CN107881217A - A kind of Moisturizer method for customizing - Google Patents

Abstract

The invention discloses a kind of Moisturizer method for customizing, comprise the steps of：Step 1, user's cell sample is gathered；Step 2, the mRNA in cell sample is extracted；Step 3, mRNA reverse transcriptions are synthesized into cDNA；Step 4, the polymorphism of moisturizing related gene and corresponding cDNA expression quantity in cDNA are detected, moisturizing related gene includes at least one of AQP3, CLDN1, JAM1；Step 5, judge moisture-retaining capacity according to gene pleiomorphism and corresponding cDNA expression quantity, and for moisture-retaining capacity selection Moisturizer composition and dosage, prepare suitable Moisturizer.This method obtains the moisture-retaining capacity of different user by measuring the expression quantity of moisturizing related gene, and prepares suitable Moisturizer according to the specific moisture-retaining capacity of user.

Description

A kind of Moisturizer method for customizing

Technical field

The present invention relates to a kind of technical field of skin care, more particularly to a kind of Moisturizer method for customizing.

Background technology

With the development of science and technology, the full genome spectrogram of the mankind can be obtained by being sequenced.Currently largely grind Study carefully and show, state and the gene of human skin have close relationship.The genotype of specific site in these moisturizing related genes (or gene mononucleotide polymorphism, SNPs, abbreviation gene pleiomorphism) can largely affect the state of skin, different Genotype be to cause the key factor of skin problems.People can have found or predict skin by detecting different genotype The problem of skin, and select suitable skin care item in this, as guidance.Further study show that in addition to genotype, led by genotype The expressing protein of cause is only the key factor for causing skin problem.

It currently there are a large amount of skin cares with difference in functionality and cosmetics select for people.But per each and every one The skin of body all has its uniqueness, for example, the moisturizing of different application on human skin is horizontal and moisture-retaining capacity differs.How to select It is exactly the major issue that people face to be adapted to the Moisturizer of oneself.The method that current consumer generally selects is exactly to adopt Moisturizer is verified if appropriate for oneself with the mode of selection-purchase-trial, and consumer not only wastes greatly in the process The time of amount and financial resources, but also it is likely to result in the discomfort of skin.Therefore the moisturizing shield for being adapted to consumer's individual how is selected Skin product, turn into a major issue urgently to be resolved hurrily.

The content of the invention

The present invention provides Moisturizer method for customizing, and this method is by measuring the polymorphism and correspondingly of moisturizing related gene CDNA expression quantity obtains the moisture-retaining capacity of different user, and prepares suitable Moisturizer according to the specific moisture-retaining capacity of user.

According to a kind of Moisturizer method for customizing of the present invention, comprise the steps of：

Step 1, user's cell sample is gathered；

Step 2, the mRNA in the cell sample is extracted；

Step 3, mRNA reverse transcriptions are synthesized into cDNA；

Step 4, the polymorphism of moisturizing related gene and corresponding cDNA expression quantity in cDNA are detected, wherein, the moisturizing Related gene includes at least one of AQP3, CLDN1, JAM1；

Step 5, the expression quantity according to the gene pleiomorphism and corresponding cDNA judges moisture-retaining capacity, and is directed to the moisturizing Ability selection component and dosage, prepare suitable Moisturizer.

Further, the step 4 includes：

Step 4a), the mRNA sequence of the moisturizing related gene is searched on NCBI, and design corresponding primer；

Step 4b), using delta-deltaCt methods, using gene GAPDH as internal reference, pass through guarantor described in the primer detection The relative expression quantity of the polymorphism of wetting phase correlation gene and corresponding cDNA.

Further, the primer includes：

GAPDH Forword：5’-CGGGAAACTGTGGCGTGATG-3’

Reverse：5’-ATGACCTTGCCCACAGCCTT-3’

AQP3 Forword：5’-CCTGGAGGCCTTCACCGT-3’

Reverse：5’-TTGACGGCATAGCCGGAGTT-3’

CLDN1 Forword：5’-CTGTGGATGTCCTGCGTGTC-3’

Reverse：5’-TGCTCAGATTCAGCAAGGAGTCA-3’

JAM1 Forword：5’-TGTCTGTTCCCAGGAGTCCTTC-3’

Reverse：5’-CTCGACTTGCGCCTTTGTCC-3’.

Further, the composition of the Moisturizer includes NMF, grease, polypeptide and auxiliary agent.

Further, the NMF includes first group of NMF and second group of NMF, first group of NMF bag Containing bright matter acid sodium, polyglutamic acid, tremella polysaccharides, panaxan, GL-B, second group of NMF include propane diols, and two Propane diols, butanediol, glycerine.

Further, the grease includes olive oil, white oil, vaseline, paraffin oil, apricot kernel oil, saualane, macadamia nut oil, Jojoba oil, the one or more of Butyrospermum parkii fruit fatty oil.

Further, it is more to include alanine/histidine/lysine polypeptide copper HCl, soybean (GLYCINEMAX) for the polypeptide Peptide, dipeptides -1, dipeptides -2, dipeptides -4, oligopeptides -1, oligopeptides -2, oligopeptides -3, oligopeptides -4, oligopeptides -5, oligopeptides -6, oligopeptides -29, widow Peptide -34, carnosine, saccharomycete polypeptide, arginine/lysine polypeptide, nonapeptide -1, poly- (tripeptides -6), ascorbyl polypeptide, withered grass Bacterium lipopeptid sodium, hexapeptide -1,2,3,5,9,11, heptapeptide -6, myristoyl hexapeptide -5, myristoyl pentapeptide -4, tripeptides -1,2,3, 10,32, the copper of tripeptides -1, decapeptide -4, tetrapeptide -1,3,4, pentapeptide -1,3,34, oat peptide, Wild soybean peptide, acetyl group octapeptide -3, second The cetyl of acyl group dipeptides -1, acetyl group hexapeptide 1,7,8, acetyl group heptapeptide -4, acetyl group tetrapeptide 2,3,5,9,11, palmityl two Peptide -7, palmityl hexapeptide -12,14,15, palmityl tripeptides -1,5,8, palmityl tetrapeptide -5,7,10, in Matrixyl -4,5 One or more of combinations.

