CN112704472A - Method for evaluating chemotactic/growth factor level in serum based on skin autofluorescence - Google Patents
Method for evaluating chemotactic/growth factor level in serum based on skin autofluorescenceDownload PDFInfo
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- A61B5/14546—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue for measuring analytes not otherwise provided for, e.g. ions, cytochromes
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- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
- A61B5/1455—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using optical sensors, e.g. spectral photometrical oximeters
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Abstract
Translated fromChinese
Description
Technical Field
The invention relates to a method for evaluating the levels of a plurality of chemokines (including MCP-1, MIP-1 alpha, MIP-1 beta and RANTES) and a growth factor G-CSF in serum based on skin autofluorescence.
Background
The body has a function of attracting leukocytes in defending and removing foreign substances such as invading pathogens, and there are substances that cause this function called chemotactic agents or chemokines, also called chemokines, chemokines or chemokines. Migration of cells to the source of the chemokine along with the signal of increasing chemokine concentration. Some chemokines control immune cell chemotaxis during immune surveillance, such as inducing lymphocytes into lymph nodes. Chemokines in these lymph nodes monitor pathogen invasion by interacting with antigen presenting cells in these tissues. These are called homeostatic chemokines, which are produced and secreted without stimulation. Some chemokines play a role in development; they can stimulate neovascularization; providing specific key signals to promote cell maturation. Other chemokines can be released by a variety of cells in response to bacterial and viral infections; it can also be released by non-infectious stimuli such as silica inhalation, urinary tract stones, etc. Release of chemokines can also stimulate the release of inflammatory cytokines. The primary role of inflammatory chemokines is to chemotaxis leukocytes (e.g., monocytes and neutrophils) from the blood circulation to the site of infection or tissue damage. Some chemokines may also promote wound healing.
MCP-1 is a chemokine regulating T cell differentiation and monocyte chemotaxis. RANTES is a chemokine that plays an important role in the migration and homing of effector and memory T cells. G-CSF controls the production, differentiation and function of granulosa cells.
The main method for detecting the levels of the above chemokines and growth factors in serum is by blood test. This method has the drawback of being invasive and requiring a professional doctor and nurse to perform the operative tests. Therefore, it is of great value to find a method and technology for non-invasive assessment of the levels of these chemokines and growth factors.
Disclosure of Invention
The invention aims to provide a noninvasive method for evaluating chemotaxis and growth factor levels in serum based on skin autofluorescence. The invention discovers that the intensity of the skin green autofluorescence is obviously and positively correlated with the levels of a plurality of chemokines (including MCP-1, MIP-1 alpha, MIP-1 beta and RANTES) and a growth factor G-CSF in serum, so that the levels of the plurality of chemokines (including MCP-1, MIP-1 alpha, MIP-1 beta and RANTES) and the growth factor G-CSF in serum can be non-invasively evaluated by non-invasively evaluating the intensity of the skin green autofluorescence.
The purpose of the invention is realized by the following technical scheme:
in a first aspect, the present invention provides a method for assessing chemokine/growth factor G-CSF levels in serum based on skin autofluorescence, in a non-diagnostic treatment, comprising the steps of:
s1, placing the skin under the excitation light with the wavelength of 450-520nm to excite the autofluorescence of the skin;
s2, detecting an autofluorescence image with the wavelength within the range of 500-620nm emitted by the skin;
s3, analyzing the intensity of the autofluorescence image to obtain the current skin autofluorescence intensity; comparing the current skin autofluorescence intensity with the previous skin autofluorescence intensity to obtain the skin autofluorescence intensity change;
and S4, evaluating the level of the chemotactic factor/growth factor G-CSF in the serum according to the obvious positive correlation between the change of the skin autofluorescence intensity and the level of the chemotactic factor/growth factor G-CSF in the serum.
The method is applied to the non-clinical field, can be used for evaluating the level of chemotactic factor/growth factor G-CSF in serum of non-disease people, and can also be used for evaluating healthy or sub-healthy people.
