[Tanshinone â ¡A activates PI3K/AKT signaling pathway to inhibit the apoptosis of mice cochlear pericytes induced by high glucose].

Objective: To investigate whether tanshinone ⅡA can protect the apoptosis of mice cochlear pericytes induced by high glucose and its specific protective mechanism, so as to provide experimental evidence for the prevention and treatment of diabetic hearing loss. Methods: C57BL/6J male mice were used to prepare type 2 diabetes model, which were divided into normal (NG) group, diabetic (DM) group, diabetic+tanshinone ⅡA (HG+tanshinone ⅡA) group and tanshinone ⅡA group. Each group had 10 animals. Primary cochlear pericytes were divided into NG group, HG group (high glucose 35 mmol/L), HG+tanshinone ⅡA (1, 3, 5 µmol/L) group, HG+Tanshinone ⅡA+LY294002 (PI3K/AKT pathway inhibitor) group, LY294002 group, tanshinone ⅡA group and DMSO group. Auditory brainstem response (ABR) was used to measure hearing threshold. Evans blue was used to detect the permeability of blood labyrinth barrier in each group. TBA methods were used to detect oxidative stress levels in various organs of mice. Morphological changes of stria vascularis were observed by hematoxylin-eosin staining (HE). Evans blue was used to detect the vascular labyrinth barrier permeability in cochlea. The expression of apoptosis protein in stria vascularis pericytes was observed by immunofluorescence. Pericytes apoptosis rate was observed by flow cytometry. DCFH-DA was combined with flow cytometry to detect intracellular ROS content, and Western blot was used to detect the expression of apoptotic proteins (Cleaved-caspase3, Bax), anti-apoptotic proteins (BCL-2) and pathway proteins (PI3K, p-PI3K, AKT, p-AKT). SPSS software was used for statistical analysis. Independent sample t test was performed, and P<0.05 was considered statistically significant. Results: Animal experiments: Tanshinone ⅡA decreased the hearing threshold of DM group [(35.0±3.5) dB SPL vs. (55.3±8.1) dB SPL] (t=4.899, P<0.01), decreased the oxidative stress level in cochlea (t=4.384, P<0.05), improved the structure disorder, atrophy of cochlea vascular lines, vacuole increased phenomenon. Tanshinone ⅡA alleviated the increased permeability of the blood labyrinth barrier [Evans blue leakage (6.84±0.27) AU vs. (8.59±0.85) AU] in the cochlea of DM mice (t=2.770, P<0.05), reversed the apoptotic protein: Caspase3 (t=4.956, P<0.01) and Bax (t=4.388, P<0.05) in cochlear vascularis. Cell experiments: Tanshinone ⅡA decreased intracellular ROS content in a concentration-dependent way (t=3.569, P<0.05; t=4.772, P<0.01; t=7.494, P<0.01); Tanshinone ⅡA decreased apoptosis rate and apoptotic protein, and increased the expression of anti-apoptotic protein, p-PI3K/PI3K and p-AKT/AKT in concentration-dependent manner (all P values<0.05); LY294002 reversed the protective effect of tanshinone ⅡA on pericytes apoptosis (all P values<0.05). Conclusion: Tanshinone ⅡA can inhibit the apoptosis of cochlear pericytes induced by high glucose by reducing oxidative stress level and activating PI3K/AKT signaling pathway under high glucose environment, thus playing a protective role in diabetic hearing loss.

[Effect of microvascular pericytes of cochlear stria vascularis on endothelial cell permeability in C57BL/6J mice].

Objective: To study the changes in the permeability of the blood labyrinth barrier of the aging cochlea in mice, and to establish a non-contact co-culture model of endothelial cells (EC) and pericytes (PC) to furtherly investigate the cochlear stria vascularis microvascular pericytes impact on the permeability of endothelial cells. Methods: C57BL/6J mice were divided into two groups, three months old as young group, 12 months old as senile group. Cell experiment was divided into four groups, EC group, EC+PC co-culture group, D-gal+EC group and D-gal+EC+PC co-culture group. Auditory brainstem response (auditory brain response, ABR) was used to detect the auditory function of the two groups of mice. Evans blue staining was applied to detect the permeability of the cochlear blood labyrinth barrier of the two groups of mice. Transmission electron microscopy was used to observe the ultrastructure of blood labyrinth barrier endothelial cells, pericytes and tight junctions in the two groups of mice. Immunohistochemistry was used to detect the expression levels of tight junction proteins in the stria vascularis of the cochlea of the two groups of mice. Transwell chamber was used to detect the permeability of endothelial cells. Western blot and immunofluorescence technology were used to detect the expression level of tight junction protein on endothelial cells. SPSS 20.0 software was used to analyze the data. Results: Compared with the young group, the ABR threshold of the aging group was significantly increased, the latency of wave I was prolonged (t=10.25, P<0.01;t=5.61, P<0.05), the permeability of the cochlear blood labyrinth barrier was increased and the expression of tight junction protein on the vascular stria was decreased (P<0.05). The cochlear ultrastructure showed that the cochlear vascular stria microvascular lumen was deformed, the basement membrane thickened and the tight junction gap between endothelium enlarged. The positive rate of ECs and PCs in primary culture was more than 95%. The cells induced by 15 g/L D-gal were determined to be senescent cells. Compared with EC group, the expression of tight junction protein in endothelial cells of D-gal+EC group decreased（t=7.42，P<0.01；t=13.19，P<0.05）and the permeability increased (t=11.17, P<0.01). In the co-culture group, the expression of tight junction protein between endothelial cells in EC+PC co-culture group and D-gal+EC+PC co-culture group increased and the permeability decreased. Conclusions: In aging mice, the permeability of cochlear blood labyrinth barrier will increase and the level of tight junction protein will decrease; in aging state, cochlear vascular stria microvascular pericytes may affect endothelial cell permeability by regulating the expression of tight junction protein.