Further, the auxiliary agent includes antioxidant, solvent, preservative, solubilizer, thickener, skin conditioning agent, Chelating agent, aromatic, anti-sensitizer and water.

Further, the step 5 includes：

Step 5a) for first group of NMF of user's skin moisture-keeping ability selection appropriate mass ratio of skin care item, institute State second group of NMF, the grease, the organic solvent, the preservative, the solubilizer, the thickener, the skin Skin conditioner, the chelating agent, the antioxidant, the polypeptide, the aromatic, the anti-sensitizer and water；

Step 5b) solubilizer, the antioxidant, the grease, the organic solvent and the aromatic are existed Hypervelocity is stirred in mulser, the mixture A of system；

Step 5c) first group of NMF be put into stirring and dissolving in agitated kettle, the mixture B of system；

Step 5d) second group of NMF, the chelating agent, the thickener are added in agitated kettle, while stirring 85-95 DEG C is warming up to, it is fully dissolved, is incubated 15-20 minutes, the mixture C of system；

Step 5e) mixture C is cooled to 40-45 DEG C, successively plus the skin conditioning agent, the anti-sensitizer and institute State polypeptide, stirring and dissolving, the mixture D of system；

Step 5f) at 40-45 DEG C of keeping temperature, add the preservative into the mixture D, the mixture A, The mixture B, it is sufficiently stirred, is down to room temperature, discharging, stands.

Further, the composition of the Moisturizer and mass ratio are：First group of NMF 3-6 parts by weight, second group NMF 4-8 parts by weight, grease 5-10 parts by weight, organic solvent 3-6 parts by weight, preservative 2-4 parts by weight, solubilizer 4-6 weights Measure part, thickener 2-4 parts by weight, skin conditioning agent 3-5 parts by weight, chelating agent 0.2-0.4 parts by weight, antioxidant 0.5-6 weights Measure part, polypeptide 0.5-2.5 parts by weight, aromatic 0.01-0.03 parts by weight, anti-sensitizer 0.5-2 parts by weight, water 50-70 weight Part.

Due to being used in upper technical scheme, the present invention has the following advantages that compared with prior art：

(1) method for customizing according to the present invention realizes that a people one is formulated, and is prepared for the specific moisture-retaining capacity of different user Moisturizer with personal exclusive formula.

(2) moisturizing related gene polymorphism and its corresponding cDNA expression quantity, which are used as, causes skin that different moisturizing shapes are presented The basic reason of state can more accurately characterize moisture-retaining capacity, of the invention according to moisturizing related gene polymorphism and its corresponding CDNA expression quantity judges moisture-retaining capacity, is obtained compared to detection methods such as graphical analysis of the prior art, facial scans Testing result it is more accurate.

(3) method for customizing according to the present invention selects Oral Mucosal Cells or skin of face cast-off cells as pattern detection The expression quantity of moisturizing related gene polymorphism and its corresponding cDNA, it is ensured that skin is from laser or the drug induced external world Stimulate, securely and reliably.

(4) according to the present invention method for customizing prepare Moisturizer use more natural animal-plant extract and polypeptide for Raw material, it is gently non-stimulated to skin, and the skin care item that Moisturizer can have other effects are used cooperatively, skin effect It is good.

(5) skin care item prepared according to the method for customizing of the present invention are directed to improving the quick ability of moisturizing of user itself, phase Than for the existing skin care item such as outside moisturizing, hyperchromic are relied on, can fundamentally solve the skin problem of user.

(6) present invention realizes new opplication of the genes such as AQP3, CLDN1, JAM1 in skin care field.

Brief description of the drawings

In specific embodiment shown in the drawings, its purpose is used to illustrate the present invention.It should be appreciated, however, that the present invention is simultaneously It is not limited to shown preferred embodiment.

In schema：

Fig. 1 is the schematic diagram according to the embodiment of Moisturizer method for customizing one of the present invention.

Embodiment

Unless otherwise defined, all meanings having in as used herein technical and scientific terms, for belonging to Technical staff is general knowledge in the technical field of the present invention.When being used herein as, following term has the meaning belonging to it, unless Separately there is special definition.

Gene expression (geneexpression) " refers to cell in life process, believes heredity in DNA sequence is stored in Breath passes through transcription and translation, is transformed into the protein molecule with bioactivity.Same gene can generate difference in different tissues Gene outcome derive from the albuminoids of different tissues, can be encoded and produced by same gene, this phenomenon be firstly because Enhancer in gene etc. is specific in a organized way, and it can be combined with the organizing specific factor in different tissues, therefore in different tissues Middle same gene can produce different transcripts and transcription post-processing effect.

" gene pleiomorphism " refers to some specific genetic locus (gene order or non-base in a colony with appropriate frequency Because of sequence) two or more phenomenon to make a variation occurs, it can be determined by Direct Analysis DNA or gene outcome.From essence On say, the generation of polymorphism is the variation on gene level, typically occur in gene order the not region of encoding proteins and There is no the region of important regulatory function.

