Liver dynamic contrast-enhanced MRI for staging liver fibrosis in a piglet model.

PURPOSE: To determine whether dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) could monitor progression of liver fibrosis in a piglet model, and which DCE-MRI parameter is most accurate for staging this disease. MATERIALS AND METHODS: Sixteen piglets were prospectively used to model liver fibrosis and underwent liver DCE-MRI followed by biopsy on the 0, 5th, 9th, 16th, and 21st weekends after modeling of fibrosis. Time of peak (TOP), time to peak (TTP), positive enhancement integral (PEI), maximum slope of increase (MSI), and maximum slope of decrease (MSD) were measured and statistically analyzed for the monitoring and staging. RESULTS: As fibrosis progresses, TOP and TTP tended to increase, whereas MSI, MSD, and PEI tended to decrease (all P < 0.05). TOP, TTP, and MSI could discriminate fibrosis stage 0 from 1-4, 0-1 from 2-4, 0-2 from 3-4, and 0-3 from 4; PEI could distinguish the above-mentioned stages except 0-3 from 4; and MSD could distinguish stage 0-3 from 4, and 0 from 1-4 (all P < 0.05). For predicting stage ≥1, ≥2, and ≥3, the area under receiver operating characteristic curve (AUC) of MSI was largest among all parameters; for stage 4 AUC of TTP was largest. CONCLUSION: DCE-MRI has the potential to dynamically stage progression of liver fibrosis.

Spleen magnetic resonance diffusion-weighted imaging for quantitative staging hepatic fibrosis in miniature pigs: An initial study.

AIM: To determine whether spleen diffusion-weighted imaging (DWI) parameters might classify liver fibrosis stage. METHODS: Sixteen miniature pigs were prospectively used to model liver fibrosis, and underwent spleen DWI by using b = 300, 500 and 800 s/mm(2) on 0, 5th, 9th, 16th and 21st weekend after the beginning of modeling. Signal intensity ratio of spleen to paraspinous muscles (S/M), spleen exponential apparent diffusion coefficient (eADC) and apparent diffusion coefficient (ADC) for each b-value were statistically analyzed. RESULTS: With increasing stages of fibrosis, S/M for all b-values showed a downward trend; and spleen eADC and ADC for b = 300 s/mm(2) showed downward and upward trends, respectively (all P < 0.05). The area under the receiver-operator curve (AUC) of spleen DWI parameters was 0.777 or more by S/M for classifying each fibrosis stage, and 0.65 or more by eADC and 0.648 or more by ADC for classifying stage ≥3 or cirrhosis. Among the spleen DWI parameters, S/M for b = 300 s/mm(2) was the best parameter in classifying stage 1 or more, 2 or more and 3 or more with AUC of 0.875, 0.851 and 0.843, respectively; and spleen eADC for b = 300 s/mm(2) was best in classifying stage 4 with an AUC of 0.988. CONCLUSION: Spleen DWI may be used to stage liver fibrosis.

Application of two-dimensional electrophoresis in the research of retinal proteins of diabetic rat.

Diabetes mellitus (DM) is a chronic disease which is associated with numerous serious health complications such as diabetic retinopathy, and is the leading cause of new cases of blindness in adults at the age of 20-74 years old. The aim of the study was to establish and optimize a two-dimensional polyacrylamide gel electrophoresis (2-DE) technique for retina proteomics to improve the resolution and reproducibility, and to observe the proteomic changes of retinal tissues in diabetic and normal rats. Proteins were extracted from retinal tissues of normal and 8 weeks diabetic SD rats and used in two-dimensional electrophoresis. Various conditions of retina proteomic 2-DE were adjusted, optimized and protein spots of differential expression were obtained through analysis of 2-DE images with PDQuest software. By choosing appropriate sample amount, using pre-cast IPG dry strips (pH 5-8) and casting 12% equal gel, satisfactory 2-DE images of retina were obtained and a steady 2-DE technique was established. In this way, we found 36 spots in 2-DE gel of diabetic retinas that exhibited statistically significant variations, including up-regulation of 5 proteins in diabetic rat retinas, down-regulation of 23, and disappearance of 8, in comparison with normal tissues. The differences of protein expression were observed in retinas between diabetic and normal rats. Our established 2-DE technique of retina proteins could be effectively applied in proteomics of retina diseases.