Knockdown of MiR-140-5 promotes osteogenesis of adipose-derived mesenchymal stem cells by targeting TLR4 and BMP2 and promoting fracture healing in the atrophic nonunion rat model.

OBJECTIVE: This study aims to assess the effect and mechanism of genetically modified adipose-derived mesenchymal stem cells (ASCs) with recombinant lentiviruses mediated knockdown of miR-140-5p in ASCs' osteogenesis in vitro and atrophic nonunion rat model. MATERIALS AND METHODS: This study included 36 male adult Sprague-Dawley (SD) rats weighing 400 g to 450 g from the experimental animal facility of our university. Approval was obtained from the University Animal Care Committee before the study. Rats' ASCs were prepared and genetically modified with lentivirus (Lv)-empty (NC) or Lv-miR-140-5p-TuD (inhibitors). After that, the expressions of RUNX2 and osteocalcin (OCN) were detected in the ASCs. To confirm the mechanisms of miR-140-5p in ASCs, we predicted the target genes by bioinformatics analysis and then the target genes were verified by luciferase reporting assay. The artificial atrophic nonunion was created in the rat's femoral bone. Animals were randomly divided into three groups according to the material implanted into bone defects space: AT scaffolds (AT group, n=12), AT scaffold with Lv-NC modified (AT+ASCs+Lv-NC group, n=12), AT scaffold with the Lv-miR-140-5p-TuD modified ASCs (AT+ASCs+Lv-miR-140-5p-TuD group, n=12). After four weeks, the rats were euthanized for the following radiographic examination, histologic study and biomechanical testing. RESULTS: MiR-140-5p was down-regulated during osteogenic differentiation of ASCs, and inhibition of MiR-140-5p promoted osteogenesis of ASCs in vitro. Inhibition of MiR-140-5p promoted osteogenesis of ASCs and enhanced fracture in the atrophic nonunion rat model: AT+ASCs+Lv-NC group, AT+ASCs+Lv-miR-140-5p-TuD group resulted in a better bone formation and higher BMD and BMC than AT group, while excellent bone formation and the highest BMD and BMC were observed in AT+ASCs+Lv-miR-140-5p-TuD group. Both AT+ASCs+Lv-NC group and AT+ASCs+Lv-miR-140-5p-TuD group presented more mature characteristics in the micro-architecture than AT group, whereas AT+ASCs+Lv-miR-140-5p-TuD group presented the highest BV/TV, Tb.Th and Tb.N as well as the lowest Tb.Sp. The peak load of the operated femur increased by 94.43% AT+ASCs+Lv-miR-140-5p-TuD group, 50.68% in AT+ASCs+Lv-NC group compared to the control AT group, respectively. The result of luciferase reporting assay showed that miR-140-5p could directly target TLR4 and BMP2. CONCLUSIONS: This study demonstrates that lentiviruses-mediated knockdown of miR-140-5p can significantly promote osteogenesis of ACSs by directly regulating its' target genes, TLR4 and BMP2, and that combined adipose scaffold with genetically modified ASCs can significantly enhance fracture-healing and bone formation in the atrophic nonunion rat model.

Imaging Validation of Drug-Eluting Beads Transarterial Chemoembolization of Hepatocellular Carcinomas in Living Donor Liver Transplantation.

