CN106511480A - Application of sweet osmanthus flower extract in preparation of composition for promoting hair growth and wound healing - Google Patents
Application of sweet osmanthus flower extract in preparation of composition for promoting hair growth and wound healing Download PDFInfo
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- CN106511480A CN106511480A CN201510573360.5A CN201510573360A CN106511480A CN 106511480 A CN106511480 A CN 106511480A CN 201510573360 A CN201510573360 A CN 201510573360A CN 106511480 A CN106511480 A CN 106511480A
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Abstract
The invention relates to application of a sweet osmanthus flower extract in preparation of compositions for promoting hair growth and wound healing. The sweet osmanthus flower extract is obtained by first extracting through pure water and then extracting through an ethanol solution. The invention non-predictably discovers the novel application of the sweet osmanthus flower extract for promoting hair growth and wound healing.
Description
Technical field
The present invention provides a kind of Flos Osmanthi Fragrantis extract is used for preparing the purposes for promoting hair to increase compositionss and Wound healing compositions.
Background technology
Flos Osmanthi Fragrantis (Osmanthus fragrans (Thunb) Lour.), for Oleaceae (Oleaceae) Osmanthuss (Osmanthus) evergreen shrubss, spend fragrance, its quintessence oil can be extracted, prepare osmanthus concrete, and can be used for the production of food and cosmetics, medically, act on warming the lung to resolve fluid-retention and dispersing cold for relieving pain.Pharmaceutically, its root or root bark can be used to treating for stomachache, have a toothache, rheumatic numbness and bones and muscles pain；Its flower can be used for resolving sputum, dissipating blood stasis, control phlegm retention cough with asthma, hemorrhoidal hamorrhage dysentery, hernia abdominal mass, toothache and halitosis；And its fruit can be used for warming the stomach, suppressing the hyperactive liver, kidney tonifying, cold expelling and control stomachache due to hyperactive liver-QI attacking the stomach.
Once there is TaiWan, China patent announcement the I341205th (patent application case the 096137207th), disclose a kind of method by anti-asthma material is extracted in Flos Osmanthi Fragrantis, the material that must have anti-asthma physiological function is extracted with ethanol, the chronic inflammation symptom of respiratory tract and pulmonary can be slowed down, reach asthma symptom of releiving, and prevention and the outbreak for reducing asthma, with obvious health-care effect.TaiWan, China patent announcement the I442929th (patent application case the 101118954th), discloses a kind of product of Flos Osmanthi Fragrantis water extraction and can be used to treat the purposes of pulmonary fibrosiss.TaiWan, China patent announcement the I411440th (patent application case the 100119436th), discloses a kind of Flos Osmanthi Fragrantis extract and is preparing prevention or treating the purposes of melancholy disease drug or health food.
Separately there is Chinese patent to disclose No. CN103768152A (patent application case the 201410043501.8th), disclose the application that a kind of Flos Osmanthi Fragrantis phenethyl alcohol glycoside extract prepares food, medicine or the health product of preventing and treating diabetes.Chinese patent is disclosed No. CN103767975A (patent application case the 201410043486.7th), discloses the application that a kind of Flos Osmanthi Fragrantis phenethyl alcohol glycoside extract prepares skin-lightening cosmetic.Chinese patent is disclosed No. CN103768151A (patent application case the 201410043488.6th), discloses the application that a kind of Flos Osmanthi Fragrantis phenethyl alcohol glycoside extract prepares antiaging agent or health product.Chinese patent is disclosed No. CN103223018A (patent application case the 201310169455.1st), discloses a kind of Flos Osmanthi Fragrantis Polyphenols extract in the application for preparing antitumor drug or health product.
The content of the invention
But have no arbitrary pertinent literature and once reported the purposes that Flos Osmanthi Fragrantis or its extract have the growth of promotion hair and wound healing.The present invention it is non-it is expected find a kind of Flos Osmanthi Fragrantis extract be used for promoting hair to increase and wound healing novel use.
One aspect of the present invention provides a kind of Flos Osmanthi Fragrantis extract is used for preparing the purposes for promoting hair to increase compositionss.
Another aspect of the present invention provides a kind of Flos Osmanthi Fragrantis extract to be used for preparing the purposes for promoting Wound healing compositions.
