Garlic and sea-buckthorn paste and application thereof in treatment of oral ulcer

Abstract

The invention provides garlic and sea-buckthorn paste and application thereof in treatment of oral ulcer. Specifically, the invention provides a specific formula and preparation of the garlic and sea-buckthorn paste, the garlic and sea-buckthorn paste comprises the following components: garlic powder, sea-buckthorn oil, vaseline, glyceryl monostearate, glyceryl distearate, cera alba, wool fat and peppermint oil, and the proportion of the components is provided. In addition, the garlic and sea-buckthorn paste has good bacteriostatic and oral ulcer repairing effects.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and more particularly, the present application relates to a garlic and sea-buckthorn paste and its application in the treatment of oral ulcer. BACKGROUND

[0002] Oral ulcer is a common and recurrent oral mucosa disease, and patients often show round or oval painful ulcers on the oral mucosa, which seriously affects eating, speaking and quality of life. At present, the clinical treatment is mainly local medication, such as using drug film, spray or gargling solution containing anti-inflammatory, analgesic and antiseptic effects, aiming to relieve pain, prevent secondary infection and promote healing. However, the existing drugs often have problems such as single effect, easy to produce drug resistance, strong irritation or unsatisfactory healing speed. Therefore, it has important clinical significance and application value to develop a multifunctional natural drug preparation with efficient antibacterial, anti-inflammatory, analgesic and significant mucosa tissue repair accelerating effects.

[0003] Garlic is a traditional medicinal and edible plant, and its strong broad-spectrum antibacterial activity has been widely proven. The main bioactive substance in garlic, allicin and its derivatives, has significant inhibition and killing effect on a variety of gram-positive bacteria, gram-negative bacteria, fungi and even viruses, which makes it an ideal natural candidate for preventing and treating oral infections.

[0004] Sea-buckthorn oil is a natural plant oil extracted from sea-buckthorn fruit, which is rich in vitamin E, carotene, flavonoids, unsaturated fatty acids and various amino acids and trace elements. Studies have shown that sea-buckthorn oil has excellent anti-inflammatory and antioxidant capacity, and can significantly promote epithelial cell growth and granulation tissue generation, accelerate wound healing, and has significant effect in treating skin burns and ulcers.

[0005] At present, there is no research on the compounding of garlic and sea-buckthorn oil to simultaneously exert the dual effects of antibacterial and healing promotion by optimizing the proportion. SUMMARY

[0006] The purpose of the present application is to provide a new garlic and sea-buckthorn paste, which optimizes the prescription ratio, utilizes the strong antibacterial effect of garlic to control wound infection and the repair characteristics of sea-buckthorn oil to promote tissue regeneration, produces a synergistic effect, realizes the comprehensive effect of "bacteria and toxin treatment, repair and promotion", effectively balances the antibacterial and healing promotion performance, and provides an innovative solution to solve the deficiencies of existing preparations.

[0007] In the first aspect of the present application, a garlic and sea-buckthorn paste is provided, which comprises or consists of the following components:

[0008] Garlic powder, sea-buckthorn oil, vaseline, glycerol monostearate and distearate, white beeswax, lanolin, and peppermint oil;

[0009] wherein the garlic powder is present in an amount of 5-15% (w / w), preferably 5-10% (w / w); and the sea buckthorn oil is present in an amount of 15-25% (w / w), preferably 18-22% (w / w).

[0010] In another preferred embodiment, the garlic sea buckthorn paste comprises the following components:

[0011] Petrolatum 55-70% (w / w), preferably 58-68% (w / w);

[0012] White beeswax 4 ± 2% (w / w);

[0013] Glyceryl monostearate 4 ± 2% (w / w);

[0014] Lanolin 2 ± 1% (w / w); and

[0015] Peppermint oil 0.5 ± 0.25% (w / w).

[0016] In another preferred embodiment, the garlic sea buckthorn paste comprises the following components:

[0017] Garlic powder 5% (w / w); and Sea buckthorn oil 20% (w / w).

[0018] In another preferred embodiment, the garlic sea buckthorn paste comprises the following components:

[0019] Garlic powder 7.5% (w / w); and Sea buckthorn oil 20% (w / w).

[0020] In another preferred embodiment, the garlic sea buckthorn paste comprises the following components:

[0021] Garlic powder 10% (w / w); and Sea buckthorn oil 20% (w / w).

[0022] In another preferred embodiment, the garlic sea buckthorn paste comprises the following components:

[0023] Garlic powder 5% (w / w);

[0024] Sea buckthorn oil 20% (w / w);

[0025] Petrolatum 64.5% (w / w);

[0026] White beeswax 4% (w / w);

[0027] Glyceryl monostearate 4% (w / w);

[0028] Lanolin 2% (w / w); and

[0029] Peppermint oil 0.5% (w / w).

