CN110221337A - A method of utilizing irradiation biological damage in α -1 antiprotease evaluation uranium ore dust - Google Patents
A method of utilizing irradiation biological damage in α -1 antiprotease evaluation uranium ore dust Download PDFInfo
- Publication number
- CN110221337A CN110221337A CN201910572069.4A CN201910572069A CN110221337A CN 110221337 A CN110221337 A CN 110221337A CN 201910572069 A CN201910572069 A CN 201910572069A CN 110221337 A CN110221337 A CN 110221337A
- Authority
- CN
- China
- Prior art keywords
- antiprotease
- uranium ore
- ore dust
- biological damage
- protein
- 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
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
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01T—MEASUREMENT OF NUCLEAR OR X-RADIATION
- G01T1/00—Measuring X-radiation, gamma radiation, corpuscular radiation, or cosmic radiation
- G01T1/02—Dosimeters
Abstract
The method that biological damage is irradiated in α -1 antiprotease evaluation uranium ore dust is utilized the present invention relates to a kind of.Utilize the characteristic very sensitive to biological damage caused by irradiation in uranium ore dust of α -1 antiprotease in WISTAR lung tissue of rats, receive the dose-effect relationship of the up-regulation rate of α -1 antiprotease relative expression quantity in the amount and lung tissue of the uranium ore dust suspension of tracheal instillation according to WISTAR rat, by the up-regulation rate of the relative expression quantity of α -1 antiprotease in WISTAR rat lung tissue, biological damage caused by irradiation in uranium ore dust is assessed.This method has the advantages that biological damage sensibility height, high specificity, good reliability caused by irradiating in uranium ore dust etc. is multiple.
Description
Technical field
The invention belongs to irradiate biological damage assessment technique field in uranium ore dust, uranium ore powder is carried out using molecular marked compound
The method of irradiation biological damage evaluation, specifically utilizes the relative expression of α -1 antiprotease in WISTAR lung tissue of rats in dirt
Amount up-regulation rate irradiates biological damage to evaluate method in uranium ore dust.
Background technique
A certain number of uranium ore dust pollutions can be all generated during uranium exploration, exploitation, hydrometallurgy and Tailings Disposal, this
Containing radionuclides such as uranium, thorium, radium in a little uranium ore dust, once it will be stranded in lung qi for a long time by human body inspiration body
In pipe, bronchus and lymph node, potential long-term interior irradiation harm is constituted, biological damage is generated.The ionising radiation master of Uranium-Mineral Dust
Will from radon and its daughter (218Po、214Pb、214Bi and214Po) α, β ray that decay generates, since the dust of sucking produces
Raw radioactive dosage is low, is difficult to show biological effect in the short time, thus is difficult to carry out detection and biological damage assessment.Cause
This, filter out in uranium ore dust irradiate caused by biological damage detection speed it is fast, sensibility is high, high specificity, good reliability
Biomarker, to having a very important significance.
Organism suck uranium ore dust after, may by change protein expression and its posttranslational modification state come
Respond ionising radiation.ITRAQ technology is emerging high throughput protein group method, passes through the amino of specific marker polypeptide
Then group carries out Tandem Mass Spectrometry Analysis, can compare the relative amount or absolute of protein in 4 kinds or 8 kinds different samples simultaneously
Content.Therefore, can be determined by the protein of differential expression in iTRAQ technology screening tissue sample in uranium ore dust
The relevant biomarker of biological damage caused by irradiation, and then made using the biomarker of screening to being irradiated in uranium ore dust
It is evaluated at biological damage.
