CN103336131A - Application of soluble CD30 as biological index for rapidly estimating ionizing radiation dosage - Google Patents
Application of soluble CD30 as biological index for rapidly estimating ionizing radiation dosage Download PDFInfo
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- CN103336131A CN103336131A CN2013102559720A CN201310255972A CN103336131A CN 103336131 A CN103336131 A CN 103336131A CN 2013102559720 A CN2013102559720 A CN 2013102559720A CN 201310255972 A CN201310255972 A CN 201310255972A CN 103336131 A CN103336131 A CN 103336131A
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Abstract
The invention belongs to a biological index for rapidly estimating the ionizing radiation dosage and particularly relates to an application of soluble CD30 (sCD30) as a biological index for rapidly estimating the ionizing radiation dosage. After radiation, the concentration of the sCD30 in blood serum or blood plasma is reduced along with the increasing of the radiation dosage; after the radiation is carried out within a certain time range, a very good dosage-effect relation exists; the rapid detection can be carried out by using a simple and feasible method through ELISA (Enzyme Linked Immunosorbent Assay), a protection chip and the like; a little amount of sample is needed by the detection and the sampling is convenient. Therefore, the sCD30 can be used as the biological index for rapidly estimating the ionizing radiation dosage; the radiation dosage of a human body and an animal after ionizing radiation is rapidly calculated by adopting the content of the sCD30 in the blood serum or the blood plasma.
Description
1 technical field
The invention belongs to ionising radiation biological dose estimation field.Be specifically related to solubility CD30 (sCD30) as the application of estimation ionizing radiation dose Biological indicators fast.
2 background technologies
The countries in the world pay attention to day by day is used nuclear energy, and China's nuclear power is strided forward to " develop actively " by " optimum development ", will play the part of very important role in the electric power supply in future.The fast development of science and technology makes radioactive source and radiotechnology in industrial or agricultural, medical science and the also widespread use of fields such as teaching even military affairs.Yet nuclear technology has also constituted potential threat to public safety and public health when promoting the well-being of mankind.Acute radiation sickness is the concentrated reflection of this threat.Acute radiation sickness refers to body once or interior gradation of short time (a few days) is subjected to the systemic disease that heavy dose of ionization radiation irradiation causes.In general, human body is subjected to whole body about 1Gy evenly or uniformly behind the ionization radiation irradiation, just acute radiation sickness may take place.Acute radiation sickness is a kind of determinacy effect that ionising radiation is caused injury, and also is the most serious radiation biological effect.Treatment scheme and the radiation sickness somatotype of acute radiation sickness are closely related, and acute radiation sickness can be divided into three types: BM form (1～10Gy), visible peristalsis visible intestinal peristalsis (10～50Gy) and the brain type (＞50Gy).In the exposure dose scope of bone marrow form acute radiation sickness, along with the increase of exposure dose, mortality ratio obviously increases, corresponding shortening of time-to-live; On the complexity of clinical manifestation, prognosis and treatment measure, also have difference.Yet, the above BM form radiation sickness of moderate (＞2Gy) there is latent phase, if accurately do not estimate actual suffered exposure dose, delay treatment easily, finally cause the loss of life.Therefore, fast, accurately estimate the radiation dose scope, be conducive to improve the radiation protection measure, also be conducive to guiding clinical treatment, prediction PD, save life.
Biological dose is that a certain biological substance or cell act on aitiogenic degree behind the body to radioactive ray in the body, is dose-dependence with radioactive dose, and is objective and reflect the whole body radioactive dose specifically.Compare with the physical dosage meter, biological dosemeter can reflect more intuitively that ionising radiation is to the degree of injury of body.In recent years, along with cell and molecular biological fast development, research to the ionising radiation biological dosemeter has both at home and abroad obtained certain progress, comprise the detection equimolecular biological analysis research of CYTOGENETIC ANALYSIS OF ONE such as chromosome aberration, lymphocyte micronucleus, precocious aggegation chromosome and fluorescence in situ hybridization and gene mutation, minisatellite DNA site mutation and mitochondrial DNA deletion, but traditional biological dosemeter such as chromosome aberration analysis method relative complex, need cell to cultivate, sense cycle is long, usually need 2～3 days, certain limitation is arranged.Therefore be necessary to seek novel, quick, reliable ionising radiation biological dose index.