Further, the chemokines include chemokine MCP-1, chemokine MIP-1 alpha, chemokine MIP-1 beta, and chemokine RANTES.
Further, in step S1, the excitation light for skin autofluorescence includes at least one of excitation using a normal continuous light output, modulation excitation using electrical modulation, or excitation using pulsed laser light.
Further, in step S1, the wavelength of the excitation light is within the range of 450-500 nm.
Further, in step S2, an autofluorescence image with a wavelength of skin autofluorescence within the range of 500-580nm is detected.
Further, in step S3, the current skin autofluorescence intensity and the past skin autofluorescence intensity are obtained by the same method.
Further, in step S3, the past skin autofluorescence intensity is the skin autofluorescence intensity measured 2 years before the current test.
Further, in step S3, the current skin green fluorescence intensity is compared with the value of the conventional skin green autofluorescence intensity.
Further, in step S4, the evaluating specifically includes: if the current skin autofluorescence intensity is higher than the previous skin autofluorescence intensity, the serum level of the existing chemotactic factor/growth factor G-CSF is higher than the previous level of the chemotactic factor/growth factor G-CSF; if the current skin autofluorescence intensity is lower than the previous skin autofluorescence intensity, the more the current chemokine/growth factor G-CSF level in the serum is lower than the previous chemokine/growth factor G-CSF level.
In a second aspect, the invention relates to a dedicated apparatus for using the aforementioned method. More particularly relates to a special device for evaluating the level of chemotactic factor/growth factor G-CSF in serum based on skin autofluorescence.
In a third aspect, the invention relates to a marker for non-invasively assessing the level of chemokine/growth factor G-CSF in serum, said marker being the skin autofluorescence intensity.
The chemokines include chemokine MCP-1, chemokine MIP-1 alpha, chemokine MIP-1 beta, and chemokine RANTES.
In a fourth aspect, the present invention relates to an evaluation model for evaluating chemokine/growth factor G-CSF in serum based on skin autofluorescence, wherein the level of chemokine/growth factor G-CSF in serum of a subject is significantly and positively correlated with the change of skin autofluorescence intensity of the subject.
Further, the subject's skin autofluorescence intensity change is measured by a method comprising the steps of:
the skin is placed under the excitation light with the wavelength within the range of 450-500nm to excite the autofluorescence of the skin;
detecting an autofluorescence image emitted by the skin with a wavelength in the range of 500-580 nm;
analyzing the intensity of the autofluorescence image to obtain the current skin autofluorescence intensity; and comparing the current skin autofluorescence intensity with the previous skin autofluorescence intensity of the subject to obtain the skin autofluorescence intensity change of the subject.
In a fifth aspect, the present invention relates to a method for constructing an evaluation model for evaluating chemokine/growth factor G-CSF in serum based on skin autofluorescence, comprising the steps of:
a1, placing the skin of the subject under the excitation light with the wavelength of 450-520nm to excite the autofluorescence of the skin;
a2, detecting the autofluorescence image of the skin with the wavelength in the range of 500-620 nm;
a3, analyzing the intensity of the autofluorescence image to obtain the current skin autofluorescence intensity; comparing the current skin autofluorescence intensity with the past skin autofluorescence intensity of the subject to obtain the skin autofluorescence intensity change of the subject;
a4, the skin autofluorescence intensity characterizes the level of chemokine/growth factor G-CSF in the serum of a subject. The chemokines include chemokine MCP-1, chemokine MIP-1 alpha, chemokine MIP-1 beta, and chemokine RANTES.
Compared with the prior art, the invention has the following beneficial effects:
1) levels of several chemokines (including MCP-1, MIP-1 α, MIP-1 β, and RANTES) and growth factor G-CSF in serum can be non-invasively assessed by non-invasively assessing skin green autofluorescence intensity;
2) provides the application of the skin autofluorescence intensity as a marker for non-invasive evaluation of a plurality of chemokines (including MCP-1, MIP-1 alpha, MIP-1 beta and RANTES) and growth factor G-CSF in serum.