Summary of the invention
For above situation, the present invention, which provides, a kind of utilizes α -1 antiprotease (Alpha-1- in WISTAR lung tissue of rats
Antiproteinase, A1AT) relative expression quantity up-regulation rate evaluate the side of biological damage caused by irradiation in uranium ore dust
Method.The present invention using α -1 antiprotease in WISTAR lung tissue of rats to being irradiated in uranium ore dust caused by biological damage very
Sensitive characteristic, according to WISTAR rat receive tracheal instillation uranium ore dust suspension (uranium ore dust and physiological saline according to
Certain proportion configuration) amount and lung tissue in α -1 antiprotease relative expression quantity up-regulation rate dose-effect relationship, pass through
The up-regulation rate of the relative expression quantity of α -1 antiprotease in WISTAR rat lung tissue, caused by the interior irradiation of assessment uranium ore dust
Biological damage.Its principle is, after inventor receives the uranium ore suspension of tracheal instillation by the discovery of iTRAQ technology screening
The relative expression quantity of α -1 antiprotease in WISTAR rat lung tissue is very sensitive to uranium ore dust, and exists centainly
Dose-effect relationship.α -1 antiprotease is the protein that plasma concentration changes in response to inflammation, participates in oxidative stress, and inflammation is anti-
Should and Apoptosis.Within the scope of certain internal dose, the internal irradiation damage that body is subject to is bigger, α -1 antiprotease
Relative expression quantity is higher, the height of α -1 antiprotease protein expression directly in uranium ore dust irradiate caused by biological damage
Degree is closely related, and this method has biological damage sensibility height, high specificity, reliability caused by irradiating in uranium ore dust
The multiple advantage such as good.
It comprises the concrete steps that:
(1) uranium ore suspension tracheal instillation WISTAR rat model is established；
(2) lung tissue is acquired；
(3) detection of α -1 antiprotease relative expression quantity up-regulation rate；
(4) biological damage overall merit is irradiated in.
Its further step is:
The specific method for establishing uranium ore suspension tracheal instillation WISTAR rat model is:
Selective body focuses on the WISTAR Adult male rats between 180-220 g, and random point 4 groups, every group of 6 mouse, control
The suspension (partial size of silica is 200 mesh) of group tracheal instillation physiological saline and silica, experimental group distinguishes tracheal instillation
The uranium ore dust suspension (partial size of uranium ore dust is 200 mesh) that concentration is 1.25,2.5 and 5.0 mg/ml, instils 0.2 every time
Ml instils 3 times, continuous 5 weeks for one week.
The specific method of the acquisition lung tissue is:
Rat is put to death with cervical dislocation, takes lung tissue after dissection, the part that classifies in three categories is organized each, falls blood with normal saline flushing
Water removes moisture, is attached in cryopreservation tube, is immediately placed into -80 DEG C of refrigerators and saves.
The detection specific method of the α -1 antiprotease relative expression quantity up-regulation rate is:
WISTAR rat lung tissue will be irradiated in the receiving of collection uranium ore dust carries out protein extraction, total protein concentration survey
Fixed, protein denaturation, transferring film, immune response, chemiluminescence reaction, gel imaging scanning analysis, finally carries out α-at gel electrophoresis
The relative expression quantity up-regulation rate of 1 antiprotease is analyzed.
Specific step is as follows for the protein extraction:
Liquid nitrogen precooler is poured into mortar, lung tissue's sample is taken out rapidly, tissue is switched to and is homogeneously disposed in mortar, is added at once
Enter liquid nitrogen, is ground.It is slowly added into 800 μ l LB lysates in mortar, cracks 30 min, vibrates 1-2 times, sufficiently splits
Solution.After cracking, the suspension of 1.5 mL is collected into EP pipe, and then at 4 DEG C, 12000 r/min of revolving speed is centrifuged 20 min
Afterwards, supernatant is collected.With the albumen in resin precipitated supernatant, 4 DEG C, 12000 r/min are centrifuged 20 min, remove supernatant, dry in the air
Dry, -80 DEG C save backup.200-250 μ L L3 lysate is added in protein agglomerate after the drying, is blown and beaten repeatedly with pipette tips
Until protein is fully dispersed.Ultrasonic dissolution assisting 4 DEG C, 12000 r/min, is centrifuged 20 min, supernatant is sucked out, is transferred to new
In EP pipe.
Specific step is as follows for the total protein concentration measurement:
The BSA standard solution of 0.2 mg/ml is added into dilution, is successively diluted to 0,5,10,20,50,100,150 and 200
The standard solution of μ g/ml.Then their absorbance values at 562 nm are measured, standard curve is drawn.Measure each sample
Solution is after the absorbance value under 562 nm, then calculates by standard curve the concentration of protein in sample.
Specific step is as follows for the protein denaturation:
5 × protein sample-loading buffer is added according to the ratio of 4:1 in protein solution, boiling water bath is denaturalized 15 min, is put into -20
DEG C refrigerator saves backup.
