Generation of functional hepatocyte-like cells from human bone marrow mesenchymal stem cells by overexpression of transcription factor HNF4α and FOXA2.

BACKGROUND: Our previous study showed that overexpression of hepatocyte nuclear factor 4α (HNF4α) could directly promote mesenchymal stem cells (MSCs) to differentiate into hepatocyte-like cells. However, the efficiency of hepatic differentiation remains low. The purpose of our study was to establish an MSC cell line that overexpressed HNF4α and FOXA2 genes to obtain an increased hepatic differentiation efficiency and hepatocyte-like cells with more mature hepatocyte functions. METHODS: Successful establishment of high-level HNF4α and FOXA2 co-overexpression in human induced hepatocyte-like cells (hiHep cells) was verified by flow cytometry, immunofluorescence and RT-PCR. Measurements of albumin (ALB), urea, glucose, indocyanine green (ICG) uptake and release, cytochrome P450 (CYP) activity and gene expression were used to analyze mature hepatic functions of hiHep cells. RESULTS: hiHep cells efficiently express HNF4α and FOXA2 genes and proteins, exhibit typical epithelial morphology and acquire mature hepatocyte-like cell functions, including ALB secretion, urea production, ICG uptake and release, and glycogen storage. hiHep cells can be activated by CYP inducers. The percentage of both ALB and α-1-antitrypsin (AAT)-positive cells was approximately 72.6%. The expression levels of hepatocyte-specific genes (ALB, AAT, and CYP1A1) and liver drug transport-related genes (ABCB1, ABCG2, and SLC22A18) in hiHep cells were significantly higher than those in MSCs-Vector cells. The hiHep cells did not form tumors after subcutaneous xenograft in BALB/c nude mice after 2 months. CONCLUSION: This study provides an accessible, feasible and efficient strategy to generate hiHep cells from MSCs.

Improvement of ablative margins by the intraoperative use of CEUS-CT/MR image fusion in hepatocellular carcinoma.

BACKGROUND: To assess whether intraoperative use of contrast-enhanced ultrasound (CEUS)-CT/MR image fusion can accurately evaluate ablative margin (AM) and guide supplementary ablation to improve AM after hepatocellular carcinoma (HCC) ablation. METHODS: Ninety-eight patients with 126 HCCs designated to undergo thermal ablation treatment were enrolled in this prospective study. CEUS-CT/MR image fusion was performed intraoperatively to evaluate whether 5-mm AM was covered by the ablative area. If possible, supplementary ablation was applied at the site of inadequate AM. The CEUS image quality, the time used for CEUS-CT/MR image fusion and the success rate of image fusion were recorded. Local tumor progression (LTP) was observed during follow-up. Clinical factors including AM were examined to identify risk factors for LTP. RESULTS: The success rate of image fusion was 96.2% (126/131), and the duration required for image fusion was 4.9 ± 2.0 (3-13) min. The CEUS image quality was good in 36.1% (53/147) and medium in 63.9% (94/147) of the cases. By supplementary ablation, 21.8% (12/55) of lesions with inadequate AMs became adequate AMs. During follow-up, there were 5 LTPs in lesions with inadequate AMs and 1 LTP in lesions with adequate AMs. Multivariate analysis showed that AM was the only independent risk factor for LTP (hazard ratio, 9.167; 95% confidence interval, 1.070-78.571; p = 0.043). CONCLUSION: CEUS-CT/MR image fusion is feasible for intraoperative use and can serve as an accurate method to evaluate AMs and guide supplementary ablation to lower inadequate AMs.

Contrast-Enhanced Sonography for Diagnosing Collateral Transformation of the Hepatic Artery After Liver Transplantation.

