CN104777306A - Blood flow volume evaluation or indirect detection method - Google Patents
Blood flow volume evaluation or indirect detection method Download PDFInfo
- Publication number
- CN104777306A CN104777306A CN201410563850.2A CN201410563850A CN104777306A CN 104777306 A CN104777306 A CN 104777306A CN 201410563850 A CN201410563850 A CN 201410563850A CN 104777306 A CN104777306 A CN 104777306A
- Authority
- CN
- China
- Prior art keywords
- interleukin
- expression
- blood flow
- alpha
- receptor antagonist
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 230000017531 blood circulation Effects 0.000 title claims abstract description 49
- 238000011156 evaluation Methods 0.000 title claims abstract description 18
- 238000001514 detection method Methods 0.000 title abstract 2
- 101001076407 Homo sapiens Interleukin-1 receptor antagonist protein Proteins 0.000 claims abstract description 55
- 229940119178 Interleukin 1 receptor antagonist Drugs 0.000 claims abstract description 55
- 102000051628 Interleukin-1 receptor antagonist Human genes 0.000 claims abstract description 55
- 239000003407 interleukin 1 receptor blocking agent Substances 0.000 claims abstract description 55
- 238000000034 method Methods 0.000 claims abstract description 31
- 108010082786 Interleukin-1alpha Proteins 0.000 claims description 50
- 102000004125 Interleukin-1alpha Human genes 0.000 claims description 50
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 16
- 208000001126 Keratosis Diseases 0.000 claims description 16
- 238000010586 diagram Methods 0.000 claims description 12
- 238000009826 distribution Methods 0.000 claims description 9
- 102000000589 Interleukin-1 Human genes 0.000 abstract description 3
- 108010002352 Interleukin-1 Proteins 0.000 abstract description 3
- 230000008326 skin blood flow Effects 0.000 abstract description 2
- 238000000691 measurement method Methods 0.000 abstract 1
- 238000004140 cleaning Methods 0.000 description 8
- 238000005259 measurement Methods 0.000 description 8
- 238000012360 testing method Methods 0.000 description 7
- 238000008157 ELISA kit Methods 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 101001002634 Homo sapiens Interleukin-1 alpha Proteins 0.000 description 3
- 239000002390 adhesive tape Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000013467 fragmentation Methods 0.000 description 3
- 238000006062 fragmentation reaction Methods 0.000 description 3
- 210000002540 macrophage Anatomy 0.000 description 3
- 238000000035 BCA protein assay Methods 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 102000000380 Matrix Metalloproteinase 1 Human genes 0.000 description 2
- 108010016113 Matrix Metalloproteinase 1 Proteins 0.000 description 2
- 230000008485 antagonism Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000003593 chromogenic compound Substances 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 238000002306 biochemical method Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000011536 extraction buffer Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000012447 hatching Effects 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
- G01N33/6869—Interleukin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
- A61B10/02—Instruments for taking cell samples or for biopsy
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Physics & Mathematics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Heart & Thoracic Surgery (AREA)
- Medical Informatics (AREA)
- Surgery (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a novel measurement/evaluation method which is not a method that a skin is obtained through a surgical operation, and the skin is stimulated, and can measure and evaluate the blood flow state simply and assuredly by means of biochemical indexes. A skin blood flow volume indirect detection/evaluation method is characterized in that the expression quantity of interleukin-1 receptor antagonist and/or interleukin-1[alpha] in the skin cuticle serves as the index.
Description
Technical field
The present invention relates to assessment of levels or the indirect determination method of skin volume of blood flow.
Background technology
As one of the reason of skin ageing phenomenon, the reduction of the ruddy degree of the colour of skin that poor circulation causes can be enumerated.The reduction phenomenon of the ruddy degree of this skin occurs as " skin is obscure " when obvious, is by the phenomenon being difficult to make up of making up.Secondly, the blood flow by improving skin can being said, can expect to improve the transparency and lightness and promoting metabolic activity, the active cell of skin histology, as its result, can tackle based on the reason of the various catabiosis of the skin of obscure reason.
Because volume of blood flow increases, metabolism is promoted thus remove refuse in skin tissue.Such as, develop by irradiate comprise according to chemiluminescent the technology (patent documentation 1:WO2005/077458 publication) that near infrared light improves skin blood flow.In patent documentation 1, use laser Doppler flowmetry to measure the change of skin volume of blood flow.
