CN105879064A - Examination indexes of SonoVue ultrasonic contrast technology in diagnosis and treatment of tumor angiogenesis mimicry - Google Patents
Examination indexes of SonoVue ultrasonic contrast technology in diagnosis and treatment of tumor angiogenesis mimicry Download PDFInfo
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Abstract
The invention discloses examination indexes of a SonoVue ultrasonic contrast technology in diagnosis and treatment of tumor angiogenesis mimicry and belongs to the technical field of clinical tumor vessel ultrasonic examination. The situation that ultrasonic examination indexes including the rising time, the time to peak and the average transition time can be used for clinical diagnosis and treatment of tumor angiogenesis mimicry is found for the first time, a basis is provided for clinical early noninvasive diagnosis of tumor angiogenesis mimicry, the grade malignancy, the blood supply mode and other characters of a tumor of a patient can be clinically diagnosed easily, and a diagnosis basis is provided for drug selection or operation treatment.
Description
Technical field:
The invention belongs to the technical field of clinical tumor vascular ultrasound, the Index for examination of non-invasive diagnosis tumor-blood-vessel growth mimicry offer in early days clinically is provided, contribute to grade malignancy and the blood supply mode feature thereof of clinical diagnosis patient tumors, and provide diagnosis basis for medicament selection or operative treatment.
Background technology:
Blood vessel occurs in tumor and has played important function in development.Blood vessel is not only tumor cell and provides oxygen and nutrient substance, transports toxic metabolic products, also provides blood passage for Nasopharyngeal neoplasms.Tumor vessel research starts from 1971, and blood vessel is contacted by Folkman first with tumor, and the proposition " Tumor vasculature targeting " of predictability is theoretical.Hereafter, along with the carrying out of tumor basic research, tumor vessel classification more refines, and specifically includes: endothelial lining vessels (EV), vasculogenic mimicry (VM), mosaic blood vessel (MV), lymphatic vessel (LV).At present, what research was relatively more is endothelial lining vessels and vasculogenic mimicry.
Maniotis in 1999 etc. are found that a kind of novel blood supply pipeline when studying melanoma at the bottom of human eye, it surrounds by tumor cell, in be lined with extracellular matrix.Fox Green Angiography associating confocal laser scanning microscope, CLSM finds there is red blood cell flow in this pipeline, shows that it is a kind of functional pipeline.This blood supply pipeline is independent of endotheliocyte, but can simulate the function of endothelial lining vessels, therefore referred to as vasculogenic mimicry.The generating process of vasculogenic mimicry need not endotheliocyte and participates in, but it can be connected with endothelial lining vessels, and this contributes to the rapid build of tumor microcirculation network.Additionally, vasculogenic mimicry and blood only one layer of cells epimatrix is separated by, tumor cell is easier to the transfer of penetration cell epimatrix generation blood.Visible, vascular mimicry plays an important role during tumor development.Clinical Retrospective Study finds, the tumor tissues such as hepatocarcinoma, glioma, ovarian cancer, astrocytoma, carcinoma of prostate all exists vasculogenic mimicry.Additionally, vasculogenic mimicry is also closely related with the grade malignancy of tumor, tumor invasion and metabasis ability, survival of patients phase and poor prognosis.At present clinically in addition to postoperative pathological examination tissue section strain, also there is no the effective ways of Noninvasive diagnosis vasculogenic mimicry.