In embodiments of the present invention, Flos Osmanthi Fragrantis extract of the present invention can promote angiogenesiss and promote collagen protein to generate.
In embodiments of the present invention, Flos Osmanthi Fragrantis extract of the present invention wherein Flos Osmanthi Fragrantis extract system is obtained so that ethanol solution is extracted after first being extracted with pure water again.
In terms of these and other of the present invention, can be able to more apparent by the description of following preferred embodiment and accompanying drawing；Even if wherein might have change or modify, but without departing substantially from disclosed herein novel concepts spirit and the scope.
Description of the drawings
The preferred embodiment diagram system presented in the present invention is for the purpose of illustrating the present invention.It is to be understood that the preferred embodiment shown in the invention is not limited in.
Figure 1A-Fig. 1 C show its trichogenous result of Flos Osmanthi Fragrantis extract of the present invention.Figure 1A shows the 0th to 28 day hair growth situation of each group.Figure 1B shows hair growth fraction (error thick stick, the SE of the 0th to 28 day；*, P<0.05).Fig. 1 C show the other skin histology coloration result of each group.
Fig. 2A-Fig. 2 B show the result of Flos Osmanthi Fragrantis extract wound healing in noninfected wounds pattern of the present invention.Fig. 2A shows wound healing efficiency (error thick stick, the SE of the 0th to 18 day；*, P<0.05).Fig. 2 B show the healing situation of the 0th to the 18 day wound.
Fig. 3 A- Fig. 3 B show the healing result of Flos Osmanthi Fragrantis extract wound in infected wound pattern of the present invention.Fig. 3 A show wound healing efficiency (error thick stick, the SE of the 0th to 18 day；*, P<0.05).Fig. 3 B show the healing situation of the 0th to the 18 day wound.
Fig. 4 A- Fig. 4 B show that Flos Osmanthi Fragrantis extract of the present invention promotes the effect of angiogenesiss in noninfected wounds pattern or infected wound pattern.Fig. 4 A be displayed in noninfected wounds pattern in each group for the impact of angiogenesiss.Fig. 4 B be displayed in infected wound pattern in each group for the impact of angiogenesiss.
Fig. 5 shows the indirect fungistatic effect of Flos Osmanthi Fragrantis extract of the present invention；Lie in noninfected wounds pattern or infected wound pattern, with infection of staphylococcus aureus wound, the wound circumference Jing after the reason of each group other places suppurate tissue containing bacterium number.
Specific embodiment
Unless otherwise defined, all used herein technical and scientific terms, to those skilled in the art, all with the same meaning known with which.
Unless Wen Zhongyou understands indicated, the singulative " " that used in this article, " one kind ", and the connotation of " being somebody's turn to do " be the plural form including " at least one ".Thus, for example, when " composition " is referred to, including a plurality of these compositions and to coordinate known to those skilled in the art.
" extract " as used herein refers to the product obtained by being extracted for a material, typically by by the be intended to material immersion for extracting or being mixed in the solution or concentrate formulation obtained in solvent.Typically, extract system is prepared from the ground or dry plant sample of fresh plant.
" wound " as used herein includes infected wound or non-infectious wound, means it is, for the injury of the injury of epidermis (such as the skin corium of skin), including skin lacerations, cut-out, or to puncture.Wound includes but is not limited to open wound, such as otch, chop wound, hurt, abrading, stabbing, traumatic skin is damaged, infiltration sexual trauma, burn；Wound caused by disease or other chronic tissue injuries；By wound caused by external injury or surgical operation；The wound that medical procedure causes, such as department of dermatologry are performed the operation.Wound also can be by corticosteroid reaction so cause skin symptom, such as allergic dermatitis.
According to the present invention, find that Flos Osmanthi Fragrantis extract has functions that to promote hair to increase in zoopery.Therefore, the present invention provides a kind of Flos Osmanthi Fragrantis extract is used for preparing the purposes for promoting hair to increase compositionss.The promotion hair increases the form that compositionss can be made into medicine, shampoo, conditioner, conditioner, hair dye, hair oil or spray.
