發(fā)布時(shí)間：2018-03-05 02:00

  本文選題：超聲造影檢查　切入點(diǎn)：Lewis肺癌　出處：《山東大學(xué)》2014年博士論文　論文類型：學(xué)位論文

【摘要】：第一部分超聲造影定量分析評(píng)價(jià)小鼠Lewis肺癌腫瘤血管的生成 研究背景及意義 腫瘤血管生成即指在腫瘤原有微血管網(wǎng)的基礎(chǔ)上通過(guò)內(nèi)皮細(xì)胞的“芽生”方式形成新生血管的過(guò)程,它是腫瘤生長(zhǎng)和轉(zhuǎn)移的一個(gè)至關(guān)重要的因素。在腫瘤內(nèi)及周圍的血管生成被認(rèn)為是惡性腫瘤細(xì)胞增殖的前提,這些新生血管不但能夠提供腫瘤細(xì)胞增殖所需的營(yíng)養(yǎng),也為腫瘤細(xì)胞在體內(nèi)的遠(yuǎn)處轉(zhuǎn)移提供了通道。所以血管生成是惡性腫瘤的重要特征,它的檢出對(duì)于腫瘤的診斷、治療與預(yù)后相當(dāng)重要。 在過(guò)去的幾年中,許多研究通過(guò)無(wú)創(chuàng)的影像學(xué)方法來(lái)評(píng)估腫瘤血管的生成,例如對(duì)比增強(qiáng)計(jì)算機(jī)斷層掃描(contrast enhanced computed tomography, CECT)、動(dòng)態(tài)對(duì)比增強(qiáng)磁共振成像(dynamic contrast enhanced magnetic resonance imaging, DCE-MRI),磁共振血管成像技術(shù)(magnetic resonance angiography, MRA）、正電子發(fā)射斷層掃描(positron emission tomography,PET)和多普勒超聲診斷技術(shù)等等。然而,以上這些方法在檢測(cè)低速血流和微小血管上,具有明顯的局限性。超聲造影明顯提高了對(duì)低速血流的檢出能力,使評(píng)價(jià)組織內(nèi)毛細(xì)血管和微血管的血流成為可能,并可提供關(guān)于腫瘤血管的詳細(xì)信息。微血管密度即平均每平方毫米腫瘤面積內(nèi)的血管數(shù),是目前評(píng)價(jià)腫瘤血管生成的公認(rèn)的參考標(biāo)準(zhǔn)。然而不同血管內(nèi)皮細(xì)胞標(biāo)記的微血管密度用于揭示不同腫瘤血管內(nèi)皮細(xì)胞的分化程度以及結(jié)構(gòu)和功能特點(diǎn)。在以往的實(shí)驗(yàn)研究中,較多使用CD31血管標(biāo)志的微血管密度,用以評(píng)價(jià)組織的血管生成情況。由于CD31微血管密度在已分化和未分化的血管內(nèi)皮細(xì)胞中均有表達(dá)；而另一常用的血管標(biāo)記物CD34則只表達(dá)于已分化的內(nèi)皮細(xì)胞中,故腫瘤血管的分化程度不同,其所代表的微血管密度亦有不同。 小鼠Lewis肺癌(Lewis lung carcinoma, LLC)是將傳代培養(yǎng)的Lewis肺癌細(xì)胞接種于C57/BL6小鼠的腋部皮下而成,已經(jīng)多次反復(fù)應(yīng)用于實(shí)驗(yàn)室的各項(xiàng)研究中,是較為成熟的動(dòng)物瘤實(shí)驗(yàn)?zāi)Ｐ汀?雖然既往已有研究證實(shí)超聲造影的定量分析參數(shù)與腫瘤的微血管密度存在一定的線性相關(guān)關(guān)系,到目前為止,還未有報(bào)道關(guān)于小鼠Lewis肺癌中不同血管標(biāo)記的微血管密度同超聲造影參數(shù)之間的相關(guān)性研究。研究目的 本研究通過(guò)建立小鼠LLC的皮下移植瘤動(dòng)物模型,應(yīng)用增強(qiáng)脈沖序列(contrast pulse sequencing, CPS)的超聲造影成像技術(shù),結(jié)合時(shí)間-強(qiáng)度曲線,研究并比較超聲造影的血流灌注參數(shù)與不同血管標(biāo)記的微血管密度在LLC動(dòng)物實(shí)驗(yàn)?zāi)Ｐ椭械南嚓P(guān)性,探討超聲造影在評(píng)價(jià)腫瘤血管生成方面的應(yīng)用價(jià)值。 研究方法 建立25只小鼠LLC皮下移植瘤模型,待腫瘤大小約1.0cm左右行超聲造影檢查,應(yīng)用超聲造影定量分析軟件進(jìn)行脫機(jī)分析,獲取峰值強(qiáng)度(peak intensity,PI)、達(dá)峰值時(shí)間(time to peak, TTP)和到達(dá)時(shí)間(arrival time,AT)等超聲造影定量分析參數(shù)。 