CN109432317A - Prevent hemorrhoid composition and its application, wet tissue - Google Patents
Prevent hemorrhoid composition and its application, wet tissue Download PDFInfo
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- CN109432317A CN109432317A CN201811515081.3A CN201811515081A CN109432317A CN 109432317 A CN109432317 A CN 109432317A CN 201811515081 A CN201811515081 A CN 201811515081A CN 109432317 A CN109432317 A CN 109432317A
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- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
- A61K8/0208—Tissues; Wipes; Patches
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4973—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with oxygen as the only hetero atom
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
- A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/14—Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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Abstract
Prevent hemorrhoid composition and its application, wet tissue the present invention relates to a kind of, wherein prevention hemorrhoid are comprised the following components in parts by weight with composition: 0.1-5 parts of big vast eucalyptus camphor tree essential oil, 0.1-5 parts of borneol camphor tree essential oil, 0.01-2 parts of tea polyphenols and 0.1-2 parts of bamboo extractive.Above-mentioned each component mutually acts synergistically, so that the composition has stronger bactericidal bacteriostasis, can play the role of preventing hemorrhoid generation.
Description
Technical field
The present invention relates to technical field of daily necessities, in particular to prevention hemorrhoid composition and its application, wet tissue.
Background technique
Hemorrhoid (Hemorrhoids), or claim hemorrhoid, are a kind of clinically most common disease of anus, can be divided into internal piles, outer
Hemorrhoid and mixed hemorrhoid.It is especially sticky in stool to be just easy in anal sphincter remained on surface, and then bacterium is caused to grow
It is raw, cause sphincters contract that pinching occurs and promotes vascular dilation, is easy to cause the generation of hemorrhoid.In addition, with rhythm of life
Accelerate, daily life system is irregular, and pressure increases, and more and more people the symptoms such as endocrine disturbance, indigestion occur, are also easy
Lead to the increase of hemorrhoid illness rate.Hemorrhoid be often accompanied by hematochezia, bleed, mole abjection, when going to the toilet with pain, the symptoms such as itch, more
Have notably, can not stand walking, seriously affect normal life, and hemorrhoid are easy to recur, it is more difficult to cure.Therefore, in daily life
In work, it should be noted that the generation of prevention hemorrhoid.Currently, the product for having more prevention hemorrhoid to generate in the market, most of is cream
Medicine or skin care item need to adhere to smear, inconvenient for use.And there are also more chemical synthesis substance, unsuitable children old man for portioned product
It uses, and is used for a long time and is also also easy to produce dependence, using limited.
Summary of the invention
Based on this, it is necessary to provide a kind of prevention hemorrhoid composition and its application, wet tissue that can prevent hemorrhoid.
It is a kind of to prevent hemorrhoid composition, it comprises the following components in parts by weight:
Above-mentioned prevention hemorrhoid are activity with big vast eucalyptus camphor tree essential oil, borneol camphor tree essential oil, tea polyphenols and bamboo extractive with composition
Component.Wherein, big vast eucalyptus camphor tree leaf is rich in cineole abundant, and mass percentage accounts for the 45%-50% of big vast eucalyptus camphor tree leaf, has
The effects of antibacterial, dispelling wind are antipyretic, clearing damp is detoxified, and it also contains the substances such as more amphene, firpene, has the work sterilized more by force
With.
Borneol is China's tradition rare traditional Chinese medicine and fine perfumery, also known as natural borneol in borneol camphor tree, is listed in successive dynasties emperor
Palace " tribute ", the function that external application has clearing away heat to and alleviating pain, anti-corrosion antipruritic can treat the illness such as sore, swelling and pain, aphtha, take orally to have and have one's ideas straightened out
The benefits of inducing resuscitation, is suitable for coma, all cards of fainting from fear.Its fragrance of borneol essential oil is more strong powerful compared with borneol, and can walk can go here and there in vivo,
Make outside the suitable granular media of poison gas, foul smell, channels and collaterals item reaches.
Tea polyphenols are the general name of Polyphenols of Tea, including flavanol compound, anthocyanin class, flavonoids, flavonols
With phenolic acid class etc..Predominantly flavanols (catechin) class, catechin account for 60~80%, and substance tea polyphenols are also known as tea tan or tea list
It is peaceful that there is disinfection, sterilizing, resisting age of skin, while itself or a kind of natural preservative, the stability of composition can be enhanced.