[0030] In another preferred embodiment, the garlic sea buckthorn cream comprises the following components:

[0031] Garlic powder 7.5% (wt%);

[0032] Sea buckthorn oil 20% (wt%);

[0033] Petrolatum 62% (wt%);

[0034] White beeswax 4% (wt%);

[0035] Glyceryl monostearate 4% (wt%);

[0036] Lanolin 2% (wt%); and

[0037] Peppermint oil 0.5% (wt%).

[0038] In another preferred embodiment, the garlic sea buckthorn cream comprises the following components:

[0039] Garlic powder 10% (wt%);

[0040] Sea buckthorn oil 20% (wt%);

[0041] Petrolatum 59.5% (wt%);

[0042] White beeswax 4% (wt%);

[0043] Glyceryl monostearate 4% (wt%);

[0044] Lanolin 2% (wt%); and

[0045] Peppermint oil 0.5% (wt%).

[0046] In another preferred embodiment, the garlic sea buckthorn cream is prepared by heating and melting the components in the formula, and then mixing them.

[0047] In another preferred embodiment, the garlic sea buckthorn cream is prepared by:

[0048] 1) Heating and melting the prescribed amount of petrolatum, white beeswax, glyceryl monostearate and lanolin at 70°C in a water bath, and mixing them;

[0049] 2) Taking out 1) and cooling it to 50°C, adding the prescribed amount of sea buckthorn oil, and mixing them;

[0050] 3) Adding the prescribed amount of garlic powder to 2), and mixing them quickly; and

[0051] 4) When 3) is cooled to room temperature, adding the prescribed amount of peppermint oil, and mixing them, to obtain the garlic sea buckthorn cream.

[0052] In another preferred embodiment, the garlic powder is obtained by drying garlic raw material.

[0053] In another preferred embodiment, the drying process includes but is not limited to freeze-drying, oven-drying.

[0054] In another preferred embodiment, the sea buckthorn oil is sea buckthorn fruit oil.

[0055] In another preferred embodiment, the sea buckthorn oil is sea buckthorn seed oil.

[0056] In another preferred embodiment, the garlic sea buckthorn paste is a yellowish to light orange paste.

[0057] In another preferred embodiment, the garlic sea buckthorn paste is capable of inhibiting the growth of Escherichia coli, Staphylococcus aureus, and / or Candida albicans.

[0058] In another preferred embodiment, the oral cure rate of the garlic sea buckthorn paste is greater than 65%.

[0059] In another preferred embodiment, the oral cure rate of the garlic sea buckthorn paste is greater than 70%.

[0060] In a second aspect of the present application, there is provided a use of a garlic sea buckthorn paste for the manufacture of a medicament for treating oral ulcer.

[0061] In a third aspect of the present application, there is provided a method of treating oral ulcer, the method comprising applying a therapeutically effective amount of the garlic sea buckthorn paste of the first aspect of the present application to the affected area of a subject.

[0062] It should be understood that, within the scope of the present application, each of the technical features described above and in the following (e.g. in the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they are not listed one by one here. BRIEF DESCRIPTION OF DRAWINGS

[0063] Figure 1 The figure shows the effect of the antibacterial ring test of the garlic sea buckthorn paste (left: garlic sea buckthorn paste; right: garlic sea buckthorn paste + gel);

[0064] Note: KB-1#: gel; KB-2#: 0% garlic powder + 0% sea buckthorn oil; KB-3#: 0% garlic powder + 20% sea buckthorn oil; D: Escherichia coli; J: Staphylococcus aureus; B: Candida albicans.

[0065] Figure 2 The figure shows the weight gain rate of rats after administration of different prescriptions.

[0066] Figure 3 The figure shows the oral ulcer surface of rats after administration of different prescriptions.

[0067] Figure 4The healing rate of oral ulcer of rats after administration of different prescriptions is shown.

[0068] Figure 5 The appearance of the garlic and sea-buckthorn paste of the present application is shown. DETAILED DESCRIPTION

[0069] The present inventors have developed a garlic and sea-buckthorn paste and its application in the treatment of oral ulcer through extensive and in-depth research. The present application finally determines a garlic and sea-buckthorn paste formula through the screening of main ingredients and excipients, which has good applicability in paste properties and skin feel use. In addition, through the proportion screening of main ingredients, the garlic and sea-buckthorn paste balances the bacteriostatic and tissue repair effects at the same time, greatly improving the healing rate of oral ulcer.

[0070] TERMS

[0071] For easier understanding of the present application, certain technical and scientific terms are defined in detail below. Unless otherwise clearly defined in this text, all other technical and scientific terms used in this text have the meanings generally understood by those skilled in the art to which the present application belongs. Before describing the present application, it should be understood that the present application is not limited to the specific methods and experimental conditions described, as such methods and conditions can vary.

[0072] As used herein, the terms "comprise", "comprising", "containing" and "containing" are used interchangeably and include not only the closed definition but also the semi-closed and open definition. In other words, the terms include "consisting of", "consisting essentially of".