On the other hand, the present invention provides effect that a kind of Flos Osmanthi Fragrantis extract promotes wound healing again.Therefore, the present invention provides a kind of Flos Osmanthi Fragrantis extract and is used for preparing the purposes for promoting Wound healing compositions.The promotion Wound healing compositions can be made into the form of medicine, skin protectant, insect-proof agent or spray.
According to the present invention, the wound can be infected wound or non-infectious wound.In a specific embodiment of the present invention, wherein the infected wound system is infected by staphylococcus aureuses.
As the embodiment of the present invention is contained, Flos Osmanthi Fragrantis extract of the present invention can promote angiogenesiss and promote collagen protein to generate.
Of the invention one implements aspect, and the Flos Osmanthi Fragrantis extract system is obtained so that ethanol solution is extracted after first being extracted with pure water again.In a wherein embodiment, after the Flos Osmanthi Fragrantis extract system takes Flos Osmanthi Fragrantis (Osmanthus fragrans) with pure water extraction filtering and concentrating, then extracted with ethanol solution, then Jing after filtering and concentrate.
For implementing the present invention, the Flos Osmanthi Fragrantis extract can be made into the form of various types of compositionss and use.Wherein said composition can be the form of a kind of solution, hydrogel, spray, drop or ointment.Additionally, also can further include composition part of other antibacterial, antibiotic medicine, cleaning or skin protection.
According to the present invention, its medicine pattern can coordinate conventional excipient or filler according to any known techniques.Also medicine can be modulated into pharmaceutically acceptable supporting agent according to the techniques or methods commonly used on general medicine, the Flos Osmanthi Fragrantis extract of the present invention containing therapeutical effective amount, assistant.
With above-mentioned invention description and the following example explanation present invention, but and it is not used to limit the scope of the present invention.
Embodiment
Embodiment 1：The preparation and its process of Flos Osmanthi Fragrantis extract
Take the pure water that 1.0kg Flos Osmanthi Fragrantis (Osmanthus fragrans) add 10-20 times of volume, heating is extracted and is filtered, and after then concentrate extract, then is extracted with ethanol solution, after filtering and concentrating, Flos Osmanthi Fragrantis extract (about 250.4g) can be obtained.
Embodiment 2：Hair growth patterns are assessed
C57BL/6 mice of the laboratory animal system of the present invention purchased from National Laboratory Animal center, age are 6 weeks.In National Defense Medical Center's animal center, its Animal House temperature maintains 18～26 DEG C to the experimental animal feeding, and relative humidity is 30%～70%, and periodicity of illumination is illumination in 12 hours, 12 hour night, and provides sufficient feedstuff and drinking-water.
For confirming that the trichogenous activity of Flos Osmanthi Fragrantis extract carries out packet transaction, wherein, positive control group is to be processed with known hair growth promoting composition Min Nuoxidi (Minoxidil).By mice be randomly divided into three groups it is as follows：Blank control group (BC；10% ethanol is as supporting agent), positive control group (PC；3% Min Nuoxidi) and Flos Osmanthi Fragrantis extract (CE；During 1% adds 10% ethanol).Each group of each 5 mice is tested.The rush of Flos Osmanthi Fragrantis extract builds method (Choi et al., the Biol Pharm Bull 37 of the activity system using part modification Choi et al. of hair growth:44-53,2014).In short, mice carefully shaves the hair of one 2cm of drosal part × 3cm areas by electric scissors.Determinand (100 μ L) system is locally applied to the skin of mice drosal part, continuous 4 weeks once a day.The hair growth of the determinand promotes activity system using drosal part skin darkening as evaluation criteria, and which represents that the hair follicle of the hair lies in trophophase.
Hair growth scoring system is performed by two independent laboratory members for not knowing embodiment.Hair growth system measures weekly once, and continuous 4 weeks, hair growth score definition was as follows：0 point=growth is not observed；1 point=reach 20% growth；2 points=reach 20-40% growths；3 points=reach 40-60% growths；4 points=reach 60-80% growths；And 5 points=reach 80-100% growths.The hair of 4 weeks is always grown digitized video system and is taken the photograph using Nikon Cool Pix P100 (Tokyo, Japan).The average hair growth index as assessment hair growth that this two are not scored.