Drawings
FIG. 1: the excitation light is 488nm to excite the skin to perform autofluorescence, and the receiving light with the wavelength range of 500-550nm is received; a correlation quantification graph of the autofluorescence intensity and the chemotactic factor MCP-1;
FIG. 2: the excitation light is 488nm to excite the skin to perform autofluorescence, and the receiving light with the wavelength range of 500-550nm is received; a quantitative graph of correlation between autofluorescence intensity and chemotactic factor MIP-1 alpha;
FIG. 3: the excitation light is 488nm to excite the skin to perform autofluorescence, and the receiving light with the wavelength range of 500-550nm is received; a quantitative graph of correlation between autofluorescence intensity and chemokine MIP-1 beta;
FIG. 4: the excitation light is 488nm to excite the skin to perform autofluorescence, and the receiving light with the wavelength range of 500-550nm is received; an autofluorescence intensity and chemokine RANTES correlation quantification graph;
FIG. 5: the excitation light is 488nm to excite the skin to perform autofluorescence, and the receiving light with the wavelength range of 500-550nm is received; a graph quantifying the correlation of autofluorescence intensity with growth factor G-CSF.
Detailed Description
The present invention will be described in detail with reference to examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that it would be apparent to those skilled in the art that several modifications and improvements can be made without departing from the inventive concept. All falling within the scope of the present invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
The term "autofluorescence" as used herein means the phenomenon in which a biomolecule, when irradiated with excitation light of an appropriate wavelength, absorbs the energy of the excitation light into an excited state and then exits the excited state to emit light of a wavelength longer than that of the excitation light.
The term "excitation light" as used in the present invention means light capable of exciting a biomolecule to undergo autofluorescence, and the wavelength should be shorter than the autofluorescence
The term "chemokine" as used herein is defined as follows: the human body has a function of attracting leukocytes in defense and elimination of foreign substances such as invading pathogens, and some substances are capable of causing the function, which is called chemotactic agents or chemokines. Migration of cells to the source of the chemokine along with the signal of increasing chemokine concentration. Some chemokines control immune cell chemotaxis during immune surveillance, such as inducing lymphocytes into lymph nodes. Chemokines in these lymph nodes monitor pathogen invasion by interacting with antigen presenting cells in these tissues. These are called homeostatic chemokines, which are produced and secreted without stimulation. Some chemokines play a role in development; they can stimulate neovascularization; providing specific key signals to promote cell maturation. Other chemokines can be released by a variety of cells in response to bacterial and viral infections; it can also be released by non-infectious stimuli such as silica inhalation, urinary tract stones, etc. Release of chemokines can also stimulate the release of inflammatory cytokines. The primary role of inflammatory chemokines is to chemotaxis leukocytes (e.g., monocytes and neutrophils) from the blood circulation to the site of infection or tissue damage. Some chemokines may also promote wound healing.
The term "MCP 1", which is used herein and is generally called "monocyte chemoattractant protein 1", is also called "CCL 2" and "chemokine (C-C motif) ligand 2". MCP1 is a subset of proteins belonging to the CC chemokine family, an inflammatory chemokine.
The term "MIP 1 α" is used herein, and is also referred to as "macrovoid in fluorescence protein 1-alpha" for short, also referred to as "CCL 3".
The term "MIP 1 β" is used in the present invention, and is also called "macroporous in-flight protein 1-beta" in English, and is also called "CCL 4" in English "
The term "RANTES" is used herein, and is also called "Chemokine (C-C motif) ligand 5" for short, or "CCL 5". The primary function is to modulate T cell activation.
The term "G-CSF" is used herein, and is also referred to as "granular-binding factor" and "binding-binding factor 3" throughout the English language. The inventors have performed a number of experiments to determine the positive correlation between the skin autofluorescence and the levels of these chemokines described above in serum.