" polymerase chain reaction (reversetranscription-polymerasechainreaction, RT-PCR) " is The technology that the polymerase chain amplification (PCR) of RNA reverse transcription (reversetranscription) and cDNA is combined.It is first The first effect through reverse transcriptase synthesizes cDNA from RNA, then using cDNA as template, amplification synthesis purpose fragment.RT-PCR technology spirit Quick and widely used, available for gene expression dose in detection cell, the content of RNA virus and Direct Cloning are specific in cell The cDNA sequence of gene.RNA as template can be the RNA products of total serum IgE, mRNA or in-vitro transcription.No matter which kind of is used RNA, it is important to ensure the pollution without RNase and genomic DNA in RNA.This method utilizes correlation technique extraction tissue or cell In total serum IgE, using mRNA therein as template, using Oligo (dT) or random primer using reverse transcriptase reverse transcription into cDNA.Expanded again using cDNA as template through performing PCR, and obtain target gene or detection gene expression.Real-time PCR (real- TimePCR), belong to quantitative PCR (Q-PCR) one kind, quantifying for DNA is carried out based on the amount of amplification of DNA in certain time Analysis.

As shown in figure 1, the skin care item method for customizing based on genetic test according to the present invention mainly comprises the steps of：

Step 1, user's cell sample is gathered.The cell sample can make Buccal mucosa cell, skin of face come off Cell or other can extract the cell of user's gene.

In an embodiment of the present invention, the specific steps for gathering Buccal mucosa cell include preparation before (1) collection：Clearly Water is gargled, it should be noted that in order to ensure that cell sample is not contaminated, gathers the half an hour before Oral Mucosal Cells, to the greatest extent Amount not drink the drink containing caffeine, the beverage containing carbonic acid or fruit juice etc.；(2) Buccal mucosa cell is gathered： Buccal swab packaging is opened, swallowing act is first done, buccal swab is put in into oral cavity afterwards, examination 20 is scraped back and forth on the inside of cheek It is secondary, buccal swab is taken out, and mark is carried out, genomic DNA is extracted immediately or is put in added with 0.5ml buccal swab genes Group DNA preservation solution (win, article No. by hundred Aurions：BTN80802 preserved in 1.5ml centrifuge tubes).

In another embodiment of the invention, the specific steps for gathering facial skin exfoliation cell include standard before (1) collection It is standby：Clear water washes one's face, it should be noted that in order to ensure that cell sample is not contaminated, testee sheds all make with makeup remover Cosmetics remain；(2) collection face comes off Skin Cell：Wait face dry after, with cell Adhesive-fetch device face repeatedly The viscous Skin Cell (if the hypocellularity of collection, several Adhesive-fetch devices collection cells can be used) for taking face.Glue after taking Properly preserve, avoid polluting.Pay attention to carrying out mark, prevent from obscuring sample.

Step 2, the mRNA in cell sample is extracted.Embodiments of the invention are using QIAGEN companies RNeasyPlusMicroKit (article No.s：74034) Total RNAs extraction is carried out, is comprised the following steps that：

A) Buccal mucosa cell sample is directed to, buccal swab is put into RNase-free 1.5ml centrifuge tubes, cut Buccal swab shank is removed, stays absorbent cotton head in centrifuge tube；For skin of face cast-off cells sample, with tweezers by cell Adhesive-fetch device On be stained with the glued membrane of Skin Cell and torn, be put into RNase-free 1.5ml centrifuge tubes and (pay attention to：Each sample uses one Individual tweezers, avoid the pollution between sample).350 μ l RLT buffer solution cell lysis is added, vortex 1min is mixed.

B) cell pyrolysis liquid in previous step is added in the centrifugal column for the removal gDNA that kit provides,>10,000rpm Centrifuge 30s.Centrifugal column is lost, retains filtered solution.

C) 70% ethanol that isometric (being usually 350 μ l) is added in filtered solution mixes.

D) sample (precipitation for including formation) of previous step is transferred to the RNeasyMinElute that is provided in kit from In stem,>10,000rpm centrifugation 15s, abandon filtered solution.

E) 700 μ l buffer solution RW1 are added into RNeasyMinElute centrifugal columns,>10,000rpm centrifugation 15s, abandon filtration Liquid.

F) 500 μ l buffer solution RPE are added into RNeasyMinElute centrifugal columns,>10,000rpm centrifugation 15s, abandon filtration Liquid.

G) 500 μ l80% ethanol are added into RNeasyMinElute centrifugal columns,>10,000rpm centrifugation 2min, abandon filtration Liquid.

H) RNeasyMinElute centrifugal columns are transferred in the new 2ml collecting pipes that kit provides, maximum turn of uncapping Speed centrifugation 5min.Abandon collecting pipe and filtered solution therein.

I) RNeasyMinElute centrifugal columns are transferred in the new 1.5ml centrifuge tubes that kit provides, to centrifugal column Center adds 14 μ l RNase-freewater, and after standing 2min, maximum speed centrifugation 1min, interior centrifuge tube is extraction RNA。

J) NanoDrop (Thermo) spectrophotometric determinations RNA concentration and quality (A260/A280) is used, is taken immediately 1 μ gRNA carry out reverse transcription or by the RNA of extraction in -80 DEG C of preservations.

Step 3, mRNA reverse transcriptions are synthesized into cDNA.Embodiments of the invention are using TOYOBO companies ReverTraAceqPCRRTMasterMixwithgDNARemover (article No.s：FSQ-301 cDNA synthesis) is carried out.Specific behaviour It is as follows to make step：

A) 4xDNMasterMix and gDNARemover mixing：Added in homogeneous tube 4xDNMasterMix (440 μ l) 8.8 μ l (1/50 amount) gDNARemover, overturn and mix.The 4xDNMasterMix after gDNARemover is added in -20 DEG C of bars The stabilization of 3 months can be at least kept under part.

B) RNA denaturation：1 μ gRNA after thermal denaturation 5 minutes, are immediately placed on cooled on ice under the conditions of 65 DEG C.

C) genomic DNA reaction (DNase reactions) is removed：Wherein, reaction solution is formulated as follows on ice.