[0073] In the case of providing a numerical range, unless the context clearly indicates otherwise, it should be understood that every integer between the values, every tenth of the value, between the upper and lower limits of the range and any other intermediate value in the specified range is included in the present application. The upper and lower limits of these smaller ranges can be independently included in the smaller range and also covered in the present application, but subject to any explicitly excluded limitations in the specified range. For example, "1 to 50" includes "2 to 25", "5 to 20", "25 to 50", "1 to 10", etc.

[0074] Garlic powder

[0075] The garlic powder described in the present application is prepared by drying the garlic, and the preparation process is as follows: the garlic is peeled, washed, dried, crushed, sieved, packaged in aluminum foil bags, and stored in a cool, dry, light-proof place. The garlic used in the present application is not limited by source and origin.

[0076] In the present application, garlic powder is used as the main component of the garlic sea buckthorn paste, which has broad-spectrum antibacterial and antiviral effects. Allicin is a natural "plant antibiotic" that can inhibit and kill a variety of bacteria, fungi, and viruses, thereby achieving an anti-inflammatory effect. In addition, garlic contains allicin and other sulfur compounds, and the active sulfur components can temporarily increase the permeability of the stratum corneum, like a "channel opener", helping other drug components in the paste to penetrate the skin barrier better, penetrate the lesion, and thus improve the overall therapeutic effect.

[0077] Sea buckthorn oil

[0078] In the present application, the sea buckthorn oil includes sea buckthorn fruit oil and sea buckthorn seed oil, preferably sea buckthorn fruit oil. Sea buckthorn oil has a unique orange appearance, thick oil texture, strong odor, and is rich in Omega-7, super-high VC, and carotenoids, etc., and has excellent repair, antioxidant, and anti-inflammatory functions. It is a very precious multifunctional natural oil.

[0079] As the main component, sea buckthorn oil can effectively promote the growth of granulation tissue and the regeneration of epithelial cells, significantly accelerate the healing of wounds, ulcers, and damaged skin; the phytosterols and fatty acids contained therein can effectively repair damaged skin barriers, prevent water loss, and keep the skin moist and soft; and can neutralize free radicals, reduce the oxidative damage of environmental factors to the skin, thereby protecting skin cells and delaying skin aging.

[0080] Garlic sea buckthorn paste

[0081] The garlic sea buckthorn paste of the present application is obtained by screening the ratio of auxiliary materials and main components, and finally a relatively optimal formula is obtained, wherein the components include garlic powder, sea buckthorn oil, vaseline, glycerol monostearate and distearate, white beeswax, lanolin, and peppermint oil. The formula has excellent paste properties and excellent bacteriostatic and tissue repair effects.

[0082] The garlic sea buckthorn paste of the present application has a higher cure rate in treating oral ulcers than oral ulcer drugs on the market (such as oral ulcer gel). In addition, the garlic sea buckthorn paste of the present application can also be used to relieve redness, itching, and burning sensation caused by mosquito bites, eczema, dermatitis, sunburn, etc.

[0083] In an embodiment, the present application also compares the formula comprising the following components:

[0084] (1) garlic powder, sea buckthorn oil, vaseline, glycerol monostearate and distearate, white beeswax, lanolin, and peppermint oil;

[0085] (2) garlic powder, sea buckthorn oil, white beeswax, glycerol monostearate and distearate, glycerol, liquid paraffin, and peppermint oil;

[0086] (3) Garlic powder, sea buckthorn oil, white beeswax, monostearate, glycerol, liquid paraffin, peppermint oil.

[0087] Comparing the properties of each formula, the paste properties and skin feel of formula (1) are better than those of formula (2) and (3), and it has more advantages in the aspect of traditional Chinese medicine.

[0088] Petroleum jelly

[0089] In the garlic sea buckthorn paste of the present application, petroleum jelly as the main base is a semi-solid mixture of hydrocarbons, which is extremely stable in nature and not hydrophilic. It can form a dense and air-tight protective film on the skin surface. This film can isolate air, bacteria, irritants and other external substances, protecting damaged skin; and can effectively prevent water evaporation from the skin, greatly softening the stratum corneum, making dry and cracked skin soft, and creating a moist environment for wound healing. In addition, petroleum jelly can well dissolve many drug ingredients, making them uniformly dispersed in the entire paste and controlling the release: drugs dissolved or mixed in the petroleum jelly base can be slowly and continuously released to the skin surface, prolonging the drug effect time.

[0090] Glyceryl monostearate

[0091] Glyceryl monostearate in garlic sea buckthorn paste serves as an emulsifying stabilizer and skin feel regulator. Compared with monostearic acid, glyceryl monostearate has stronger emulsifying ability, can form a film at the interface, and at the same time construct a three-dimensional network structure in the liquid phase, greatly improving the stability of the paste. And make the paste more delicate, smooth, white, with a silk-like luster, and have good spreading property, moisturizing but not greasy comfortable skin feel.