超聲檢查結(jié)束后經(jīng)頸椎脫位處死小鼠,完整切除腫瘤,免疫組織化學(xué)法分別檢測(cè)腫瘤的CD31及CD34微血管密度的表達(dá)情況,并分別計(jì)算出微血管密度值(microvessel density,MVD)。 結(jié)果 1.接種后7天出現(xiàn)肉眼可見的腫瘤,模型建立成功。腫瘤的平均直徑約0.9cm,最大直徑為1.3cm,大多呈橢圓形,成瘤率達(dá)100%,且無(wú)一例復(fù)種。 2.25例小鼠LLC的超聲造影增強(qiáng)模式均表現(xiàn)為環(huán)狀強(qiáng)化,即腫瘤組織周邊部明顯強(qiáng)化,而中央部呈現(xiàn)低強(qiáng)化或無(wú)強(qiáng)化。在時(shí)間-強(qiáng)度曲線上上述LLC腫瘤均呈現(xiàn)“快進(jìn)、慢出”的表現(xiàn),即上升支陡直,迅速達(dá)峰,下降支相對(duì)較緩。 3.在小鼠的LLC腫瘤中,存在兩種代表血管內(nèi)皮細(xì)胞不同分化程度的血管標(biāo)記物：CD31和CD34. 4.超聲造影的灌注參數(shù)峰值強(qiáng)度(PI)與CD31微血管密度呈正相關(guān)(r=0.428;P0.05)與CD34微血管密度無(wú)線性相關(guān)。 5.在小鼠的LLC腫瘤中,CD31微血管密度與CD34微血管密度呈負(fù)相關(guān)(r=-0.474;P0.05)。 結(jié)論 1.本實(shí)驗(yàn)成功建立了小鼠Lewis肺癌的腫瘤模型,進(jìn)一步證明了該模型是研究組織腫瘤的可靠實(shí)用的動(dòng)物實(shí)驗(yàn)工具,并具有較好的可重復(fù)性。 2.在小鼠Lewis肺癌腫瘤模型中,存在兩種不同分化程度的腫瘤血管：分別是代表未分化和已分化成熟血管內(nèi)皮細(xì)胞的CD31微血管密度和只代表分化成熟的血管內(nèi)皮細(xì)胞的CD34微血管密度。 3.超聲定量分析參數(shù)與腫瘤微血管密度的相關(guān)性提示：超聲造影(團(tuán)注法)可以通過(guò)監(jiān)測(cè)腫瘤的血流灌注參數(shù)來(lái)有效評(píng)價(jià)腫瘤血管生成；其中超聲造影參數(shù)中的峰值強(qiáng)度可以在一定程度上評(píng)價(jià)腫瘤未分化的血管生成,指導(dǎo)臨床應(yīng)用抗血管治療以及評(píng)估腫瘤的分級(jí)和預(yù)后上有著重要的臨床實(shí)用價(jià)值。 第二部分超聲造影定量分析評(píng)價(jià)恩度對(duì)小鼠Lewis肺癌腫瘤的治療療效 研究背景及意義 抗腫瘤血管生成治療是目前治療腫瘤的研究熱點(diǎn)和重點(diǎn)。由于抗腫瘤血管生成治療的靶向作用于腫瘤內(nèi)異常的新生血管或不成熟血管,其治療會(huì)引起腫瘤內(nèi)血容量、血流灌注速度及其它血流動(dòng)力學(xué)指標(biāo)的改變,監(jiān)測(cè)這些血流動(dòng)力學(xué)指標(biāo)的改變成為臨床上評(píng)價(jià)抗血管生成治療的有效指標(biāo)。 既往多種影像學(xué)檢查方法(如CT、 MRI及超聲等)都曾來(lái)評(píng)價(jià)腫瘤血管的生成和抗腫瘤血管生成治療的療效。超聲檢查尤其是彩色多普勒超聲由于具有實(shí)時(shí)、費(fèi)用低、重復(fù)性好以及能直接評(píng)價(jià)腫瘤的血流灌注等優(yōu)點(diǎn),在評(píng)價(jià)腫瘤血管生成及抗血管生成治療中起著重要的作用。但彩色多普勒超聲對(duì)微血管的顯示及評(píng)價(jià)微血管的血流灌注有一定的局限性,近年來(lái)超聲造影更多的應(yīng)用于臨床,使對(duì)腫瘤內(nèi)微小血管內(nèi)血流的評(píng)價(jià)成為可能。在一定濃度范圍內(nèi),超聲造影的信號(hào)強(qiáng)度與血液內(nèi)微泡濃度存在線性關(guān)系,故可利用時(shí)間-強(qiáng)度曲線軟件對(duì)超聲造影過(guò)程進(jìn)行定量分析。 恩度(endostar)即重組人血管內(nèi)皮抑素,是一種血管生成抑制藥物,其可能作用機(jī)理是通過(guò)抑制血管內(nèi)皮細(xì)胞的增殖、遷移達(dá)到抑制腫瘤新生血管的生成,阻斷了腫瘤細(xì)胞的營(yíng)養(yǎng)供給,從而達(dá)到抑制腫瘤的目的。臨床主要應(yīng)用于非小細(xì)胞肺癌的抗腫瘤血管生成治療。 由于血管生成抑制藥物主要作用于腫瘤內(nèi)的異常新生血管,治療后不僅會(huì)引起組織內(nèi)血管數(shù)目的變化,更重要的是會(huì)引起腫瘤血管的形態(tài)、以及功能的改變,上述改變往往繼發(fā)了腫瘤內(nèi)血流動(dòng)力學(xué)的異常,因此監(jiān)測(cè)此異常變化可以成為臨床上評(píng)估抗血管生成治療的一個(gè)可靠手段。 