The therapeutic scheme of tumor patient is selected by early discovery, treatment endpoint results is most important.Although existing kinds of tumors early diagnosis technology, such as tumor marker inspection, needle aspiration biopsy inspection, NMR (Nuclear Magnetic Resonance)-imaging, computed tomography and radionuclide scanning etc., but due to shortcomings such as side effect are big, sensitivity is low, check cost is high, these technology do not play intended effect in early diagnosis of tumor.And SonoVue ultrasonic contrast (Contrast-enhanced ultrasound) is one of Main Diagnosis technology being currently used in evaluation parenchymatous organ's blood perfusion situation.SonoVue ultrasonic contrast uses SonoVue microbubble contrast agent.SonoVue contrast agent is a kind of phospholipid microbubble including sulfur hexafluoride noble gas, and good stability shakes under low acoustic pressure effect and do not breaks, has good harmonic characterisitic.It addition, SonoVue contrast agent particle diameter is about 2-5 μm, suitable with red blood cell diameter, it is a kind of well blood pool contrast agent, exists only in Ink vessel transfusing and does not enter interstice, can the blood perfusion state of response organization exactly.Ultrasonic contrast instrument and SonoVue contrast agent are combined, can dynamically, the capillary vessel that clearly displays in parenchymatous organ and blood perfusion situation, accordingly histoorgan pathological changes situation is carried out etiologic diagnosis.Additionally, ultrasonic contrast also has multiple advantages such as non-invasive, safety, real-time, inspection fee be relatively low.
Ultrasonic contrast is the vascularity situation of substantive tissue, blood flow characteristics, perfusion feature and vascular morphology for the main theory foundation of clinical tumor diagnosis, and main detection content is the reinforced foam plastics of contrast agent, enhancing level, distribution characteristics and enhancement mode.The blood perfusion situation that time-activity curve (TIC) is organized during can reflecting ultrasonic contrast, areas (AUC) etc. under wherein ultrasonic contrast quantitative parameter mainly includes rise time (RT), peak time (TTP), peak strength (IMAX), Mean Transit Time (mTT) and time-density curve, they can the hemoperfusion situation of response organization's organ from different perspectives.At present, the overall blood supply situation of tumor tissues can only be evaluated by ultrasonic contrast, still endothelial lining vessels in tumor tissues and vasculogenic mimicry can not be carried out qualitative differentiation and quantitative analysis, which greatly limits the development and application in benign from malignant tumors diagnoses of the SonoVue ultrasonic contrast.
Through investigation, having no pertinent literature report (from CNKI, tieing up general CNKI) of SonoVue contrast-enhanced ultrasound technique Index for examination in diagnosis and treatment tumor-blood-vessel growth mimicry, therefore this block research field is still in the blank stage.
Summary of the invention:
The invention provides SonoVue contrast-enhanced ultrasound technique Index for examination in diagnosis and treatment tumor-blood-vessel growth mimicry, including rise time, peak time and Mean Transit Time, these Index for examinations can be used for clinical diagnosis and the treatment of tumor-blood-vessel growth mimicry.
Accompanying drawing explanation
Fig. 1 is mouse tumor ultrasonic contrast figure (A: mouse tumor conventional Ultrasound；B: color doppler ultrasonography；C, D:SonoVue ultrasonic contrast detects).
Detailed description of the invention:
Embodiment 1:
The preparation of 1.1H22 tumor source Mus
Take out frozen mice H22 cell strain, quick-thawing at bath temperature 39 DEG C from-80 DEG C of ultra cold storage freezers, aseptically use normal saline dilution cell suspension, count under optical microscope, adjust cell concentration to 2 × 107-4×107Individual/mL.Draw 0.3mL cell diluent with 1mL syringe and be inoculated into ICR mouse peritoneal, conventional raising, make H22 tumor source Mus.
1.2H22 the preparation of bearing mouse model
After H22 tumor source Mus raises 7d, choosing the abdominal part obvious mice of protuberance, de-neck is put to death.Reject the tumor source Mus having bloody ascites.Aseptically cut mouse part skin open with shears, peritoneum is cut a duck eye, then extracts ascites with 1mL needleless injector, gained ascites is placed in the centrifuge tube of ice bath.Dilute with physiological saline solution, count under optical microscope, adjust cell concentration to 2 × 107-4×107Individual/mL.Two people coordinate, and a people is responsible for drawing cell diluent, and a people binds mice and inoculates.Right side of mice axillary fossa is subcutaneous, every mouse inoculation 0.2mL cell diluent, and inoculum concentration is 4 × 106-8×106Individual/only.Seeded process notices inserting needle position and angle, it is to avoid cell diluent spills.