Observation mice process after result, when the 16th day, it is found that Flos Osmanthi Fragrantis extract group organizes compared with other that its wound outward appearance is more complete, wound tissue's epidermal area is restored completely, corium Rotating fields are more similar to normal mouse.Each group mice from process after the 0-28 days wound Photograph image as shown in Figure 1A, compared to blank control group and positive control group, Flos Osmanthi Fragrantis extract group mouse back skin from the 14th day has grown hair, and its hair follicle development, piliation area and hair amount were only all remarkably higher than other groups to 28 days from the 16th day.Its quantized result is presented with hair growth index, and such as Figure 1B shows, with the hair growth of Flos Osmanthi Fragrantis extract process, compared to other groups, there were significant differences (referring to Figure 1B).By skin histology dyeing, skin histology stained photographs image after each group mice treatment of wounds as shown in Figure 1 C, understands that its hair follicle development of Flos Osmanthi Fragrantis extract group is obvious and has hair growth compared to blank control group and positive control group.Shown by result above, Flos Osmanthi Fragrantis extract has trichogenous effect.
From embodiment result, Flos Osmanthi Fragrantis extract provided by the present invention can be obviously promoted the effect of hair growth.
Embodiment 3：Wound healing assay zootype is assessed
1. laboratory animal and pattern
C57BL/6J Narl Strains of Mouse of the laboratory animal system of this experiment purchased from National Laboratory Animal center, 40～45g of body weight, age are 8 weeks.In National Defense Medical Center's animal center, its Animal House temperature maintains 18～26 DEG C to the experimental animal feeding, and relative humidity is 30%～70%, and periodicity of illumination is illumination in 12 hours, 12 hour night, and provides sufficient feedstuff and drinking-water.
The group of each experiment model is as shown in the table：
Wherein, positive control group is Min Nuoxidi (Minoxidil).
2. treatment of wounds
Mice is given 0.6g/kg Avertin in lumbar injection mode by operation consent, wherein the Avertin systems are with 0.6g 2,2,2- tribromoethanol (2,2, add 2-Tribromoethanol) and after the mixing of 2 butanol (2-Methyl-2-butanol) of 1.5ml 2- methyl normal saline solution to 50ml, filtered with 0.2 μm of filter paper, and preserved with 4 DEG C.After mouse anesthesia, by its back hair cutting.Paper card is fixed on into mouse back and determines wound site, 1.2x 1.2cm are depicted with Mike's pen2Area.The skin in this area is cut using surgical scissorses, form wound.In infected wound pattern, staphylococcus aureuses are diluted to into 105CFU/150 μ l, and 150 μ l bacterium solutions are added in every mice, bacterium solution is fully penetrated in skin about 5 minutes (noninfected wounds pattern is not required to carry out this step).Picked after appropriate medicine to be tested, by its uniform application on wound using Cotton swab.Wound is covered using waterproof and breathable dressing (Tegaderm, 3M), waterproof and breathable dressing is fixed with breathable adhesive tape, it is to avoid waterproof and breathable dressing comes off and other wound infections.
3. staphylococcus aureuses culture and calculating
With the single S. aureus colonies of inoculating loop picking, it is inoculated on LB agar dish (LB agar plate) with subregion setting-out method, and cultivates about 16 hours in 37 DEG C of incubators.With the single S. aureus colonies of inoculating loop picking, the antibacterial culturing Tube propagation about 16 hours containing LB culture medium is put it into.Bacterium solution in test tube is taken out, is centrifuged 20 minutes under 4 DEG C, 7,000rpm.Supernatant is removed, and centrifugation gained staphylococcus aureuses thalline is diluted to into 1x 10 with LB culture medium7CFU/ml is standby.
By bacterium solution with serial dilution to 10-9.100 μ l bacterium solutions are taken out from different extension rate test tubes and add LB agar dish uniform application.In 37 DEG C of incubator cultures about 16 hours and note down clump count.Bacterium number (CFU/ml) contained by per milliliter in original bacterium solution=clump count * extension rates * 10.
4. antibacterial ring test
Take out 100 μ l staphylococcus aureuses liquid from different extension rate test tubes, dripped in agar dish uniform application respectively, after bacterium solution is by agar suction, with tweezers by filter paper smooth on agar, every filter paper is instilled into 50 μ l Flos Osmanthi Fragrantis extracts.After cultivating about 16 hours in 37 DEG C of incubators, liquid bacterium diameter is measured.Antibacterial ring size scope=inhibition zone diameter (mm)-filter paper diameter (mm).