研究目的 本研究擬采用Lewis肺癌荷瘤小鼠模型應(yīng)用腹腔內(nèi)注射抗血管藥物恩度治療后與對(duì)照組的超聲造影血流灌注參數(shù)進(jìn)行對(duì)照性研究,并將超聲造影參數(shù)與腫瘤的微血管密度進(jìn)行相關(guān)性比較,旨在探討超聲造影對(duì)評(píng)價(jià)抗腫瘤血管生成藥物療效的作用。 研究方法 建立30只小鼠LLC皮下移植瘤模型,隨機(jī)分為2組,各模型自建立次日起分別經(jīng)腹腔注射生理鹽水鈉(normal saline, NS)、恩度(5mg/kg體重),每天給藥,連續(xù)給藥14天。用藥后每天觀察兩組小鼠的一般情況,并每隔一天測(cè)定小鼠腫瘤的大小。 末次給藥24h后行超聲造影檢查,并應(yīng)用超聲造影定量分析軟件進(jìn)行脫機(jī)分析,獲取峰值強(qiáng)度(PI)、達(dá)峰時(shí)間(TTP)和到達(dá)時(shí)間(AT)等參數(shù)。超聲檢查結(jié)束后頸椎脫位處死小鼠,并完整切除腫瘤,免疫組織化學(xué)檢測(cè)腫瘤CD31的表達(dá)情況,并計(jì)算腫瘤的微血管密度(MVD)。 1.接種后7天出現(xiàn)肉眼可見的腫瘤,模型建立成功,腫瘤的平均直徑約為0.9cm,最大直徑1.3cm,大多呈橢圓形,成瘤率達(dá)100%,且無(wú)一例復(fù)種。 2.對(duì)小鼠一般情況的影響 用藥后生理鹽水組小鼠隨著移植瘤結(jié)節(jié)的逐漸長(zhǎng)大,進(jìn)食、飲水、活動(dòng)均較前減少;恩度組小鼠進(jìn)食、飲水較前無(wú)明顯變化,活動(dòng)正常,持續(xù)到實(shí)驗(yàn)結(jié)束。 實(shí)驗(yàn)結(jié)束后,恩度藥物干預(yù)組小鼠LLC皮下移植瘤體積(1762.72±53.08mm3)明顯低于生理鹽水對(duì)照組(2503.72±64.78mm3),兩組之間有統(tǒng)計(jì)學(xué)差異(p0.05)。 3.對(duì)移植瘤微血管密度的影響 以CD31作為血管內(nèi)皮細(xì)胞的標(biāo)志物,血管內(nèi)皮細(xì)胞的胞漿被染成棕黃色。NS組及恩度組CD31MVD計(jì)數(shù)平均值分別為98.50±13.53、56.21±5.10。與NS組比較,恩度藥物干預(yù)組MVD值減低且差異具有顯著性(p0.05)。 4.超聲造影軟件定量分析 實(shí)驗(yàn)結(jié)束后,恩度治療組的峰值強(qiáng)度(PI)明顯低于生理鹽水對(duì)照組(p0.05),其峰值強(qiáng)度分別為50.74±5.82及88.56±25.99。 5.超聲造影的定量分析參數(shù)與腫瘤微血管密度之間的關(guān)系 實(shí)驗(yàn)結(jié)束后恩度組超聲造影參數(shù)峰值強(qiáng)度(PI)與腫瘤的CD31微血管密度呈正相關(guān)(r=0.532,p0.05)。 結(jié)論 1.重組人血管內(nèi)皮抑素(恩度)能夠有效抑制小鼠Lewis肺癌皮下移植瘤的生長(zhǎng)。 2.超聲造影定量分析(團(tuán)注法)可以通過(guò)監(jiān)測(cè)腫瘤的血流灌注參數(shù)的改變來(lái)評(píng)價(jià)恩度抗血管生成治療的效果。 3.超聲造影定量分析參數(shù)中的峰值強(qiáng)度可以有效反映恩度抗血管生成治療引起的腫瘤血流灌注的改變。
[Abstract]:The first part of the quantitative analysis of contrast-enhanced ultrasound in the evaluation of tumor angiogenesis in Lewis lung cancer in mice
Research background and significance
Refers to the process of tumor angiogenesis and angiogenesis through endothelial cells "budding" based on the original tumor microvascular network, it is an essential factor for tumor growth and metastasis. In tumor angiogenesis in and around is considered to be the prerequisite for malignant tumor cell proliferation, these new blood vessels the proliferation of tumor cells can not only provide the necessary nutrition, but also provides a channel for the distant metastasis of tumor cells in vivo. Therefore angiogenesis is an important feature of malignant tumors, its detection for tumor diagnosis, treatment and prognosis is very important.
