Metastatic Lung Cancer Blood Supply And By The Treatment Of Pulmonary Embolism Research

Part One:Hematogenous pulmonary metastases of VX2 carcinoma in rabbit:establishing model and observing its biological featuresPurpose: To establish hematogenous pulmonary metastases of VX2 carcinoma in rabbit experimental model, and to observe its biological features. Material and method: The first section 20 New Zealand white rabbits were implanted with VX2 single cell suspensions acquired from a previous tumor-bearing rabbit. All the rabbits were randomly divided into 2 groups, 1.5ml cell suspension was injected into ear vein in group A and 0.5 ml was injected in group B, respectively. Multi-slices spiral computed tomography (MSCT) were performed to detect tumor nodules on 28th day after implantation. All the rabbits were sacrificed after MSCT scan , the trachea and lung were removed together for pathological examination. The nodules diameter were measured with digital slide gaud as scheduled .The second section 1.5ml VX2 single cell suspensions was injected into right ear vein in another 10 New Zealand white rabbits, respectively. One rabbit was randomly sacrificed on the 14th day, 21st day, 28th day after implantation in turn. The trachea and lung were kept to observe tumor growth. The other 7 rabbits were killed to obtain pathological examination (HE stain) on the 32nd day after implantation as scheduled. Result: The first section VX2 carcinoma was successfully implanted to lung fields in 19 rabbits, the other one died of lobar pneumonia on the 21^th day. Successful inoculation rate was 95%. The fresh specimens showed tumor nodules diffusively distributed on the lung. Microscopic findings showed tumor spread along interstitial tissue, infiltration into arterioles, bronchial wall, and diffusive emphysema as well. MSCT demonstrated signs of diffusive nodules in the lung parenchyma, and irregularly increased lung markings. The two groups had significant difference in tumor nodule diameter. The second section VX2 carcinoma implantation were all succeeded in 10 rabbits. All specimens showed tumor nodules diffusively distributed on the lung surface with rapidly increasing in diameter and density. Extrapulmonary infiltration occurred with breeding persisting with time. Interstitial tumor spread, infiltration into arterioles, bronchial wall, and diffusive emphysema were showed by microscopic fingdings. Conclusions: The pulmonary metastases of VX2 carcinoma had been establishedsuccessfully. Formation and growth of metastatic nodules was directly related to the number of invasive tumor cells. Collagenase IV would be essential to dissove tumor interstitial tissue, it might be contributive to tumor infiltration, and infection should be prevented in order to simultaneously obtain large number of animal models. VX2 tumor is a malignant neoplasm similar to that in human. Establishing pulmonary metastatic models will be helpful to prevention and treatment of pulmonary metastases.Part Two:Application of selective pulmonary arterial chemoembolization inexperimental pulmonary metasatasesPurpose : To investigate feasibility and efficacy of pulmonary artery chemoembolization in experimental lung metastases, and to elucidate pulmonary arterial supply to metastases using lipiodol as a tracerMaterials and methods: 14 New Zealand white rabbits were implanted with VX2 single cell suspensions acquired from a previous tumor-bearing rabbit. 1.5ml cell suspension was injected into ear vein in each rabbit, respectively. Multi-detecter spiral computed tomography (MDCT) were performed to confirm establishment of pulmonary metastases in the third weekend after implantation, then all the rabbits were randomly divided into 3 groups including 4 rabbits in group A, 4 rabbits in group B and 6 rabbits in group C. Group A served as control who accepted saline (5ml per rabbit), group B and C were assigned as observation. Group B received anticancer drug infusion including Pharmorubicin (2mg/Kg) and carboplatin (15mg/Kg) simultaneously. Unilateral pulmonary chemoembolization (UPCE) combined with infusion of carboplatin (15mg/Kg) was carried out in group C, and UPCE was performed by injecting a emulsion which was mixed by Pharmorubicin (2mg/Kg) and lipiodol (lml). All 14 rabbits were catheterized to the left pulmonary artery through right femoral vein using 3 Fr SP microcatheters under fluoroscopy and digital subtraction angiography (DSA) guidance. Unfortunately, 2 rabbits were suffered from sudden death because of pulmonary oil embolism due to withdrawal of SP microcatheter to pulmonary trunk during chemoembolization, and then the two lungspecimens were obtained and fixed with formaldehyde solution. The other four survival rabbits were sacrificed 3 weeks after treatment by an overdose of anesthesia, the trachea and lung were removed together for pathological examination, respectively. The nodule diameters were measured with digital slide gaud as scheduled. Cryostat sections from different size of metastases were dyed with Sudan III in group C.Results: The average volume of metastatic nodules in group A (130.56 ± 9. 