OBJECTIVE: The aim of this study is to determine whether post-transarterial chemoembolization imaging (computed tomography or magnetic resonance imaging) could accurately predict the tumors' necrosis on pathologic specimens. BACKGROUND: Transarterial chemoembolization with drug-eluting beads has been proven to be an effective way to bridge patients with hepatocellular carcinomas to liver transplantation. MATERIALS AND METHODS: From September 2012 to June 2017, 59 patients with a total of 78 hepatocellular carcinomas, who received transarterial chemoembolization with drug-eluting beads before liver transplantation in Kaohsiung Chang Gung Memorial Hospital, were included in the study. All patients and hepatocellular carcinomas have pre-transarterial chemoembolization and post-transarterial chemoembolization images (computed tomography or magnetic resonance imaging) and pathological findings for correlation. Tumor response was evaluated according to modified Response Evaluation Criteria in Solid Tumors. The ranges of necrotic percentage are 100%, 91-99%, 51-90%, and <50%. RESULTS: The accuracy rate between the imaging and pathology correlation was 40% for computed tomography and 42% for magnetic resonance imaging. The recurrent rate of the complete respond group is 11.5%, the partial respond group is 16.0%, and the stationary group is 28.6%. CONCLUSION: Computed tomography and magnetic resonance imaging sensitivity is not satisfactory for microscopic evaluation of residual tumors after transarterial chemoembolization with drug-eluting beads. However, survival is good after liver transplantation no matter what the microscopic findings were.

Acoustic Radiation Force Impulse Elastography in Post-transplant Recurrent Hepatitis C in Living Donor Liver Transplantation.

OBJECTIVE: The aim of the study was to evaluate the efficacy of liver fibrosis (LF) with acoustic radiation force impulse (ARFI) elastography for post-transplant (post-LT) HCV recurrence. PATIENTS AND METHODS: We enrolled 89 adult recipients of living donor liver transplantation (LDLT) who had HCV, with or without post-LT HCV recurrence and treated or not treated. The post-LT HCV recurrence was diagnosed on the basis of RNA viral load present. ARFI examination was performed every 3 months for all patients, with shear wave velocity (SWV) obtained quantitatively in m/s and correlated with histopathologic fibrosis scoring of liver biopsy (LB). RESULTS: There were 50 (50 of 89) patients without HCV recurrence and 39 (39 of 89) with post-LT recurrence in the 89 patients studied. The recurrent group had significantly higher median SWVs (1.87 ± 0.52 vs 1.37 ± 0.52 m/s, P < .0001), in which 18 (18 of 39) patients had antiviral drug treatment and obtained significant improvement with SWVs from 1.83 ± 0.49 to 1.68 ± 0.56 m/s, P = .043. The correlations of LF staging between ARFI elastography and Ishak histopathologic LF scores showed great significance, P = .045. The HCV RNA titer after antiviral treatment decreased from 3,831,750 to 0, P < .0001, but the RNA titer of nontreated patients remained high and the median SWV increased. The Ishak LF staging in the nontreated group progressed from stage 1 to 2, P = .012 and SWV increased from 1.69 ± 0.54 to 1.91 ± 0.66 m/s, P = .085 at 1-year follow-up. CONCLUSION: ARFI elastography has efficient quantitative LF monitoring correlated with histopathologic staging for post-LT HCV recurrence. It could be an alternative, noninvasive method for frequent LB in the disease follow-up.

Impact of Different Dominant Hepatic Vein Patterns in Left-Lobe Donors of Adult Living Donor Liver Transplantation.

OBJECTIVE: Optimal hepatic venous tributary flow is correlated with liver function and regeneration. In left-lobe graft living donor liver transplantation, the stump of segment 5 and 8 hepatic veins (S5V and S8V) are ligated without performing hepatic tributary reconstruction. The aim of this article was to evaluate the different dominate hepatic vein patterns that affect left-lobe living donor safety. MATERIALS AND METHODS: A total of 44 donors who underwent left-lobe hepatectomy were divided into 2 groups, middle hepatic vein (MHV) dominance (group 1) and right hepatic vein (RHV) dominance (group 2), according to the dominant venous territory drainage from S5V and S8V or RHV. The clinical pathological data, postoperative laboratory data, complication, remnant liver volume and remnant liver regeneration rate at 6 months after surgery were compared. RESULTS: No difference was found in blood loss, postoperative liver function such as alanine transaminase value, complications, and hospital stays between groups. Group 1 had slightly higher total bilirubin level than group 2 (1.99 vs 1.79; P = .49). Group 2 had significantly better remnant liver regeneration rate than group 1 (89.2% vs 82.5%; P = .026). CONCLUSION: It is important to recognize the dominant MHV group. Ligation large S5V and S8V in dominant MHV donors led to lower remnant liver regeneration in our series. This might be critical in extremely small RHV territory and potential large remnant liver congestion donors. Adjusting surgical planning, such as hepatic vein reconstruction, in this kind of donor might be appropriate for donor safety.