In the past few years, many studies by noninvasive imaging methods to evaluate tumor angiogenesis, such as contrast enhanced computed tomography (contrast enhanced computed tomography, CECT), dynamic contrast-enhanced magnetic resonance imaging (dynamic contrast enhanced magnetic resonance imaging, DCE-MRI), magnetic resonance angiography (magnetic resonance angiography, MRA), positron emission tomography (positron emission tomography, PET) and Doppler ultrasound technology and so on. However, these methods to measure low flow and small blood vessels in the inspection, has obvious limitations. CEUS can improve the detection capability of low flow, so that capillary tissue and microvascular evaluation the blood becomes possible, and can provide detailed information about tumor angiogenesis. MVD is the average per square millimeter of tumor area The number of blood vessels, is currently the evaluation of tumor angiogenesis as a reference standard. However, the microvessel density of different vascular endothelial cell markers to reveal the degree of differentiation and the structural and functional characteristics of different tumor endothelial cells. In previous experimental studies, more microvascular density by CD31 vascular markers, with vascular production evaluation organization. Because the CD31 microvessel density in differentiated and undifferentiated vascular endothelial cells expressed; and another common vascular marker CD34 is only expressed in the differentiated endothelial cells, the tumor vascular differentiation, microvascular density is also different.
Lewis Lewis lung cancer (lung carcinoma, LLC) is a subcutaneous injection of subcultured Lewis lung cancer cells into the axilla of C57/BL6 mice. It has been repeatedly applied to various studies in the laboratory, and is a relatively mature animal tumor experimental model.