Histological study of vas deferens following intravasal laser irradiation.

AIM: To study the histologic changes of the vas deferens following Nd: YAG laser irradiation. METHODS: Intravasal laser irradiation was given to (i) 52 segments of rabbit (laser dosage: 2 seconds at 40 W approximately 50 W) and 16 segments of human (3 seconds at 45 W approximately 55 W) vas deferens in vitro, (ii) 25 rabbit vasa (2 seconds approximately 2.5 seconds at 40 W approximately 45 W) in vivo and (iii) 2 human vasa (3 seconds at 55 W) in vivo. Segments of vasa were removed from the in vivo irradiated vasa deferentia 15 days approximately 180 days (rabbit) or 15 days (man) after the exposure. All vas segments were embedded in methacrylate resin. Serial sections (thickness 25 microm approximately 30 microm) were obtained and observed under a light microscope. RESULTS: (i) Laser-induced damage reached the muscularis layer in 27% and 94% of the rabbit and human vas segments in vitro, respectively. (ii) Fourteen of the 25 in vivo rabbit vasa were completely occluded by fibrous tissue and the longer the time interval after treatment, the more likely was the vas occluded. Those unoccluded vasa had either a normal histology or a mucosal damage. (iii) One in vivo human vas was almost completely occluded by the fibrous tissue but the other had a relatively large lumen packed with sperm granulomatous tissue and partial destruction of the smooth muscle layer. CONCLUSION: Laser irradiation can induce long-term vas occlusion; for rapid occlusion, laser doses just completely destroying the mucosal layer will be advisable.

Liver lobe volumes and the ratios of liver lobe volumes to spleen volume on magnetic resonance imaging for staging liver fibrosis in a minipig model.

OBJECTIVE: To investigate liver lobe volumes and the ratios of liver lobe volumes to spleen volume measured with magnetic resonance imaging (MRI) for quantitatively monitoring and staging liver fibrosis. METHODS: Animal study was approved by Institutional Animal Care and Use Committee. Sixteen minipigs were prospectively used to model liver fibrosis, and underwent abdominal gadolinium-enhanced MRI on 0, 5(th), 9(th), 16(th) and 21(st) weekend after modeling this disease staged by biopsy according to METAVIR classification system. On MRI, volume parameters including left lateral liver lobe volume (LLV), left medial liver lobe volume (LMV), right liver lobe volume (RV), caudate lobe volume (CV), and spleen volume (SV) were measured; and LLV/SV, LMV/SV, RV/SV and CV/SV were calculated. Statistical analyses were performed for staging this fibrosis. RESULTS: LLV and CV increased with increasing stage of fibrosis (r = 0.711, 0.526, respectively; all P < 0.05). RV and LMV increased from stage 0 to 2 and decreased from 2 to 4; and RV/SV decreased from 0 to 1, increased from 1 to 2, and decreased from 3 to 4 (all P > 0.05). LLV/SV, LMV/SV and CV/SV decreased from stage 0 to 4 (r = -0.566, -0.748 and -0.620, respectively; all P < 0.05). LLV, CV, LLV/SV, LMV/SV, RV/SV, and CV/SV could distinguish stage 0-1 from 2-4 and 0-2 from 3-4 (all P < 0.05). Among these parameters, LLV and LMV/SV could best classify stage ≥2 and ≥3, respectively (area under receiver operating characteristic curve = 0.893 and 0.946, respectively). CONCLUSION: LLV and LMV/SV complement each other in staging liver fibrosis, and both parameters should be used to stage this disease.

Spleen dynamic contrast-enhanced magnetic resonance imaging as a new method for staging liver fibrosis in a piglet model.

OBJECTIVE: To explore spleen hemodynamic alteration in liver fibrosis with dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), and to determine how to stage liver fibrosis with spleen DCE-MRI parameters. MATERIALS AND METHODS: Sixteen piglets were prospectively used to model liver fibrosis staged by liver biopsy, and underwent spleen DCE-MRI on 0, 5th, 9th, 16th and 21st weekend after modeling this disease. DCE-MRI parameters including time to peak (TTP), positive enhancement integral (PEI), maximum slope of increase (MSI) and maximum slope of decrease (MSD) of spleen were measured, and statistically analyzed to stage this disease. RESULTS: Spearman's rank correlation tests showed that TTP tended to increase with increasing stages of liver fibrosis (r = 0.647, P<0.001), and that PEI tended to decrease from stage 0 to 4 (r = -0.709, P<0.001). MSD increased slightly from stage 0 to 2 (P>0.05), and decreased from stage 2 to 4 (P<0.05). MSI increased from stage 0 to 1, and decreased from stage 1 to 4 (all P>0.05). Mann-Whitney tests demonstrated that TTP and PEI could classify fibrosis between stage 0 and 1-4, between 0-1 and 2-4, between 0-2 and 3-4, or between 0-3 and 4 (all P<0.01). MSD could discriminate between 0-2 and 3-4 (P = 0.006), or between 0-3 and 4 (P = 0.012). MSI could not differentiate between any two stages. Receiver operating characteristic analysis illustrated that area under receiver operating characteristic curve (AUC) of TTP was larger than of PEI for classifying stage ≥1 and ≥2 (AUC = 0.851 and 0.783, respectively). PEI could best classify stage ≥3 and 4 (AUC = 0.903 and 0.96, respectively). CONCLUSION: Spleen DCE-MRI has potential to monitor spleen hemodynamic alteration and classify liver fibrosis stages.

Magnetic resonance-based total liver volume and magnetic resonance-diffusion weighted imaging for staging liver fibrosis in mini-pigs.

AIM: To determine whether and how magnetic resonance imaging (MRI)-based total liver volume (TLV) and diffusion weighted imaging (DWI) could predict liver fibrosis. METHODS: Sixteen experimental mature mini-pigs (6 males, 10 females), weighing between 20.0 and 24.0 kg were prospectively used to model liver fibrosis induced by intraperitoneal injection of 40% CCl(4) dissolved in fat emulsion twice a week for 16 wk, and by feeding 40% CCl(4) mixed with maize flour twice daily for the subsequent 5 wk. All the survival animals underwent percutaneous liver biopsy and DWI using b = 300, 500 and 800 s/mm(2) followed by abdominal gadolinium-enhanced MRI at the 0, 5th, 9th, 16th and 21st weekend after beginning of the modeling. TLV was obtained on enhanced MRI, and apparent diffusion coefficient (ADC) was obtained on DWI. Hepatic tissue specimens were stained with hematoxylin and Masson's trichrome staining for staging liver fibrosis. Pathological specimens were scored using the human METAVIR classification system. Statistical analyses were performed to determine whether and how the TLV and ADC could be used to predict the stage of liver fibrosis. RESULTS: TLV increased from stage 0 to 2 and decreased from stage 3 (r = 0.211; P < 0.001). There was a difference in TLV between stage 0-1 and 2-4 (P = 0.03) whereas no difference between stage 0-2 and 3-4 (P = 0.71). TLV could predict stage ≥ 2 [area under receiver operating characteristic curve (AUC) = 0.682]. There was a decrease in ADC values with increasing stage of fibrosis for b = 300, 500 and 800 s/mm(2) (r = -0.418, -0.535 and -0.622, respectively; all P < 0.001). Differences were found between stage 0-1 and 2-4 in ADC values for b = 300, 500 and 800 s/mm(2), and between stage 0-2 and 3-4 for b = 500 or 800 s/mm(2) (all P < 0.05). For predicting stage ≥ 2 and ≥ 3, AUC was 0.803 and 0.847 for b = 500 s/mm(2), and 0.848 and 0.887 for b = 800 s/mm(2), respectively. CONCLUSION: ADC for b = 500 or 800 s/mm(2) could be better than TLV and ADC for b = 300 s/mm(2) to predict fibrosis stage ≥ 2 or ≥ 3.