The Contrast-enhanced ultrasonography of 1.3 mouse tumors
3d after inoculation, according to right side of mice skin of axillary fossa protuberance situation, rejects the mice that modeling is failed.Then after mouse inoculation H22 tumor the 6th, 9,12,15d totally 4 time points, 5 mices of random choose carry out ultrasonic contrast to ultrasound medicine section of Wuxi City the People's Hospital respectively.Before ultrasonic contrast, with depilatory solution, right side of mice axillary fossa tumor locus hair is removed in advance, it is to avoid when ultrasonic, produce Image noise.
Instrument and equipment: Philips iU22 diasonograph, 12L5 pops one's head in row routine examination, and 9L3 pops one's head in row ultrasonic contrast.
Photo condition is arranged: mechanical index M10.10, focus point is positioned at directly over tumor.All radiography parameters are arranged in research process and all keep constant.
Acoustic contrast agent: SonoVue contrast agent (Bu Laike company of Italy), experiment forms microvesicle suspension after adding injection normal saline 5mL shaking before starting, now with the current.
Ultrasonic contrast step is as follows:
(1) under waking state, mice adhesive tape is fixed on cystosepiment so that it is in dorsal position, fixes mice extremity and mouth, in case mice tampers in angiographic procedure or hurts sb.'s feelings.
(2) detect tumor locus with conventional Ultrasound before radiography, measure the length and width of Subcutaneous tumor, thick three radial lines.Determine, with color Doppler, the big blood vessel that blood flows into, observe the echo of inside tumor, blood supply and downright bad situation.
(3) 0.1mL injector for medical purpose is used to connect No. 5 half scalp intravenous needles, through mouse tail vein injection 0.02mL contrast agent.Contrast agent starts video recording while injecting, observe contrast agent mobility status in tumor, until contrast agent dissipates, terminates video recording, and storage radiography overall process is for off-line analysis.
(4) mice finishing ultrasonic contrast is taken back laboratory, put to death through de-neck, strip tumor, weigh.First with normal saline, the blood of tumor surface being cleaned, being fixed with 10% formaldehyde, in case carrying out the experiment of embodiment 5 afterwards.
1.4 ultrasonic contrast graphical analyses
(1) being derived from Philips iU22 diasonograph in dicom format by ultrasonic contrast video, quarter, dish was standby.
(2) video recording of DICOM format radiography is imported Sonoliver software, select the time period interested according to contrast agent intensity.
(3) with reference to conventional ultrasound images, selecting exceptions area (tumor area) and normal district (check plot) in ultrasonic contrast image successively is region of interest (region of interesting, ROI), avoid the highlight bar produced because of skin, demarcation line as far as possible.
(4) carry out motion compensation, eliminate the noise that is mobile and that produce that tampers because of mice in angiographic procedure or pop one's head in.
(5) quantitative analysis obtains original time-activity curve (TIC), fit time intensity curve, and every mice replicate analysis three times, degree of fitting (quality of fit, QOF) >=75% is valid data.
(6) derivation includes the EXCEL file of the quantitative parameters such as rise time (RT), peak time (TTP), peak-peak intensity (IMAX), Mean Transit Time (mTT) and includes the JPG file of time-activity curve, preserves whole data handling procedure.
1.5 tumor tissues embedding and microsection manufactures
Specific experiment step is as follows:
(1) washing the formaldehyde of inside tumor off: tumor taken out from 10% formaldehyde fixative, flowing water rinses 1h.
(2) gradient alcohol dehydration: 70% ethanol overnight, 80% ethanol 1h, 90% ethanol 20min, a time 95% ethanol 20min, two passage 95% ethanol 20min, a time 100% ethanol 30min, two passage 100% ethanol 30min.
(3) dimethylbenzene is transparent: a time 100% dimethylbenzene 20min, two passage 100% dimethylbenzene 20min.
(4) tissue waxdip: 60 DEG C of the left box of wax cylinder leaching 40min, 60 DEG C of the right box of wax cylinder leaching 40min.
(5) organization embedding: use plastic embedding box and supporting rustless steel embedded box base, 65 DEG C of wax liquid embeddings.
(6) section: make 4-6 μm paraffin section with microtome.
(7) stand sheet: 40 DEG C of thermostat water bath stand sheets, rejecting exhibition sheet is incomplete, organize crannied section.