Specific step is as follows for the gel electrophoresis:
30% acrylamide, 1.5 M TRIS-HCl, 10%SDS, 10% ammonium persulfate and TEMED are matched according to a certain percentage
10% separation gel and 5% concentration glue is made；After gelling to be separated is solid, electrophoresis liquid is added in electrophoresis tank, by the egg after denaturation
White matter sample is added in electrophoresis hole, carries out electrophoresis.Concentrate glue voltage is adjusted to 75 V, separation gel voltage is adjusted to 100 V, and electrophoresis is extremely
Bromophenol blue has just been run to separation gel bottom, and electrophoresis can be terminated, and carries out transferring film.
Specific step is as follows for the transferring film:
First pvdf membrane is activated using methanol, then places the filter paper of 5-6 7 × 9 cm in the bottom of film, is together placed
In transferring film instrument, transferring film liquid, 300 mA constant current transferring film, 30 min is added.In During migration, transferring film slot is placed in ice water and is dropped
Temperature.
Specific step is as follows for the immune response:
5% skim milk (0.5% TBST dilution) of the film to take a turn for the better is closed, 1 h is mixed on decolorization swinging table.Using TBST
Dissolution 5% skim milk to α -1 antiprotease primary antibody (Alah3al) carry out 1:1000 concentration dilution, then by its with close
Film mixed, be incubated overnight at 4 DEG C.It is cleaned with the film that 2% TBST is incubated overnight to α -1 antiprotease primary antibody, room
Clean 3 times under temperature on decolorization swinging table, every time 5 min.8000 times finally are diluted to secondary antibody (goat-anti rabbit) with TBST, is incubated at room temperature
After educating 1.5 h, cleaned on decolorization swinging table three times, every time 10 min.
Specific step is as follows for the chemiluminescence reaction:
It is by two kinds of reagents of ECLA and ECLB in the medium volume mixture of centrifuge tube in darkroom, the albumen of pvdf membrane is face-up, add
Enter after the ECL solution mixed sufficiently reacts 1-2 min, is put into gel imaging system and is scanned analysis.
Specific step is as follows for the gel imaging scanning analysis:
It is taken pictures using gel imaging system to protein band, then using the gray value of Image J software analysis object tape.It is logical
It crosses to scan α -1 antiprotease albumen gray value and obtains α -1 antiprotease expressing quantity, to internal reference Protein G APDH albumen ash
Angle value scanning obtains GAPDH expressing quantity；By α -1 antiprotease expressing quantity divided by internal reference Protein G APDH protein expression
Amount obtains the relative expression quantity of α -1 antiprotease.
Specific step is as follows for the relative expression quantity up-regulation rate analysis of α -1 antiprotease:
α -1 antiprotease (A1AT) relative expression quantity up-regulation rate is calculated by formula (1):
(1)
The specific method of the interior irradiation biological damage overall merit is:
Biological damage caused by irradiating in uranium ore dust is evaluated using the relative expression quantity up-regulation rate of α -1 antiprotease.When
The relative expression quantity up-regulation rate of α -1 antiprotease in WISTAR rat lung tissue shows low dosage gamma when within 30%
X ray irradiation x is to inanimate object damage effect；When the relative expression quantity of α -1 antiprotease in WISTAR rat lung tissue raises
Rate is greater than 30% when less than 100%, and biological damage grade caused by irradiation is I grade in uranium ore dust at this time；When WISTAR is big
The relative expression quantity up-regulation rate of α -1 antiprotease in mouse lung tissue be greater than 100% and when less than 200%, uranium ore powder at this time
Biological damage grade caused by irradiation is II grade in dirt；When the opposite table of α -1 antiprotease in WISTAR rat lung tissue
When being greater than 200% up to amount up-regulation rate, biological damage grade caused by irradiation is III grade in uranium ore dust at this time.When uranium ore dust
When biological damage grade caused by interior irradiation reaches I grade, occupational staff should reinforce radiation protection；When irradiation causes in uranium ore dust
Biological damage grade when reaching II grade, occupational staff should rest a period of time in time；It is raw caused by being irradiated in uranium ore dust
When object impairment scale reaches III grade, occupational staff should be transferred from work position, and receive appropriate treatment.