After reaction solution is lightly stirred, incubated 5 minutes under the conditions of 37 DEG C.

D) reverse transcription reaction：Wherein, reaction solution is formulated as follows on ice.

After reaction solution is lightly stirred, reacted by temperature below.

After reaction terminates, preserved under the conditions of -20 DEG C.During Real-timeRTPCR, added after diluting 10 times as template.

Step 4, the polymorphism of moisturizing related gene and corresponding cDNA expression quantity in cDNA are detected.

In an embodiment of the present invention, moisturizing related gene can include one kind in the genes such as MAQP3, CLDN1, JAM1 Or several combination.

AQP3 (aquaporin 3) is moisturizing related gene, and it is located at Chromosome 9 9q13.3 positions.AQP3 genes Total length 36.46kb, share 6 extrons.AQP3 mRNA total length 1870nt, encode the egg being made up of 292 amino acid residues In vain.Gene function：Aquaporin 3 is a kind of transport protein of transdermal delivery water on cell membrane, contributes to cell quick regulation Own vol and internal penetration pressure, and maintain a key factor of skin hydration effect, i.e. the expression quantity of the gene is got over Height, illustrate that the risk of the dry water shortage of skin appearance is lower.

CLDN1 (albumen 1 of combining closely) is moisturizing related gene, and it is located at No. 3 chromosome 3q28 position, full length gene 16.77kb, share 4 extrons.CLDN1 mRNA total length 3452nt, encode the albumen being made up of 211 amino acid residues. Gene function：Claudin1 albumen is that epidermis maintains the essential structure of normal physiological function.Studies have found that Claudin Albumen is relevant with the barrier function of epidermal tissue, i.e. the application on human skin moisture-retaining capacity of the genotype is high, and skin occurs drying water shortage Risk is low.

JAM1 (link adhesion molecule 1) is moisturizing related gene, and it is located at No. 1 chromosome 1q23.3 position, and gene is complete Long 26.14kb, share 10 extrons.JAM1 mRNA total length 4835nt, encode the egg being made up of 299 amino acid residues In vain.Gene function：JAM1 is the I type memebrane proteins with transmembrane structure, is distributed mainly on epithelial cell and endothelial cell Tight junctions, main function is between mediated cell or the interaction between cell and extracellular matrix, maintain skin environment and The stabilization of organismic internal environment, i.e. the application on human skin moisture-retaining capacity of the genotype is high, and the risk that water shortage occurs drying in skin is low.

Embodiments of the invention are comprised the following steps that using the amount of real-timeRTPCR detection related genes：

A) mRNA sequence of related gene is searched on NCBI, and designs corresponding primer.

Embodiments of the invention are using gene GAPDH as internal reference, i.e. GAPDH gene expression amounts are set as 1, using delta- DeltaCt methods carry out relative quantitative assay to above-mentioned moisturizing related gene.The primer sequence of GAPDH and moisturizing related gene such as table Shown in 1：

Table 1

The amplified fragments sequence of each gene shown in table 1 is as shown in table 2：

Table 2

B) using the SYBRGreenRealtimePCRMasterMix (article No.s of TOYOBO companies：QPK-201) carry out Detection, using Roche companiesMachine testing on 96.

The preparation program of reaction solution is as shown in table 3：

Table 3

PCR cycling condition is as shown in table 4：

Table 4

C) data analysis：Solubility curve is analyzed to be unimodal, the specificity for illustrating primer very well, can be used for subsequent experimental； If solubility curve is non-unimodal, illustrate the specific bad, it is necessary to redesign primer of primer；Each sample, each pair primer enter Row repeats three times, reduces the error in operating process；Using GAPDH as internal reference, i.e. set GAPDH gene expression amounts as 1, use Delta-deltaCt methods carry out relative quantitative assay.

Step 5, the expression quantity according to moisturizing related gene judges skin moisture-keeping ability, and is directed to user's skin moisture-keeping ability Skin care item composition and dosage are selected, prepares suitable skin care item.

The effect composition of certain content can regulate and control to different types of gene expression amount in Moisturizer, so as to real Now improve the purpose of skin quality.For example, the NMF regulated and controled for the moisturizing related gene expression amount such as AQP3, CLDN1, JAM1 Comprising Sodium Hyaluronate, polyglutamic acid, tremella polysaccharides, panaxan, GL-B, propane diols, DPG, butanediol are sweet One or more in oil；Grease type includes olive oil, white oil, vaseline, paraffin oil, apricot kernel oil, saualane, macadamia nut oil, suddenly Bar oil suddenly, the one or more of Butyrospermum parkii fruit fatty oil.And polypeptide then can by adjust in skin with moisturizing gene expression phase The signal path of pass realizes the purpose of modulin expression.

Based on above-mentioned principle, the Moisturizer according to the present invention totally includes NMF, grease, polypeptide and auxiliary agent. Wherein, auxiliary agent can include antioxidant, solvent, preservative, solubilizer, thickener, skin conditioning agent, chelating agent, fragrance Agent, anti-sensitizer and water.Totally first group of NMF 3-6 parts by weight is included for the skin care item that different user is prepared, second group NMF 4-8 parts by weight, grease 5-10 parts by weight, organic solvent 3-6 parts by weight, preservative 2-4 parts by weight, solubilizer 4-6 weights Measure part, thickener 2-4 parts by weight, skin conditioning agent 3-5 parts by weight, chelating agent 0.2-0.4 parts by weight, antioxidant 0.5-6 weights Measure part, polypeptide 0.5-2.5 parts by weight, aromatic 0.01-0.03 parts by weight, anti-sensitizer 0.5-2 parts by weight, water 50-70 weight Part.Wherein,

NMF includes Sodium Hyaluronate, polyglutamic acid, tremella polysaccharides, panaxan, GL-B, propane diols, dipropyl Glycol, butanediol, the wherein one or more of glycerine, NMF are divided into first group according to the difference of order of addition in preparation process NMF and second group of NMF；

