| Part Ⅰ The utility of 99m Tc-tetrofosmin SPECT tumour tomography in assessment of chemoradiotherapy in the nasopharyngeal carcinomaObjective: To observe the detection ability of 99m Tc-tetrofosmin(99mTc-TF)in the patients with nasopharyngeal carcinoma（ NPC） by using 99m Tc-TFSPECT tomography in different periods of the treatment, and to research its ability to observe therapeutic effect of NPC by observing the concentration of 99m Tc-TF in the tumor location in different periods after radiation and chemotherapy of NPC, thus provides research foundation for the clinical application of 99m Tc-TF in the diagnosis of NPC and observing the efficacy of radiotherapy and chemotherapy.Methods:1. 20 patients were confirmed by pathology for NPC from Yancheng NO.1people’s hospital and all of them accepted 99m Tc-TF SPECT tomography. During the same period, 5 patients with SPECT myocardial perfusion imaging were collected as the control group. 2. Patients accepted 99m Tc-TF SPECT tomography respectively in different periods such as before treatment and early of treatment（< 11.5 Gy）, medium-term of treatment（more than 5 times in radiotherapy after initial imaging） and after treatment（total dose radiation > 66 Gy）. 3. Calculate the T/N ratio of early imaging as 99m Tc-TF uptake index（MUI）, and get the 99m Tc-TF delay imaging uptake index（DUI） from the 2 hours delay imaging, at the same time calculated retention rate（RI）. RI =（DUI-MUI） / MUI×100%.Using receiver operating characteristics（ROC） curve analysis to determine the MUI discriminant threshold in order to evaluate the value of the assessment of 99m Tc-TF SPECT tomography for primary NPC early treatment efficacy. 4. The experimental results are showed as mean ± standard deviation（x±S）, data analysis uses the SPSS19.0 software,and using one-way analysis of variance（ANOVA） or T test, P < 0.05 for the difference was statistically significant.Results:1.11 patients accepted 99m Tc-TF SPECT tomographybefore treat-ment, and the other 9 patients accepted the imaging in the period of early treatment. The MUI of the11 patients before treatment was 8.74±2.43, and the MUI of early treatment patients was6.78±3.17,there was no statistically significant difference between them（P>0.05）;the DUI of the 11 patients before treatment was 10.41±2.36 and DUI of other 9 patients was8.39±3.49, also there was no statistically signifi-cant difference between them（P>0.05）.The MUI of the 20 patients with NPC in before and early treatment was 7.86±2.89, and the DUI was 9.50±3.02, the DUI increased obviously, the difference was statistically significant（P<0.01）. The ratio of early imaging between NPC and the lateral nasal pharyngeal was 1.57±0.27, and this ratio in the control group was 1.14±0.09, the difference was statistically significant（P<0.01）; the ratio of delay imaging between NPC and the lateral nasa-l pharyngeal was 1.66±0.25, and was higher than early imaging; the difference was statistically significant between this two groups（P<0.05）. 2. The MUI ofmedium-term SPECT imaging in 18 NPC patients was 5.92±1.55, and the DUI was 7.29±2.11. The delayed imaging index obviously higher than early imaging in the 18 NPC patients who received both early imaging and delayed imaging, the difference was statistically significant（P<0.01）. The early imaging MUI of 18 patients in the treatment of medium-term is lower than before treatment, the difference was statistically significant（P<0.01）. The DUI of 19 patients in the treatment of medium-term was also significantly reduced compared to the early treatment, the difference was statistically significant（P<0.01）. 3. The MUI of 18 patients after the treatment was 5.01±1.95 and the DUI was 6.11±2.70, the delayed imaging index obviously higher than early imaging and the difference was statistically significant（P<0.01）. Compared to the imaging of early treatment, the MUI and DUI were obviously reduced, the difference was statistically significant（P<0.01）. And compared to the imaging of medium-term treatment, there was no statistically significant difference（P>0.05）. 