Magnetic Resonance Spectroscopy in Living-Donor Liver Transplantation.

OBJECTIVE: The purpose of this study is to evaluate the accuracy rate of the one breath-hold single voxel hydrogen-1 magnetic resonance spectroscopy (MRS) in comparison with intraoperative biopsy for liver fat quantification in living-donor liver transplantation. MATERIALS AND METHODS: A total of 80 living liver donors participated in this study. Each patient underwent both MRS and intraoperative biopsy for evaluation of liver fatty content. MRS was performed using 1.5-T magnetic resonance imaging and placed in segments 2-4, 5-8, and left lateral segment for each donor. Accuracy was assessed through receiver operating characteristic curve analysis. Sensitivity and specificity of MRS fat fractions were also calculated. RESULTS: Eighty living-donor liver transplantation donors were enrolled in this study. There was no fatty liver in 59 subjects (73.8%), 5% to 10% fatty liver in 17 subjects, 11% to 15% fatty liver in 3 subjects, and >16% fatty liver in 1 subject. MRS fat fraction showed excellent parameters to predict between normal liver and fatty liver groups (1.85% ± 0.98, 8.13% ± 3.52, respectively; P < .0001). Linear regression between MRS fat fraction and pathology grading showed high correlation (R(2) = 0.7092). Pearson correlation revealed high correlation between MRS and pathology results (r = 0.936), poor correlation between body mass index and pathology results (r = 0.390). The sensitivity and specificity for detection of liver steatosis in MRS fat fraction were 95.2% and 98.3%, respectively. CONCLUSION: (1)H MRS fat fraction is a highly precise and accurate method in quantification of hepatic steatosis for the living donor and can be finished in a single breath-hold.

Impact of Graft Type in Living Donor Liver Transplantation: Remnant Liver Regeneration and Outcome in Donors.

OBJECTIVES: Liver regeneration and donor safety in right-lobe (RL) and left-lobe (LL) grafts are essential for donors in living donor liver transplantation (LDLT). Our aim was to compare the liver regeneration rate and postoperative outcome between different donor graft types in LDLT. MATERIALS AND METHODS: A total of 95 donors were divided into 2 groups: RL (n = 42) and LL (n = 53). The remnant liver of LL donors were subdivided into 3 subgroups according to the different hepatic venous drainage pattern that dominates from right hepatic vein (dominant RHV; n = 34), middle hepatic vein (dominant MHV; n = 10), and include MHV for left lateral segment (LLS) graft (n = 9). The demographic data, postoperative laboratory data, complications, remnant liver volume (RLV), and remnant liver regeneration rate (RLRR) 6 months after surgery were compared. RESULTS: The postoperative total bilirubin (TB), prothrombin time (PT), and intensive care unit (ICU) stays of the LL group were lower than the RL group (P < .05). The LL group has no significant better regeneration rate 6 months after surgery than the RL group. However, dominant RHV and LLS groups have significantly better RLRR than the RL group (89.2% vs 86% and 95.1% vs 86%, respectively, P < .05), but no significance in the dominant MHV group. CONCLUSION: In conclusion, different hepatic venous drainage patterns of remnant liver grafts may affect the regeneration rate in LL LDLT, especially with dominant RHV donors, may have more comparable outcomes with that of RL, and should be a favorable option during donor selection.

Drug-Eluting Bead Transarterial Chemoembolization as Bridge Therapy for Hepatocellular Carcinoma Before Living-Donor Liver Transplantation.

BACKGROUND: The majority of hepatocellular carcinoma (HCC) patients is diagnosed in late stages and therefore becomes ineligible for potentially curative treatment such as resection or liver transplantation. Transarterial chemoembolization (TACE) with drug-eluting beads (DC beads, Biocompatibles, Farnham, United Kingdom) has been proven with less side effects and better efficacy than conventional TACE, especially among patients with poor liver function. PURPOSE: The aim of this study is to evaluate outcomes of HCC patients who received TACE with DC beads, which resulted to eligibility for liver transplantation. METHODS AND MATERIALS: From January 2012 to June 2015, 60 patients with HCC received pre-liver transplantation evaluation whose cases were managed with TACE using DC beads at Kaohsiung Chang Gung Memorial Hospital were included in the study. DC beads loaded with doxorubicin were used. RESULTS: Forty percent of the patients had complete tumor response. Thirty-three percent of the patients had partial tumor response, of which 15% showed stable disease, 11.7% exhibited disease progression including 3 with portal vein thrombosis, 1 with both hepatic vein and portal vein thrombosis, and 3 with increase in tumor size. Twenty-three patients were beyond University of California, San Francisco (UCSF) criteria initially. The successful downstage rate was 73.9% (17 of 23). Thirty-seven patients fit the USCF criteria initially. The 3-, 6- and 12-month drop rates of these patients were 0%, 3.9%, and 16.8%, respectively. Twenty-four (40%) patients successfully underwent liver transplantation. Three patients (12.5%) demonstrated recurrent HCC after liver transplantation. CONCLUSION: TACE with DC bead can effectively induce tumor necrosis and appears to be a successful approach as bridge therapy for patients with advanced HCC and poor liver function.