(8) paster: 60 DEG C of constant temperature blast drying ovens, is placed into section setting on section frame, 4-5h paster.
(9) section preserves: the section left at room temperature over night in tissue patch jail, 4 DEG C of preservations.
The double dye of the CD31 SABC-PAS of 1.6 tumor tissue sections
Specific experiment step is as follows:
(1) roasting sheet: 60 DEG C of constant temperature blast drying ovens, 30min.
(2) dimethylbenzene dewaxing: a time 100% dimethylbenzene 10min, two passage 100% dimethylbenzene 15min, three passage 100% dimethylbenzene 20min.
(3) graded ethanol aquation: 100% ethanol 10min, 100% ethanol 5min, 95% ethanol 5min, 90% ethanol 5min, 80% ethanol 5min, 70% ethanol 5min.Deionized water embathes aquation 5min, three times.
(4) antigen retrieval: citric acid repair liquid preheats in advance, 90-100 DEG C of water-bath 20min.After reparation, the citric acid repair liquid being soaked with tissue slice is naturally cooled to room temperature.PBS embathes 5min, three times.
(5) peroxide enzyme-deactivating: to tissue dropping 3% hydrogen peroxide, lucifuge reaction 15min, reaction temperature 37 DEG C.PBS embathes 5min, four times.
(6) BSA closes: dropping doctor's moral test kit BSA confining liquid, reacts 20min, reaction temperature 37 DEG C.
(7) one anti-hatch: with PBS by the anti-Anti-CD31antibody of 1:40 dilution proportion one, 4 DEG C of preservations.Getting rid of BSA confining liquid, dropping one resists, 4 DEG C of refrigerator overnight.
(8) reclaim one to resist: equilibrium at room temperature 30min, reclaim one and resist.PBS embathes 5min, five times.
(9) two anti-hatch: the anti-solution of doctor's moral test kit two hatches 20min, incubation temperature 37 DEG C.PBS embathes 5min, four times.
(10) SABC is hatched: doctor's moral test kit SABC solution hatches 20min, incubation temperature 37 DEG C.PBS embathes 5min, four times.
(11) DAB colour developing: DAB nitrite ion A liquid and B liquid equal-volume are mixed, now with the current.In darkroom, drip DAB nitrite ion, lucifuge reaction 1-2min.Microscopy under microscope, terminates reaction with water in time.PBS embathes 5min, twice.
(12) PAS-A liquid dyeing: thunder root biology A liquid, lucifuge is hatched 1min, is terminated reaction with water in time.Flowing water rinses 7min, PBS and embathes 5min, once.
(13) PAS-B liquid dyeing: thunder root biology B liquid, lucifuge is hatched 1min, is terminated reaction with water in time.Flowing water rinses 7min.
(14) haematoxylin is redyed: the haematoxylin that 4 DEG C preserve is shifted to an earlier date equilibrium at room temperature, contaminates 1min, terminates reaction with water in time.Flowing water rinses anti-blue 7min.
(15) hydrochloride alcohol differentiation: break up 5 seconds, terminate reaction with water in time.Flowing water rinses anti-blue 7min.
(16) microscopy: observe dyeing depth degree, it may be judged whether need again to dye or break up.
(17) gradient alcohol dehydration: 70% ethanol 5min, 80% ethanol 5min, 90% ethanol 5min, 95% ethanol 10min, a time 100% ethanol 20min, a time 100% ethanol 30min.
(18) dimethylbenzene is transparent: a time 20min, two passages 20min.
(19) mounting: neutral gum mounting.Microscopy: observe and have bubble-free, if need mounting again.
1.7 vasculogenic mimicries, endothelial lining vessels analysis of accounts
Vascular counts is carried out with reference to Weidner method.First under low power lens (100 ×), observe whole section, choose the hot spot region that vasculogenic mimicry density is the highest, under high power lens (200 ×), then shoot 10 different hot spot region photos.Finally with the vasculogenic mimicry number of Image-Pro Plus 6.0 every photo of software statistics, result represents with Mean ± SD.Vasculogenic mimicry inclusion criteria: first, CD31 antibody staining feminine gender-PAS stained positive is prerequisite.Secondly, tube chamber close and in have erythrocyte.Finally, without the most downright bad and inflammatory infiltration around tube chamber.