Bamboo extractive contains the substances such as flavones, phenolic acid, anthraquinone, lactones, polysaccharide, amino acid, microelement, has relatively strong
Antibacterial, anti-oxidant, fatigue-resisting function, especially its bamboo-leaves flavones rich in, be a kind of stronger antibacterial, disease-resistant
Malicious, deodorization substance.And to show that bamboo extractive takes LD50 to mouse oral equal for the toxicological test that does of Zhejiang Medical science institutes
Greater than 10g/kg weight, true border is nontoxic, and micronucleus test, sperm shape abnormality test and Salmonella reversion test testing result are feminine gender.Greatly
Mouse acute toxicity test also indicates that the true border of bamboo extractive is nontoxic；Each dosage group has no obvious to 30 days feeding trials of rat
Toxic reaction show that its unmanned selection dosage is 5.0g/kg weight, has higher-security.
Above-mentioned prevention hemorrhoid composition is by being aided with tea polyphenols and bamboo using big vast eucalyptus camphor tree essential oil and borneol camphor tree essential oil as monarch drug in a prescription
Leaf extract, and by optimization each group distribution ratio, so that above-mentioned composition has stronger sterilization, bacteriostasis, it can be effective
Ground prevents the generation of hemorrhoid, and above-mentioned each raw material is from a wealth of sources, non-toxic and safe, has wider application range, equally applicable
In old man and child.
In one embodiment, in the prevention hemorrhoid composition, the flood eucalyptus camphor tree essential oil is 0.5-1 parts, institute
Stating borneol camphor tree essential oil is 0.5-1 parts, and the tea polyphenols are 0.1-0.5 parts, and the bamboo extractive is 0.1-0.5 parts.
In one embodiment, in the prevention hemorrhoid composition, the borneol camphor tree essential oil is d-Bomeol camphor tree
Essential oil.
D-Bomeol is the main active component of natural borneol camphor tree, and drug effect is that borneolum syntheticum is incomparable, Ke Yixian
Write the curative effect of enhancing composition.
Above-mentioned prevention hemorrhoid composition is preparing the application in paper handkerchief.
Since above-mentioned prevention hemorrhoid composition has stronger bactericidal bacteriostasis, and safe without toxic side effect, it is suitable for
It is used to prepare paper handkerchief, so that anal sphincter surface is cleared up and sterilized during routine use paper handkerchief,
Being effectively prevented from residue leads to bacteria breed, easily and effectively achievees the purpose that prevent hemorrhoid.
A kind of preparation method for preventing hemorrhoid composition, comprising the following steps:
Big vast eucalyptus camphor tree essential oil, borneol camphor tree essential oil, tea polyphenols and bamboo extractive are mixed to get prevention hemorrhoid combination
Object.
A kind of to prevent hemorrhoid wet tissue, including wet paper immersion and carrier, the wet paper immersion includes that above-mentioned prevention hemorrhoid are used
Auxiliary material and daily necessities acceptable solvent are subjected in composition, daily necessities.
Above-mentioned prevention hemorrhoid include the wet paper immersion and carrier being mainly made of above-mentioned prevention hemorrhoid with composition with wet tissue,
By the way that wet paper immersion to be supported on carrier, wet tissue is made, routine use can be played conveniently and efficiently pre- in daily life
The effect of anti-hemorrhoid, and have no toxic side effect, it is applicable in relatively wide.
In one embodiment, the volume ratio of the wet paper immersion and the moisture paper tissue papers is (2-3): 1.
In one embodiment, the mass percentage of the wet paper immersion is 60%-90%.
In one embodiment, in the wet paper immersion, the quality percentage of the above-mentioned prevention hemorrhoid composition
Content is 0.5%-20%.
In one embodiment, the auxiliary material includes moisturizer, surfactant and acid-base modifier.
In one embodiment, in the prevention hemorrhoid wet tissue, in terms of mass percentage, the moisturizer is
1%-6%, the surfactant are 1%-4%, and the acid-base modifier adjusting wet paper immersion to pH is 5.5-7.
In one embodiment, the moisturizer is glycerol, propylene glycol, sorbierite, polyvinyl alcohol, plant polyose extraction
One of object is a variety of；And/or
The surfactant is one of Crodaret, ethoxylated dodecyl alcohol, polysorbate
Or it is a variety of；And/or
The acid-base modifier is one of citric acid and sodium citrate or a variety of.
In one embodiment, the carrier be non-woven fabrics, the non-woven fabrics with a thickness of 40g/ ㎡ -60g/ ㎡.