Combined Hepatocellular Carcinoma and Cholangiocarcinoma: Diagnosis and Prognosis After Resection or Transplantation.

BACKGROUND: Primary liver malignancy is the leading cause of cancer death worldwide, with hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC) representing the majority. Combined HCC-CC, in contrast, accounts for less than 5% of these liver cancers and has not been clearly characterized by imaging, making diagnosis and management difficult. MATERIALS AND METHODS: This retrospective study investigated 32 patients with early-stage combined HCC-CC tumor who underwent hepatectomy (n = 24) or liver transplantation (n = 8). Preoperative imaging and pathologic reports were retrospectively reviewed and correlated. Survival and recurrence rates were then analyzed. RESULTS: Twelve patients with more than 50% CC component showed typical CC enhancement, whereas 17 patients with less than 50% CC component exhibited typical HCC enhancement. Those with equivocal imaging findings resulted near equal tumor component. The majority demonstrated either heterogeneous or peripheral enhancement. Considering the major tumor component, 66% of the images were consistent with histopathology. The over-all 3-year recurrent rate was 59%, with a mean time to recurrence of about 7 months. The 3-year survival rate of combined tumor after hepatectomy was 76% and after transplant was 75%, regardless of major tumor component. However, patients with more than 50% CC component showed a decrease in 3-year survival rate to 50% when transplantation was performed. CONCLUSION: The overall survival rate for combined tumor after either hepatectomy or transplantation seems to be satisfactory but carries a high risk of recurrent when compared to pure HCC. On the other hand, a major CC component tumor after transplantation is associated with poor survival outcome; thus, liver transplantation has no role and is not a good management option.

Magnetic resonance fat quantification in living donor liver transplantation.

OBJECTIVE: Hepatic steatosis can cause substantial problems for both donors and recipients in living donor liver transplantation (LDLT). The aim of this study is to evaluate the accuracy of the magnetic resonance IDEAL (iterative decomposition of water and fat with echo asymmetry and least squares estimation) sequence in quantifying the liver fat during LDLT. MATERIALS AND METHODS: A total of 63 liver donors (29 men and 34 women ranging from 18 to 47 years old with a mean age of 30) who received both magnetic resonance imaging (MRI) and intraoperative liver biopsy were enrolled in this study. MR IDEAL IQ sequences were performed by 1.5-T MRI (Discovery 450; GE Healthcare, Milwaukee, Wis, United States) to estimate the liver fatty content. Accuracy was assessed through linear regression between fat fraction image and pathology grading. Sensitivity and specificity of MR IDEAL IQ fat fractions were also calculated. RESULTS: A total of 63 LDLTs were performed and with pathology grading. No fatty content was found in 48 donors (76.2%; group 1), 5% to 10% fatty liver in 11 donors (17.4%; group 2), 11% to 15% fatty liver in 2 donors (3.2%; group 3), and >16% fatty change in 2 donors (3.2%; group 4). MR IDEAL fat fraction results were excellent in prediction of the normal and fatty content and with good correlation with the pathology grading (2.9 ± 0.9, 8.3 ± 4.2, P < .0001). Linear regression between IDEAL image and pathology grading indicated a high accuracy rate (R(2) = 0.813, R(2) = 0.9286) for all 4 groups. The sensitivity and specificity for detection of liver steatosis in MRI fat fraction image were 100% and 77.1% (P < .0001, 95% confidence interval 0.000-1.000). CONCLUSION: MR IDEAL IQ sequencing is a highly precise and accurate method in quantifying hepatic steatosis for the living donor.

The acoustic radiation force impulse elastography evaluation of liver fibrosis in posttransplantation dysfunction of living donor liver transplantation.