41mm³) was significant larger than that in group B (69.47±10. 00 mm³ ) and C (19.73 ± 3.54 mm3), respectively. Meanwhile, there was also significant difference between group B and C. The average volume in group C was smaller than that in group B. lipiodol drops were detected in different size of metastases under microscopy after Sudan III dying. Those the diameter of which were no more than 2 milimeters presented "+", those between 2 and 4 milimeters in diameter showed "++", and those diameter more than 4 milimeters in diameter took on "+++". Even 3 weeks after treatment, lipiodol drops could be detected in both interstitial and parenchymal fields of metastases. Conclusion: This was a successful experimental study of chemoembolization applied to pulmonary metastases. Compared with chemotherapy such as intravenous therapy and transarterial infusion, chemoembolization associated with transpulmonary infusion was more effective without serious complications. But more attention should be paid to avoid pulmonary embolism during procedure. If chemoembolization is applied clinically, catheters should be inserted more close to metastasis in case of catheter's withdrawal. On the other hand, it has been comfirmed that pulmonary artery is the main component of blood supply to micrometastases by means of Sudan III dying for lipiodol drops. The blood supply will become more and more abundant with growth of metastases. Part Three:Appraisal of tumor-associated neoangiogenesis in pulmonarymetastases with anti-105 /anti-34 immunohistochemical stainingPurpose: To appraise tumor-associated neoangiogenesis of pulmonary metastatic nodules with CD34, CD 105 (Endoglin) immunohistochemical staining, and to evaluate significance of microvessel density (MVD) for endoglin immunostaining inpulmonary metastases from colorectal adenocarcinoma compared with nodules from primary hepatocellular carcinoma (HCC).Methods: Surgical specimens from 18 patients with pulmonary metastatic nodules were immunostained with anti-CD34 mAb, anti-CD 105 mAb, respectively. All 18 cases included 8 specimens bloodbored from HCC and 10 cases originated from colorectal adenocarcinoma which consist of 8 cases moderately differentiated and 2 cases poorly differentiated tumor. Positively stained microvessels were counted in densely vascular foci (Hotspots), and microvessel morphological analysis was reviewed at X 400 fields in each specimen under Motic Microscopes System, respectively.Result: MVD of CD34 immunostaining showed significant difference compared with that of endoglin staining (5.27±1.47 vs 1.19 ± 0.53, P<0.05). Tumor vessels might be divided into several components such as entrapped normal blood vessels, matured tumor vessels, mosaic vessels, and activated proliferating vessels, et al. CD34 staining was apt to all kinds of vessels. Nevertheless, endoglin significantly demonstrated more proliferating neoplastic microvessels including activated tumor vessel and mosaic vessel. At the same time, there was no significance in MVD of pulmonary metastases for endoglin staining between the two different nodules originated from colorectal adenocarcinoma and HCC( 1.25± 0.68 vs 1.16 ± 0.42, P>0.05). Additionally, morphylogical analysis from magnification fields displayed several characteristic phenotypes including branch-like, sinusoidal, sprout-like, round, mosaic vessels and so on. CD34 staining fields showed more branch-like, sinusoidal and sprout-like types of vessels, whereas endoglin staining presented more round and mosaic vessels. CD34 usually stained large amount of bulky vessels in diameter's calibration than endoglin. Conclusion: Different original pulmonary metastases equally presented large amount of tumor vessels due to tumor-associated angiogenesis. Endoglin immunostaining could demonstrate more activated proliferating tumor vessels than CD34 staining. Endoglin might be one of specific markers aimed at activated neoplastic vessels in pulmonary metastatic nodules, and could monitor angiogenesis in secondary metastatic site.Part four:Functional CT used to evaluate blood