Although there is a linear relationship between the microvessel density in previous studies have confirmed the quantitative parameters of contrast-enhanced ultrasound and tumor, so far, there is no reports of related research on different vascular markers in mice with Lewis lung cancer in microvessel density with contrast-enhanced ultrasound parameters. The purpose of the study
This study by subcutaneous transplantation tumor animal model of mouse LLC, application of enhanced pulse sequence (contrast pulse sequencing, CPS) of the ultrasound contrast imaging technology, combined with time intensity curve, the correlation study of microvessel density and blood perfusion parameters comparison of contrast-enhanced ultrasound with different vascular markers in LLC animal models, to explore the application value of contrast-enhanced ultrasound in the evaluation of tumor angiogenesis.
research method
The establishment of 25 mice transplanted LLC tumor model, the tumor size is about 1.0cm underwent contrast-enhanced ultrasound, contrast-enhanced ultrasound quantitative analysis software for off-line analysis, to obtain the peak intensity (peak intensity, PI), time to peak (time to, peak, TTP) and time of arrival (arrival time, AT) and other parameters of quantitative analysis ultrasound contrast.
After the end of ultrasound examination, the mice were killed by cervical dislocation, and the tumor was removed completely. The expression of CD31 and CD34 microvessel density was detected by immunohistochemistry, and the microvessel density (microvessel density, MVD) was calculated.
Result
1., 7 days after inoculation, the tumor was visible to the naked eye. The model was successfully established. The average diameter of the tumor was about 0.9CM, the largest diameter was 1.3cm. Most of the tumors were oval and the tumor formation rate was 100%, and no case was repeated.
2.25 cases of ultrasound contrast enhancement pattern of LLC mice showed ring enhancement, in which the tumor periphery was significantly enhanced, while the central part showed low enhancement or no enhancement. The time intensity curve on the LLC tumor showed "fast, slow out" performance, increases rapidly to peak, a steep, descending relatively slow.
3. in the LLC tumor of mice, there are two kinds of vascular markers representing different degree of differentiation of vascular endothelial cells: CD31 and CD34.
The peak intensity of perfusion parameters (PI) was positively correlated with CD31 microvascular density (r=0.428; P0.05), and there was no linear correlation with CD34 microvascular density (r=0.428).
5. in the LLC tumor of mice, the CD31 microvessel density was negatively correlated with the CD34 microvascular density (r=-0.474; P0.05).
conclusion
1., we successfully established a tumor model of Lewis lung cancer in mice, which further proved that this model is a reliable and practical animal experimental tool for studying tissue tumors, and has good repeatability.
2., in the mouse Lewis lung cancer model, there are two kinds of differentiated tumor vessels: CD31 microvessel density representing undifferentiated and differentiated vascular endothelial cells, and CD34 microvessel density representing only mature vascular endothelial cells.
3. correlation analysis of ultrasonic quantitative parameters and microvessel density of tumor tip: ultrasound contrast (bolus) can effectively evaluate tumor angiogenesis through the perfusion parameters monitoring tumor angiogenesis; the peak intensity in the parameters of CEUS can to some extent in the evaluation of tumor differentiation, clinical classification and prognosis guidance anti angiogenesis therapy and evaluation of tumor has important clinical value.
Therapeutic effect evaluation of endostatin on mouse Lewis lung carcinoma ultrasound contrast quantitative analysis of the second part
Research background and significance
Anti angiogenesis therapy is currently the research hotspot in tumor therapy. By targeting angiogenesis within the tumor abnormal or immature vascular antiangiogenesis therapy, the treatment will cause the tumor blood volume, blood perfusion rate and other hemodynamic index changes, monitoring the hemodynamic parameters change becomes effective index for clinical evaluation of anti angiogenesis therapy.
Check the previous method of multiple imaging (such as CT, MRI and ultrasound) have to evaluate the curative effect of tumor angiogenesis and anti angiogenesis therapy. Especially the ultrasound color Doppler ultrasound with real-time, low cost, good repeatability and can directly evaluate the tumor perfusion and other advantages, in the generation and the important role of anti angiogenesis therapy in evaluation of tumor blood vessels. But the blood perfusion of the evaluation of color Doppler ultrasound on microvascular angiogenesis have certain limitations, in recent years more for the clinical application of contrast-enhanced ultrasound, the evaluation of blood flow within the tumor in the tiny blood vessels become possible. Within a certain concentration range the signal intensity and contrast enhanced ultrasound microbubble concentration in the blood has a linear relationship, it can use the time intensity curve software for the quantitative analysis of the process of contrast-enhanced ultrasound.
