| Tumor necrosis therapy (TNT) is a new targeted cancer treatment accompanied by the development of monoclonal antibody.TNT has a characteristic of broad spectrum and specificity。Researchers have confirmed that TNT can bind to nuclear antigen of tumor cell in Mammals include mouse. There are spontaneous necrosis regions in tumor tissues,131I-chTNT can be combined with the nuclear antigen in the necrosis region. However, spontaneous necrosis in tumor tissue is limited;the expansion of tumor necrosis can increase 131I-chTNT binding sites, increasing the distribution of drug in the tumor. Image-guided PMCT(percutaneous microwave coagulation therapy) is a new technology,developed in the late 20th century, local high-temperature induced by microwave can lead to tumor necrosis, so PMCT is a effective methods to increase the region of tumor necrosis. Thus, we designed the experiment, combine microwave coagulation therapy and treatment of 131I-chTNT。First we did microwave coagulation therapy to the tumors,resulting in necrosis area, and then injected 131I-chTNT. We research the effects of Microwave coagulation therapy on distribution of 131I-chTNT in mice,Meanwhile,whether PMCT effects the diffusion of drug in tumor tissue, wether the injection in the center of microwave coagulation area (hereinafter referred to as central injection) or outside the scope of tumor coagulation injection (hereinafter referred to as periphery injection) is better,is also a clinical application problems, so we compared central injection and periphery injection, provided a reference for clinical application.Chapter 1 the effects of microwave coagulation on the distribution of 131I-chTNT in mice bearing H22 hepatoma and imaging studyPurposeStudy the change of necrosis region after microwave coagulation therapy followed by intratumoral injection of 131I-chTNT in tumor-bearing mice; study the effects on distribution of drug in mice and imaging.Method1. A mouse tumor model was established by subcutaneous injection with H22 tumor cells:0.1ml suspension containing 1.2×106 H22 tumor cells was injected in the right side of the back skin in 70 5-week-old Kunming mice. About two weeks, when the maximum tumor diameter reach to 15mm,they can be used in the experiment.2. Group and treatment:When the tumor grew to a suitable size,51 tumor-bearing mice were selected to used in experiments in accordance with the principles of tumor size。The 51 mice were divided into 3 groups:there were 17mice in each group,12 mice in each group were used to measure the drug distribution in tissues and organs and the remaining 5 in each group were used for SPECT imaging. Group A, single-drug group, only with the injection of 131I-chTNT drugs,18.5MBq/0.05ml,; Group B, PMCT and central injection of drug (hereinafter referred to as central injection), microwave coagulation therapy were implement 3 days prior drug injections, then injected with 131I-chTNT。The methods and quantity of drug injection were the same as Group A;Group C,PMCT and periphery injection of drug (hereinafter referred to as periphery injection), microwave coagulation therapy were implemented 3 days prior drug injections, then injected with drugs。The methods and quantity of drug injection were the same as Group A。3. Drug distribution in mice:1,3,5,7 days after drug injection,three mice were killed in each groups each time, got the interested organs and useγcounter to weight determination of radioactivity uptake per gram of tissue (%ID/g,).4. The imaging study and the T/NT study:1,3,5,7 days after injection, five mice in A, B, C three groups were implemented SPECT imaging.Observe the difference in images of each group situation and using imaging techniques ROI (region of interest) to measure and calculate T/NT(toumor/nomal tissues)ratios.5. Statistical analysis:The sample data are analyzed statistically by SPSS 13.0 softwear. The uantity datas were represented by the way of "Mean±SD"; The datas of completely random design were analyzed with one-way analysis of variance (One-way ANOVO); As the ratio of T/NT of antibody in mice, and the comparison between two groups use LSD Test while Welch or Dunnett'3 Test was with equal variances not assumed. P＜0.05 was defined statistically significant.Results1. The tissue distribution in mice:There were no significant difference of the drug distribution in the normal tissue wile the combined groups (Group B,C) was significantly higher than single-drug group in tumor tissue.There were no difference between central injection group and periphery injection