Like this, in order to evaluate the health status of skin, need the volume of blood flow measuring skin.Same with described laser-Doppler formula blood flowmeter, the multi-channel type blood flowmeter (patent documentation 2: Jap.P. No. 3783491 publication) of use laser, adding pressure type blood flowmeter (patent documentation 3: Japanese Unexamined Patent Publication 11-285476 publication) are also used to the measuring of blood flow of skin.But, need the direct blood flowmeter to measure experimenter when measuring, in order to measure the skin volume of blood flow of distant place experimenter, unless determinator to be arranged on the setting place that the residential neighborhood of experimenter or experimenter are directly on loan to determinator, otherwise just can not measure.
Prior art document
Patent documentation
Patent documentation 1WO2005/077458 publication
Patent documentation 2 Jap.P. No. 3783491 publication
Patent documentation 3 Japanese Unexamined Patent Publication 11-285476 publication
Summary of the invention
As mentioned above, in order to the blood flow state of distant place experimenter indirectly can be evaluated, expect to use the cuticula gathered from experimenter indirectly measure or evaluate volume of blood flow.
It is not that surgery is won skin and skin brought to the method for stimulation etc. that problem of the present invention is to provide, but can be easy and positively measure or evaluate the mensuration of novelty or the evaluation method of the use biochemical index of blood flow state.
The present invention is following formation.
(1) indirect determination of DBF or an evaluation method, is characterized in that, using the expression of the interleukin-1 receptor antagonist in keratoderma and/or interleukin-1 alpha as index.
(2) method Gen Ju (1), is characterized in that, for measure or the keratoderma of evaluation method is gathered by tape stripping method.
(3) a kind of method, it is characterized in that, the relational expression made from the correlativity of the expression based on the interleukin-1 receptor antagonist the volume of blood flow measured in advance and keratoderma and/or interleukin-1 alpha carrys out the volume of blood flow of indirect determination skin.
(4) a kind of Indirect Determination of DBF, it is characterized in that, be made up of following operation: gather cuticular collection operation, measure the interleukin-1 receptor antagonist in the cuticula gathered in described collection operation, the mensuration operation of the expression of interleukin-1 alpha and the interleukin-1 receptor antagonist will measured in described mensuration operation, the expression of interleukin-1 alpha is converted into the operation of DBF by following regression equation, described regression equation is from the interleukin-1 receptor antagonist the collective's cuticula measured in advance, what the expression distribution of interleukin-1 alpha was obtained obtains the relevant regression equation of volume of blood flow.
(5) a kind of evaluation method of DBF, it is characterized in that, the scatter diagram made with the correlativity of the expression based on the interleukin-1 receptor antagonist in the volume of blood flow measured in advance and keratoderma and/or interleukin-1 alpha carries out contrasting evaluating.
(6) a kind of evaluation method of SkBF, it is characterized in that, be made up of following operation: gather cuticular collection operation, measure the cuticula that gathers in described collection operation in interleukin-1 receptor antagonist, the mensuration operation of expression of interleukin-1 alpha and the contrast operation carrying out contrasting that the expression of the interleukin-1 receptor antagonist measured in described mensuration operation, interleukin-1 alpha and the expression of the interleukin-1 receptor antagonist in the collective's cuticula measured in advance, interleukin-1 alpha are distributed.
According to method of the present invention, mensuration or the expression evaluated by the interleukin-1 receptor antagonist in mensuration cuticula or interleukin-1 alpha of SkBF indirectly measure or evaluate.
In addition, because method of the present invention gathers evaluation sample by easy methods of operating such as tape stripping methods, so the burden of experimenter is few, no matter who all can be evaluated simply.
Owing to particularly using the sample gathered from distant place experimenter, such as, can the easy state indirectly evaluating skin volume of blood flow or blood flow by the cuticula of tape stripping method, so do not need experimenter directly to carry out test site.Like this, become the applied widely of blood flow determination object, can diagnose easily or the state of evaluating skin.
And then, do not need directly to measure experimenter with the blood flowmeter, even if as long as just can measure easily or evaluating skin volume of blood flow for the experimenter in a distant place can obtain cuticula.