OBJECTIVES: To determine the contrast-enhanced sonographic features of hepatic artery collateral transformation in patients with hepatic artery complications after liver transplantation. METHODS: Ninety-nine liver transplant recipients who underwent contrast-enhanced sonography were recruited from April 2004 to May 2014. The reference standards were conventional angiography and computed tomographic angiography. The contrast-enhanced sonographic features of the hepatic artery in patients with and without collateral arteries were retrospectively analyzed. RESULTS: All 15 patients with hepatic artery collateral transformation had hepatic artery thrombosis (10 of 15) or hepatic artery stenosis (5 of 15). The collateral artery detection rate on contrast-enhanced sonography was 100%. The peripheral hepatic artery could not be visualized by contrast-enhanced sonography in most of the patients with hepatic artery collateral transformation (14 of 15). Additionally, many small tortuous collateral arteries in the porta hepatis region were visualized during the arterial and early portal phases, showing reticulated/patchy (15 of 15) and striped (3 of 15) enhancement patterns on contrast-enhanced sonography. CONCLUSIONS: Collateral transformation of the hepatic artery in patients with hepatic artery complications after liver transplantation appears to have characteristic features on contrast-enhanced sonography, especially a reticulated or patchy enhancement pattern in the porta hepatis region during the arterial and early portal phases combined with the absence of the peripheral hepatic artery. Contrast-enhanced sonography may be a novel method for diagnosing hepatic artery collateral transformation, which may be a highly specific sign of hepatic artery thrombosis or stenosis.

[Clinical observation of transcatheter arterial chemoembolization plus sorafenib in the treatment of advanced hepatocellular carcinoma with different types of portal vein tumor thrombosis].

OBJECTIVE: To explore the efficacy and safety of transcatheter arterial chemoembolization (TACE) plus sorafenib in the treatment of advanced hepatocellular carcinoma with different types of portal vein tumor thrombosis. METHODS: A total of 32 patients of advanced hepatocellular carcinoma with tumor thrombosis in portal vein were retrospectively analyzed. All of them took oral sorafenib after TACE. They were divided into 3 groups according to imaging examinations of tumor thrombosis in portal vein. Tumor thrombosis in main portal vein was group A, tumor thrombosis in right/left portal branch group B and tumor thrombosis in the second branch of portal vein group C. Tumor response rate, disease control rate (DCR), overall survival (OS) and time to tumor progression (TTP) was followed up. Liver functions were compared with the pre-treatment level. The occurrences of adverse events were recorded. RESULTS: DCR was 20.0% (Group A), 70.0% (Group B) and 91.7 % (Group C) at 2 months post-treatment. DCR in groups B and C had significant differences with group A (P < 0.05). The median OS was 3 (Group A), 9 (Group B) and 14 months (Group C) and the median TTP 0 (Group A), 3 (Group B) and 6 months (Group C) respectively. The median OS and median TTP were significantly longer in Groups B and C than those in Group A (P < 0.05). Liver function at 2 months post-treatment had no statistical difference with the baseline. The most common adverse effects included hand foot skin reaction (n = 23, 3 cases of grade 3), hypertension (n = 3), diarrhea (n = 25, 3 cases of grade 3), hair loss (n = 12), oral ulcers (n = 1) and gastrointestinal bleeding (n = 2). CONCLUSION: The combined use of TACE and sorafenib is both safe and efficacious in the treatment of advanced hepatocellular carcinoma with tumor thrombosis in portal vein. And it may prolong OS and TTP in hepatocellular carcinoma with tumor thrombosis in right/left portal vein and second branch of portal vein.

[Evaluation of graft perfusion in patients with ischemic-type of biliary lesions after liver transplantation].

OBJECTIVE: To investigate the value of 320-rows CT perfusion (CTP) imaging in the study of hepatic hemodynamic characters in ischemic-type biliary lesions (ITBL) after liver transplantation. METHODS: A total of 11 ITBL patients received 320-slice CT angiography (CTA) and CTP after liver transplantation scheduled at 5-10 min away. Four patients underwent liver biopsy While 7 patients with normal liver after transplantation were selected as the control group. The parameters of hepatic artery perfusion (HAP), portal vein perfusion (PVP), total hepatic perfusion (TLP) and hepatic arterial perfusion index (HPI) were measured and compared for all patients. And the blood perfusion characters of liver with ITBL after transplantation were analyzed. RESULTS: (1) In 11 ITBL patients, 3 patients had no vascular complications on CTA, 1 with simple hepatic artery stenosis (HAS), 1 with HAS and arterioportal shunt (APS), 2 with HAS and portal vein stenosis/right hepatic vein stenosis (PVS/RHVS), 1 with simple APS, 2 with simple PVS and 1 with portal vein thrombosis and cavernous transformation of portal vein (PVT and CTPV). And 4/11 patients underwent liver biopsy, 2 in which confirmed mild acute rejection and 2 confirmed biliary obstruction associated with ascending biliary infection.(2) HAP of the ITBL and control groups were (66 ± 38) and (40 ± 8) ml×min(-1)·(100 ml)(-1), PVP (128 ± 35) and (163 ± 21) ml×min(-1)·(100 ml)(-1), TLP (194 ± 58) and (203 ± 19) ml×min(-1)·(100 ml)(-1), HPI 34% ± 14% and 21% ± 4% respectively. The differences in the value of HAP, PVP and HPI between the groups were statistically significant (P < 0.05) excluding TLP. CONCLUSION: Various liver perfusion abnormalities of ITBL may be evaluated objectively by CTP. ITBL might occurred when HAP and HPI increased with a decreased of PVP.