group.2. The image study in experimental animals:There were obviously enrichment in tumor tissue Id after injection. As time continued, the normal tissue radioactivity began to decrease,but the presence of radioactive in tumor tissue maintained at a high level until 7 days after injection.3.T/NT ratio:After microwave coagulation,Group B, group C had higher T/NT than the single drug group A, while the central injection group (B) and the periphery injection group (C group) have no significant difference in T/NT.After injection, T/NT ratio has been increased, reached a peak in the fifth day.Conclusions1.Microwave coagulation treapy prior 131I-chTNT injection can increase uptake of drug in tumor tissue;2.Central injection and the periphery injection were no significant different.Chapter 2 The study of how microwave coagulation effect the distribution of 131I-chTNT and the ability to inhibit tumor growthObjectiveTo study the Synergy Effect of PMCT and 131I-chTNT to inhibit tumor growthMethod1. Establish a mouse tumor model with 60 mice, the method were the same as the chapter 1;2. Group and treatment:When the tumor grew to a suitable size,40 tumor-bearing mice were selected to used in experiments in accordance with the principles of tumor size。The 40 mice were divided into 5 groups.Group A,single-drug group,10 mice, only with the injection of 131I-chTNT drugs,18.5MBq/0.05ml;Group B, PMCT and central injection of drug (hereinafter referred to as central injection),10 mice, microwave coagulation therapy were implemed 3 days prior drug injection,then injected with 131I-chTNT。The methods and quantity of drug injection were the same as Group A; Group C, PMCT and periphery injection of drug (hereinafter referred to as periphery injection),10 mice,microwave coagulation therapy were implement 3 days prior drug injection.The methods and quantity of drug injection were the same as Group A。Group D, single-microwave group,5 mice were given only microwave coagulation; Group E, control group,5 mice, only with the intratumoral injection of 0.05ml saline3. Autoradiography and observation:Five mice were killed 3 days after 131I-chTNT injection in group A,B,C,got tumor tissue and normal mouse liver and kidney, did routine pathological examination, two consecutive slices of each tissue were performed autoradiography and HE staining. Observation and compare the images of HE staining and autoradiography,study the different tumor necrosis and radioactive distribution in groups.4. Study the ability to inhibit tumor growth:observe appetite, movement, coat and mental changes of mice every day after injection,measured the long-diameter(a) and the short diameter(b) every 3 days until the 28 days. According to volume formula V =(a×b2)/2 calculated tumor volume and draw the tumor volume growth curves in mice.5. Statistical analysis:The sample data are analyzed statistically by SPSS 13.0 softwear. The uantity datas were represented by the way of'Mean±SD"; The datas of completely random design were analyzed in one-way analysis of variance (One-way ANOVO); As the tumor volume, Repeated Measure ANOVA is used and the comparison between two groups use LSD Test while Welch or Dunnett'3 Test was with equal variances not assumed.P<0.05 was defined statistically significant.Results 1. Intratumoral distribution.Both autoradiography and HE staining show that:single drug group had smller tumor necrosis region,and there were also less drug distribution in the tumor,while the regions of tumor necrosis in combined groups significantly greater than the single drug group,the distribution of drugs in the tumor were also more. There were no drugs distributions in normal tissues such as liver and kidney.2. Drugs specific binding verification:Compare autoradiography and HE staining images, we can see that only necrosis had drug distributions. There is no drug distributions in non-necrotic area.3. Observation of the effects of different treatment groups:tumor growth in different group were different. Combined treatment groups significantly slowed tumor growth, the tumor volum were significantly smaller than the control group,single-drug group, single microwave coagulation, P<0.05.Conclusions1.131I-chTNT is specific of binding in the tumor necrosis area;2.Microwave coagulation prior injection of 1311-chTNT to microwave treatment, can increase drug uptake within the tumor and have Synergy Effect to inhibit tumor growth;3.Central injection and the periphery injection were no significant different。...
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