BACKGROUND: The acoustic radiation force impulse elastography (ARFI) is a new technology of elastography integrated into B-mode ultrasonography. It has been a reliable method to evaluate liver fibrosis of chronic liver disease in recent years, but less applied in the posttransplantation liver. The aim of the study was to evaluate liver fibrosis by the ARFI with correlation of pathological stages in living donor liver transplantation (LDLT). MATERIALS AND METHODS: From August 2010 to August 2012, there were 57 LDLT patients with liver biopsy (LB) due to posttransplantation dysfunction; all patients also received posttransplantation ARFI liver stiffness measurement (LSM) after transplantation for liver fibrosis staging. The ARFI elastography was performed using a Siemens Acuson S2000 ultrasound system with 4V1 transducers (Acusion, Siemens Medical Systems Co. Ltd. Erlangen, Germany). The ARFI LSM value was presented by shear wave velocity (SWV, m/s). The fibrosis staging as F0 to F4 was in accordance with the Metavir scoring system. RESULTS: A total of 57 patients had both posttransplantation LB and effective ARFI fibrosis staging for correlation. The ARFI LSM value increased with severity of liver fibrosis and had significant linear correlation with the results of histological fibrosis staging. The ARFI LSM sensitivities (Se), specificities (Sp), and cutoff values based on receiver-operator characteristic curve were F0: 0.75 m/s (Se: 93.8%, Sp: 4%), F1: 1.06 m/s (Se: 95.5%, Sp: 25.7%), F2: 1.81 m/s (Se: 50%, Sp: 83.6%) and F3: 2.33 m/s (Se: 100%, Sp: 92.9%). Predictive value of ARFI LSM reported a significant difference between early fibrosis stage (F0-F1) and advanced fibrosis stage (F ≧ 2) (P < .05). CONCLUSION: In this study, ARFI demonstrated a strong linear correlation and severity of liver fibrosis with LB pathologic staging. ARFI can be an alternative and compensatory method for frequent LB in the posttransplantation liver.

Magnetic resonance cholangiography in living donor liver transplantation: comparison of preenhanced and post-gadolinium-enhanced methods.

OBJECTIVE: The purpose of this study was to evaluate the image quality and diagnostic accuracy of postgadolinium complex of diethylenetriaminepentaacetic acid (GD-DTPA)-enhanced magnetic resonance cholangiography (MRC) in donor selection. MATERIALS AND METHODS: Donors (n=228) with both preoperative MRC and intraoperative cholangiography (IOC) were enrolled in this study. MRC pre- and post-GD-DTPA enhancement were performed using 1.5-T magnetic resonance imaging. The signal-to-noise ratio (SNR) of liver parenchyma and contrast-to-noise ratio of bile duct, as well as the contrast between bile duct and liver parenchyma, were calculated. The biliary anatomy correlation with the IOC during hepatectomy and patient prognosis were also evaluated. RESULTS: Quantitative results of the SNR of the liver parenchyma post-GD-DTPA were statistically significantly lower than preenhanced MRC (2.69 times reduced from the preenhanced MRC). The contrast of the bile duct and liver parenchyma in post-GD-DTPA were significantly higher than the preenhancement MRC. The anatomic diagnostic accuracy rate of post-GD-DTPA MRC was 92.9%. The sensitivity and specificity of GD-PTPA MRC were 85% and 96%, respectively. GD-DTPA-enhanced MRC has higher accuracy than the preenhanced MRC (92.9% vs 75%). The concurrence between GD-DTPA-enhanced MRC and IOC were commendable (kappa=0.9). The posttransplant biliary complication rate was 5.5%, and the 3-year survival rate was 91.2% in the recipients. CONCLUSION: GD-DTPA, a paramagnetic metal, can shorten the T1 and T2 relaxation values of surrounding protons. This decreases the signal of the liver parenchyma and brightens the biliary anatomy. It can improve the image quality of MRC and increase the diagnostic accuracy of the biliary tract classification. It is mandatory in the "donor and recipient surgery during the LDLT".

Hepatocellular carcinoma downstaging in liver transplantation.

BACKGROUND: Hepatocellular carcinoma (HCC) is the leading malignant tumor in Taiwan. The majority of HCC patients are diagnosed in late stages and therefore in eligible for potentially curative treatments. Locoregional therapy has been advocated as an effective treatment for patients with advanced HCCs. PURPOSE: The aim of this study was to evaluate the outcomes of HCC downstaged patients after locoregional therapy to allow eligibility for liver transplantation. METHODS AND MATERIALS: From January 2004 to June 2010, 161 patients with HCCs underwent liver transplantation including 51 (31.6%) who exceeded the University of California-San Francisco (UCSF) who had undergone successful locoregional therapy to be downstaged within these criteria. Among the downstaged patients, 48 (94.1%) underwent transarterial embolization; 7 (13.8%), percutaneous ethanol injection; 24 (47.1%), radiofrequency ablation; 15 (29.4%), surgical resection, and 34 (66.7%), combined treatment. RESULTS: The overall 1- and 5-year survival rates of all HCC patients (n=161) were 93.2% and 80.5%. The overall 1- and 5-year survival rates of downstaged (n=51) versus non-downstaged (n=110) subjects were 94.1% versus 83.7% and 92.7% versus 78.9%, respectively (P=.727). There are 15 (9.2%) HCC recurrences. The overall 1- and 5-year tumor-free rates of all HCC patients were 94.8% and 87.2%. The overall 1- and 5-year tumor-free rates between downstaged versus non-downstaged patients were 93.9% and 90.1% versus 95.2% and 86.0%, respectively (P=.812). CONCLUSION: Patients with advanced HCC exceeding the UCSF/Milan criteria can be downstaged to fit the criteria using locoregional therapy. Importantly, successfully downstaged patients who are transplanted show excellent tumor-free and overall survival rates, similar to fit-criteria group.