A kind of preparation method for preventing hemorrhoid wet tissue, comprising the following steps:
Big vast eucalyptus camphor tree essential oil, borneol camphor tree, tea polyphenols and bamboo extractive are mixed, added water to cook, is filtered, concentration obtains dense
Contracting liquid；Auxiliary material and the concentrate are mixed, are uniformly dispersed to get the wet paper immersion；
The carrier is immersed in above-mentioned wet paper immersion, is packed up to the prevention hemorrhoid wet tissue.
Specific embodiment
To facilitate the understanding of the present invention, below will to invention is more fully described, and give it is of the invention compared with
Good embodiment.But the invention can be realized in many different forms, however it is not limited to embodiment described herein.Phase
Instead, purpose of providing these embodiments is makes the disclosure of the present invention more thorough and comprehensive.
Unless otherwise defined, all technical and scientific terms used herein and belong to technical field of the invention
The normally understood meaning of technical staff is identical.Term as used herein in the specification of the present invention is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention.Term as used herein "and/or" includes one or more phases
Any and all combinations of the listed item of pass.
Specific embodiment is set forth below, and the present invention will be described.
Embodiment 1
Raw material (totally 100 parts): big vast 0.5 part of eucalyptus camphor tree essential oil, 0.5-1 parts of borneol camphor tree essential oil, 0.1 part of tea polyphenols, the leaf of bamboo extract
0.1 part of object, 2 parts of glycerol, 2 parts of propylene glycol, 2 parts of Crodaret, deionized water surplus, citric acid tune pH to 5-
6.5。
Preparation method: above-mentioned big vast eucalyptus camphor tree essential oil, borneol camphor tree essential oil and Crodaret are mixed, obtained water-soluble
Essential oil mixed liquor；
Deionized water is added in obtained soluble fine mixed liquor and glycerol, propylene glycol, tea polyphenols, bamboo extractive
In be uniformly dispersed, adding citric acid tune pH to 5-6.5 obtains wet paper immersion；
It takes with a thickness of the non-woven fabrics of 40g/ ㎡, sterilize and is immersed in above-mentioned wet paper immersion, pack up to the pre- of embodiment 1
Anti- hemorrhoid wet tissue.
Embodiment 2
Raw material (totally 100 parts): big vast 0.5 part of eucalyptus camphor tree essential oil, 1 part of borneol camphor tree essential oil, 0.2 part of tea polyphenols, bamboo extractive 0.5
Part, 2 parts of glycerol, 2 parts of propylene glycol, 2 parts of Crodaret, deionized water surplus, citric acid tune pH to 5-6.5.
Preparation method: above-mentioned big vast eucalyptus camphor tree essential oil, borneol camphor tree essential oil and Crodaret are mixed, obtained water-soluble
Essential oil mixed liquor；
Above-mentioned soluble fine mixed liquor and glycerol, propylene glycol, tea polyphenols, bamboo extractive are added in deionized water and are divided
It dissipates uniformly, adding citric acid tune pH to 5-6.5 obtains wet paper immersion；
It takes with a thickness of the non-woven fabrics of 40g/ ㎡, sterilize and is immersed in above-mentioned wet paper immersion, pack up to the pre- of embodiment 2
Anti- hemorrhoid wet tissue.
Embodiment 3
Raw material (totally 100 parts): big vast 1 part of eucalyptus camphor tree essential oil, borneol camphor tree essential oil 1,0.5 part of tea polyphenols, 0.8 part of bamboo extractive,
2 parts of glycerol, 2 parts of propylene glycol, 2 parts of Crodaret, deionized water surplus, citric acid tune pH to 5-6.5.
Preparation method: above-mentioned big vast eucalyptus camphor tree essential oil, borneol camphor tree essential oil and Crodaret are mixed, obtained water-soluble
Essential oil mixed liquor；
Above-mentioned soluble fine mixed liquor and glycerol, propylene glycol, tea polyphenols, bamboo extractive are added in deionized water and are divided
It dissipates uniformly, adding citric acid tune pH to 5-6.5 obtains wet paper immersion；
It takes with a thickness of the non-woven fabrics of 40g/ ㎡, sterilize and is immersed in above-mentioned wet paper immersion, pack up to the pre- of embodiment 3
Anti- hemorrhoid wet tissue.
Embodiment 4
Raw material (totally 100 parts): big vast 1 part of eucalyptus camphor tree essential oil, 1 part of borneol camphor tree essential oil, 0.5 part of tea polyphenols, bamboo extractive 0.5
Part, 2 parts of glycerol, 2 parts of propylene glycol, 2 parts of Crodaret, deionized water surplus, sodium citrate tune pH to 7.