Endostar (Endostar) recombinant human endostatin, is a kind of anti angiogenesis drugs, the possible mechanism is the inhibition of endothelial cell proliferation, migration to inhibition of tumor angiogenesis, blocking the nutrition supply of tumor cells, so as to achieve the purpose of suppressing tumor. Anti angiogenesis therapy in clinical application in non small cell lung cancer.
Due to the abnormal angiogenesis anti angiogenesis drugs mainly contributes to the tumor, after the treatment will not only cause the change of the number of blood vessels within the organization, more important is to cause tumor vascular morphology and function changes, these changes often secondary to abnormal tumor blood flow dynamics, so the changes can become the clinical monitoring on the evaluation of a reliable method for anti angiogenesis therapy.
research objective
This study used Lewis lung carcinoma bearing mice model by intraperitoneal injection of ultrasound contrast perfusion parameters of anti angiogenic drugs after Endostar treatment and control groups compared, and the parameters of contrast-enhanced ultrasound and microvessel density of tumor were compared, to explore the ultrasound contrast effect on the efficacy of anti angiogenic drugs.
research method
The establishment of 30 mice transplanted LLC tumor models were randomly divided into 2 groups, each model since the establishment of the next day respectively by intraperitoneal injection of saline sodium (normal saline, NS), Endostar (5mg/kg body weight), daily administration, continuous administration for 14 days. To observe the general situation of two groups of mice every day after administration. And every other day were measured tumor size.
At the dose of 24h after contrast-enhanced ultrasound, and the application of ultrasound contrast quantitative analysis software for off-line analysis, gets the peak intensity (PI), time to peak (TTP) and time of arrival (AT) and other parameters. Ultrasonography after cervical dislocation mice were sacrificed, and complete resection of the tumor, the expression of immunohistochemical detection of CD31 tumor microvessel density, and calculate the tumor (MVD).
1., 7 days after inoculation, the tumor was visible to the naked eye. The success of the model was established. The average diameter of the tumor was about 0.9CM, the largest diameter was 1.3cm. Most of the tumors were oval and the tumor formation rate was 100%, and no case was repeated.
The effect of 2. on the general condition of mice
After administration of saline group of mice with transplanted tumornodules gradually grew up, eating, drinking, eating activities were decreased; Endostar group mice, compared with the previous drinking water had no obvious change, the activity is normal, until the end of the experiment.
After the experiment, Endostar drug intervention LLC mice subcutaneous transplantation tumor volume (1762.72 + 53.08mm3) was significantly lower than the normal saline control group (2503.72 + 64.78mm3), there were significant differences between the two groups (P0.05).
Effect of 3. on microvascular density of transplanted tumor
Using CD31 as a marker of endothelium, vascular endothelial cells, the cytoplasm was stained brown.NS group and Endostar group CD31MVD count average value were 98.50 + 13.53,56.21 + 5.10. and NS group, Endostar drug intervention group MVD value decreased and the difference was significant (P0.05).
Quantitative analysis of 4. ultrasound contrast software
After the end of the experiment, the peak intensity of Endostar in the treatment group (PI) was significantly lower than the normal saline control group (P0.05), the peak intensities were 50.74 + 5.82 and 88.56 + 25.99.
The relationship between quantitative analysis parameters of 5. contrast-enhanced ultrasound and tumor microvessel density
After the experiment, Endostar group the parameters of CEUS peak intensity (PI) was positively correlated with tumor microvessel density CD31 (r=0.532, P0.05).
conclusion
1. recombinant human endostatin (endostatin) can effectively inhibit the growth of subcutaneous transplanted tumor of Lewis lung cancer in mice.
Analysis of 2. CEUS (bolus) can monitor tumor perfusion parameters to evaluate the change of Endostar antiangiogenic therapy.
3. ultrasound contrast quantitative analysis of the peak intensity in the parameters can effectively reflect the tumor perfusion caused by Endostar anti angiogenic treatment changes.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R445.1;R734.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Study of angiogenesis induced by metastatic and non-metastatic liver cancer by corneal micropocket model in nude mice[J];World Journal of Gastroenterology;1999年02期

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本文編號(hào)：1568232