Grease includes olive oil, white oil, vaseline, paraffin oil, apricot kernel oil, saualane, macadamia nut oil, jojoba oil, butter The one or more of fruit tree fruit fatty oil；

Organic solvent includes dodecanol, hexadecanol, octadecanol, tadenan, the one or more of cetanol；

Preservative includes Nipasol, iodine propilolic alcohol butyl mephenesin Carbamate, 2- phenoxetols, one kind of methyl hydroxybenzoate It is or a variety of；

Solubilizer include stearic acid, PEG20 rilanit specials, PEG20 stearines, PEG60 rilanit specials, PEG60 iso stearic acid of glycerine ester, Natrulon H-10 stearate, the one or more of PEG/PPG-17/6 copolymers；

Thickener includes hydroxyethyl cellulose, carboxymethyl cellulose, HPG, polymeric acrylic acid esters thickener One or more；

Skin conditioning agent includes Gotu Kola P.E, lycopene, ceramide, lecithin, propolis extract, yellow chamomile Extract, ginseng root extract, yeast extract, the one or more of houttuynia extract；

Chelating agent includes EDETATE SODIUM；

It is more that antioxidant includes brown alga extract, ginseng extract, panaxoside, nutgall extractive, Tea Polyphenols, ganoderma lucidum Sugar, algal polysaccharides, raspberry extract, resveratrol, avocado oil, grape-kernel oil, astaxanthin, forulic acid, Co-Q10, N- acetyl Cysteine, curcumin, tanshinone IIA, glabridin, the one or more of lipoic acid；

Anti-sensitizer includes dipotassium glycyrrhizinate, hanfangchin A, camomile extract, one kind in Gotu Kola P.E or more Kind；

Aromatic includes the one or more in yulan water extract, aloe water extract, rose water extract；

Polypeptide, the purpose of modulin expression, bag are realized by adjusting signal path related to gene expression in skin Containing alanine/histidine/lysine polypeptide copper HCl, soybean (GLYCINEMAX) polypeptide, dipeptides -1, dipeptides -2, dipeptides -4, widow Peptide -1, oligopeptides -2, oligopeptides -3, oligopeptides -4, oligopeptides -5, oligopeptides -6, oligopeptides -29, oligopeptides -34, carnosine, saccharomycete polypeptide, essence Propylhomoserin/lysine polypeptide, nonapeptide -1, poly- (tripeptides -6), ascorbyl polypeptide, sodium bacillus subtilis lipopeptide, hexapeptide -1,2,3,5,9, 11st, heptapeptide -6, myristoyl hexapeptide -5, myristoyl pentapeptide -4, tripeptides -1,2,3,10,32, the copper of tripeptides -1, decapeptide -4, four Peptide -1,3,4, pentapeptide -1,3,34, oat peptide, Wild soybean peptide, acetyl group octapeptide -3, the cetyl of acetyl group dipeptides -1, acetyl group six Peptide 1,7,8, acetyl group heptapeptide -4, acetyl group tetrapeptide 2,3,5,9,11, palmityl dipeptides -7, palmityl hexapeptide -12,14,15, palm fibre Palmitic acid acyl tripeptides -1,5,8, palmityl tetrapeptide -5,7,10, one or more of combinations in Matrixyl -4,5.

The preparation method of skin care item totally comprises the steps of：

Step 5a) for the NMF of user's skin moisture-keeping ability selection appropriate mass ratio of skin care item, grease, You Jirong Agent, preservative, solubilizer, thickener, skin conditioning agent, chelating agent, antioxidant, polypeptide, aromatic, anti-sensitizer and Water；

Step 5b) solubilizer, antioxidant, grease, organic solvent and aromatic exceed the speed limit in mulser to be stirred, The mixture A of system；

Step 5c) first group of NMF be put into stirring and dissolving in agitated kettle, the mixture B of system；

Step 5d) second group of NMF, chelating agent, thickener are added in agitated kettle, warming while stirring to 85-95 DEG C, it is fully dissolved, be incubated 15-20 minutes, the mixture C of system；

Step 5e) mixture C is cooled to 40-45 DEG C, successively plus skin conditioning agent, anti-sensitizer and polypeptide, stirring and dissolving, The mixture D of system；

Step 5f) at 40-45 DEG C of keeping temperature, add preservative into mixture D, mixture A, mixture B, fully Stirring, room temperature is down to, discharging, is stood.

Embodiment 1

The embodiment of the present invention 1 for 39 years old volunteer 1 carry out genetic test, moisturizing related gene AQP3, CLDN1, JAM1.According to testing result the skin care item of formula are formulated as follows for it：Wherein, skin care item gross mass is common 50Kg,

First group of NMF：Polyglutamic acid 1Kg, tremella polysaccharides 1Kg

Second group of NMF：DPG 1Kg, glycerine 1Kg

Grease：Saualane 1Kg, macadamia nut oil 1Kg, jojoba oil 1Kg, Butyrospermum parkii fruit fatty oil 1Kg

Organic solvent：Hexadecanol 1Kg, octadecanol 2Kg,

Preservative：2- phenoxetols 2Kg

Solubilizer：PEG20 stearines 1Kg, PEG60 rilanit special 1Kg,

Thickener：Polymeric acrylic acid esters (Acritamer 940) 1Kg

Skin conditioning agent：Propolis extract 1Kg, chamomile extract 0.5Kg

Chelating agent：EDETATE SODIUM 0.1Kg

Antioxidant：Grape-kernel oil 1Kg, curcumin 1Kg, Co-Q10 0.6Kg

Polypeptide：Carnosine 1Kg, Matrixyl 0.2Kg

Anti-sensitizer：Dipotassium glycyrrhizinate 1Kg

Aromatic：Yulan water extract 0.01Kg

Other：Water surplus

Volunteer 1 continues to carry out genetic test again after the skin care item using above-mentioned formula in 100 days, and it uses above-mentioned skin care Gene relative expression quantity before and after product is as shown in table 5：

Table 5

Gene name	AQP3	CLDN1	JAM1	GAPDH
					Before use	0.329877	0.990801	1.023374	1
After use	0.351111	1.071773	1.122462	1
					Reference example	0.536127	1.569819	1.326094	1

Wherein, embodiment 1-4 reference example is 100 volunteers each gene relative expression quantity of the age at -30 years old 20 years old Average value.