And then, the effect of skin state, cosmetics etc. can be evaluated objectively according to method of the present invention.
Accompanying drawing explanation
The measurement result of the interleukin-1 receptor antagonist (IL-1RA) in keratoderma and the relative output value proportional with DBF (relative output amount) (V) is made scatter diagram and has obtained the figure of regression straight line by Fig. 1.
The measurement result of the interleukin-1 alpha (IL-1 α) in keratoderma and the relative output value (V) proportional with DBF is made scatter diagram and has obtained the figure of regression straight line by Fig. 2.
Embodiment
The present invention relates to the indirect determination of DBF or the invention of evaluation method, it is characterized in that, using the expression of the interleukin-1 receptor antagonist in keratoderma and/or interleukin-1 alpha as index.
Below, be described in detail about the present invention.
The mensuration of volume of blood flow of the present invention or the feature of evaluation method are, (below, are called IL-1RA with the interleukin-1 receptor antagonist in keratoderma.) or interleukin-1 alpha (following, be called IL-1 α.) expression as index.
Volume of blood flow in the present invention refers to the volume of blood flow measured with Laser Doppler flowmetry in fixed capillary of skin.It is relevant to the expression of IL-1RA or the IL-1 α in keratoderma that discovery Laser Doppler flowmetry measures fixed volume of blood flow.Therefore, by measuring the expression in the keratoderma of IL-1RA or IL-1 α, can indirectly evaluate DBF or SkBF state.
IL-1RA is the cell factor of the anti-inflammatory that monocyte, macrophage etc. produce, and increases and make skin normalization during inflammation.IL-1RA Antagonism ground hinders the function of interleukin 1.When cellular level IL-1RA increases, as decomposing and the Fibroblast collagenase minimizing of the enzyme of fragmentation collagen.
IL-1 α is the cell factor of pro-inflammatory that monocyte, macrophage etc. produce, activated lymphocyte, macrophage causing inflammation.IL-1RA Antagonism ground hinders the function of interleukin 1.When known IL-1 α increases, as decomposing and the Fibroblast collagenase increase of the enzyme of fragmentation collagen.
Cuticula is the tissue being positioned at the skin the superiors, has protection skin not by the function coming exogenic foreign matter, stimulation.
As long as the mensuration in the present invention or evaluation object position can obtain cuticular part, then also can comprise any part, as main position, face, neck, upper arm parts can be enumerated.The cuticula from above-mentioned area skin can be obtained according to method in the past.But as previously mentioned, the method winning skin etc. due to surgery can bring burden to user, so preferably tape stripping method, scraping etc. can obtain cuticular method easily.Particularly preferably tape stripping method.
The expression of IL-1RA or the IL-1 α in each sample of such preparation can measure by the past known method.Such as, the method for carrying out the enzymoimmunoassay, radioimmunoassay, Western blotting assays etc. reacted based on the antibody with IL-1RA or IL-1 α can be used.Such mensuration means have commercially available, the representatively ELISA measuring reagent kit of IL-1RA, can illustrate the IL-1RA Human ELISA Kit of Human IL-1RA/IL-1F3Quantikine ELISA Kit, LifeTechnologies Inc. of R & D Systems.
In addition, the representatively ELISA measuring reagent kit of IL-1 α, can illustrate Human IL-1 α/IL-1F1Quantikine ELISA Kit of R & DSystems, the Human IL-1alpha ELISA Kit of Thermo Scientific Inc..
From each sample, IL-1RA, IL-1 α is passed through himself known biochemical method, such as freeze-thaw method, ultrasonic fragmentation, homogeneous method etc. are extracted as soluble component.Promptly measure after extraction.
Because the IL-1RA expression in the cuticula of the many people of DBF is higher than the IL-1RA expression in the cuticula of the few people of DBF, so can using the number of the IL-1RA expression in cuticula as index, the number of mensuration or evaluating skin volume of blood flow easily.
In addition, because the IL-1 alpha expression amount in the cuticula of the many people of DBF is lower than the IL-1 alpha expression amount in the cuticula of the few people of DBF, so can using the number of the IL-1 alpha expression amount in cuticula as index, the number of evaluating skin volume of blood flow easily.