CD30 finds as the surface antigen of the hodgkin's cells in the hodgkin's lymphoma and Reed-sternberg cell the earliest.The human CD30cDNA of coding was successfully cloned in 1992, and was included into the TNF receptor superfamily.CD30 is that molecular weight is the I type transmembrane glycoprotein of 120Kda, finds first that CD30 existed with trimeric form in 2000, and the N of CD30 after birth outskirt holds and the C end is adjacent to each other, and exists with membrane-bound trimeric form, forms a style structure.The CD30 of cell surface is activated after its part CD30L is combined, and activates the outer part of its born of the same parents of back and is had no progeny by proteolytic cleavage that to have formed molecular weight be the soluble molecule of 85Kd---sCD30.SCD30 content and CD30
+Cell quantity does not have obvious correlativity, and in virus infections and some autoimmune diseases, the sCD30 abnormal expression acts on not clear so far yet which kind of sCD30 brings into play in disease.In addition, sCD30 content and ionizing radiation dose correlativity do not have report so far.SCD30 content can be by ELISA and protein chip method fast detecting, so if be dosimeter for slit radiographic apparatus with sCD30, can judge radioactive dose fast by serum or blood plasma ELISA or protein chip method, has certain theory innovation and method innovation.
3 summary of the invention
The object of the invention is to provide the application of solubility CD30 (sCD30) as the ionising radiation biological dosemeter.
In order to achieve the above object, the technical solution used in the present invention is: sCD30 is as the application of ionising radiation biological dosemeter in the serum.
Step 1: sample collection: certain hour behind the irradiation, get blood, retain serum or plasma sample.
Step 2: pattern detection: detect sCD30 content in the sample.
Step 3: dosage estimation: according to sCD30 content, according to the back time point, the estimation ionizing radiation dose.
In the technique scheme, serum or plasma sample derive from mammal.
In the technique scheme, radiation dose is as the criterion with integral dose.
In the technique scheme, the sCD30 content detection can be passed through ELISA, chip or other biological detection means and realize.
In the technique scheme, sCD30 content can be united other index, estimates radiation dose jointly.
Compared with the prior art the present invention has following notable feature:
(1) sample collection is convenient, only needs serum or blood plasma to get final product.
(2) the sample requirement is few, and detecting only needs 100 μ l serum or blood plasma.
(3) sample is preserved conveniently, can preserve in short-term at 4 ℃, also can be-80 ℃ of long preservation.
(4) lack detection time, can finish ELISA or cell factor chip detection in the 15h, be fit to estimation ionizing radiation dose fast.
(5) result is objective, but by microplate reader or chip scanner quantitative test sCD30 content, does not need artificial subjective counting or judgement.
4 description of drawings
Fig. 1 sCD30 chip detection typical curve
The change of Fig. 2 different time points sCD30 content in normal serum
Fig. 3 different time points sCD30 content changes and variation tendency with ionizing radiation dose
5 embodiments
Below in conjunction with specific embodiment, further illustrate the present invention.Should be understood that these embodiments only to be used for explanation the present invention and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example is usually according to normal condition.
Embodiment one: foundation and the sample collection of mouse irradiation model
Animal used as test: male C57BL/6 mouse, 7 ages in week, body weight 19 ± 1g, available from Shanghai Slac Experimental Animal Co., Ltd., animal quality conformity certification number: 2007000534826, animal used as test production licence number SCXK (Shanghai) 2007-0005.
Serum is prepared: mouse irradiation exists
60Carry out at cobalt irradiation center, and 80 mouse are divided into four groups at random, 20 every group: wherein control group is 1 group, 4 groups of radiation groups.Radiation group mouse is accepted the disposable full-body exposure of 1Gy, 3Gy, 5Gy and 10Gy respectively, and dose rate is 0.78Gy/min.According to back 24h, 48h and 72h, mouse is got blood through eyeball, and whole blood is in 37 ℃ of blood coagulation 1h, and 3000rpm * 15min is centrifugal, separates upper serum, is sub-packed in the centrifuge tube with 100 μ l/ pipes.Divide the serum that installs to be stored in-80 ℃.12h before detecting takes out serum from-80 ℃ of refrigerators, places 4 ℃ of thawings.
Embodiment two: sCD30 content detection in the mice serum
(1) sCD30 content detection in the radiation murine serum not
Test material and method:
Adopt the cytokine antibodies chip detection.The chip name is called:
Custom Mouse
The Array chip customizes the company in RayBiotech.
The chip detection operation steps is as follows:
(1) placed 30 minutes in the chip bag room temperature, tear the envelope film off, tagging chip, dry 2 hours of super-clean bench;
(2) sample is prepared: 60 μ l serum+60 μ l dilutions mix;
(3) preparation of standard items: get 10000pg/ml titer (std1), 6 times (std2～7) of three times of dilutions, least concentration is 13.7pg/ml (std7).