Invention is the albumen for quantifying iTRAQ technology analysis differential expression entirely using proteomics, identifies α -1
Biomarker of the antiprotease as biological damage caused by irradiation in uranium ore dust, then utilizes the phase of α -1 antiprotease
Dose-effect relationship between biological damage caused by irradiating in expression quantity and uranium ore dust irradiates to evaluate in uranium ore dust
Caused by biological damage.Compared to other biomarkers and evaluation method, this method, which has, to be made to irradiating in uranium ore dust
At biological damage sensibility height, high specificity, the multiple advantage such as good reliability.
Specific embodiment
Embodiment 1
Selective body focuses on the WISTAR Adult male rats between 180-220 g, and random point 4 groups, every group of 6 mouse, control
The suspension (partial size of silica is 200 mesh) of group tracheal instillation physiological saline and silica, experimental group distinguishes tracheal instillation
The uranium ore dust suspension (partial size of uranium ore dust is 200 mesh) that concentration is 1.25 mg/ml, instil 0.2 ml every time, instils within one week
3 times, continuous 5 weeks.Rat is put to death with cervical dislocation, takes lung tissue after dissection, the part that classifies in three categories is organized each, uses physiological saline
Watery blood is rinsed out, moisture is removed, is attached in cryopreservation tube, -80 DEG C of refrigerators is immediately placed into and saves.By the receiving of collection low dosage electricity
WISTAR rat lung tissue from x ray irradiation x carries out protein extraction, total protein concentration measurement, protein denaturation, gel electricity
Swimming, transferring film, immune response, chemiluminescence reaction, gel imaging scanning analysis, finally according to formula (1) carry out α -1 antiprotease
Relative expression quantity up-regulation rate analysis.
WISTAR Adult male rats tracheal instillation concentration was the uranium ore dust suspension of 1.25 mg/ml after 5 weeks, lung's group
The relative expression quantity up-regulation rate for knitting middle α -1 antiprotease is 85.6%, at this time biological damage grade caused by irradiation in uranium ore dust
It is I grade, occupation work personnel should enhance radiation protection.
Embodiment 2
Selective body focuses on the WISTAR Adult male rats between 180-220 g, and random point 4 groups, every group of 6 mouse, control
The suspension (partial size of silica is 200 mesh) of group tracheal instillation physiological saline and silica, experimental group distinguishes tracheal instillation
The uranium ore dust suspension (partial size of uranium ore dust is 200 mesh) that concentration is 2.5 mg/ml, instil 0.2 ml every time, instillation 3 in one week
It is secondary, continuous 5 weeks.Rat is put to death with cervical dislocation, takes lung tissue after dissection, the part that classifies in three categories is organized each, is rushed with physiological saline
Watery blood is washed off, moisture is removed, is attached in cryopreservation tube, -80 DEG C of refrigerators is immediately placed into and saves.The receiving of collection low dosage is ionized
The WISTAR rat lung tissue of x ray irradiation x carry out protein extraction, total protein concentration measurement, protein denaturation, gel electrophoresis,
Transferring film, chemiluminescence reaction, gel imaging scanning analysis, finally according to formula (1) carries out α -1 antiprotease at immune response
The analysis of relative expression quantity up-regulation rate.
WISTAR Adult male rats tracheal instillation concentration was the uranium ore dust suspension of 2.5 mg/ml after 5 weeks, lung tissue
Middle α -1 antiprotease relative expression quantity up-regulation rate is 158.5 %, and biological damage grade caused by irradiation is in uranium ore dust at this time
II grade, occupation work personnel should rest a period of time in time.
Embodiment 3
Selective body focuses on the WISTAR Adult male rats between 180-220 g, and random point 4 groups, every group of 6 mouse, control
The suspension (partial size of silica is 200 mesh) of group tracheal instillation physiological saline and silica, experimental group distinguishes tracheal instillation
The uranium ore dust suspension (partial size of uranium ore dust is 200 mesh) that concentration is 5.0 mg/ml, instil 0.2 ml every time, instillation 3 in one week
It is secondary, continuous 5 weeks.Rat is put to death with cervical dislocation, takes lung tissue after dissection, the part that classifies in three categories is organized each, is rushed with physiological saline
Watery blood is washed off, moisture is removed, is attached in cryopreservation tube, -80 DEG C of refrigerators is immediately placed into and saves.The receiving of collection low dosage is ionized
The WISTAR rat lung tissue of x ray irradiation x carry out protein extraction, total protein concentration measurement, protein denaturation, gel electrophoresis,
Transferring film, chemiluminescence reaction, gel imaging scanning analysis, finally according to formula (1) carries out α -1 antiprotease at immune response
The analysis of relative expression quantity up-regulation rate.