1.8 data statisticss and analysis
Using SPSS 18.0 statistical software to carry out analysis of experimental data, all results all represent with Mean ± SD.Correlation analysis uses Pearson inspection, and between many groups, mean compares employing one factor analysis of variance, and between two groups, mean compares employing independent samples t test.Statistically significant with P < 0.05 and P < 0.01 for difference.
1.9 experimental result
1.9.1 the two-dimensional ultrasound of H22 tumor-bearing mice and SonoVue Contrast-enhanced ultrasonography result
As shown in Fig. 1 .A, before radiography, first detect tumor locus with conventional Ultrasound, measure and record the length and width of mice H22 tumor, thick three radial lines.Then, choosing, with color Doppler, the big blood vessel that blood flows into, the blood supply of gross examination of skeletal muscle inside tumor, downright bad situation, as shown in Fig. 1 .B.Finally, SonoVue contrast agent injects and starts simultaneously at video recording, observes contrast agent mobility status in tumor, during until contrast agent dissipates, terminates video recording and stores.As shown in Fig. 1 .C, with reference to conventional ultrasound images, ultrasonic contrast image selects exceptions area (tumor area, green circle labelling) and (check plot, normal district successively, yellow circle labelling) it is region of interest (ROI), avoid the highlight bar produced because of skin, demarcation line.As shown in Fig. 1 .D, obtain original time-activity curve (the upper figure in Fig. 1 .D), fit time intensity curve (figure below in Fig. 1 .D) by quantitative analysis.The quantitative parameters such as rise time (RT), peak time (TTP), peak-peak intensity (IMAX), Mean Transit Time (mTT) can be obtained from figure.
1.9.2 vasculogenic mimicry density and the correlation analysis of SonoVue ultrasonic contrast quantitative parameter
This research, by H22 bearing mouse model, arranges multiple time point, is obtained the Changing Pattern of vasculogenic mimicry by cytological stains and microscopic counting, and result is as shown in table 1.With mice H22 tumor growth, vasculogenic mimicry density in tumor tissues is in steady statue after first increasing, vasculogenic mimicry is formed and occurs in 6d after H22 tumor inoculation the earliest, 6-9d after mice H22 tumor inoculation is obvious ascendant trend, shows that vasculogenic mimicry is formed all relatively more active.There is angiogenesis and mean that tumor tissues severe depletion of oxygen, mice H22 tumor 1-9d tumor after inoculation heavily gathers way more slow, just demonstrates this point.After mice H22 tumor inoculation, 9-15d vessel density ascendant trend slows down, and shows that the anaerobic condition of tumor tissues is improved, and the blood that i.e. in unit are, blood capillary is supplied be enough to maintain the existence of tumor cell to grow.9-15d after mice H22 tumor inoculation, mouse tumor heavily gathers way and substantially accelerates, just demonstrates this point.
The experimental result of table 1 mice H22 tumor-blood-vessel growth mimicry density
Peak strength (IMAX) refers to the maximum intensity of contrast agent in tumor, the vessel density in reflection certain cross section of tumor.Rise time (RT) refers to that contrast agent in tumor is by the time required for upstroke 10%IMAX to upstroke 90%IMAX.Peak time (TTP) refers to that in tumor, contrast agent is injected into the time required for 100%IMAX.Rise time (RT) and peak time (TTP) reflect the inflow velocity of tumor imaging agent jointly.Mean Transit Time (mTT) refers to that contrast agent in tumor is by the time required for upstroke 50%IMAX to decent 50%IMAX, the elution speed of reflection contrast agent.The analysis result of H22 tumor-bearing mice ultrasonic contrast quantitative parameter is as shown in table 2.
The analysis result of table 2 H22 tumor-bearing mice ultrasonic contrast quantitative parameter
Checking with P through statistical analysis, result is as shown in table 3, and research finds the vasculogenic mimicry density in mice H22 tumor and rise time, peak time, Mean Transit Time significant positive correlation.