[0092] White beeswax, lanolin, and peppermint oil

[0093] White beeswax is a product obtained by bleaching and refining of beeswax, which is white in color and has the functions of thickening and hardening. Beeswax is solid at room temperature, which can significantly improve the melting point and hardness of the paste, preventing the paste from softening and flowing due to temperature changes during storage and use.

[0094] Lanolin is an oil extracted from sheep wool, which is similar in composition to human sebum and is an excellent "moisturizing and penetration enhancer". Lanolin has super water absorption (can absorb more than 2 times its own weight of water), and can effectively prevent water evaporation, thereby deeply softening the stratum corneum and moisturizing dry and cracked skin. Promote drug penetration: as a good transdermal absorption enhancer, it can help the effective ingredients (including peppermint oil and other drugs) in the ointment penetrate the skin barrier better and reach the lesion directly.

[0095] Peppermint oil is a volatile oil extracted from the peppermint plant. The menthol in peppermint oil can stimulate the cold receptors on the skin, producing an instant and strong cooling sensation. Through the cooling sensation and mild local anesthesia, it can effectively relieve itching, mild neuralgia and muscle pain.

[0096] Compared with the prior art, the main advantages of the present application include:

[0097] 1. The garlic and sea buckthorn paste of the present application has an optimized proportion of excipients, and the paste has uniform texture, moderate oiliness, luster, and good skin feel.

[0098] 2. The garlic and sea buckthorn paste of the present application has an optimized proportion of main components, which can balance the bacteriostatic and tissue repair effects, and has excellent oral ulcer repair effect.

[0099] The present application will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and not used to limit the scope of the present application. The experimental methods in the following examples, if not specified, are usually carried out according to the conventional conditions, such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are weight percentages and weight parts.

[0100] Example 1. Prescription screening

[0101] 1. Experimental materials: garlic powder, vaseline, sea buckthorn fruit oil, white beeswax, glycerol monostearate, lanolin, peppermint oil.

[0102] 2. Preparation of garlic powder: peel the garlic cloves, wash, dry, crush, pass through a 100-mesh sieve, seal in an aluminum foil bag, and store in a cool, dry, and dark place.

[0103] 3. Preparation of prescription

[0104] Weigh the prescribed amount (Table 1) of vaseline, white beeswax, glycerol monostearate and lanolin, heat to melt in a 70℃ water bath, mix well; take out, cool to 50℃, add the prescribed amount of sea buckthorn oil, mix well; add the prescribed amount of garlic powder, mix quickly; cool to room temperature, add the prescribed amount of peppermint oil, mix well, and it is ready.

[0105] Table 1. Amount of each component in the prescription

[0106]

[0107]

[0108] The blank matrix prescription is to replace garlic powder with an equal proportion of vaseline under the established prescription, and the proportions of other components remain unchanged.

[0109] 4. Evaluation index of paste body of garlic sea buckthorn paste

[0110] 4.1 Viscosity (Table 2): Stir the paste in the sample jar with a glass rod at a uniform speed for 10 times, and feel the overall resistance.

[0111] Table 2 Viscosity evaluation grade and standard

[0112] Score Grade Sensory Description 1 Too thin Almost no resistance, like stirring emulsion, feels thin. 2 Thinner Slight resistance, feels soft, easy to stir. 3 Moderate Moderate resistance, smooth stirring feeling, good cream followability. 2 Thicker Obvious resistance, need to stir slightly. 1 Too thick Very large resistance, stirring is difficult, cream is heavy.

[0113] 4.2 Flowability (Table 3): Tilt the wide-mouth bottle or flat plate containing the ointment to an angle of 45° and hold for 30 seconds, and observe the flow and deformation of the paste surface.

[0114] Table 3 Flowability evaluation grade and standard

[0115]

[0116] 4.3 Skin feel

[0117] Spreadability (Table 4): Take a certain amount of paste (such as a pea size) on the back of the hand or the inside of the forearm, and spread it out with the palm of the hand with moderate force, and feel the process of pushing it away.

[0118] Table 4 Spreadability evaluation grade and standard

[0119] Score Grade Sensory Description 1 Very difficult to spread Severe skin pulling, large resistance, difficult to spread. 2 More difficult to spread Difficult to spread, not smooth enough, need to repeat several times. 3 Moderate spread Smooth spread. 2 Easy to spread Spread easily, can quickly cover the skin. 1 Very easy to spread One wipe, instant uniform spread, smooth and no resistance.

[0120] Stickiness (Table 5): After the paste is evenly applied for about 30 seconds, gently touch the applied area with the fingertips, and then gently lift it up.