WISTAR Adult male rats tracheal instillation concentration was the uranium ore dust suspension of 5.0 mg/ml after 5 weeks, lung tissue
Middle α -1 antiprotease relative expression quantity up-regulation rate is 245.6 %, and biological damage grade caused by irradiation is in uranium ore dust at this time
III grade, occupation work personnel should be transferred from former work position, and receive appropriate treatment.
It is above only better embodiment of the invention, according to the above-mentioned design, those skilled in the art
Can also to this various modification can be adapted and transformation.For example, the type of transformation uranium ore suspension concentration, contaminating mode, transformation rat model,
Convert irradiation damage appraisement system etc..However, these similar transformation and modification belong to essence of the invention.
Claims (10)
1. it is a kind of using the method for irradiating biological damage in α -1 antiprotease evaluation uranium ore dust, received according to WISTAR rat
Dosage-effect of the up-regulation rate of α -1 antiprotease relative expression quantity in the amount and lung tissue of the uranium ore dust suspension of tracheal instillation
It should be related to, by the up-regulation rate of the relative expression quantity of α -1 antiprotease in WISTAR rat lung tissue, assess in uranium ore dust
Biological damage caused by irradiation, which is characterized in that comprise the concrete steps that:
(1) uranium ore suspension tracheal instillation WISTAR rat model is established；
(2) lung tissue is acquired；
(3) detection of α -1 antiprotease relative expression quantity up-regulation rate；
(4) biological damage overall merit is irradiated in；
The specific method for establishing uranium ore suspension tracheal instillation WISTAR rat model is:
Selective body focuses on the WISTAR Adult male rats between 180-220 g, and points 4 groups, every group of 6 mouse, control group gas
The suspension of pipe instillation physiological saline and silica, it is 1.25,2.5 and 5.0 mg/ml's that experimental group, which distinguishes tracheal instillation concentration,
Uranium ore dust suspension, instil 0.2 ml every time, instils 3 times, continuous 5 weeks within one week；
The specific method of the acquisition lung tissue is:
Rat is put to death with cervical dislocation, takes lung tissue after dissection, the part that classifies in three categories is organized each, falls blood with normal saline flushing
Water removes moisture, is attached in cryopreservation tube, is immediately placed into -80 DEG C of refrigerators and saves；
The detection specific method of the α -1 antiprotease relative expression quantity up-regulation rate is:
WISTAR rat lung tissue will be irradiated in the receiving of collection uranium ore dust carries out protein extraction, total protein concentration survey
Fixed, protein denaturation, transferring film, immune response, chemiluminescence reaction, gel imaging scanning analysis, finally carries out α-at gel electrophoresis
The relative expression quantity up-regulation rate of 1 antiprotease is analyzed；
The specific method of the interior irradiation biological damage overall merit is:
When the relative expression quantity up-regulation rate of α -1 antiprotease in WISTAR rat lung tissue is when within 30%, show low dose
Radiated by gamma-ray is measured to inanimate object damage effect；As the relative expression of α -1 antiprotease in WISTAR rat lung tissue
It measures up-regulation rate and is greater than 30% when less than 100%, biological damage grade caused by irradiation is I grade in uranium ore dust at this time；When
The relative expression quantity up-regulation rate of α -1 antiprotease in WISTAR rat lung tissue be greater than 100% and when less than 200%, at this time
Uranium ore dust in irradiation caused by biological damage grade be II grade；α -1 antiprotease in WISTAR rat lung tissue
Relative expression quantity up-regulation rate when being greater than 200%, biological damage grade caused by irradiation is III grade in uranium ore dust at this time.