Biliary complications and management in pediatric living donor liver transplantation for underlying biliary atresia.

BACKGROUND: Biliary complications are a major problem in pediatric liver transplantation. The aim of this study was to evaluate the management and outcomes of biliary complication after pediatric living donor liver transplantation (LDLT). METHODS: From 1994 to 2010, 157 pediatric LDLT due to biliary atresia were performed in our center. Doppler ultrasound was initially performed daily for 2 weeks postoperatively to evaluate biliary and vascular complications. Computed tomography and or magnetic resonance cholangiography were performed when complications were suspected. They were treated using radiological or surgical interventions. RESULTS: Among the 157 cases, we observed 10 (6.3%) biliary complications, which were divided into three groups: bile leakage (n=3); biliary stricture without vascular complication (n=4); and biliary stricture with vascular complication (n=3). The three cases bile leakages recovered after interventional procedures. The seven biliary strictures underwent percutaneous transhepatic cholangial drainage (PTCD). All cases without vascular complications were completely cured after PTCD or a subsequent surgical re-anastomosis. In the vascular complication group, early recorrection of the HA occlusion with successful PTCD treatment were performed in two cases, but one other case with diffuse ischemic biliary destruction had a poor result. CONCLUSION: Successful interventional radiographic approaches are effective for anastomotic biliary complications but with poor results in diffuse ischemic biliary destruction.

1,4-diarylpiperazines and analogs as anti-tubercular agents: synthesis and biological evaluation.

Despite progress in modern chemotherapy to combat tuberculosis, the causative pathogen Mycobacterium tuberculosis (M.tb.) is far from eradicated. Bacillary resistance to anti-mycobacterial agents, bacillary persistence and human immunodeficiency virus (HIV) co-infection hamper current drug treatment to completely cure the infection, generating a constant demand for novel drug candidates to tackle these problems. A small library of novel heterocyclic compounds was screened in a rapid luminometric in vitro assay against the laboratory M.tb. strain H37Rv. A group of amidines was found to have the highest potency and was further evaluated for acute toxicity against C3A hepatocytes. Next, the most promising compounds were evaluated for activity against a multi-drug resistant clinical isolate. The group of amidines was also tested for their ability to kill intracellular M.tb. residing in mouse J774A.1 macrophages. Finally, we report on a correlation between the structural differences of the compounds and their anti-mycobacterial activity.

Superparamagnectic iron oxide-enhanced magnetic resonance for tumor surveillance in cirrhotic liver before liver transplantation with explanted liver correlation.

OBJECTIVE: The aim of this study was to determine the accuracy of superparamagnetic iron oxide (SPIO)-enhanced magnetic resonance imaging (MRI) in the detection of liver tumors in patients with liver cirrhosis before liver transplantation (LT). PATIENTS AND METHODS: Twenty-one patients with moderate to severe liver cirrhoses (Child-Pugh score B or C) underwent SPIO-enhanced MRI before LT. SPIO-enhanced MRI was composed of SPIO-enhanced T1-weighted images, T2-weighted turbo spin-echo, and T2*-weighted sequences. The detection of malignant liver tumor was based on hepatic lesions that remain with hyper-signal intensity on SPIO-enhanced delayed T2 and T2*-weighted images. The imaging was compared with findings during histopathologic evaluation of the explanted liver. RESULTS: Forty-two liver nodules were found in 16 of the 21 patients. Thirty-six malignant hepatic neoplasms were detected at histopathologic evaluation. SPIO-enhanced MRI depicted 30 of 36 malignant hepatic neoplasms. Patient-based analysis showed that the sensitivity, specificity, and accuracy of MRI were 93.75%, 100%, and 95.2%, respectively. For lesion-based analysis, the sensitivity, specificity, and the area under the receiver operating characteristic (ROC) curve were 90%, 100%, and 0.95, respectively, for lesions >2 cm. CONCLUSIONS: SPIO-enhanced MRI can be used as an additional diagnostic tool for the detection and characterization of malignant tumors in cirrhotic livers. This diagnostic evaluation is accurate to triage patients for LT.