Example 1
Male C57 mice were used and were housed in an animal house at 22-24℃ for 12 hours light/dark cycles and were allowed free access to water. Injecting lipopolysaccharide into the abdominal cavity of the mouse, and performing non-invasive real-time imaging on the skin of the mouse treated by the lipopolysaccharide by using a laser confocal microscope after 1 day and 3 days. The excitation wavelength of the confocal laser scanning microscope is 450-520nm, and 488nm is selected in this embodiment. The receiving band is 500-620nm, and two receiving bands, 500-550nm and 575-620nm, are selected in this embodiment.
After 1 day and 3 days, the mouse serum was quantitatively assayed for chemokines.
The correlation results of the mouse skin autofluorescence intensity and the serum chemokine MCP-1 level are shown in FIG. 1, the correlation results of the mouse skin autofluorescence intensity and the serum chemokine MIP-1 alpha level are shown in FIG. 2, the correlation results of the mouse skin autofluorescence intensity and the serum chemokine MIP-1 beta level are shown in FIG. 3, and the correlation results of the mouse skin autofluorescence intensity and the serum chemokine RANTES level are shown in FIG. 4 after 3 days of treatment of the mice with lipopolysaccharide.
As can be seen from FIGS. 1-4, the skin fluorescence intensity allows the detection of chemokine concentrations. Meanwhile, the fluorescence intensity of the skin is found to be in positive correlation with the levels of MCP-1, MIP-1 alpha, MIP-1 beta and RANTES in serum. The higher the fluorescence intensity of the skin, the higher the level of these chemokines in the serum. The higher the fluorescence intensity of the skin, the higher the level of these factors in the serum.
Example 2
Male C57 mice were used and were housed in an animal house at 22-24℃ for 12 hours light/dark cycles and were allowed free access to water. Injecting lipopolysaccharide into the abdominal cavity of the mouse, and performing non-invasive real-time imaging on the skin of the mouse treated by the lipopolysaccharide by using a laser confocal microscope after 1 day and 3 days. The excitation wavelength of the confocal laser scanning microscope is 450-520nm, and 488nm is selected in this embodiment. The receiving band is 500-620nm, and two receiving bands, 500-550nm and 575-620nm, are selected in this embodiment.
Growth factors were quantitatively determined inmouse serum 1 day and 3 days later.
The results of correlation between the skin autofluorescence intensity of mice and the level of growth factor G-CSF in serum 3 days after treatment of mice with lipopolysaccharide are shown in FIG. 5; the skin fluorescence intensity was found to detect chemokine concentrations. Meanwhile, the fluorescence intensity of the skin is found to be positively correlated with the level of G-CSF in the serum. The higher the fluorescence intensity of the skin, the higher the level of G-CSF in the serum. The higher the fluorescence intensity of the skin, the higher the level of G-CSF in the serum.
The above experiments show that the autofluorescence of mice treated with lipopolysaccharide changes, and the change is positively correlated with the above-mentioned G-CSF level in serum. Thus, skin autofluorescence can be used as a marker for non-invasive assessment of G-CSF levels in serum.
Based on the discovery, the invention establishes a method for noninvasive evaluation of serum chemokine concentration based on skin autofluorescence, which comprises the following steps:
1. a method for assessing levels of several chemokines (including MCP-1, MIP-1 alpha, MIP-1 beta and RANTES) and growth factor G-CSF in serum based on skin autofluorescence comprises:
(1) the skin is placed under excitation light with a wavelength in the range of 450-520nm to excite the autofluorescence of the skin.
(2) Detecting an autofluorescence image emitted by the skin with the wavelength within the range of 500-620 nm;
(3) analyzing the intensity of the autofluorescence image, and comparing the intensity of the autofluorescence with the past fluorescence intensity of the subject;
(4) the serum levels of several chemokines (including MCP-1, MIP-1 α, MIP-1 β and RANTES) and the growth factor G-CSF were evaluated in subjects according to the following methods: the levels of several chemotactic factors (including MCP-1, MIP-1 alpha, MIP-1 beta and RANTES) and growth factor G-CSF in the serum of the tested person are in obvious positive correlation with the change of the skin autofluorescence intensity of the tested person. If the present autofluorescence intensity of the tested person is higher than the previous autofluorescence intensity of the tested person, the higher the level of the cytokines present in the serum of the tested person is. If the existing autofluorescence intensity of the tested person is lower than the former autofluorescence intensity of the tested person, the cytokine level existing in the serum of the tested person is lower than the former cytokine level of the tested person.