2. a kind of method using irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1,
Phase is characterized in that,
Specific step is as follows for the protein extraction:
Liquid nitrogen precooler is poured into mortar, lung tissue's sample is taken out rapidly, tissue is switched to and is homogeneously disposed in mortar, is added at once
Enter liquid nitrogen, ground, 800 μ l LB lysates are slowly added into mortar, crack 30 min, vibrates 1-2 times, sufficiently split
Solution after cracking, collects the suspension of 1.5 mL into EP pipe, and then at 4 DEG C, 12000 r/min of revolving speed is centrifuged 20 min
Afterwards, supernatant is collected, with the albumen in resin precipitated supernatant, 4 DEG C, 12000 r/min are centrifuged 20 min, remove supernatant, dry in the air
Dry, -80 DEG C save backup, and 200-250 μ L L3 lysate is added in protein agglomerate after the drying, is blown and beaten repeatedly with pipette tips
Until protein is fully dispersed, ultrasonic dissolution assisting 4 DEG C, 12000 r/min, is centrifuged 20 min, supernatant is sucked out, is transferred to new
In EP pipe.
3. a kind of method using irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1,
Phase is characterized in that,
Specific step is as follows for the total protein concentration measurement:
The BSA standard solution of 0.2 mg/ml is added into dilution, is successively diluted to 0,5,10,20,50,100,150 and 200
Then the standard solution of μ g/ml measures their absorbance values at 562 nm, draw standard curve, measure each sample
Solution is after the absorbance value under 562 nm, then calculates by standard curve the concentration of protein in sample.
4. a kind of method using irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1,
Phase is characterized in that,
Specific step is as follows for the protein denaturation:
5 × protein sample-loading buffer is added according to the ratio of 4:1 in protein solution, boiling water bath is denaturalized 15 min, is put into -20
DEG C refrigerator saves backup.
5. a kind of method using irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1,
Phase is characterized in that,
Specific step is as follows for the gel electrophoresis:
30% acrylamide, 1.5 M TRIS-HCl, 10%SDS, 10% ammonium persulfate and TEMED are matched according to a certain percentage
10% separation gel and 5% concentration glue is made；After gelling to be separated is solid, electrophoresis liquid is added in electrophoresis tank, by the egg after denaturation
White matter sample is added in electrophoresis hole, carries out electrophoresis, concentrate glue voltage is adjusted to 75 V, separation gel voltage is adjusted to 100 V, and electrophoresis is extremely
Bromophenol blue has just been run to separation gel bottom, i.e. termination electrophoresis, and transferring film is carried out.
6. a kind of method using irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1,
Phase is characterized in that,
Specific step is as follows for the transferring film:
First pvdf membrane is activated using methanol, then places the filter paper of 5-6 7 × 9 cm in the bottom of film, is together placed
In transferring film instrument, addition transferring film liquid, 300 mA constant current transferring film, 30 min,
In During migration, transferring film slot is placed in ice water and is cooled down.
7. a kind of method using irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1,
Phase is characterized in that,
Specific step is as follows for the immune response:
5% skim milk of the film to take a turn for the better is closed, 1 h is mixed on decolorization swinging table,
1:1000 concentration dilution is carried out to α -1 antiprotease primary antibody using 5% skim milk of TBST dissolution, then by itself and envelope
The film closed is mixed, and is incubated overnight at 4 DEG C, is carried out clearly with the film that 2% TBST is incubated overnight to α -1 antiprotease primary antibody
It washes, cleans 3 times on decolorization swinging table at room temperature, every time 5 min,
8000 times finally are diluted to secondary antibody with TBST, after being incubated for 1.5 h at room temperature, is cleaned on decolorization swinging table three times, every time 10
min。
8. a kind of method using irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1,
Phase is characterized in that,
Specific step is as follows for the chemiluminescence reaction:
It is by two kinds of reagents of ECLA and ECLB in the medium volume mixture of centrifuge tube in darkroom, the albumen of pvdf membrane is face-up, add
Enter after the ECL solution mixed sufficiently reacts 1-2 min, is put into gel imaging system and is scanned analysis.
9. a kind of method using irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1,
Phase is characterized in that,
Specific step is as follows for the gel imaging scanning analysis:
It is taken pictures using gel imaging system to protein band, then using the gray value of Image J software analysis object tape, led to
It crosses to scan α -1 antiprotease albumen gray value and obtains α -1 antiprotease expressing quantity, to internal reference Protein G APDH albumen ash
Angle value scanning obtains GAPDH expressing quantity；By α -1 antiprotease expressing quantity divided by internal reference Protein G APDH protein expression
Amount obtains the relative expression quantity of α -1 antiprotease.