[The diagnosis and treatment of isolated celiac and superior mesenteric artery dissection: 2 cases report and literature review].

OBJECTIVE: To investigate the diagnosis and treatment of isolated celiac artery (CA) dissection and superior mesenteric artery (SMA) dissection. METHODS: Integrating clinical data of 119 cases with isolated dissection of the visceral arteries (IDVA) reported in literature and 2 patients with spontaneous isolated dissections of both CA and SMA treated in the Third Affiliated Hospital of Sun Yat-sen University, the diagnosis and treatment of IDVA were analyzed retrospectively. RESULTS: Among 119 cases reported in the literature, 69 cases were symptomatic. All of the cases were diagnosed by contrast-enhanced abdominal CT or MRI. After IDVA was discovered, surgical treatment and endovascular stent placement was performed in 8 and 5 patients respectively, although the remaining 106 patients were managed conservatively with good results. In our 2 cases, the diagnosis of CA and SMA dissection was established by contrast-enhanced CT and confirmed by conventional angiography. One patient was treated with anticoagulation and antihypertension, and the other patient was treated with endovascular stenting. Both of the patients didn't have discomfort during the follow-up period of 12 and 3 months respectively. CONCLUSIONS: Contrast-enhanced abdominal CT is the main tool for detection of IDVA. Most of the patients with IDVA can be managed conservatively, but close surveillance with imaging studies is necessary for early recognition of dissection progression. Patients with persistent or relapsed symptoms, and dissection progression, should undergo surgical or endovascular treatment.

[Construction and validation of dual fusion reporter gene expression vector for molecular imaging study].

OBJECTIVE: To construct dual fusion reporter gene expression vector containing enhanced green fluorescence protein (EGFP) and human transferrin receptor (TfR), and validate the reconstructed plasmid, which will provide experimental foundation for in vivo dual-modality optical/Magnetic Resonance (MR) imaging. METHODS: Clone TfR into the pEGFP-C1 vector to construct pEGFP-C1-TfR plasmid.pEGFP-C1-TfR plasmid was transfected into 293T cells for 48 h, then investigate EGFP expression under a fluorescence microscope; detect TfR expression through PT-PCR; inspect the subcellular location of EGFP-TfR fusion protein through Confocal Scanning Laser Microscopy; evaluate the function of EGFP-TfR fusion protein through Tf probe uptake and competition assays. RESULTS: DNA sequencing analysis confirmed that EGFP-TfR gene sequence was correct, and there was no mutation and deletion. After transfecting the reconstructed plasmid into 293T cells, fluorescence microscope observation and RT-PCR results demonstrated that EGFP and TfR were expressed efficiently. EGFP-TfR fusion protein was located predominantly in the cellular membrane, and could specifically mediate internalization of Tf. CONCLUSION: EGFP-TfR dual fusion reporter gene expression vector has been successfully constructed, and could be expressed efficiently with functional features. Thus, the expression vector could be applied for in vivo dual-modality optical/Magnetic Resonance (MR) imaging.

[Labeling of liver cancer cell for fluorescence imaging study by far-red fluorescence protein reporter gene mKate2].