Correlation between imaging and pathologic findings in explanted livers of hepatocellular carcinoma cases.

PURPOSE: We sought investigate the accuracy of preoperative computed tomography angiography (CTA) and magnetic resonance imaging (MRI) to evaluate tumor-related prognostic factors, including tumor size, number, portal vein (PV) thrombosis, and bile duct invasion. MATERIALS AND METHODS: From March 2006 to October 2008, we enrolled 57 patients with hepatocellular carcinoma (HCC) who were undergoing liver transplantation at our institute. Imaging was performed with multidetector 64 slice CTA and MRI within 1 month preoperatively. Imaging findings including tumor size, number, PV thrombosis, and bile duct invasion were correlated with histopathologic features from the explanted livers. RESULTS: We included 128 HCCs in 57 patients in this study. The sensitivities to detect tumor number and size were 83.6% and 88.8% for CTA, and 75.8% and 88.7% for MRI. In addition, CTA and MRI were both accurate to determine whether patients were beyond or within the Milan and UCSF criteria. Fifteen patients with PV microthrombosis and 1 with bile duct microinvasion were not found on CTA or MRI preoperatively; negative predictive value of PV microthrombosis and bile duct microinvasion were 73.68% and 98.25%, respectively. During follow-up, 2 patients experienced recurrence, one of which was associated with PV microthrombosis, and 4 patients died of causes unrelated to HCC. CONCLUSION: CTA and MRI were both accurate to determine whether patients fit within the Milan or UCSF criteria, but CTA was slightly better than MRI to evaluate tumor number and size. However, pretransplant diagnostic pitfalls were PV microthrombosis and bile duct microinvasion.

Early modulation of portal graft inflow in adult living donor liver transplant recipients with high portal inflow detected by intraoperative color Doppler ultrasound.

Splenic flow clearly contributes to portal hyperperfusion in end-stage liver disease patients with hypersplenism. Early modulation of portal graft inflow can relieve the partial graft from the deleterious effects of this portal overflow. Color Doppler ultrasound may play an important role in the intraoperative detection of patients with high recipient portal inflow. Among 138 cases, portal inflow increased after liver transplantation, namely recipient portal inflow (rPVF) of 60 to 554 mL/min/100 g graft weight (mean=156 mL/min/100 g). There were eight cases in which the rPVF was >250 mL/min/100 g graft weight. In the early period of this study, there were two patients with high portal inflow (rPVF>250 mL/min/100 g graft) who did not undergo modulation of the portal flow. Small-for-size syndrome occurred in two patients. Later in the series, six patients underwent intraoperative splenic arterial ligation or splenectomy; only one of them displayed prolonged hyperbilirubinemia. The incidence of small-for-size syndrome between these two groups was significant (P=.048). No complication occurred related to the splenic artery ligation or the splenectomy. In conclusion, early modulation of portal graft inflow can protect the graft from portal overflow damage. Intraoperative Doppler ultrasound measurement is an optimal modality for detection of high rPVF.

Does hepatic graft weight affect the reduction of spleen size after living donor liver transplantation?

OBJECTIVE: Our aim was to evaluate whether the reduction in spleen volume at 6 months after living donor liver transplantation (LDLT) was affected by the size of the right lobe liver graft. PATIENTS AND METHODS: We analyzed 87 adult recipients of right lobe liver grafts who displayed preoperative splenomegaly: spleen volume>500 cm3 by computed tomographic (CT) volumetry. The recipients were grouped according to the graft weight-to-recipient weight ratio: GRWR>1 versus GRWR<1. The 2 groups were compared at 6 months after LDLT for mean postoperative spleen volume (SV) and mean SV change ratio 5, which was defined as [(SVpreop-SV6m)/SVpreop]x100%, where SVpreop and SV6m represent SV calculated based on CT examinations preoperatively and at 6 month follow-up after LDLT, respectively. RESULTS: The GRWR ranged from 0.77 to 1.66. There were 53 patients with GRWR>1 and 34 with GRWR<1. Our analysis showed significant hepatic graft volume regeneration and SV reduction at 6 months after LDLT. The SV change ratio weakly but significantly correlated with the transplanted liver graft weight (Pearson correlation coefficient, r=0.274; P<.009). In the group GRWR>1, the mean postoperative SV and the mean SV change ratio were 632+/-220 cm3 and decreased by 32+/-11%, respectively. The mean postoperative SV and the mean SV change ratio in group GRWR<1 were 598+/-188 cm3 and decreased by 34+/-13%, respectively. There were no differences in mean postoperative SV and mean SV change ratios between the 2 groups. CONCLUSION: LDLT using a right lobe graft resulted in a significant reduction of SV at 6 months after surgery, but there were no significant differences between recipients who received different sized right lobe liver grafts.