The excitation of skin autofluorescence with excitation light according to the present invention includes at least one of excitation using a normal continuous light output, modulation excitation using electrical modulation, or excitation using pulsed laser.
It will be apparent to those skilled in the art that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention. Therefore, the detailed description and examples of the invention should not be construed as limiting the scope of the invention. The invention is limited only by the appended claims. All documents cited in this application are incorporated herein by reference in their entirety.
Claims (10)
1. A method for non-diagnostic treatment for assessing chemokine/growth factor G-CSF levels in serum based on skin autofluorescence, comprising the steps of:
s1, placing the skin under the excitation light with the wavelength of 450-520nm to excite the autofluorescence of the skin;
s2, detecting an autofluorescence image with the wavelength within the range of 500-620nm emitted by the skin;
s3, analyzing the intensity of the autofluorescence image to obtain the current skin autofluorescence intensity; comparing the current skin autofluorescence intensity with the previous skin autofluorescence intensity to obtain the skin autofluorescence intensity change;
and S4, evaluating the level of the chemotactic factor/growth factor G-CSF in the serum according to the obvious positive correlation between the change of the skin autofluorescence intensity and the level of the chemotactic factor/growth factor G-CSF in the serum.
2. The method of claim 1, wherein said chemokine comprises the chemokine MCP-1, the chemokine MIP-1 α, the chemokine MIP-1 β, and the chemokine RANTES.
3. The method according to claim 1, wherein the excitation of the skin autofluorescence with the excitation light in step S1 comprises at least one of excitation with a common continuous light output, modulated excitation with electrical modulation, or excitation with pulsed laser light.
4. The method as claimed in claim 1, wherein in step S1, the wavelength of the excitation light is in the range of 450 nm and 500 nm.
5. The method as claimed in claim 1, wherein in step S2, an autofluorescence image of skin autofluorescence is detected at a wavelength in the range of 500-580 nm.
6. The method according to claim 1, wherein in step S3, the current skin autofluorescence intensity and the past skin autofluorescence intensity are obtained based on the same method; the past skin autofluorescence intensity refers to the skin autofluorescence intensity measured within 2 years before the current test.
7. The method according to claim 1, wherein in step S3, the current skin green fluorescence intensity is compared with the previous values of skin green autofluorescence intensity.
8. A dedicated apparatus for use with the method of any one of claims 1 to 7.
9. A marker for non-invasively assessing levels of chemokine/growth factor G-CSF in serum, wherein said marker is skin autofluorescence intensity.
10. A construction method of an evaluation model for evaluating chemokine/growth factor G-CSF in serum based on skin autofluorescence, the construction method comprising the steps of:
a1, placing the skin of the subject under the excitation light with the wavelength of 450-520nm to excite the autofluorescence of the skin;
a2, detecting the autofluorescence image of the skin with the wavelength in the range of 500-620 nm;
a3, analyzing the intensity of the autofluorescence image to obtain the current skin autofluorescence intensity; comparing the current skin autofluorescence intensity with the past skin autofluorescence intensity of the subject to obtain the skin autofluorescence intensity change of the subject;
a4, the skin autofluorescence intensity characterizes the level of chemokine/growth factor G-CSF in the serum of a subject.
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- 2019
- 2019-10-25CNCN201911021972.8Apatent/CN112704472A/enactivePending
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| US20020151774A1 (en)* | 2001-03-01 | 2002-10-17 | Umass/Worcester | Ocular spectrometer and probe method for non-invasive spectral measurement |
| CN104939806A (en)* | 2008-05-20 | 2015-09-30 | 大学健康网络 | Device and method for fluorescence-based imaging and monitoring |
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