10. a kind of side for utilizing irradiation biological damage in α -1 antiprotease evaluation uranium ore dust according to claim 1
Method, phase be characterized in that,
Specific step is as follows for the relative expression quantity up-regulation rate analysis of α -1 antiprotease:
α -1 antiprotease (A1AT) relative expression quantity up-regulation rate is calculated by formula (1):
(1)。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910572069.4A CN110221337B (en) | 2019-06-28 | 2019-06-28 | Method for evaluating irradiation biological damage in uranium ore dust by using alpha-1 protease resistance |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910572069.4A CN110221337B (en) | 2019-06-28 | 2019-06-28 | Method for evaluating irradiation biological damage in uranium ore dust by using alpha-1 protease resistance |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110221337A true CN110221337A (en) | 2019-09-10 |
| CN110221337B CN110221337B (en) | 2022-07-01 |
Family
ID=67815245
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910572069.4A Active CN110221337B (en) | 2019-06-28 | 2019-06-28 | Method for evaluating irradiation biological damage in uranium ore dust by using alpha-1 protease resistance |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110221337B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112858692A (en) * | 2021-01-24 | 2021-05-28 | 南华大学 | BeSO4Method for detecting and analyzing lung tissue differential expression protein of infected rat |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006308533A (en) * | 2005-05-02 | 2006-11-09 | Mcbi:Kk | New hepatoma biomarker and hepatoma detection method using same |
| TW200726976A (en) * | 2005-11-18 | 2007-07-16 | Chi Mei Foundation Hospital | Biomarker for determining predisposition and/or prognosis of hepatocellular carcinoma |
| EP1963521A2 (en) * | 2005-12-06 | 2008-09-03 | Cynthia Bristow | Therapeutic use for alpha 1 proteinase inhibitor in hematopoiesis |
| US20100179067A1 (en) * | 2007-05-18 | 2010-07-15 | Patz Jr Edward F | Serum biomarkers for the early detection of lung cancer |
| CN102844661A (en) * | 2010-02-11 | 2012-12-26 | 香港理工大学 | Biomarkers of gastric cancer and use thereof |
| US20150105293A1 (en) * | 2009-05-14 | 2015-04-16 | The Chancellor, Masters And Scholars Of The University Of Oxford | Clinical diagnosis of hepatic fibrosis using a novel panel of low abundant human plasma protein biomarkers |
| JP2016048242A (en) * | 2008-08-28 | 2016-04-07 | アスチュート メディカル，インコーポレイテッド | Methods and compositions for diagnosis and prognosis of renal injury and renal failure |
| WO2017212348A1 (en) * | 2016-06-06 | 2017-12-14 | Uvic Industry Partnerships Inc. | Assay for quantitation of proteins and peptides using stable isotope standards |
-
2019
- 2019-06-28 CN CN201910572069.4A patent/CN110221337B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006308533A (en) * | 2005-05-02 | 2006-11-09 | Mcbi:Kk | New hepatoma biomarker and hepatoma detection method using same |
| TW200726976A (en) * | 2005-11-18 | 2007-07-16 | Chi Mei Foundation Hospital | Biomarker for determining predisposition and/or prognosis of hepatocellular carcinoma |
| EP1963521A2 (en) * | 2005-12-06 | 2008-09-03 | Cynthia Bristow | Therapeutic use for alpha 1 proteinase inhibitor in hematopoiesis |
| US20100179067A1 (en) * | 2007-05-18 | 2010-07-15 | Patz Jr Edward F | Serum biomarkers for the early detection of lung cancer |
| JP2016048242A (en) * | 2008-08-28 | 2016-04-07 | アスチュート メディカル，インコーポレイテッド | Methods and compositions for diagnosis and prognosis of renal injury and renal failure |
| US20150105293A1 (en) * | 2009-05-14 | 2015-04-16 | The Chancellor, Masters And Scholars Of The University Of Oxford | Clinical diagnosis of hepatic fibrosis using a novel panel of low abundant human plasma protein biomarkers |
| CN102844661A (en) * | 2010-02-11 | 2012-12-26 | 香港理工大学 | Biomarkers of gastric cancer and use thereof |