[0121] Table 5 Stickiness evaluation grade and standard

[0122] Score Grade Sensory Description 1 Severe stickiness Finger and skin are severely connected, feel very unclean. 2 Obvious stickiness There is a clear sense of adhesion, feel not clean. 3 Light stickiness There is a slight stickiness, but it is within an acceptable range. 4 Almost no stickiness Very slight stickiness, close to clean. 5 Clean and no stickiness No stickiness at all, skin feels smooth and dry.

[0123] 5. Evaluation results of paste body of different prescriptions

[0124] 5.1 Appearance

[0125] The product is a uniform cream with a light yellow to light orange yellow color, with a smooth and delicate texture, and is rich in luster. The paste is moderately oily, with no visible particles, impurities or liquid separation phenomenon. Its odor is characterized by the characteristic cooling aroma of peppermint oil as the leading aroma, which is in harmony with the natural fruit aroma inherent in sea buckthorn oil and the inherent aroma of garlic extract, with no odor. The overall appearance is uniform, delicate and stable in quality.

[0126] 5.2 Prescription score results (Table 6)

[0127] Table 6 Prescription evaluation score results

[0128]

[0129] 6. Prescription determination

[0130] According to the product viscosity, fluidity, skin feel and other indicators, the final prescription is as follows (Table 7), and the appearance of the paste is as shown in Figure 5 .

[0131] Table 7 Garlic and sea buckthorn paste prescription

[0132]

[0133]

[0134] Example 2. Evaluation of the bacteriostatic effect of garlic and sea buckthorn paste

[0135] 1. Experimental materials

[0136] Table 8 Experimental instruments

[0137] Name Model Manufacturer Constant temperature incubator BPX-272 Shanghai Buxun Industry Co., Ltd. Biochemical incubator SPX-250B-Z Shanghai Buxun Industry Co., Ltd. Vertical pressure sterilization pot YXQ-LS-50A Shanghai Buxun Industry Co., Ltd. Biological safety cabinet BHC-1300ⅡA2 Wujiang Huadu Purification Equipment Co., Ltd. Electric heating constant temperature water tank SSW-600-2S Shanghai Buxun Industry Co., Ltd. Multi-purpose test tube stirrer TM-2N Shanghai Donggomei Trade Development Co., Ltd. Bacterial turbidity meter JC-XJZD1A Qingdao Jingcheng Instrument and Equipment Co., Ltd. Electronic balance YP30001 Shanghai Yueping Scientific Instrument Co., Ltd.

[0138] Test strains: Staphylococcus aureus (ATCC6538), Escherichia coli (8099 or ATCC25922), and Candida albicans (ATCC10231).

[0139] Bacteriostatic agent carrier (5mm diameter round qualitative filter paper, sterilized by pressure steam, then baked at 120°C for 2h, and stored for use).

[0140] Culture medium: nutrient agar medium and sabouraud agar medium.

[0141] 2. Ointment preparation

[0142] Ointment (0% garlic powder + 0% sea buckthorn oil, 5% garlic powder + 0% sea buckthorn oil, 7.5% garlic powder + 0% sea buckthorn oil, 10% garlic powder + 0% sea buckthorn oil, 0% garlic powder + 20% sea buckthorn oil, 5% garlic powder + 20% sea buckthorn oil, 7.5% garlic powder + 20% sea buckthorn oil, 10% garlic powder + 20% sea buckthorn oil) and gel (aqueous base), specific prescription as shown in Table 9.

[0143] Table 9 Ointment prescription

[0144]

[0145] 3. Experimental method

[0146] According to "2.1.8.2 Inhibition Zone Test" under "2.1.8 Antimicrobial Test" in "Disinfection Technical Specification" (2002 Edition), the inhibition test of blank group, ointment group and ointment + gel group was investigated.

[0147] 3.1 Specific operation steps

[0148] (1) Preparation of bacteriostatic pieces: Take Xinhua No. 1 qualitative filter paper pieces, and use a puncher to make 5mm diameter round pieces. After pressure steam sterilization treatment, bake at 120℃ for 2h, and store for standby use.

[0149] (2) Sample preparation: Different addition ratios of garlic and sea buckthorn paste (5% garlic powder + 20% sea buckthorn oil, 7.5% garlic powder + 20% sea buckthorn oil, 10% garlic powder + 20% sea buckthorn oil) were prepared in a sterile laboratory safety biological cabinet. The paste (garlic and sea buckthorn paste alone, garlic and sea buckthorn paste + gel mixed) with a thickness not exceeding 4mm was placed on the bacteriostatic carrier of (1), and each group had 4 pieces (blocks).

[0150] (3) Preparation of negative control sample pieces: Gel, 0% garlic powder + 0% sea buckthorn oil, and 0% garlic powder + 20% sea buckthorn oil were used to prepare sample pieces (blocks) with the same size as the test group.