Preparation method: above-mentioned big vast eucalyptus camphor tree essential oil, borneol camphor tree essential oil and Crodaret are mixed, obtained water-soluble
Essential oil mixed liquor；
Above-mentioned soluble fine mixed liquor and glycerol, propylene glycol, tea polyphenols, bamboo extractive are added in deionized water and are divided
It dissipates uniformly, adding citric acid tune pH to 5-6.5 obtains wet paper immersion；
It takes with a thickness of the non-woven fabrics of 40g/ ㎡, sterilize and is immersed in above-mentioned wet paper immersion, pack up to the pre- of embodiment 4
Anti- hemorrhoid wet tissue.
Embodiment 5
Raw material (totally 100 parts): big vast 2 parts of eucalyptus camphor tree essential oil, 2 parts of borneol camphor tree, 0.8 part of tea polyphenols, 1 part of bamboo extractive, glycerol 2
Part, 2 parts of propylene glycol, 2 parts of Crodaret, deionized water surplus, sodium citrate tune pH to 5-6.5.
Preparation method: above-mentioned big vast eucalyptus camphor tree essential oil, borneol camphor tree essential oil and Crodaret are mixed, obtained water-soluble
Essential oil mixed liquor；
Above-mentioned soluble fine mixed liquor and glycerol, propylene glycol, tea polyphenols, bamboo extractive are added in deionized water and are divided
It dissipates uniformly, adding citric acid tune pH to 5-6.5 obtains wet paper immersion；
It takes with a thickness of the non-woven fabrics of 40g/ ㎡, sterilize and is immersed in above-mentioned wet paper immersion, pack up to the pre- of embodiment 5
Anti- hemorrhoid wet tissue.
Comparative example 1
It is substantially the same manner as Example 3, it is without place, does not contain borneol camphor tree.
Comparative example 2
It is substantially the same manner as Example 3, the difference is that, do not contain tea polyphenols.
Comparative example 3
It is substantially the same manner as Example 3, the difference is that, do not contain bamboo extractive.
Comparative example 4
It is substantially the same manner as Example 3, the difference is that, only contain auxiliary material, i.e.,
Raw material (totally 100 parts): 2 parts of glycerol, 2 parts of propylene glycol, 2 parts of Crodaret, deionized water surplus, lemon
Lemon acid tune pH to 7.
Bacteriostatic experiment
Referring to " GB15979-2002 disposable sanitary articles sanitary standard " appendix C " product bactericidal property, bacteriostasis property with
Stability test method " antibiotic rate test is carried out to the wet tissue of embodiment 1-5 and comparative example 1-4, action time is 5 minutes, knot
Fruit is shown in Table 1.
It takes 0.5 gram of wet paper immersion in surface plate, adds 1 × 108The bacteria suspension of cfu/ml, 49ml antibacterial water are being shaken
After swinging on device concussion 5,15,30min, stands after 30min takes 1ml treated bacteria suspension to make 10 times of gradient dilutions, take 1ml and training
It supports in ware, 40 DEG C or so of culture medium is melted and is cooled in addition, shakes up, and in 37 DEG C of culture 72h, calculates bacteriostasis rate, test result
Such as the following table 1:
Table 1
Wherein "-" indicates substantially without bactericidal effect.
As it can be seen from table 1 the wet paper immersion of embodiment 1- embodiment 5 has stronger bactericidal effect, and its sterilization effect
Fruit is apparently higher than comparative example 1- comparative example 4.
Typical case
Case 1: Lee, female 30 years old, burning sensation of the anus, used prepared by the embodiment of the present invention 3 wet tissue 1 day often during breaking-out
Afterwards, hence it is evident that feel that burning sensation of the anus mitigates, detection uses 1 week, and symptom disappears substantially, hereafter continues to use the wet tissue of embodiment 3, disease
Shape does not recur.
Case 2: Wu, female 25 years old, burning sensation of the anus often, anal area itch, and stool has pain, uses during breaking-out
Behind prepared by the embodiment of the present invention 3 wet tissue 1 day, hence it is evident that feel that burning sensation of the anus mitigates, adhere to using 1 week, scorching hot and pruritis
It disappears, continues to use 3 months, good lifestyle habits are kept during use, does not recur during 3 months, stops after 3 months
This product is only used, does not occur sense of discomfort.