| US20130071857A1 (en) * | 2010-02-11 | 2013-03-21 | Samuel Chun-Lap Lo | Biomarkers Of Gastric Cancer And Use Thereof |
| WO2017212348A1 (en) * | 2016-06-06 | 2017-12-14 | Uvic Industry Partnerships Inc. | Assay for quantitation of proteins and peptides using stable isotope standards |
Non-Patent Citations (3)
| Title |
|---|
| ARTURO ORTEGA ET AL.: "Treatment of experimental acute uranium poisoning by chelating agents", 《PHARMACOLOGY & TOXICOLOGY》 * |
| VIJAY K SINGH ET AL.: "Use of biomarkers for assessing radiation injury", 《EXPERT REVIEWS》 * |
| 顾小利 等: "雾化吸入柠檬酸对贫铀致大鼠肺成纤维细胞增生的影响", 《中国药房》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112858692A (en) * | 2021-01-24 | 2021-05-28 | 南华大学 | BeSO4Method for detecting and analyzing lung tissue differential expression protein of infected rat |
| CN112858692B (en) * | 2021-01-24 | 2024-03-12 | 南华大学 | BeSO 4 Method for detecting and analyzing differential expression protein of lung tissue of infected rat |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110221337B (en) | 2022-07-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0064484A2 (en) | Method for fluorescence spectroscopic determination of a biologically active substance | |
| Nicolaieff et al. | Detection of rotavirus by serological trapping on antibody-coated electron microscope grids | |
| JPS58501637A (en) | Allergy screening device and its manufacturing and usage methods | |
| CN102023211A (en) | Immunochromatographic test strip for full quantitative detection of C-reactive protein and preparation method thereof | |
| CN111024956A (en) | Time-resolved fluorescence immunochromatography kit for detecting PTX3 | |
| CN104880393B (en) | A kind of device and method for measuring particular place PM2.5 | |
| Ashton et al. | Serum albumin and transferrin polymorphism in East African cattle | |
| CN108351358A (en) | Liver cancer test method | |
| CN111999508A (en) | Diagnostic marker and application thereof in COVID-19 diagnosis and coronavirus past infection detection | |
| CN107056891A (en) | One group of polypeptide and its application for preparation system lupus erythematosus diagnosis product | |
| CN110441296A (en) | A kind of preparation method based on Ru@MOF-5 bipotential Ratio-type Electrochemiluminescsensor sensor | |
| CN110221337A (en) | A method of utilizing irradiation biological damage in α -1 antiprotease evaluation uranium ore dust | |
| JP3362054B2 (en) | Squamous cell carcinoma and its separation method | |
| Vihko et al. | Rapid radioimmunoassay for prostate-specific acid phosphatase in human serum. | |
| CN206038688U (en) | Immunity chromatography detection test strip's fluorescent quantitation spectral detection system | |
| SAKAI et al. | Coproantigen detection in a survey of Echinococcus multilocularis infection among red foxes, Vulpes vulpes schrencki, in Hokkaido, Japan | |
| WO2024001044A1 (en) | Biomarker combination related to lung cancer, kit containing same, and use thereof | |
| CA1190853A (en) | Method and compositions for carcinoma diagnosis | |
| CN110031527A (en) | A kind of double reading biosensors of human thyroglobulin | |
| CN1873410A (en) | Method for detecting id marks related to tumor in blood serum from patient of oesophagus cancer | |
| Buffone et al. | Characterization and evaluation of immunochemical methods for the measurement of fecal α 1-antitrypsin | |
| CN110196320A (en) | A method of the biological damage of low dosage radiated by gamma-ray is evaluated using isocitric dehydrogenase 2 | |
| Rhodes et al. | Comparison of results using electron microscope, immunodiffusion and fluorescent antibody analyses to detect rotavirus in diarrheic fecal samples of calves. | |
| CN110108886A (en) | For the PRDX2 antibody colloidal gold test strip of on-site quick screening chronic obstructive pneumonia patient | |
| Louvet et al. | Method for extraction of glycoprotein hormones from plasma for use in radioimmunoassays |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |