The Experimental And Clinical Studies Of Using Synchrotron Radiation X-ray Fluorescence Facilities And Susceptibility-weighted MR Imaging To Quantitatively Assess The Hepatic Iron Deposition In Liver Fibrosis

PartⅠ The experimental study of using3.0T susceptibility-weighted MR imaging to quantitatively assess the hepatic iron deposition in rat liver fibrosis modelObjectives:To assess the value of abdominal susceptibility-weighted MR imaging (SWI) to quantitatively assess the hepatic iron deposition (HID) in a high-dose injection of dimethylnitrosamine (DMN) induced rat liver fibrosis model.Materials and Methods:Experimental group consisted with SD rats (n=50) with liver fibrosis accompanied by iron deposition induced by a single intraperitoneal injection of a high dose DMN (50mg/kg of body weight).10saline-injected SD rats (using the same dose of DMN) were used as control group. All rats in both experimental and control groups were underwent MR abdominal SWI examinations, liver phase values measurement and histopathological assessment for detecting HID and evaluating its grade. At five timepoints (day5,7,10,14and28after DMN injections),10rats were randomized taken to examinations in experimental group.Results:A total of47rats in experimental group and10rats in control group were included in the final data analysis and the other3rats in experimental group died on day3after DMN injection for acute submassive hemorrhagic necrosis. The mean liver phase values in histopathological iron deposition grade0(n=10), Ⅰ (n=42) and Ⅱ(n=5) were respectively0.0183±0.0697,-0.0302±0.0158and-0.0656±0.0062. Significant differences were found among these groups and any two of them (F=70.057,p<0.001). A high negative correlation was detected between phase values and histopatho logical iron deposition grades (r=-0.843, p<0.01). Furthermore, the mean phase values in experimental group at the five timepoints (day5,7,10,14, and28after DMN injections) were respectively-0.0059±0.0087,-0.0234±0.0068,-0.0424±0.0061,-0.0589±0.0082,-0.030718±0.0078and in control group were0.0183±0.0697. There were significant differences among these six groups (F=132.743,p<0.001).Conclusion:Abdominal SWI was a new sensitive method for quantitatively assessing HID in DMN induced rat liver fibrosis, which was highly correlated with the histopathological results. Part Ⅱ The experimental study of using synchrotron radiation X-ray fluorescence facilities to quantitatively assess the hepatic iron deposition in rat liver fibres is modelObjectives:To discuss the value of synchrotron radiation X-ray fluorescence (SRXRF) facilities to quantitatively assess hepatic iron deposition (HID) in rat liver fibrosis model induced by single high-dose injection of dimethylnitrosamine (DMN).Materials and Methods:Experimental group consisted with SD rats (n=50) with liver fibrosis accompanied by iron deposition induced by a single intraperitoneal injection of a high dose DMN (50mg/kg ofbody weight).10saline-injected SD rats (using the same dose of DMN) were used as control group. All rats in experimental and control groups were received both SRXRF and histopathological examinations for assessing HID. All samples are fully dried before SRXRF scanning. The energy of monochromatic light in SRXRF examination was12.5KeV and the spot size was2mm x2mm. The peak of iron-specific fluorescence located at6.40keV. Every time10rats at day5,7,10,14and28after DMN injections in experimental groups were received SRXRF and histopathological examinations. The photons of iron in SRXRF were measured and analyzed. Correlation was detected between the photons of iron and histopathological iron deposition grades.Results:A total of47rats in experimental group and all rats in control group were included in the final data analysis. The other3rats in experimental group died on day3after DMN injection for acute submassive hemorrhagic necrosis. The mean photons of iron in control and experimental groups were (4.10±0.91) and (10.01±4.03) x104Fe photons/g dry liver tissue respectively and significant statistically differences were found between them. Additionally, according to the results of pathological Prussian blue stain, HID divided into three groups (grade0(n=10), Ⅰ (n=42) and Ⅱ (n=5)) and the mean photons in the three groups were (4.10±0.91),(9.16±3.15) and (17.16±3.63) x104Fe photons/g dry liver tissue respectively. Significant statistically differences were also observed among them (F=33.174, p<0.01). A moderate positive correlation was found between the photons of iron and histopathological iron deposition grades (r=0.722， p<0.01). Furthermore, the mean photons of iron at the five timepoints (day5,7,10,14and28after DMN injections) were (5.03±1.23),(8.85±2.66),(10.39±1.73),(15.10±4.25) and (9.17±1.77) x104Fe photons/g dry liver tissue respectively. When compared the photons of iron in these five groups and the controls, significant differences were observed among them (F=25.412,p<0.01).Conclusion:SRXRF was a new nondestructive analytical technique that could be used to quantitatively assess liver iron concentration in rat liver fibrosis. The mean photons of iron had significant differences in different grade of HID and a moderate positive correlation was detected between iron photons and histopathological iron deposition grades. PartⅢ-1The clinical study of3.0T abdominal susceptibility-weighted MR imaging to quantitatively assess hepatic iron deposition in chronic hepatitis B patientsObjectives:To assess the value of abdominal susceptibility-weighted MR imaging (SWI) with three TE values (10,5and2.5ms) to quantitatively assess hepatic iron deposition (HID) in patients with chronic hepatitis B (CHB) and to investigate the correlations between HID and the grade of liver fibrosis and inflammatory activity.Materials and Methods:A total of57CHB patients were prospectively included in this study. All patients were underwent both abdominal SWI and histopatho logical examinations. Liver phase values were measured on the images with three different TE values, which were measured containing the entire right liver avoiding large vessels or artifacts. Histopatho logical evaluations were included semi-quantitative Pearls’Prussian blue stains for iron deposition grade, liver fibrosis scores and inflammatory activity. ROC curves were used to predict HID.Results:Final data analysis included51patients.36patients were determined with HID based on the results of Pearls’Prussian blue stains. There were15,12,12,8and4patients respectively according the histopatho logical iron deposition grade0to4. The sensitivities and specificities of SWI for the different three TE values (10,5and2.5ms) were88.89%/66.67%,86.11%/60%and75%/80%, respectively. There were statistically significant differences of the liver mean phase values among the histopathological iron deposition grades (F=115.01,41.86and31.167respectively;p<0.001of all) and high negative correlations were observed between them (r=-0.904,-0.856,-0.799; p<0.001). When the phase values were respectively greater than-0.0453,-0.0127and0.0125for the three different TE values, the sensitivities were all100%and the specificities were86.1%,80.6%and83.3%, respectively. Furthermore, low negative correlations were found between the inflammatory activity and phase values in both10and5ms TE values (r=-0.394,-0.368; p=0.004,0.008). However, there were no correlations between phase values and liver fibrosis in all the three different TE values (p>0.05).Conclusion:Abdominal SWI was able to be used to quantitatively detect HID in CHB patients and the liver phase values had significant differences in different histopathological iron deposition grade particularly when using a longer TE value (TE=10ms). HID was associated with inflammatory activity rather than liver fibrosis. PartⅢ-2Comparative study of3.0T abdominal susceptibility-weighted imaging and T2*-weighted gradient-echo imaging in quantitatively assessing hepatic iron deposition in patients with chronic hepatitis BObjectives:To compare the ability of3.0T abdominal susceptibility-weighted imaging (SWI) and multi-echo T2*-weighted gradient-echo imaging (T2*WI) sequences in quantitatively assessing hepatic iron deposition (HID) in patients with chronic hepatitis B (CHB).Materials and methods:A total of43patients were prospectively included in this study. All of them were consecutive underwent SWI, T2*WI examinations and liver histopathological examination. Phase values were measured in phase images of SWI sequence and T2*values in T2*map. The phase and T2*values were measured contained the entire right liver avoiding large vessels or artifacts. Two pathologists (R1and R2) consulted together in determining the iron deposition grade by Pearls’Prussian blue staining.Results:39patients (male/female,29/10) aged27-72(51.69±11.57) years were included in the final data analysis. The case numbers were11,8,9,7,4patients respectively according to histopatho logical iron deposition grade0to4and the liver mean phase values were sequentially-0.0213±0.0094,-0.05063±0.0010,-0.0589±0.0117,-0.0940±0.0110,-0.1568±0.0119. The mean T2*values were19.65±1.31,15.53±1.96,15.34±1.91,8.36±1.07,4.60±1.50, respectively. Significant statistically differences were observed in both phase and T2*values among these five different histopatho logical iron grades (F=139.60,93.91;p<0.001). Both phase values and T2*values were found having negative correlations with the