supply of pulmonary metastases in vivoObjective: Functional CT used to evaluate blood supply of pulmonary metastases in vivo.Meterials and methods: Nineteen patients according to inclusive criteria were enrolled in this study including 14 males and 5 females. Mean age was 54.37±6.78 years. Selective bronchial arterial catheterization and superselective pulmonary arterial catheterization were carried out simultaneously in each patient under digital subtraction angiography (DSA) guidance before CT scan, respectively. And then patients were moved smoothly to multi-detecter spiral CT scanner . During breathhold single location dynamic CT sequence was performed to acquire contrast enhancement imaging in pulmonary metastasis, firstly at a rate of 1.5ml/s and total amount of 8ml contrast medium by bronchial arterial catheter infusion. Then ten minutes later at a rate of 3ml/s and total amount of 40 ml contrast material through pulmonary arterial catheter infusion. Density-time curves were evaluated from three different regions of interest (ROI) in each patient: a large ROI comprising the whole tumor area and two smaller ROI placed over the area with the highest and lowest enhancement, respectively. Perfusion and peak enhancement index (PEI) were calculated using functional CT software retrieved from MDCT instrument in each patient, and the largest diameter was measured through axial CT images.Results: Values of perfusion of metastasis through pulmonary arterial enhancement was significantly higher than that through bronchial arterial enhancement ( 206. 67 + 217. .62 ml/min. ml vs 19. 27 + 33. 68 ml/min. ml). There was significant difference in PEI between pulmonary arterial enhancement and bronchial arterial enhancement (223.85+234.54 Hu vs 17.59±22.02 Hu) as well. Values of perfusion and PEI measured from the three ROIs through pulmonary enhancement were significantly larger than those obtained through bronchial artery enhancement, respectively (PO.05). Perfusion was not correlated with tumor diameter under the two different enhancement patterns (P>0.05). The correlation coefficients were —0.167 and 0.104, respectively. Otherwise, PEI of metastasis was correlated with tumor diameters simultaneously under the two different enhancement ways (P<0.05). The correlation coefficients were 0.04 and 0.405, respectively. Conclusion: Blood supply of pulmonary metastases whatever due to HCC orcolorectal carcinoma mostly originated from pulmonary artery, bronchial artery mightpartially participate in the blood supply to pulmonary metastases at the same time.Pulmonary arterial and bronchial arterial blood supply would be increasing with themetastasis growth. Functional CT has played vital role in evaluation of angiogenesisin pulmonary metastases in vivo.Part fiveClinical evaluation of efficacy in treating pulmonary metastases withsubsegmental pulmonary chemoembolization (SPCE)Objective: The purpose of this study was to investigate efficacy of SPCE in treating pulmonary metastases.Materials and methods: Nineteen patients corresponding with inclusive criteria were enrolled in this study including 14 males and 5 females. Mean age was 54.37 + 6.78 years. According to localization of metastases in thoracic CT images, subsegmental pulmonary arterial catheterization targeting on metastases were done in all patients under DSA guidance. And then all patients received chemoembolization. The different dosage of lipiodol was administrated according to the size of metastasis, lml lipiodol for 2³cm metastasis, and 2ml for metastasis >3cm in diameter. Chemoembolization was performed under fluoroscopy by injection an emulsion which was mixed by same amount of contrast media and lipiodol, lidocaine 20mg, Pharmorubicin 10-20mg . Blood-gas analysis was measured to monitor lung function 30min earlier before procedure and lh after treatment, respectively. Noenhancement CT was performed 24-48h after intervention to observe the lipiodol accumulation in metastastic nodule and surrounding normal lung fields. And CT scan and thoracic plain film were repeated to monitor lipiodol deposit every 1 month. The maximium diameter of metastasis was measured through transverse CT images, and volume of metastasis was calculated using volume formula. The volume of metastatic nodules before intervention were compared with those measured 4-6 months after embolization. Result: Successful subsegmental catheterizing rate was 100%. No fatal and major complications related to the treatment were observed. Only 2 patients had minor complications as follows: cough and expectoraton of sputum. Major indexes of...