Cuticula used in the present invention is by such the gathering by the straightforward procedure that cuticula adhesive tape only gathers cuticular surface part of tape stripping method.Therefore can not bring burden to experimenter, the DBF of experimenter or the blood flow state of evaluating skin can be measured.
The assay method of DBF of the present invention is made up of following operation: gather cuticular collection operation, measure the cuticula that gathers in described collection operation in IL-1RA, IL-1 α expression mensuration operation and the expression of IL-1RA, IL-1 α measured in described mensuration operation is converted into the operation of DBF by following regression equation, that namely obtains from the expression distribution of IL-1RA, IL-1 α the collective's cuticula measured in advance obtains the relevant regression equation of volume of blood flow.
The evaluation method of DBF of the present invention is made up of following operation: gather cuticular collection operation, measure the cuticula that gathers in described collection operation in the mensuration operation of expression of IL-1RA, IL-1 α and the contrast operation carrying out contrasting that the expression of IL-1RA, IL-1 α measured in described mensuration operation and the expression of IL-1RA, IL-1 α in the collective's cuticula measured in advance are distributed.Compared with the IL-1RA expression of experimenter distributes with the IL-1RA expression in the collective's cuticula measured in advance, when expression is many, is judged to be that the volume of blood flow of skin is large, when expression is few, is judged to be that the volume of blood flow of skin is little.The IL-1 alpha expression amount of experimenter when expression is many, is judged to be that the volume of blood flow of skin is little compared with distributing with the IL-1 alpha expression amount in the collective's cuticula measured in advance, when expression is few, is judged to be that the volume of blood flow of skin is large.
Below, be described about specific embodiment, but the present invention is not limited to following embodiment.
Embodiment
Test example 1
The measurement result of the DBF of tested collective is relevant to the distribution of IL-1RA expression measurement result.
< experimenter >
The cuticula of 75 healthy women cheek is gathered by tape stripping method.
< cuticula sample extraction method, quantification of protein >
Stick cuticula test pieces (ASAHIBIOMED Inc.) at the cheek of experimenter and gather cuticula.(Tissue Protein ExtRAction Reagent Thermo Scientific system (product#78510) carries out the extraction of cuticula protein to use the T-PER Extraction buffer of 500 μ l to grind method (add tested material in a reservoir, method that beaded glass that diameter is about 2mm and extract carry out mechanical shaking extraction) from cuticula test pieces with pearl.The albumen quality of each sample is measured by Pierce BCA proteinAssay Kit (Thermo Scientific#23225).During mensuration, in cuticula extracting solution of protein described in 10 μ l, add the reaction reagent liquid of 200 μ l Pierce BCA protein Assay Kit, after hatching 30 minutes at 60 DEG C, measure the absorbance at 562nm place.Meanwhile, with BSA (bovine serum albumin(BSA)) production standard curve, calculate albumen quality from the value of absorbance.
The expression of the IL-1RA in < cuticula measures >
The ELISA kit [Human IL-1RA/IL-1F3Quantikine ELISAKit] of R & D systems is similarly used by the IL-1RA expression in the cuticula extracting solution of protein extracted from adhesive tape to measure.First, the concentration of being specified by the immobilized antibody kit becoming initial antibody in 96 orifice plates (#3590) of costar is spent the night at 20 DEG C and is carried out immobilization.Then, place at 37 DEG C with 1%BSA and carry out closed culture plate in 1 hour, and after cleaning culture plate with 0.01%Tween-PBS, the standard items and the sample that each 100 μ l are become typical curve carry out application of sample, hatch 2 hours at 25 DEG C.After cleaning culture plate with 0.01%Tween-PBS, 2 antibody kit prescribed concentration are hatched 2 hours at 25 DEG C.And then, after cleaning culture plate with 0.01%Tween-PBS, Streptavidin-HRP kit prescribed concentration is hatched 30 minutes at 25 DEG C.Finally, after cleaning culture plate with 0.01%Tween-PBS, TMB solution (the Promega as chromogenic substrate is added according to 100 μ l/well, G7431)), after the time that kit is specified, add 100 μ l stop buffers (0.5M sulfuric acid), the absorbance measuring 450nm place calculates the expression of IL-1RA.