Case 3: Mr. Wang, male 45 years old, suffer from hemorrhoid for many years, be after diagnosing external piles, often have hematochezia, hemorrhoid symptom outstanding, and such as
There is apparent pain when lavatory, can not be move freely when serious, is often used plaster treatment, but recurrence frequency is higher.During breaking-out after
The continuous plaster using treatment hemorrhoid, while the wet tissue prepared using the embodiment of the present invention 3, affected part no longer aches and itches unbearably after 2 days, and 8
It fully recovers after it, plaster is stopped using after recovery from illness, continue to use the wet tissue of embodiment 3 12 months, during which hemorrhoid do not recur.
Case 4: Liu, male 57 years old, suffer from hemorrhoid for many years, and always recurrent exerbation after each medication treatment makes during breaking-out
Affected part no longer ache and itches unbearably behind wet tissue 1 day prepared with the embodiment of the present invention 3, fully recovers after 7 days, continues after recovery from illness using the present invention
Wet tissue 12 months, during which hemorrhoid do not recur.
Case 5: the Qin, female 37 years old, suffer from hemorrhoid for many years, and always recurrent exerbation after each medication treatment uses during breaking-out
Affected part, which is no longer ached, behind wet tissue 2 days of the preparation of the embodiment of the present invention 3 itches unbearably, fully recovers, continues after recovery from illness real using the present invention after 5 days
Apply the wet tissue of example 3 15 months, during which hemorrhoid do not recur.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of prevent hemorrhoid composition, which is characterized in that comprise the following components in parts by weight:
2. prevention hemorrhoid composition according to claim 1, which is characterized in that the flood eucalyptus camphor tree essential oil is 0.5-1 parts,
The borneol camphor tree essential oil is 0.5-1 parts, and the tea polyphenols are 0.1-0.5 parts, and the bamboo extractive is 0.1-0.5 parts.
3. prevention hemorrhoid composition according to claim 1 or 2, which is characterized in that the borneol camphor tree essential oil is dextrorotation
Borneol camphor tree essential oil.
4. the described in any item prevention hemorrhoid compositions of claim 1-3 are preparing the application in daily necessities.
5. a kind of prevent hemorrhoid wet tissue, which is characterized in that including wet paper immersion and carrier, the wet paper immersion includes that right is wanted
It seeks the described in any item prevention hemorrhoid compositions of 1-3, be subjected to auxiliary material and daily necessities acceptable solvent in daily necessities.
6. prevention hemorrhoid wet tissue according to claim 5, which is characterized in that the body of the wet paper immersion and the carrier
Product is than being (2-3): 1.
7. prevention hemorrhoid wet tissue according to claim 6, which is characterized in that in the wet paper immersion, the prevention hemorrhoid
The mass percentage of sore composition is 0.5%-20%.
8. prevention hemorrhoid wet tissue according to claim 7, which is characterized in that the auxiliary material includes moisturizer, surface work
Property agent and acid-base modifier；
In the prevention hemorrhoid wet tissue, in terms of mass percentage, the moisturizer is 1%-6%, the surface-active
Agent is 1%-4%, and the acid-base modifier adjusting wet paper immersion to pH is 5.5-7.
9. prevention hemorrhoid wet tissue according to claim 8, which is characterized in that the moisturizer is glycerol, propylene glycol, mountain
One of pears alcohol, polyvinyl alcohol, plant polyose extract are a variety of；And/or
The surfactant is one of Crodaret, ethoxylated dodecyl alcohol, polysorbate or more
Kind；And/or
The acid-base modifier is one of citric acid and sodium citrate or a variety of.
10. -9 described in any item prevention hemorrhoid wet tissues according to claim 1, which is characterized in that the carrier is non-woven fabrics,
The non-woven fabrics with a thickness of 40g/ ㎡ -60g/ ㎡.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811515081.3A CN109432317A (en) | 2018-12-12 | 2018-12-12 | Prevent hemorrhoid composition and its application, wet tissue |
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| CN1840013A (en) * | 2006-01-25 | 2006-10-04 | 徐凤鸣 | Wet pad for treating piles |
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| CN108834597A (en) * | 2018-08-31 | 2018-11-20 | 洪江市金土地生态农业有限责任公司 | A kind of flood eucalyptus camphor tree cuttage and seedling culture method |
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| CN1840013A (en) * | 2006-01-25 | 2006-10-04 | 徐凤鸣 | Wet pad for treating piles |
| CN105310922A (en) * | 2014-08-04 | 2016-02-10 | 李天德 | Compound essential oil and wet tissue for haemorrhoids care |
| CN108834597A (en) * | 2018-08-31 | 2018-11-20 | 洪江市金土地生态农业有限责任公司 | A kind of flood eucalyptus camphor tree cuttage and seedling culture method |
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