Table 3 vasculogenic mimicry density and the correlation analysis of SonoVue ultrasonic contrast quantitative parameter
The significance degree of two correlation of variables, * be P < 0.05, * * be P < 0.01
The present invention combines most preferred embodiment and is described, but after having read the foregoing of the present invention, those skilled in the art can appreciate the fact that making many in the embodiment disclosed changes and also can obtain same or similar result, and without departing from the design of the present invention, spirit and scope.More specifically, it is clear that some alternative reagent disclosed herein of chemical and physiological related reagent and obtain same or similar result.All similar replacements and modification are for a person skilled in the art, obviously being regarded as in the spirit of the present invention, scope and spirit and right, the most all these equivalent form of values above-mentioned and all improvement to technological parameter and variation all also belong to claims of the present invention limited range.
Claims (3)
1.SonoVue contrast-enhanced ultrasound technique Index for examination in diagnosis and treatment tumor-blood-vessel growth mimicry.
The most according to claim 1, these Index for examinations include rise time, peak time and Mean Transit Time.
The most according to claim 1, rise time, peak time and Mean Transit Time are used for facing as the Index for examination of tumor-blood-vessel growth mimicry
Bed Clinics and Practices.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110458834A (en) * | 2019-02-25 | 2019-11-15 | 腾讯科技（深圳）有限公司 | A kind of tumor of breast image processing system, method and device |
| CN111047580A (en) * | 2019-12-14 | 2020-04-21 | 四川大学华西医院 | Acoustic image quantitative analysis and comprehensive quantitative analysis method for ultrasonic molecular image research |
| CN111798432A (en) * | 2020-07-07 | 2020-10-20 | 刘军 | Dynamic video image processing method for angiography |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101829326A (en) * | 2009-12-19 | 2010-09-15 | 刘政 | Use of microbubbles with high-intensity cavitation generated by combining ultrasonic in preparation of anti-tumor neovascularization medicaments |
| US20140161732A1 (en) * | 2002-03-01 | 2014-06-12 | Bracco Suisse S.A. | Kdr and vegf/kdr binding peptides and their use in diagnosis and therapy |
| CN104189918A (en) * | 2014-07-21 | 2014-12-10 | 香港中文大学深圳研究院 | Application of dynein light chain roadblock-type 2 in preparation of medicine treating kidney failure and the medicine thereof |
-
2016
- 2016-04-08 CN CN201610216415.1A patent/CN105879064A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140161732A1 (en) * | 2002-03-01 | 2014-06-12 | Bracco Suisse S.A. | Kdr and vegf/kdr binding peptides and their use in diagnosis and therapy |
| CN101829326A (en) * | 2009-12-19 | 2010-09-15 | 刘政 | Use of microbubbles with high-intensity cavitation generated by combining ultrasonic in preparation of anti-tumor neovascularization medicaments |
| CN104189918A (en) * | 2014-07-21 | 2014-12-10 | 香港中文大学深圳研究院 | Application of dynein light chain roadblock-type 2 in preparation of medicine treating kidney failure and the medicine thereof |
Non-Patent Citations (1)
| Title |
|---|
| 姜玉新 等: "《浅表器官及组织超声诊断学》", 31 January 2010 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110458834A (en) * | 2019-02-25 | 2019-11-15 | 腾讯科技（深圳）有限公司 | A kind of tumor of breast image processing system, method and device |
| US11928816B2 (en) | 2019-02-25 | 2024-03-12 | Tencent Technology (Shenzhen) Company Limited | Image processing method, apparatus, and system, electronic device, and storage medium |
| CN111047580A (en) * | 2019-12-14 | 2020-04-21 | 四川大学华西医院 | Acoustic image quantitative analysis and comprehensive quantitative analysis method for ultrasonic molecular image research |
| CN111047580B (en) * | 2019-12-14 | 2022-08-23 | 四川大学华西医院 | Acoustic image quantitative analysis and comprehensive quantitative analysis method for ultrasonic molecular image research |
| CN111798432A (en) * | 2020-07-07 | 2020-10-20 | 刘军 | Dynamic video image processing method for angiography |
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