The mensuration > of < DBF
Laser Doppler flowmetry meter (Lisca Ab PIMII Laser Doppler PerfusionImager) is used to measure the relative output value (V) proportional with the volume of blood flow of experimenter's cheek.
< result >
X-axis is IL1-RA expression, Y-axis is that measurement result is made scatter diagram by the relative output value (V) proportional with volume of blood flow, obtains straight-line regression formula based on this scattered data.Scatter diagram and regression straight line, related coefficient, relational expression are as shown in Figure 1.
When X-axis is IL1-RA value, Y-axis is relative output value (V) proportional with volume of blood flow, regression equation represents Y=0.005110+0.4434, R
2=0.0703, the straight-line regression of related coefficient 0.27.
From the regression straight line shown in Fig. 1 or regression equation, the IL1-RA of mensuration can be converted into the relative output value (V) proportional with volume of blood flow.In addition, the distribution of the relative output value (V) proportional with the DBF of standard is distinguished from scatter diagram.As long as based on this scatter diagram and regression equation, by measuring the expression of the IL-1RA in keratoderma, the volume of blood flow of skin and known, and also whether can evaluate the volume of blood flow of experimenter from distribution normal.
Test example 2
The measurement result of relative output value (V) proportional to the DBF of tested collective is relevant with the distribution of the expression measurement result of IL-1 α.
< experimenter >
The cuticula of 73 healthy women cheek is gathered by tape stripping method.
< cuticula sample extraction method, quantification of protein >
Measure in the same manner as the mensuration of IL-1RA.
The mensuration > of the IL-1 α in < cuticula
The ELISA kit of R & D systems [Human IL-1 α/IL-1F1QuantikineELISA Kit] IL-1 alpha expression amount in the cuticula extracting solution of protein extracted from adhesive tape is used to measure in the same manner as test example 1.First, the concentration of being specified by the immobilized antibody kit becoming initial antibody in 96 orifice plates (#3590) of costar is spent the night at 20 DEG C and is carried out immobilization.Then, place at 37 DEG C with 1%BSA and carry out closed culture plate in 1 hour, and after cleaning culture plate with 0.01%Tween-PBS, the standard items and the sample that each 100 μ l are become typical curve carry out application of sample, hatch 2 hours at 25 DEG C.After cleaning culture plate with 0.01%Tween-PBS, 2 antibody kit prescribed concentration are hatched 2 hours at 25 DEG C.And then, after cleaning culture plate with 0.01%Tween-PBS, Streptavidin-HRP kit prescribed concentration is hatched 30 minutes at 25 DEG C.Finally, after cleaning culture plate with 0.01%Tween-PBS, TMB solution (the Promega as chromogenic substrate is added according to 100 μ l/well, G7431), after the time that kit is specified, add 100 μ l stop buffers (0.5M sulfuric acid), measure the absorbance at 450nm place and calculation expression amount.
The mensuration > of < DBF
Same with test example 1, use Laser Doppler flowmetry meter (Lisca Ab PIMII LaserDoppler Perfusion Imager) to measure the relative output value (V) proportional with the volume of blood flow of experimenter's cheek.
< result >
X-axis is IL-1 alpha expression amount, Y-axis is that measurement result is made scatter diagram by volume of blood flow, obtains straight-line regression formula based on this scattered data.Scatter diagram and regression straight line, related coefficient, relational expression are as shown in Figure 2.
When X-axis is IL-1 α value, Y-axis is relative output value (V) proportional with volume of blood flow, regression equation represents Y=-0.1873X+0.5681, R
2=0.0912, the negative straight-line regression of related coefficient 0.30.
From the regression straight line shown in Fig. 1 or regression equation, the IL-1 α of mensuration can be converted into volume of blood flow.In addition, the distribution of the DBF of standard has been distinguished from scatter diagram.As long as based on this scatter diagram and regression equation, by measuring the expression of the IL-1 α in keratoderma, the volume of blood flow of skin and known, and also whether can evaluate the volume of blood flow of experimenter from distribution normal.
Claims (6)
1. the indirect determination of DBF or an evaluation method, is characterized in that, using the expression of the interleukin-1 receptor antagonist in keratoderma and/or interleukin-1 alpha as index.