The expression quantity of gene shows shown in table 5, volunteer 1 using Gene A QP3, CLDN1 before Moisturizer, JAM1 expression quantity is relatively low, skin moisture-keeping ability, easily dry and cracked.After anti-aging skin care product using above-mentioned formula, Gene A QP3, CLDN1, JAM1 expression quantity substantially reduce, and correspondingly, moisture-retaining capacity increases.

Embodiment 2

The embodiment of the present invention 2 for 46 years old volunteer 2 carry out genetic test, moisturizing related gene AQP3, CLDN1, JAM1.According to testing result the skin care item of formula are formulated as follows for it：Wherein, skin care item gross mass is common 50Kg,

First group of NMF：Sodium Hyaluronate 0.5Kg, polyglutamic acid 0.5Kg, tremella polysaccharides 0.5Kg

Second group of NMF：Propane diols 3Kg, butanediol 1Kg

Grease：Apricot kernel oil 3Kg, saualane 1Kg, macadamia nut oil 0.5Kg,

Organic solvent：Dodecanol 1Kg, tadenan 0.5Kg

Preservative：2- phenoxetols 1.2Kg

Solubilizer：PEG60 rilanit specials 0.6Kg, PEG60 iso stearic acid of glycerine ester 1.4Kg

Thickener：HPG 1Kg

Skin conditioning agent：Ginseng root extract 1Kg, yeast extract 0.5Kg, propolis extract 0.5Kg

Chelating agent：EDETATE SODIUM 0.1Kg

Antioxidant：Ginseng extract 0.6Kg, panaxoside 0.5Kg, curcumin 0.1Kg

Polypeptide：The 0.1Kg of oligopeptides -5, the 0.1Kg of pentapeptide -1, the 0.1Kg of palmityl tetrapeptide -5

Anti-sensitizer：Dipotassium glycyrrhizinate 0.1Kg

Aromatic：Rose water extract 0.01Kg

Other：Water surplus

Volunteer 2 continues to carry out genetic test again after the skin care item using above-mentioned formula in 100 days, and it uses above-mentioned skin care Gene relative expression quantity before and after product is as shown in table 6：

Table 6

Gene name	AQP3	CLDN1	JAM1	GAPDH
					Before use	0.502316	1.319508	1.025741	1
After use	0.55421	1.53621	1.12242	1
					Reference example	0.536127	1.569819	1.326094	1

The expression quantity of gene shows shown in table 6, volunteer 1 using Gene A QP3, CLDN1 before Moisturizer, The landing of JAM1 expression quantity is low, and skin moisture-keeping ability is poor, easily dry and cracked.After anti-aging skin care product using above-mentioned formula, gene AQP3, CLDN1, JAM1 expression quantity substantially reduce, and correspondingly, moisture-retaining capacity increases.

Embodiment 3

The embodiment of the present invention 3 carried out genetic test, moisturizing related gene AQP3, CLDN1 for the volunteer 3 of 45 years old.According to According to testing result the skin care item of formula are formulated as follows for it：Wherein, skin care item gross mass is common 50Kg,

First group of NMF：Tremella polysaccharides 1Kg, panaxan 1Kg, GL-B 1Kg

Second group of NMF：DPG 1Kg, glycerine 3Kg

Grease：Jojoba oil 1Kg, Butyrospermum parkii fruit fatty oil 1Kg, apricot kernel oil 1Kg

Organic solvent：Hexadecanol 1Kg, octadecanol 0.4Kg, tadenan 1.2Kg

Preservative：Nipasol 1Kg, methyl hydroxybenzoate 1Kg

Solubilizer：Natrulon H-10 stearate 1Kg, PEG/PPG-17/6 copolymer 1 Kg

Thickener：Polymeric acrylic acid esters thickener 1Kg

Skin conditioning agent：Lecithin 1Kg, propolis extract 1Kg

Chelating agent：EDETATE SODIUM 0.1Kg

Antioxidant：Astaxanthin 0.05Kg, forulic acid 0.5Kg, Co-Q10 0.2Kg, N-acetylcystein 0.3Kg

Polypeptide：The 0.2Kg of myristoyl pentapeptide -4, the 0.2Kg of tripeptides -2, the 0.2Kg anti-sensitizers of acetyl group hexapeptide 8：Chamomile Extract 0.2Kg, Gotu Kola P.E 0.2Kg

Aromatic：Rose water extract 0.01Kg

Other：Water surplus

Volunteer 3 continues to carry out genetic test again after the skin care item using above-mentioned formula in 100 days, and it uses above-mentioned skin care Gene relative expression quantity before and after product is as shown in table 7：

Table 7

Gene name	AQP3	CLDN1	GAPDH
				Before use	0.331405	1.200249	1
After use	0.551421	1.43521	1
				Reference example	0.536127	1.569819	1

The expression quantity of gene shows shown in table 7, volunteer 1 using Gene A QP3, CLDN1 before Moisturizer, JAM1 expression quantity is relatively low, skin moisture-keeping ability, easily dry and cracked.After anti-aging skin care product using above-mentioned formula, Gene A QP3, CLDN1, JAM1 expression quantity substantially reduce, and correspondingly, moisture-retaining capacity increases.

