-03-06 |
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Inventor after: Zhao Ya Inventor after: Xiang Leihong Inventor after: Chen Mo Inventor after: Zhu Le Inventor after: Sun Yi Inventor after: Yan Shuxian Inventor after: Ren Jie Inventor after: Cheng Kang Inventor after: Xiong Weiguo Inventor after: Fang Cheng Inventor before: Zhao Ya Inventor before: Xiang Honglei Inventor before: Chen Mo Inventor before: Zhu Le Inventor before: Sun Yi Inventor before: Yan Shuxian Inventor before: Ren Jie Inventor before: Cheng Kang Inventor before: Xiong Weiguo Inventor before: Fang Cheng |
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Free format text: CORRECT: INVENTOR; FROM: ZHAO YA CHEN MO ZHU LE SUN YI YAN SHUXIAN REN JIE CHENG KANG XIONG WEIGUO FANG CHENG XIANG HONGLEI TO: ZHAO YA CHEN MO ZHU LE SUN YI YAN SHUXIAN REN JIE CHENG KANG XIONG WEIGUO FANG CHENG XIANG LEIHONG |
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