2. method according to claim 1, is characterized in that, for measure or the keratoderma of evaluation method is gathered by tape stripping method.
3. a method, is characterized in that, the relational expression made from the correlativity of the expression based on the interleukin-1 receptor antagonist the volume of blood flow measured in advance and keratoderma and/or interleukin-1 alpha carrys out the volume of blood flow of indirect determination skin.
4. an Indirect Determination for DBF, is characterized in that, is made up of following operation:
Gather cuticular collection operation,
Measure the mensuration operation of expression of the interleukin-1 receptor antagonist in the cuticula gathered in described collection operation, interleukin-1 alpha, and
The expression of the interleukin-1 receptor antagonist measured in described mensuration operation, interleukin-1 alpha is converted into the operation of DBF by following regression equation, described regression equation be obtain from the distribution of the expression of the interleukin-1 receptor antagonist the collective's cuticula measured in advance, interleukin-1 alpha obtain the relevant regression equation of volume of blood flow.
5. the evaluation method of a DBF, it is characterized in that, the scatter diagram made with the correlativity of the expression based on the interleukin-1 receptor antagonist in the volume of blood flow measured in advance and keratoderma and/or interleukin-1 alpha carries out contrasting evaluating.
6. an evaluation method for SkBF, is characterized in that, is made up of following operation:
Gather cuticular collection operation,
Measure the mensuration operation of expression of the interleukin-1 receptor antagonist in the cuticula gathered in described collection operation, interleukin-1 alpha, and
Distribute the expression of the interleukin-1 receptor antagonist measured in described mensuration operation, interleukin-1 alpha and the expression of the interleukin-1 receptor antagonist in the collective's cuticula measured in advance, interleukin-1 alpha the contrast operation carrying out contrasting.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2014-002663 | 2014-01-09 | ||
| JP2014002663A JP5731678B1 (en) | 2014-01-09 | 2014-01-09 | Blood flow evaluation or indirect measurement method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104777306A true CN104777306A (en) | 2015-07-15 |
| CN104777306B CN104777306B (en) | 2018-05-01 |
Family
ID=53486841
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410563850.2A Expired - Fee Related CN104777306B (en) | 2014-01-09 | 2014-10-21 | The evaluation of blood flow or indirect determination method |
Country Status (4)
| Country | Link |
|---|---|
| JP (1) | JP5731678B1 (en) |
| CN (1) | CN104777306B (en) |
| HK (1) | HK1209483A1 (en) |
| TW (1) | TWI638990B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1231173A (en) * | 1997-12-18 | 1999-10-13 | 尤尼利弗公司 | Cosmetic skin or hair care compositions containing fluorocarbons infused with carbon dioxide |
| JPH11285476A (en) * | 1998-04-01 | 1999-10-19 | Advance Co Ltd | Method of measuring blood flow and rheometer |
| JP2001112742A (en) * | 1999-10-21 | 2001-04-24 | Kao Corp | Method and apparatus for evaluating skin condition |
| CN1917919A (en) * | 2004-02-13 | 2007-02-21 | 株式会社芳珂 | Method of chemiluminescence-utilizing makeup and beautification, luminant for skin irradiation beautification and makeup/ beautification equipment |
| WO2012135597A2 (en) * | 2011-03-30 | 2012-10-04 | The Johns Hopkins University | Novel, protective, anti-inflammatory receptor and its use in preservation of mitochondrial function, wound healing and repair |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3590708B2 (en) * | 1997-02-07 | 2004-11-17 | 株式会社資生堂 | Evaluation method of skin quality |
| WO2011087525A1 (en) * | 2010-01-17 | 2011-07-21 | The Procter & Gamble Company | Biomarker-based methods for formulating compositions that improve skin quality and reduce the visible signs of aging in skin for individuals in a selected population |
| JP5653673B2 (en) * | 2010-07-23 | 2015-01-14 | 日本メナード化粧品株式会社 | Skin vascular function evaluation method |