Embodiment 4

The embodiment of the present invention 4 carried out genetic test, moisturizing related gene AQP3, CLDN1 for the volunteer 4 of 41 years old.According to According to testing result the skin care item of formula are formulated as follows for it：Wherein, skin care item gross mass is common 50Kg,

First group of NMF：Polyglutamic acid 1Kg, tremella polysaccharides 0.5Kg, panaxan 0.5Kg

Second group of NMF：DPG 1Kg, butanediol 0.5Kg, glycerine 2Kg

Grease：Paraffin oil 1Kg, apricot kernel oil 1Kg, macadamia nut oil 0.2Kg, jojoba oil 0.4Kg

Organic solvent：Hexadecanol 1.3Kg, octadecanol 1.7Kg

Preservative：2- phenoxetols 1.4Kg

Solubilizer：PEG20 rilanit specials 1.2Kg, PEG60 iso stearic acid of glycerine ester 1.4Kg

Thickener：HPG 1.7Kg

Skin conditioning agent：Cer EOS Kg, lecithin 1Kg, propolis extract 0.5Kg

Chelating agent：EDETATE SODIUM 0.2Kg

Antioxidant：Tea Polyphenols 0.2Kg, brown alga extract 0.1Kg, Co-Q10 0.05Kg, curcumin 0.05Kg, the red sage root Ketone IIA0.02Kg

Polypeptide：The 0.1Kg of acetyl group tetrapeptide 11, the 0.1Kg of decapeptide -4, the 0.1Kg of Matrixyl -4

Anti-sensitizer：Dipotassium glycyrrhizinate 0.8Kg, hanfangchin A 0.1Kg

Aromatic：Yulan water extract 0.015Kg

Other：Water surplus

Volunteer 4 continues to carry out genetic test again after the skin care item using above-mentioned formula in 100 days, and it uses above-mentioned skin care Gene relative expression quantity before and after product is as shown in table 8：

Table 8

The expression quantity of gene shows shown in table 8, volunteer 1 using Gene A QP3, CLDN1 before Moisturizer, JAM1 expression quantity is relatively low, skin moisture-keeping ability, easily dry and cracked.After anti-aging skin care product using above-mentioned formula, Gene A QP3, CLDN1, JAM1 expression quantity substantially reduce, and correspondingly, moisture-retaining capacity increases.

For the art, personnel can understand, above-mentioned specific embodiment can be in the generalized concept without departing substantially from the present invention Change down.Therefore, it should be apparent that the present invention is not limited to disclosed certain specific embodiments, but this is intended to comprising belonging to this Various change in the spirit and scope of invention, i.e., such as appended claim.

SEQUENCE LISTING

<110>Du Libo

<120>A kind of Moisturizer method for customizing

<130> 1

<160> 12

<170> PatentIn version 3.5

<210> 1

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223>GAPDH primers 1

<400> 1

cgggaaactg tggcgtgatg 20

<210> 2

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223>GAPDH primer 2s

<400> 2

atgaccttgc ccacagcctt 20

<210> 11

<211> 18

<212> DNA

<213> Artificial Sequence

<220>

<223>AQP3 primers 1

<400> 11

cctggaggcc ttcaccgt 18

<210> 12

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223>AQP3 primer 2s

<400> 12

ttgacggcat agccggagtt 20

<210> 13

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223>CLDN1 primers 1

<400> 13

ctgtggatgt cctgcgtgtc 20

<210> 14

<211> 23

<212> DNA

<213> Artificial Sequence

<220>

<223>Primer 2

<400> 14

tgctcagatt cagcaaggag tca 23

<210> 15

<211> 22

<212> DNA

<213> Artificial Sequence

<220>

<223>JAM1 primers 1

<400> 15

tgtctgttcc caggagtcct tc 22

<210> 16

<211> 20

<212> DNA

<213> Artificial Sequence

<220>

<223>JAM1 primer 2s

<400> 16

ctcgacttgc gcctttgtcc 20

<210> 9

<211> 87

<212> DNA

<213> Artificial Sequence

<220>

<223>GAPDH amplified fragments

<400> 9

cgggaaactg tggcgtgatg gccgcggggc tctccagaac atcatccctg cctctactgg 60

cgctgccaag gctgtgggca aggtcat 87

<210> 10

<211> 78

<212> DNA

<213> Artificial Sequence

<220>

<223>AQP3 amplified fragments

<400> 10

cctggaggcc ttcaccgtgg gcctggtggt cctggtcatt ggcacctcca tgggcttcaa 60

ctccggctat gccgtcaa 78

<210> 11

<211> 76

<212> DNA

<213> Artificial Sequence

<220>

<223>CLDN1 amplified fragments

<400> 11

ctgtggatgt cctgcgtgtc gcagagcacc gggcagatcc agtgcaaagt ctttgactcc 60

ttgctgaatc tgagca 76

<210> 12

<211> 76

<212> DNA

<213> Artificial Sequence

<220>

<223>JAM1 amplified fragments

<400> 12

tgtctgttcc caggagtcct tcggcggctg ttgtgtcggg agcctgatcg cgatggggac 60

aaaggcgcaa gtcgag 76

Claims (10)

1. a kind of Moisturizer method for customizing, it is characterised in that comprise the steps of：

Step 1, user's cell sample is gathered；

Step 2, the mRNA in the cell sample is extracted；

Step 3, mRNA reverse transcriptions are synthesized into cDNA；

Step 4, the polymorphism of moisturizing related gene and corresponding cDNA expression quantity in cDNA are detected, wherein, the moisturizing is related Gene includes at least one of AQP3, CLDN1, JAM1；

Step 5, the expression quantity according to the gene pleiomorphism and corresponding cDNA judges moisture-retaining capacity, and is directed to the moisture-retaining capacity Selection component and dosage, prepare suitable Moisturizer.