histopathological iron deposition grades (r=-0.937,-0.898; p<0.001) and the correlations of histopathological iron grades with phase values were higher than that with T2*values.Conclusion:Both abdominal SWI and T2*WI could be used to quantitatively assess HID in CHB patients and SWI appeared to have higher sensitivity. Part IV The clinical study of synchrotron radiation X-ray fluorescence in quantitatively assessing hepatic iron depositionObjectives:To use synchrotron radiation X-ray fluorescence (SRXRF) facilities to quantitatively assess hepatic iron deposition (HID) and to investigate the relationship between HID and the grade of liver fibrosis and inflammatory activity.Materials and Methods:A total of64patients were prospectively included in this study (partial liver excision (n=58), liver transplantation (n=6)). All the specimens were selected in the liver parenchyma away from tumor masses and all the required specimens received both SRXRF and histopathological examinations. The receiver operating characteristics (ROC) curves were determined to predict patients with HID and significant iron deposition (histopathological iron grade3and4).Results:35patients were determined with HID based on the results of Pearls’Prussian blue stains in all the included64patients and there were respectively15,9,7and4patients according to the histopathological iron deposition grade1to4.29patients were not found iron deposition (histopathological iron deposition grade0). The photons of iron assessed by SRXRF in five histopathological iron deposition grades were respectively (1.30±0.48),(2.12±0.55),(3.23±1.44),(4.08±0.88) and (6.06±1.51) x105photons/g dry liver tissue and significantly statistically differences were observed among them (F=44.166, p<0.001). High positive correlation was obtained between histopathological iron grades and the amounts of iron photons (r=0.855,p<0.001). When the photons of iron were more than1.95x105photons/g dry liver tissue, the sensitivity and specificity to predict HID were82.9%and89.7%. Meanwhile, when the photons of iron were more than3.43x105photons/g dry liver, the sensitivity and specificity to predict patients with significant HID were90.9%and91.7%. Furthermore, weak positive correlation was observed between the amounts of photons of iron and inflammatory activity (r=0.252, p=0.045). However, no correlation was found between it with liver fibrosis (p-0.706).Conclusion:SRXRF was a powerful nondestructive analytical technique that could be used to quantitatively assess HID. In our clinical study, HID was found to associate with inflammatory activity rather than liver fibrosis. PartV Value of3.0T abdominal susceptibility-weighted MR imaging on quantitatively assessing hepatic iron deposition and to determine its correlations with serum iron markers in chronic hepatitis B patientsObjectives:Using abdominal susceptibility-weighted MR Imaging (SWI) technique to quantitatively assess hepatic iron deposition (HID) in chronic hepatitis B (CHB) patients and to determine its correlations with serum iron markers (serum iron, ferritin, transferrin and transferrin saturation).Materials and Methods:Patients (n=327) with HBV infection and healthy controls (n=50) were prospectively enrolled. All patients underwent abdominal SWI and were assessed for serum iron markers. Phase values were measured and five grades of hepatic iron deposition were described by SWI.Results:303patients and47controls were included in the final data analysis. In total,77(25.4%) of patients with HBV infection had HID as determined by SWI. The mean phase values in patients with HID, without HID and normal controls were respectively-0.0710±0.047,-0.0152±0.0103and-0.0141±0.0147. There were significant differences between patients with HID and patients without (p<0.001). However, no significant statistically differences were observed between patients without HID and the normal controls (p=0.254). Phase values were also significantly different across different grades of HID (F=84.044, p<0.001). Levels of serum iron, ferritin, transferrin and transferrin saturation were significantly higher in patients with compared with those without HID (p<0.001). Only serum ferritin levels were significantly different across different grades of HID (x2=8.519,p=0.036) and there was a low negative correlation between serum ferritin levels and phase values (r=-0.365, p=0.007).Conclusion:Abdominal SWI could be used to quantitatively assess HID in CHB patients. Only serum ferritin levels in serum iron markers were significantly different across different grades of HID and low negative correlation was found between serum ferritin levels and phase values. SWI may represent a more powerful tool to assess HID.