(4) preparation of negative control: get dilution 120 μ l (CNTRL)
(5) chip sealing and sample are hatched: every hole adds the 100Rl confining liquid, room temperature sealing 0.5h, and jog adds 100 μ l negative controls, standard items and sample, and 4 spend night hatches jog.According to kit explanation preparation washing lotion I, II, each 600ml, every hole adds washing lotion I150 μ l, and room temperature was washed 5 minutes, washed 5 times, and every hole adds washing lotion II150 μ l then, and room temperature was washed 5 minutes, washed 2 times.
(6) detect hatching and cleaning of mixtures of antibodies: the by specification requirement adds 1.4ml sample diluent in Detection antibody pipe, the mixing dissolving.Every hole adds 80 μ l and detects mixtures of antibodies.Incubated at room 2 hours, jog.Every hole adds washing lotion I150 μ l, and room temperature was washed 5 minutes, washes 5 times.Every hole adds washing lotion II150 μ l, and room temperature was washed 5 minutes, washes 2 times.
(7) the hatching and cleaning of Cy3 mark Streptavidin: by specification requires to add 1.4ml sample diluent in Cy3equivalent dye-conj μ gated streptavidin pipe, the mixing dissolving.Every hole adds 80 μ l Cy3 mark Streptavidin solution, and incubated at room 1-2 hour, every hole added washing lotion I150 μ l, and room temperature was washed 5 minutes, washed 5 times.Unload slide, washing lotion I room temperature lucifuge washing 15min in the Washer/Dryer slide pipe.Washing lotion II room temperature lucifuge washing 5min in the Washer/Dryer slide pipe.Chip is washed dried instrument and is dried.
(8) fluoroscopic examination.
Testing result: along with standard items concentration increases, antigen-antibody reaction strengthens, and obtains sCD30 typical curve (see figure 1) and sCD30 detectability, is (32.4～4000pg/ml); According to typical curve, obtain the not irradiation serum sCD30 concentration that different time points detects, see the following form and Fig. 2.As seen from table, do not shine mice serum sCD30 expression and stablize, concentration range is: 98.7～152.9.1pm/ml.
Table 1 normal mouse serum sCD30 expression
(2) according to back 24h, the various dose ionising radiation is to the influence of mice serum sCD30 content.
Test material and method: with embodiment one.
Testing result: the result is shown in Fig. 3 and table 2, and according to back 24 hours, when radiation dose was 1Gy, sCD30 concentration namely dropped to 82.9pg/ml by 121.2pg/ml, and when radiation dose was 3Gy～5Gy, its expression dropped to 52.6～62.3pg/ml; During 7Gy, expression more drops to 42.1pg/ml, above presentation of results, when 24h was described behind the irradiation, the variation of serum sCD30 content was relevant with radiation dose, increases its expression with radiation dose and reduces, wherein, when radiation dose was 3Gy～5Gy, serum sCD30 content was more stable.
24h after table 2 acute exposure, serum sCD30 expression changes
(3) according to back 48h, the various dose ionising radiation is to the influence of mice serum sCD30 content.
Animal used as test: with embodiment one.
Chmice acute irradiation and serum are prepared: mouse irradiation exists
60Carry out chamber, cobalt source, and 80 mouse are divided into four groups at random, 20 every group: wherein control group is 1 group, 3 groups of radiation groups.Radiation group mouse is accepted the disposable full-body exposure of 1Gy, 3Gy, 5Gy and 7Gy respectively, and dose rate is 0.78Gy/min.According to back 48h, mouse is got blood through eyeball, and whole blood is in 37 ℃ of blood coagulation 1h, and 3000rpm * 15min is centrifugal, separates upper serum, is sub-packed in the centrifuge tube with 100 μ l/ pipes.Divide the serum that installs to freeze in-80 ℃.12h before detecting takes out serum from-80 ℃ of refrigerators, places 4 ℃ of thawings.
Test material and method: with embodiment one.
Testing result: the result is shown in Fig. 3, table 3, according to back 48 hours, when radiation dose is 1Gy, the concentration of serum sCD30 is reduced to 76.8pg/ml, when dosage was respectively 3Gy, 5Gy and 7Gy, the sAXL mean concentration reduced to 72.0,29.0 and 32.7pg/ml, and compared according to back 24h, when radiation dose 〉=5Gy, the sCD30 mean concentration descends obviously.