5. wound tissue's bacterial content test
After infected wound pattern mouse experiment the 7th day, the suppuration tissue of wound mouse back wound is collected.Tissue shear is fragmentated, and tissue is smashed with ultrasound cell disruptor.After being centrifuged 8 minutes with 1,500rpm at 4 DEG C, supernatant is collected.The supernatant is carried out into serial dilution, after bacterium solution uniform application is on the agar, after cultivating about 16 hours in 37 DEG C of incubators, clump count is calculated.Bacterium number contained by per gram of tissue (CFU/g)=(clump count * extension rates * 10)/tissue grams.
6. Skin slice and hematoxylin and Yihong (Hematoxylin＆Eosin, H＆E) dyeing
Appropriate OCT (optimum cutting temperature) embedding medium is added in aluminium foil tissue embedding box, the fresh skin histology removed is put in embedded box, add OCT embedding mediums to cover whole tissue, and be placed on liquid nitrogen after quick freezing, preserve as -70 DEG C.Appropriate OCT embedding mediums are added on aluminum alloy organization's oblect stand, pretreated tissue as on oblect stand, then oblect stand is placed on microtome freezing stage and is freezed, and oblect stand is placed on microtome and is locked.Then, piece of tissue is made to form plane slightly to cut pattern (20 μm), recycling is frittered pattern (5～7 μm) and starts section.Microscope slide is attached at into tissue slice, and slide is soaked in acetone, be placed in -20 DEG C, slide is placed on into -70 DEG C of preservations after 30 minutes.
After Tissue slides are immersed in PBS about 3～4 minutes, by slide with brazilwood extract dyeing 10 minutes.Slide is cleaned after three times with pure water, then is soaked in pure water, respectively 5,5,5,10 minutes.To slide with eosin stains 15 seconds.Slide is soaked in 95%, 95%, 100% respectively, 100% ethanol, each 1 minute.Dimethylbenzene (xylene) is soaked in again 1 minute.With 70% ethanol by slide back side wiped clean, sample is waited to be dried.Finally with coverslip mounting (mountant：Dimethylbenzene=2：1).
7. result
In noninfected wounds pattern, mice wound cosmetic variation is estimated.First, after C57BL/6 mouse back skins start wound, wound was cleared up per two days and medicine is changed.As shown in Figure 2 A, compared to other groups, after non-infectious wound is processed with Flos Osmanthi Fragrantis extract, its wound has preferably healing efficiency.As shown in Figure 2 B, when the 4th to 8 day, blank control group and its wound healing of vaseline group be not obvious.Compare down, Flos Osmanthi Fragrantis extract group its wound outward appearance has substantially started healing.Shown by result, after the treatment of Flos Osmanthi Fragrantis extract is given, non-infectious wound can heal for about 4 to 8 days in advance.
And in infected wound pattern, with infection of staphylococcus aureus wound, after infection the 7th day, cleared up wound per two days and change medicine.As shown in Figure 3A, compared to other groups, after infected wound is processed with Flos Osmanthi Fragrantis extract, its wound has the trend being reduced significantly.At the 6th day, with the wound of Flos Osmanthi Fragrantis extract process, more than 60% wound healing has been reached.As shown in Figure 3 B, when the 6th day, blank control group and vaseline group its wound not yet start healing.Compare down, Flos Osmanthi Fragrantis extract group its wound outward appearance has significantly started healing.Shown by result, after giving the treatment of Flos Osmanthi Fragrantis extract in infected wound pattern, wound can heal for about 8 to 12 days in advance.
No matter in noninfected wounds pattern (Fig. 4 A) or infected wound pattern (Fig. 4 B), its wound skin appearance can find, compared to blank control group and vaseline group, Flos Osmanthi Fragrantis extract group its have remarkably promote vessel area, length and branch development situation, show Flos Osmanthi Fragrantis extract significantly have functions that promotion angiogenesiss.
Because the synthesis of collagen protein contributes to wound healing, when further inspecting wound healing, Flos Osmanthi Fragrantis extract vinegar promotes the effect of collagen protein synthesis.For noninfected wounds pattern, in mice wound the 21st day, and for infected wound pattern, then at the 27th day, periwound skin tissue is taken, the situation of collagen protein synthesis in skin histology is observed with Masson ' s trichrom stainings, collagen protein is dyed to blue presentation.Observation experiment result, it is found that no matter in non-infectious or contaminated wound pattern, under comparing with normal mouse, in blank control group and vaseline group, organization internal collagen protein synthesis quantity is few and loosely organized；And in the group processed with Flos Osmanthi Fragrantis extract, organization internal collagen protein synthesis quantity and close structure degree are all similar to normal mouse.Shown by result above, Flos Osmanthi Fragrantis extract has functions that to promote collagen protein synthesis.
Wound infection is the one of the reason for causing wound effectively cannot heal, therefore whether has antibacterial activity with antibacterial ring size experimental test Flos Osmanthi Fragrantis extract.After result of the test shows staphylococcus aureuses with drug treating, the antibacterial phenomenon of discovery after comparing with matched group, is had no, show Flos Osmanthi Fragrantis extract without directly antibacterial effect.
Further mouse back skin is started after wound, with infection of staphylococcus aureus wound and with drug treating, after infecting 7 days, wound circumference suppuration tissue is collected, is analyzed bacteria containing amount.As shown in figure 5, compared to blank control group and vaseline group, being significantly reduced with its wound surface pus bacterium number after the process of Flos Osmanthi Fragrantis extract.As a result learn, Flos Osmanthi Fragrantis extract has indirect bacteriostatic activity.
From result above, Flos Osmanthi Fragrantis extract of the present invention can promote wound healing, have functions that to promote wound blood vessels newborn and promote collagen protein to generate.
Claims (10)
1. a kind of Flos Osmanthi Fragrantis extract is used for preparing the purposes for promoting hair growth stimulant compositions.
2. purposes according to claim 1, wherein the Flos Osmanthi Fragrantis extract system first with pure water extraction after
Obtain so that ethanol solution is extracted again.
3. a kind of Flos Osmanthi Fragrantis extract is used for preparing the purposes for promoting Wound healing compositions.
4. the purposes as described in claims 3, wherein the Flos Osmanthi Fragrantis extract system first with after pure water extraction again with
Ethanol solution is extracted and is obtained.
5. purposes according to claim 4, wherein the Flos Osmanthi Fragrantis extract can promote angiogenesiss.
6. purposes according to claim 4, wherein the Flos Osmanthi Fragrantis extract can promote collagen protein to give birth to
Into.
7. purposes according to claim 1, wherein said composition make medicine, shampoo, profit
The form of silk essence, conditioner, hair dye, hair oil or spray.
8. purposes according to claim 3, wherein said composition make medicine, skin protectant, anti-
Worm agent or the form of spray.
9. the purposes according to claim 1 or 3, wherein said composition be solution, hydrogel,
The form of spray, drop or ointment.
10. the purposes according to claim 1 or 3, wherein said composition can further include other
Composition part of antibacterial, antibiotic medicine, cleaning or skin protection.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113018336A (en) * | 2019-12-06 | 2021-06-25 | 罗胥恩 | Externally applied traditional Chinese medicine composition for chronic wounds and application thereof |
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| JPH08268844A (en) * | 1995-03-29 | 1996-10-15 | Nonogawa Shoji Kk | Hair nourishing agent |
| CN101203277A (en) * | 2005-04-28 | 2008-06-18 | 因德纳有限公司 | Hair tonic |
| CN104887661A (en) * | 2014-03-06 | 2015-09-09 | 兰亭生物科技有限公司 | Medicinal composition for promoting wound healing, and use thereof |
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2015
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH08268844A (en) * | 1995-03-29 | 1996-10-15 | Nonogawa Shoji Kk | Hair nourishing agent |
| CN101203277A (en) * | 2005-04-28 | 2008-06-18 | 因德纳有限公司 | Hair tonic |
| CN104887661A (en) * | 2014-03-06 | 2015-09-09 | 兰亭生物科技有限公司 | Medicinal composition for promoting wound healing, and use thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113018336A (en) * | 2019-12-06 | 2021-06-25 | 罗胥恩 | Externally applied traditional Chinese medicine composition for chronic wounds and application thereof |
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Application publication date: 20170322 |