| PART ONE SD Rats Liver Cirrhosis, Hepatocellular Carcinoma Model Induced by Diethylnitrosamine(DEN)OBJECTIVE:To study the success rate and characteristics of SD rats liver cirrhosis,hepatocellular carcinoma model induced by diethylnitrosamine(DEN).MATERIALS AND METHODS:128SD rats of4-5weeks,male,150-180g were included. DEN,with99.9%Purity,was injected intraperitoneally with20mg/kg and3times every week. The weight of rats was measured every week.Every rat was carried out by Ultrasound(US) on beginning day and in8th,12th and14th week.Five rats were killed and rat liver was obtained for pathology after US.Data was analysed by SPSS19.0statistical software.RESULTS:72rat liver tumor models were obtained after14weeks and the success rate was56.25%. Rat liver pathology achieved approximately prophase cirrhosis, advanced cirrhosis and hepatocellular carcinoma in8th±1week,12th±1week and14±1week.The change of rat weight was significant(F=54.79, P<0.001) between beginning day and in8th,12th and14th week.CONCLUSION:Stable and ideal cirrhosis and hepatocellular carcinoma model can be establish by DEN and the development of rat liver pathology also can be reflected approximately by the change in body weight. PART TWO Characteristics on Diffusion-weighted Imaging(DWI), Susceptibility Weighted Imaging(SWI) of SD Rats Liver Cancer Model before and after High Intensity Focused Ultrasound(HIFU)OBJECTIVE:To study characteristics of SD rats prophase cirrhosis, advanced cirrhosis and hepatocellular carcinoma model and after HIFU on DWI and SWI and study the value of DWI after HIFU.MATERIALS AND METHODS:72rat liver tumor models were obtained after14weeks by injecting DEN intraperitoneally for128SD rats with99.9%purity,20mg/kg and3times every week. Every rat was dynamically carried out by MRI,DWI and SWI on beginning day and in8th,12th and14th week and after HIFU.Data was analysed between40complete data on DWI before HIFU, between10complete data on SWI before and after HIFU and between23complete data on DWI after HIFU and47complete data on DWI non-HIFU.Some rats were killed and rat liver or tumor were obtained for pathology on beginning day and in8th,12th and14th week and10th after HIFU. Data was analysed by SPSS19.0statistical software.RESULTS:The signal of liver normal parenchyma, prophase cirrhosis, advanced cirrhosis and hepatocellular carcinoma increased gradually on DWI. Part of viable cell tumor manifested high signal and zone of necrosis manifested low signal. ADC values of liver normal parenchyma, prophase cirrhosis, advanced cirrhosis and hepatocellular carcinoma were2645.18±369.97,2008.65±691.87,1679.50±206.16and2453.65±861.64mm2/s and the distinction was significant (F=4.34, p<0.05) when b value was50s/mm, which the distinction was significant (t=-2.82, p<0.05) between advanced cirrhosis and hepatocellular carcinoma. ADC values of liver normal parenchyma, prophase cirrhosis, advanced cirrhosis and hepatocellular carcinoma were2504.90±320.38,2185.90±455.91,1708.40±416.91and2284.30±223.18mm2/s and the distinction was not significant (F=3.23, p=0.08) when b value was100s/mm2.ADC values of hepatocellular carcinoma were2190.52±778.65and2115.39±633.08mm2/s and the distinction was not significant (t=0.35, p>0.05) between before and after HIFU when b value was50s/mm2and its were2047.26±487.88,1889.33±547.17mm2/s and the distinction was not significant (Z=-0.88, p>0.05) when b value was100s/mm2. Data of blood vessel counting of liver normal parenchyma,advanced cirrhosis and hepatocellular carcinoma on SWI was14±2.9,25.6±6.0,34.6±21.0by turns and the distinction was not significant(p>0.05)CONCLUSION:Manifestation of Rat liver cirrhosis and hepatocellular carcinoma is characteristic on DWI and it is important and potential for DWI to detect,diagnose, tracking hepatocellular carcinoma as well as to assess therapeutic effect after HIFU. PART THREE Characteristics of Flow Cytometry (FCM) and Proteomics on SD Rats Liver Cancer Model after High Intensity Focused UltrasoundOBJECTIVE:To study the immune and protein mechanism of rat hepatocellular carcinoma HIFU therapy.MATERIALS AND METHODS:72rat hepatocellular carcinoma models were obtained after14weeks by injecting DEN intraperitoneally.23single neoplasms models were treated one or more by HIFU and all tumor samples were obtained on10day after HIFU and12rat blood preparation of them were obtained,while23tumor samples were obtained and6rat blood preparation of them from non-HIFU rat hepatocellular carcinoma models. All blood preparation were detected respectively by flow cytometry and all tumor samples were detected respectively by proteomics. Data was analysed by SPSS19.0statistical software.RESULTS:The distinction of CD4,CD8,CD28,CD25of FoxP3r-IFN,IL4,CD4/8, Thl/Th2was not significant(P>0.05) between the factor expression of rat blood preparation after HIFU and non-HIFU.26protein spots of differential expression were detected by proteomics of them,15nonredundant protein were discriminated further. Compared with hepatocellular carcinoma protein spectrum,5proteins were down-regulated in cells and21proteins were up-regulated. CONCLUSION:The distinction of protein expression is significant between HIFU hepatocellular carcinoma and non-HIFU,which its protein active components is similar either raised or lowered, and residual viable cell tumor must be accept further treatments as early as possible because its ability of invasion and metastasis rised. |
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