[0151] (4) Preparation of bacterial suspension: Staphylococcus aureus and Escherichia coli were inoculated into TSB and mixed, and cultured in a 36℃ constant temperature incubator for 24 hours. Candida albicans was inoculated into SDB and mixed, and cultured in a 25℃ constant temperature incubator for 48 hours. 1mL of Staphylococcus aureus, Escherichia coli and Candida albicans culture was taken and diluted with 0.85% NaCl solution by 10 times gradient to the appropriate concentration, i.e. the medium suitability and counting method verification bacterial suspension concentration was not more than 100cfu / ml; the bacteriostatic efficacy check test bacterial solution was about 108cfu / ml.

[0152] (5) Inoculation of test bacteria: A sterile cotton swab was dipped in the test bacterial suspension with a concentration of 5.0×105cfu / ml-5.0×106cfu / ml, and the surface of the nutrient agar culture medium plate was evenly smeared 3 times. Each time of smearing, the plate should be rotated by 60°, and finally the cotton swab was smeared around the edge of the plate. Cover the plate and dry at room temperature for 5min.

[0153] (6) Bacteriostatic agent-like sample placement: 1 plate of each test was placed with 4 pieces of test sample, 1 piece of negative control sample, a total of 5 pieces. The sample was placed on the surface of the plate with sterile forceps (the ointment side down, the filter paper disc on top, even if the ointment directly contacts the bacterized plate). The centers of each sample were separated by more than 25 mm, and the edge of the plate was more than 15 mm apart. After placement, the sample was lightly pressed with sterile forceps to make the ointment adhere to the surface of the plate. Cover the plate and place it in a 37°C incubator for 16-18 h. Measure the diameter of the bacteriostatic ring with a vernier caliper and record it.

[0154] When measuring the bacteriostatic ring, a uniform and completely sterile growth bacteriostatic ring should be selected. The diameter should be measured with the outer edge of the bacteriostatic ring as the boundary.

[0155] 3.2 Evaluation criteria for bacteriostasis

[0156] (1) Judgment of bacteriostatic effect: If the diameter of the bacteriostatic ring is greater than 7 mm, it is judged to have bacteriostatic effect; if the diameter of the bacteriostatic ring is less than or equal to 7 mm, it is judged to have no bacteriostatic effect.

[0157] (2) If all 3 repeated tests have bacteriostatic effect, it is judged to be qualified.

[0158] (3) The negative control group should have no bacteriostatic ring. Otherwise, the test is invalid.

[0159] 3.3 Bacteriostatic effect of different prescriptions

[0160] Table 10 Bacteriostatic test results

[0161]

[0162] The results showed that the blank group (KB-1#, KB-2#, KB-3#) and the garlic and sea buckthorn ointment group ( Figure 1 left) had no bacteriostatic effect on E. coli, S. aureus and C. albicans; the garlic and sea buckthorn ointment + gel group ( Figure 1 right) had a bacteriostatic ring diameter greater than 7 mm on E. coli, S. aureus and C. albicans, indicating that the garlic and sea buckthorn ointment + gel had bacteriostatic effect on E. coli, S. aureus and C. albicans. Among them, the 10% garlic powder had the largest bacteriostatic ring on E. coli, S. aureus and C. albicans (Table 10), indicating that the garlic and sea buckthorn ointment with 10% garlic powder had the best bacteriostatic effect on the three bacteria; the garlic ointment + gel group without sea buckthorn oil also had bacteriostatic effect on the three bacteria (Table 10), and overall the bacteriostatic ring of the garlic and sea buckthorn ointment + gel group was slightly larger than that of the garlic ointment + gel group, indicating that garlic powder and sea buckthorn oil had a certain synergistic bacteriostatic effect.

[0163] Example 3. Oral ulcer repair effect of garlic and sea buckthorn ointment

[0164] 1. Experimental materials

[0165] Glacial acetic acid, sodium pentobarbital, normal saline, garlic and sea buckthorn paste (5%, 7.5%, 10% garlic powder + 24% sea buckthorn oil), garlic ointment (5% garlic powder), medical oral ulcer gel, SD rats.

[0166] 2. Experimental method

[0167] 2.1 Establishment of rat oral ulcer model

[0168] (1) Select 54 male SD rats and adaptively feed them for 2 days to ensure that they can adapt to the new experimental environment and facilitate subsequent experimental operations.

[0169] (2) When conducting animal experiments, anesthetize the rats with sodium pentobarbital at a concentration of 0.4-0.5 ml / 100 g of body weight, and use forceps to fix the rat's mouth to fully expose the lower lip mucosa, providing a good view for subsequent operations.

[0170] (3) Cut a 5mm diameter circular filter paper, immerse it in glacial acetic acid for 10s, press it onto the inner side of the lower lip mucosa for 30s, and use a cotton swab to wipe off the excess glacial acetic acid. Observe the mucosa at the pressed site, which is damaged and turns bright red.