2. Moisturizer method for customizing as claimed in claim 1, it is characterised in that the step 4 includes：

Step 4a), the mRNA sequence of the moisturizing related gene is searched on NCBI, and design corresponding primer；

Step 4b), using delta-deltaCt methods, using gene GAPDH as internal reference, pass through moisturizing phase described in the primer detection The relative expression quantity of the polymorphism of correlation gene and corresponding cDNA.

3. Moisturizer method for customizing as claimed in claim 2, it is characterised in that the primer includes：

GAPDH Forword：5’-CGGGAAACTGTGGCGTGATG-3’

Reverse：5’-ATGACCTTGCCCACAGCCTT-3’

AQP3 Forword：5’-CCTGGAGGCCTTCACCGT-3’

Reverse：5’-TTGACGGCATAGCCGGAGTT-3’

CLDN1 Forword：5’-CTGTGGATGTCCTGCGTGTC-3’

Reverse：5’-TGCTCAGATTCAGCAAGGAGTCA-3’

JAM1 Forword：5’-TGTCTGTTCCCAGGAGTCCTTC-3’

Reverse：5’-CTCGACTTGCGCCTTTGTCC-3’.

4. Moisturizer method for customizing as claimed in claim 1, it is characterised in that the composition of the Moisturizer includes NMF, grease, polypeptide and auxiliary agent.

5. Moisturizer method for customizing as claimed in claim 4, it is characterised in that the NMF includes first group of moisturizing Agent and second group of NMF, first group of NMF include bright matter acid sodium, polyglutamic acid, tremella polysaccharides, panaxan, ganoderma lucidum Polysaccharide, second group of NMF include propane diols, DPG, butanediol, glycerine.

6. Moisturizer method for customizing as claimed in claim 4, it is characterised in that the grease includes olive oil, white oil, Vaseline, paraffin oil, apricot kernel oil, saualane, macadamia nut oil, jojoba oil, the one or more of Butyrospermum parkii fruit fatty oil.

7. Moisturizer method for customizing as claimed in claim 4, it is characterised in that the polypeptide includes alanine/group ammonia Acid/lysine polypeptide copper HCl, soybean (GLYCINEMAX) polypeptide, dipeptides -1, dipeptides -2, dipeptides -4, oligopeptides -1, oligopeptides -2, widow Peptide -3, oligopeptides -4, oligopeptides -5, oligopeptides -6, oligopeptides -29, oligopeptides -34, carnosine, saccharomycete polypeptide, arginine/lysine are more Peptide, nonapeptide -1, poly- (tripeptides -6), ascorbyl polypeptide, sodium bacillus subtilis lipopeptide, hexapeptide -1,2,3,5,9,11, heptapeptide -6, Pork and beans Cool acyl hexapeptide -5, myristoyl pentapeptide -4, tripeptides -1,2,3,10,32, the copper of tripeptides -1, decapeptide -4, tetrapeptide -1,3,4, pentapeptide -1, 3,34, oat peptide, Wild soybean peptide, acetyl group octapeptide -3, the cetyl of acetyl group dipeptides -1, acetyl group hexapeptide 1,7,8, acetyl group seven Peptide -4, acetyl group tetrapeptide 2,3,5,9,11, palmityl dipeptides -7, palmityl hexapeptide -12,14,15, palmityl tripeptides -1,5,8, Palmityl tetrapeptide -5,7,10, one or more of combinations in Matrixyl -4,5.

8. Moisturizer method for customizing as claimed in claim 4, it is characterised in that the auxiliary agent includes antioxidant, machine Solvent, preservative, solubilizer, thickener, skin conditioning agent, chelating agent, aromatic, anti-sensitizer and water.

9. Moisturizer method for customizing as claimed in claim 8, it is characterised in that the step 5 includes：

Step 5a) for first group of NMF of user's skin moisture-keeping ability selection appropriate mass ratio of skin care item, described the Two groups of NMFs, the grease, the organic solvent, the preservative, the solubilizer, the thickener, the skin are adjusted Manage agent, the chelating agent, the antioxidant, the polypeptide, the aromatic, the anti-sensitizer and water；

Step 5b) solubilizer, the antioxidant, the grease, the organic solvent and the aromatic are being emulsified Hypervelocity is stirred in machine, the mixture A of system；

Step 5c) first group of NMF be put into stirring and dissolving in agitated kettle, the mixture B of system；

Step 5d) second group of NMF, the chelating agent, the thickener are added in agitated kettle, warming while stirring To 85-95 DEG C, it is fully dissolved, be incubated 15-20 minutes, the mixture C of system；

Step 5e) mixture C is cooled to 40-45 DEG C, successively plus the skin conditioning agent, the anti-sensitizer and described more Peptide, stirring and dissolving, the mixture D of system；

Step 5f) at 40-45 DEG C of keeping temperature, add the preservative into the mixture D, it is the mixture A, described Mixture B, it is sufficiently stirred, is down to room temperature, discharging, stands.

10. Moisturizer method for customizing as claimed in claim 9, it is characterised in that the composition of the Moisturizer and Mass ratio is：First group of NMF 3-6 parts by weight, second group of NMF 4-8 parts by weight, grease 5-10 parts by weight, organic solvent 3-6 parts by weight, preservative 2-4 parts by weight, solubilizer 4-6 parts by weight, thickener 2-4 parts by weight, skin conditioning agent 3-5 weight Part, chelating agent 0.2-0.4 parts by weight, antioxidant 0.5-6 parts by weight, polypeptide 0.5-2.5 parts by weight, aromatic 0.01- 0.03 parts by weight, anti-sensitizer 0.5-2 parts by weight, water 50-70 parts by weight.