| JP6022260B2 (en) * | 2012-08-22 | 2016-11-09 | ロート製薬株式会社 | Evaluation method of skin condition |
-
2014
- 2014-01-09 JP JP2014002663A patent/JP5731678B1/en active Active
- 2014-10-21 CN CN201410563850.2A patent/CN104777306B/en not_active Expired - Fee Related
- 2014-11-03 TW TW103138015A patent/TWI638990B/en not_active IP Right Cessation
-
2015
- 2015-10-15 HK HK15110139.4A patent/HK1209483A1/en not_active IP Right Cessation
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1231173A (en) * | 1997-12-18 | 1999-10-13 | 尤尼利弗公司 | Cosmetic skin or hair care compositions containing fluorocarbons infused with carbon dioxide |
| JPH11285476A (en) * | 1998-04-01 | 1999-10-19 | Advance Co Ltd | Method of measuring blood flow and rheometer |
| JP2001112742A (en) * | 1999-10-21 | 2001-04-24 | Kao Corp | Method and apparatus for evaluating skin condition |
| CN1917919A (en) * | 2004-02-13 | 2007-02-21 | 株式会社芳珂 | Method of chemiluminescence-utilizing makeup and beautification, luminant for skin irradiation beautification and makeup/ beautification equipment |
| WO2012135597A2 (en) * | 2011-03-30 | 2012-10-04 | The Johns Hopkins University | Novel, protective, anti-inflammatory receptor and its use in preservation of mitochondrial function, wound healing and repair |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104777306B (en) | 2018-05-01 |
| HK1209483A1 (en) | 2016-04-01 |
| JP2015132482A (en) | 2015-07-23 |
| JP5731678B1 (en) | 2015-06-10 |
| TW201527737A (en) | 2015-07-16 |
| TWI638990B (en) | 2018-10-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| König et al. | Cytokine profiles in nasal fluid of patients with seasonal or persistent allergic rhinitis | |
| Morita et al. | Stratum corneum TARC level is a new indicator of lesional skin inflammation in atopic dermatitis | |
| CN103175972B (en) | Atopic dermatitis marker and technique of using the same | |
| CN112067764B (en) | Method for evaluating skin inflammation relieving or repairing effect of cosmetic | |
| Glinos et al. | Optical coherence tomography for assessment of epithelialization in a human ex vivo wound model | |
| US20150141332A1 (en) | Methods for diagnosing osteoarthritis | |
| Caporizzo et al. | Strain-rate dependence of elastic modulus reveals silver nanoparticle induced cytotoxicity | |
| Salem et al. | Study of serum levels and skin expression of S100B protein in psoriasis | |
| CN104777306A (en) | Blood flow volume evaluation or indirect detection method | |
| CN104345146A (en) | Cosmetic selection device | |
| CN104777292B (en) | The evaluation method of skin texture | |
| CN104007057A (en) | Skin stress volume evaluating method | |
| CN104007056A (en) | Ultraviolet ray damage resistance evaluating method | |
| CN105807067A (en) | Dry-type immunofluorescence kit for detecting NCAM2 (neural cell adhesion molecules 2) of patient suffering from alzheimer's syndromes and preparation method | |
| Rich et al. | Science-in-brief: What is needed to prevent tendon injury in equine athletes? A conversation between researchers and industry stakeholders | |
| JP2012501660A (en) | Cell culture system for determining sensitization, allergenic and / or stimulatory effects of substances | |
| CN103808940A (en) | Evaluation method for rough skin | |
| Rosińska-Więckowicz et al. | Selected immunological parameters in clinical evaluation of patients with atopic dermatitis | |
| Eid et al. | Serum levels of interleukin-33 in alopecia areata: relation to disease severity and chronicity | |
| Dzimira et al. | Immunohistochemical diagnostic of hibernoma in dog | |
| RU2594250C2 (en) | Method for diagnosis of psoriasis | |
| EP4083629A1 (en) | Non-invasive diagnostic prediction of treatment success in allergen specific immunotherapy | |
| Yamanaka et al. | A new biomarker for psoriasis. | |
| Shee Tan | Laser-Scribed Graphene Electrodes as Inflammatory Biosensors (LASERFLAME) | |
| Hernández-Aguilar et al. | Rapid generation of an RBL cellular model to study proteins that cause allergenic reactions in vitro |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 1209483 Country of ref document: HK |
|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: GR Ref document number: 1209483 Country of ref document: HK |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180501 |