| Part I Comparative Study on Implanted Intrahepatic, Intramuscular and Subcutaneous VX-2Tumor Model of RabbitObjective:To find out the most suitable model of tumor-bearing rabbit for percutaneous interventional therapy by comparison of intrahepatic, intra-muscular and subcutaneous inoculation of VX-2tumor. Methods:54New Zealand rabbits were randomly divided into three groups (A, B and C):group A were inoculated with VX-2tumor intra-hepaticly, group B intra-muscularly and group C subcutaneously. On12th,16th and20th day after inoculation the rabbits were perfusion-scanned with multi-planar CT to determine the size of tumors and6rabbits from each group were killed every time respectively for pathological observation. The operation time for modeling, tumor size and modeling-related complications of the three groups were compared. Results:The VX-2tumor of group A grew fastest and VX-2tumor of group C grew slowest and statistic significance showed among these three groups. Tumors of group A might grow to the size of about1cm in diameter on12th day after inoculation while tumors of group C would on20th day and tumors of group B would on16th day. The operation time needed for modeling for group A, B and C were,5.54±0.51min,1.02±0.20min and0.98±0.21min respectively, showing markedly long for group A. Except for group A, of which several rabbits developed ascites, intrahepatic metastases or abdominal adhesive, no obvious complications affected VX-2tumor model were observed for group B and C. Conclusions: Compared to intrahepatic and subcutaneous VX-2tumor model, intra-muscular VX-2tumor mode of rabbit is easier to be established with high tumor-formation rate and proper growing rate, so it is most suitable for percutaneous interventional study. Part II:Study on the Ultra-microstructural Changes of Transplanted VX-2Tumor of Rabbit Caused by Intra-tumorous Injection of OzoneObjective:To investigate the changes of ultra-microstructure of transplanted VX-2tumor of rabbit caused by intra-tumorous injection of different concentrations of O3in order to determine proper concentration of O3which would be safely and effectively used experimentally or clinically and time interval for possible repeated injections of O3. Methods:20rabbits bearing VX-2tumor on hind legs were divided into4groups:2for control (group C), of which the tumors were punctured only,6for20%O3、40%O3and60%O3injection respectively (group T20, T40and T60), of which the tumors were punctured and injected2times the volume of20%O3、40%O3and60%O3under direct vision.2rabbits from each group were killed immediately after puncture/injection and the tumors were removed for fixation. The remaining rabbits from group T20, T40and T60were killed on the4th (2rabbits) and8th day (2rabbits) respectively after injection of03and the tumors were resected, fixed and made sections for transmission electron microscopic observation. Results:Group T20:mild injuries to tumor cells might be seen immediately after O3injection, which were shown as slight lysis of ridges of mitochondia and sporadic swelling of mitochondria, dilation of endoplasmic reticulum, a little widening of perinucleic gap in some cells, but the space among tumor cells were normal. Group T40and T60:obvious damage to tumor cells were observed immediately after O3injection, which manifested as obvious swelling of mitochondria, dilation of endoplasmic reticulum, swelling of nucleus, widening of perinucleic gap, breakup of cellular membrane and collapse of tissue structure. On the4th day, massive necrosis appeared with infiltration of inflammatory cells. Other changes such as incomplete cellular membrane, mitochondria-lysosome formation (presented as high electron-density particle) or lysis of lysosome, dilation of endoplasmic reticulum, partial lysis of cytoplasm, irregular nucleus and widening of perinucleic gap occurred and even more seriously in group T60. Vessel endothelial cells appeared to be injured too. All these changes might be observed on8th day after treatment but recovered gradually. Conclusions:Intra-tumorous injection of20%-60%O3can cause injuries to VX-2tumor cells and intra-tumorous vessel endothelial cells of rabbit even to death. The injuries caused by20%O3are relatively mild but more severe by40%and60%O3. In consideration of safety and effectiveness, perhaps40%O3is more suitable for experimental or clinical use than60%O3. Definite injuries to tumor cells can still be observed on8th day after treatment in spite of partial recovery, so we suppose that the time interval of4-8days for repeated injection of O3is reasonable. Part Ⅲ Study on Transplanted VX-2Tumor Effect of Different Concentrations of Ozone Therapy of the Tumor of RabbitObjective:Study on the different concentrations of03therapy efficacy of percutaneous injection of VX-2tumor of rabbit. Materials and Methods:New Zealand110rabbits, randomly divided into4groups,20in the control group (group A), treatment group with30in each group:B group, C group,20ug/ml concentration of O340ug//ml concentration of O3and D concentration of60ug/ml O3in tumor group,2weeks after transplantation in treatment, A group only simulation puncture, B, C, D three groups according to tumor volume3times volume of different concentrations of O3multiphase injected directly into the tumor in vivo, underwent CT perfusion scanning,4th,8th days after operation, CT perfusion scanning, measuring the size of tumors, and on the8th day after all were sacrificed after CT perfusion of tumor pathological observation. Results:(1)Volume:ozone injection before A, B, C, D four groups of average volume were:0.53±0.46,1.02±1.10,0.71±0.62,0.70±0.39; the average volume of ozone injection after fourth days in the A group, B group, C group, D group:0.91±0.60,1.15±0.95,1.03±1.03,1.12±0.81; the average volume of eighth days in the A group, B group, C group, D group after the injection of ozone,1.75±1.31,2.18±2.18,1.84±1.71,1.54±1.02;(2) The growth rate of tumor:A, B, C, D four groups of V4/V0were:2.19±1.21,1.43±0.80,1.54±0.82,1.62±0.92; A, B, C, D four groups of V8/V4were:2.47±2.31,1.80±0.72,1.98±1.28,1.56±0.86; A, B, C, D four groups of V8/V0were:5.14±3.67,2.64±1.59,2.93±1.56,2.27±1.34. V4∨V0results showed that A, B two groups of P=0.05, P>0.05other groups; comparison between groups P>0.05V8/V4; V8W0results showed A, B two groups of P=0.002, A, C two groups of P=0.006, A, D two groups of P=0.000, B, C, D each other P>0.05. Conclusions:(1) Percutaneous intratumoral injection of20%,40%and60%concentrations of03could effectively inhibit the growth of VX-2tumor in rabbits.(2)20%,40%and60%different concentrations of03on rabbit VX-2tumor inhibitory effect of no significant difference, but with the increasing of concentration, the effect of longer duration, the effector of inhibiting tumor growth more obviously. |
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