48h after table 3 acute exposure, serum sCD30 expression changes
(4) according to back 72h, the various dose ionising radiation is to the influence of mice serum sCD30 content.
Animal used as test: with embodiment one.
Chmice acute irradiation and serum are prepared: mouse irradiation exists
60Carry out chamber, cobalt source, and 80 mouse are divided into four groups at random, 20 every group: wherein control group is 1 group, 3 groups of radiation groups.Radiation group mouse is accepted the disposable full-body exposure of 1Gy, 3Gy, 5Gy and 7Gy respectively, and dose rate is 0.78Gy/min.According to back 48h, mouse is got blood through eyeball, and whole blood is in 37 ℃ of blood coagulation 1h, and 3000rpm * 15min is centrifugal, separates upper serum, is sub-packed in the centrifuge tube with 100 μ l/ pipes.Divide the serum that installs to freeze in-80 ℃.12h before detecting takes out serum from-80 ℃ of refrigerators, places 4 ℃ of thawings.
Test material and method: with embodiment one
Testing result: the result is shown in Fig. 3 and table 4, and according to back 72 hours, when radiation dose was 1Gy, the concentration of serum sCD30 was 100.21pg/ml, returns to normal level, and during but exposure dose 〉=3Gy, sCD30 decline degree and 48h are similar.Illustrate that along with time lengthening behind the irradiation, sCD30 that low dose irradiation causes descends and can be repaired, during but dosage 〉=3Gy, sCD30 descends and is not repaired.
72h after table 4 acute exposure, serum sCD30 expression changes
Embodiment three: mouse irradiation dose estimation
According to serum sAXL concentration and exposure time, estimation mouse radioactive dose, evaluation method sees Table 4～5.
Table 4 is estimated the mouse radioactive dose according to back 0～36h
Table 5 is estimated the mouse radioactive dose according to back 36～60h
Table 6 is estimated the mouse radioactive dose according to back 60～84h
The present invention, those skilled in the art can obviously can not break away from content of the present invention, the spirit and scope by using for reference this paper, and appropriate change animal model, radiation dose and detection method realize this application.Special needs to be pointed out is that the replacement that all are similar and change all are regarded as in spirit of the present invention, scope and content.
Claims (7)
1. solubility CD30 (sCD30) is as the application of estimation ionizing radiation dose Biological indicators fast.
2. sCD30 is as the application of estimation ionizing radiation dose Biological indicators fast in the claim 1, and it is characterized in that: the sCD30 expression is relevant with irradiation dose in the certain hour scope; Identical irradiation dose, sCD30 expression and photograph back time correlation.
3. irradiation dose is characterized as integral dose described in the claim 2.
4.sCD30 as the application of estimation ionizing radiation dose Biological indicators fast, may further comprise the steps:
Step 1: sample collection: certain hour behind the irradiation, get blood, retain serum or plasma sample.
Step 2: pattern detection: detect sCD30 content in the sample.
Step 3: dosage estimation: according to sCD30 content, according to the back time point, the estimation ionizing radiation dose.
5. the detection of the content of sCD30 described in the claim 4 can be passed through ELISA, protein chip or the realization of other biological detection means.
6. the content of sCD30 described in the claim 4 also can be united other indexs, estimates radiation dose jointly.
7. serum described in the claim 4 or plasma sample derive from mammal.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105968188A (en) * | 2016-06-19 | 2016-09-28 | 苏州普罗达生物科技有限公司 | CD30 immunogen polypeptide and application thereof |
| CN105968190A (en) * | 2016-06-19 | 2016-09-28 | 苏州普罗达生物科技有限公司 | CD30 immunogen polypeptide and application thereof |
| CN106084034A (en) * | 2016-06-19 | 2016-11-09 | 苏州普罗达生物科技有限公司 | A kind of CD30 immunogen polypeptide and application thereof |
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2013
- 2013-06-25 CN CN2013102559720A patent/CN103336131A/en active Pending
Non-Patent Citations (1)
| Title |
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| 江雪等: "紫甘薯多糖对辐射的防护作用", 《食品与生物技术学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105968188A (en) * | 2016-06-19 | 2016-09-28 | 苏州普罗达生物科技有限公司 | CD30 immunogen polypeptide and application thereof |
| CN105968190A (en) * | 2016-06-19 | 2016-09-28 | 苏州普罗达生物科技有限公司 | CD30 immunogen polypeptide and application thereof |
| CN106084034A (en) * | 2016-06-19 | 2016-11-09 | 苏州普罗达生物科技有限公司 | A kind of CD30 immunogen polypeptide and application thereof |
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Application publication date: 20131002 |