[0171] (4) Observe the local swelling and ulceration of the buccal mucosa at the treated site 24 hours later. If the model is successful, record it as D1.

[0172] 2.2 Grouping and administration

[0173] The oral ulcer group is randomly divided into a model group, a base 1# group, a base 2# group, a garlic ointment group, a low-dose administration group, a medium-dose administration group, a high-dose administration group, and a positive drug group, with 6 rats in each group. The normal control (blank) group has 6 rats. Start wound administration on the second day of modeling, and flush the wound with normal saline in each group.

[0174] The blank group and the model group are not given any medication.

[0175] The base 1# group is locally smeared with 0% garlic powder + 0% sea buckthorn oil ointment.

[0176] The base 2# group is locally smeared with 0% garlic powder + 20% sea buckthorn oil ointment.

[0177] The garlic ointment group is administered with 5% garlic powder + 0% sea buckthorn oil ointment.

[0178] The low-dose administration group is locally smeared with garlic and sea buckthorn paste (5% garlic powder + 20% sea buckthorn oil).

[0179] The medium-dose administration group is locally smeared with garlic and sea buckthorn paste (7.5% garlic powder + 20% sea buckthorn oil).

[0180] High-dose administration group with garlic sea buckthorn paste (10% garlic powder + 20% sea buckthorn oil) topical application;

[0181] The positive drug group is applied with medical oral ulcer gel topically;

[0182] Each time the drug is used to completely cover the ulcer, and the drug is administered once a day.

[0183] 2.3 Clinical evaluation

[0184] (1) D1 starts every day to take pictures of the oral ulcer of the rat, and the ruler is placed on the edge of the ulcer for contrast when taking pictures.

[0185] (2) Each photo is analyzed with image analysis software (ImageJ) to quantitatively measure the ulcer area. Calculate the ulcer healing degree: ulcer healing degree = (A1-An) / A1*100%, where A0 is the original ulcer area on D1 day, and An is the ulcer area that has not healed after Dn day.

[0186] 2.4 Oral ulcer degree and ulcer healing grading standard

[0187] 0 level: no ulcer formation, normal mucosa, ulcer damage repair score is 100 points;

[0188] Grade I: ulcer formation, no obvious pseudomembrane on the surface of the ulcer, no tissue necrosis, ulcer damage repair score is 80 points;

[0189] Grade II: ulcer formation, a thin layer of yellow-white pseudomembrane on the surface of the ulcer, with a small amount of tissue necrosis, ulcer damage repair score is 60 points;

[0190] Grade III: ulcer formation, the ulcer surface is covered with a thick pseudomembrane, inflammatory edema appears around the ulcer, with more tissue necrosis, ulcer damage repair score is 40 points;

[0191] Grade IV: ulcer formation, the ulcer surface is covered with a thick pseudomembrane, with obvious inflammatory edema around and a large amount of tissue necrosis, ulcer damage repair score is 20 points.

[0192] 3. Oral ulcer repair effect

[0193] 3.1 Effect on body weight of oral ulcer model rats

[0194] During the experiment, the body weight and food intake of rats in each group increased, and the body weight gain rate was higher than that of the model group (18.8%), indicating that the oral ulcer induced by ice acetic acid burn had a certain effect on the body weight of rats, and the administration could reduce the effect of oral ulcer on the food intake of rats Figure 2 ).

[0195] 3.2 Effect on the area of oral ulcer in rats

[0196] After 24 h of modeling, the oral mucosa of each group of rats showed a concave defect in the modeling area, with red and swollen tissue around the concave, and a yellow-white pseudomembrane covering the surface. On the third day of administration, the lower lip of the model group and each administration group was swollen, and the ulcer surface pseudomembrane was thickened. On the fifth day of administration, the lower lip of the model group and the matrix 1# group was swollen, and the ulcer surface was obvious, with obvious inflammatory edema and hyperemia around the tissue; the administration group had a thin pseudomembrane, reduced inflammatory edema and hyperemia, and reduced ulcer area. On the seventh day of administration, the symptoms of the model group and each administration group were reduced, and the model group still had mild lower lip swelling; in the administration group, the ulcer surface pseudomembrane was removed, with mild inflammatory edema and hyperemia, and the ulcer surface was significantly reduced, and the repair score of the garlic and sea buckthorn ointment administration group (low, medium and high doses) was higher than that of the other administration groups (Tables 10 and 11). Figure 3 and Table 11).