17 patients recevied both early imaging and delayed imaging in all of the treatment period, the retention index in different periods were0.22±0.05, 0.19±0.07 and 0.19±0.08, There was no statistically significant difference among this three groups.Conclusion:The radioactivity uptake in the nasopharyngeal tumor location is higher than the other side of the nasopharynx and brain,with the extension of time, the tumor site uptake also increases. Compared to before treatment and early of treatment, the radioactivity uptake in the tumor site of medium-term treatment imaging was reduced, but there was no significant difference between medium-term treatment group with after treatment group. This study confirmed that the 99m Tc-TF can be used for SPECT imaging of NPC and could early and accurately evaluate the curative effect of the treatment.Part Ⅱ The experimental study of 99m Tc-tetrofosmin combined with CNE-1 cells to early evaluate radiotherapy curative effect of nasopharyngeal carcinoma.Objective: To verify the feasibility of 99m Tc-TF in observing the NPC radiotherapy curative effect from cytology level by measuring cells conjugation rate of 99m Tc-TF before radiotherapy and after different dose radiotherapy, and comparing analyses with tumor proliferation index.Methods: Cultured CNE-1 cells were dealed with trypsin digestion, count and immunization, then abandoned supernatant fluid after cultivating 24 hours, add 3.7KBq（50μL） in the culture dish, and the cells were incubated in a modular chamberflushed with a complex air of 5 % CO2 at 37 °C for 100 minutes, then put the supernatant fluid in the supernatant fluid tube. The cells were digested by trypsin and collected cells into the cell tube. Using gamma measurement instrument to measure the CPM count of the cells and supernatant. Calculate the cells conjugation rate of 99m Tc-TF. 2. The CNE-1 cells were cultured in the cell culture bottle for 24 hours, and then exposed to different doses of radiation（0, 2, 4, 6 or 8Gy）. 24 hours after radiation, calculate the cells conjugation rate of99mTc-TF of each bottle. 3. The cells were seeded into four 96-wells plates for 24 hours,and then exposed to different doses of radiation（0, 2, 4, 6 or 8Gy）. Using methyl thiazolyl tetrazolium（MTT） assay to detect optical density（OD） value on the enzyme mark analyzer in 492 nm wavelength and calculate cell growth inhibition rate,. 4. The cells cultured for24 hours, each groups were exposed to different doses of radiation（0, 2, 4, 6 or 8Gy）,subculture the cells for 24 hours and then digest and collect the cells, using flow cytometry instrument to detect cell early apoptosis rate. 5. The experimental data was showed by mean ± standard deviation（x±s）, data analysis uses the SPSS19.0 software, and using completely random design of single factor variance analysis, Exposure dose and other related parameters using spearman correlation test to analyze, P < 0.05 for the difference was statistically significant.Result: 24 hours after exposure to different doses, compared with control group, the irradiation group’s OD values were falling, cell growth inhibition rate of 2, 4 and 8 Gy group were（7.14±1.86）% 、（17.44±2.21）% and（33.54±3.51）%, the differences were statistically significant between the three groups（P<0.05）. Cell growth inhibition rate increased along with the increase of irradiation dose, the two were positively correlated（r=0.945,P<0.01）. 2. Different dose irradiation to CNE-1 cells after 24 hours,compared with control group, the cell early apoptosis rate measured by flow cytometry instrument was increased obviously in the irradiation groups, the differences were statistically significant between thses groups（P<0.01）. Early cell apoptosis rate increased along with the increase of irradiation dose, both were positively correlated（r=0.969 P<0.01）. 3.Different dose irradiation to CNE-1 cells after 24 hours, the cells conjugation rate of 99m Tc-TF in both 4Gy group and 8Gy group were reduced compared to control group,the difference was statistically significant（P<0.01）. Compared to 2Gy group and 4Gy group, the cells conjugation rate of 99m Tc-TF in 8 Gy group was reduced, the difference was statistically significant（P<0.05）.The cells conjugation rate of 99m Tc-TF decreased along with the increase of irradiation dose, both negatively correlated（r=-0.805,P<0.01）.Conclusion: 24 hours after exposure to different doses, the growth of CNE-1 cells was restrained, both cell early apoptosis rate and cell growth inhibition rate increased along with the increase of irradiation dose, and the cells conjugation rate of 99m Tc-TF decreased,negatively correlated with cell growth inhibition, to verify the 99Tcm-TF could observe the NPC radiotherapy curative effect from cytology level. |
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