[0197] Table 11. Ulcer injury repair score

[0198] Group Day 1 Day 3 Day 5 Day 7 Model 20 20 40 60 Matrix 1# 20 40 40 60 Matrix 2# 20 40 60 60 Garlic ointment group 20 40 60 60 Garlic and sea buckthorn ointment low-dose group 20 40 60 80 Garlic and sea buckthorn ointment medium-dose group 20 40 60 80 Garlic and sea buckthorn ointment high-dose group 20 40 60 80 Positive drug 20 40 60 60

[0199] In addition, the wound healing rate of the garlic and sea buckthorn ointment administration group was significantly higher than that of the matrix 1# group, the model group and the positive drug group; compared with the matrix 1# group (0% garlic powder + 0% sea buckthorn oil), the healing rate of the matrix 2# group (0% garlic powder + 20% sea buckthorn oil) was higher; the wound healing rate of the low-dose garlic and sea buckthorn ointment group was 76.39% after seven days of administration, and the healing rate of the garlic ointment group without adding sea buckthorn oil was 62.94%, which was lower than that of the garlic and sea buckthorn ointment group (Table 12). Figure 4 and Table 12).

[0200] The above results show that the garlic and sea buckthorn ointment (low, medium and high doses) prepared in the present application can reduce the effect of oral ulcer on the weight gain of rats, and effectively promote the healing of oral ulcer in rats, and garlic powder and sea buckthorn oil have a synergistic effect.

[0201] Table 12. Wound healing rate (%)

[0202] Group Day 3 Day 5 Day 7 Model 0.62 34.91 38.01 Matrix 1# 5.79 33.78 41.73 Matrix 2# 12.24 37.18 47.58 Garlic ointment group 19.39 45.31 62.94 Garlic and sea buckthorn ointment low-dose group 24.61 58.89 76.39 Garlic and sea buckthorn ointment medium-dose group 22.92 40.58 72.68 Garlic and sea buckthorn ointment high-dose group 26.05 54.20 65.17 Positive drug 5.84 26.00 40.51

[0203] All the documents mentioned in the present application are incorporated herein by reference as if each document was individually incorporated by reference. In addition, it should be understood that various changes and modifications can be made to the present application by those skilled in the art after reading the above description of the present application, and these equivalent forms also fall within the scope defined by the claims attached hereto.

Claims

1. A garlic and sea buckthorn paste, characterized in that, The garlic and sea buckthorn paste comprises or is composed of the following components: Garlic powder, sea buckthorn oil, petrolatum, glyceryl mono- and di-stearates, white beeswax, lanolin, and peppermint oil; The garlic powder content is 5-15% (by weight); the sea buckthorn oil content is 15-25% (by weight).

2. The garlic and sea buckthorn paste as described in claim 1, characterized in that, The garlic and sea buckthorn paste comprises the following components: Vaseline 55-70% (by weight); White beeswax 4±2% (by weight); Glyceryl monostearate and glyceryl distearate 4 ± 2% (wt%); Lanolin 2 ± 1% (wt%); and Peppermint oil 0.5 ± 0.25% (by weight).

3. The garlic and sea buckthorn paste as described in claim 1, characterized in that, The garlic and sea buckthorn paste comprises the following components: Garlic powder 5% (by weight); and sea buckthorn oil 20% (by weight).

4. The garlic and sea buckthorn paste as described in claim 1, characterized in that, The garlic and sea buckthorn paste comprises the following components: Garlic powder 7.5% (by weight); and sea buckthorn oil 20% (by weight).

5. The garlic and sea buckthorn paste as described in claim 1, characterized in that, The garlic and sea buckthorn paste comprises the following components: Garlic powder 10% (by weight); and sea buckthorn oil 20% (by weight).

6. The garlic and sea buckthorn paste as described in claim 1, characterized in that, The garlic and sea buckthorn paste comprises the following components: Garlic powder 5% (by weight); Sea buckthorn oil 20% (by weight); Vaseline 64.5% (by weight); 4% (by weight) white beeswax; Glyceryl monostearate and glyceryl distearate 4% (wt%); Lanolin 2% (by weight); and Peppermint oil 0.5% (by weight).

7. The garlic and sea buckthorn paste as described in claim 1, characterized in that, The garlic and sea buckthorn paste comprises the following components: Garlic powder 7.5% (by weight); Sea buckthorn oil 20% (by weight); Vaseline 62% (by weight); 4% (by weight) white beeswax; Glyceryl monostearate and glyceryl distearate 4% (wt%); Lanolin 2% (by weight); and Peppermint oil 0.5% (by weight).

8. The garlic and sea buckthorn paste as described in claim 1, characterized in that, The garlic and sea buckthorn paste comprises the following components: Garlic powder 10% (by weight); Sea buckthorn oil 20% (by weight); Vaseline 59.5% (by weight); 4% (by weight) white beeswax; Glyceryl monostearate and glyceryl distearate 4% (wt%); Lanolin 2% (by weight); and Peppermint oil 0.5% (by weight).

9. The garlic and sea buckthorn paste as described in claim 1, characterized in that, The garlic and sea buckthorn extract can inhibit the growth and reproduction of Escherichia coli, Staphylococcus aureus, and / or Candida albicans.

10. The use of the garlic and sea buckthorn extract according to claim 1, characterized in that, Used to prepare drugs for treating oral ulcers.

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