OBJECTIVE: To create far-red fluorescence protein reporter gene mKate2 lentivirus, label human liver cancer cell line HepG2 with lentivirus and explore the feasibility of in vitro fluorescence imaging of labeled tumor cells so as to provide experimental rationales for in vivo fluorescence tumor imaging. METHODS: mKate2 gene was amplified from pmKate2-N plasmid. Then the fragment was inserted into the lentivirus expression vector pLenti6.3/V5-DEST. The expression plasmids pLenti6.3-mKate2 and the packaging plasmids were cotransfected into 293T cells. The biological titer of lentivirus was determined. HepG2 cells were infected with mKate2 lentivirus at a MOI (virus multiplicity of infection) of 6 for 96 hours. The infection efficiency was assayed through fluorescence microscope and fluorescent-activated cell scanning (FACS). And 2 × 10(6) mKate2-HepG2 cells were collected for fluorescence imaging through an optical imaging system. And the optimal imaging parameters were determined. RESULTS: DNA sequencing analysis confirmed that mKate2 gene sequence was correct and there was no mutation or deletion. The biological titer of produced mKate2 lentivirus was 1.6 × 10(6) TU/ml. At 96 hours after mKate2 lentivirus infection, fluorescence microscope showed that mKate2 was expressed in a large percentage of cells. FACS assay showed that the mKate2 positive rate was 93.8% ± 0.4%. Excitation light 530 ± 15 nm and emission light 710 ± 28 nm were the optimal imaging parameters for mKate2-HepG2 cells. CONCLUSION: Lentivirus can mediate efficiently the mKate2 reporter gene labeling of human liver cancer cell line HepG2. The mKate2-labeled HepG2 cells can be detected through in vitro fluorescence imaging. Further tracing studies of in vivo tumor fluorescence imaging are technically feasible.

Chemoembolization with lobaplatin mixed with iodized oil for unresectable recurrent hepatocellular carcinoma after orthotopic liver transplantation.

PURPOSE: To determine whether chemoembolization can benefit patients with unresectable recurrent hepatocellular carcinoma (HCC) after orthotopic liver transplantation (OLT). MATERIALS AND METHODS: Twenty-eight of 71 patients (39%) with unresectable recurrent HCC following OLT and without contradictions to chemoembolization were included: 14 patients received chemoembolization after OLT (chemoembolization group) and 14 matched control subjects who did not receive chemoembolization (non-chemoembolization group). Tumor response was determined with follow-up computed tomography after each chemoembolization procedure and classified into four grades according to Response Evaluation Criteria in Solid Tumors. Overall survival was evaluated from OLT and from the diagnosis of recurrent HCC. RESULTS: Within a median follow-up of 14.5-months, 12 of the 14 patients in the chemoembolization group (86%) and 13 of the 14 in the non-chemoembolization group (93%) developed new recurrences. Eight of the 14 patients in the chemoembolization group (57%) showed partial tumor response (>30% reduction in the size of target lesions). Moreover, patients who underwent chemoembolization had a significantly longer overall survival after OLT (P = .0133) and after the diagnosis of HCC recurrence (P = .0338) compared to those who did not. No severe complications developed in patients receiving chemoembolization during follow-up. CONCLUSIONS: Lobaplatin-based chemoembolization may elicit effective tumor response for recurrent HCCs and improve the overall survival of patients with unresectable HCC recurrence following OLT.

[Clinical application of prospective electrocardiogram-gated 320-detector computed tomography coronary angiography].

OBJECTIVE: to compare the success rate, radiation dose, image quality and diagnosis of prospective electrocardiogram(ECG)-gated 320-detector computed tomography coronary angiography (CTCA) versus retrospective ECG-gated CTCA. METHODS: patients suspected coronary artery disease were divided into two groups which underwent 320-detector CTCA with prospective ECG-gated and retrospective ECG-gated scanning (n = 240 each, HR < 65 bpm). Curved-planar reconstruction (CPR), maximum intensity projection (MIP) and volume rendering (VR) were performed to demonstrate the coronary arteries. The image quality was defined as excellent, good and poor by motion and stair-step artifacts. Effective radiation exposure dose was estimated from the dose-length product. Effective radiation dose, image quality and diagnosis were evaluated. RESULTS: the success rate of examination was 100% in prospective ECG-gated group and retrospective ECG-gated group. The mean effective radiation dose of prospective ECG-gated CTCA [(3.3 ± 1.3) mSv] was significantly lower than that of retrospective ECG-gated CTCA [(13.0 ± 1.6) mSv, P < 0.01]. Segments of diagnostic image quality (95.42%, 3435/3600) and non-diagnostic coronary segments (4.58%, 165/3600) in prospective ECG-gated group were similar as those of retrospective ECG-gated group (95.81%, 3449/3600 and 4.19%, 151/3600, all P > 0.05). Compared with CAG, the sensitivity, specificity, false positive and false negative value in prospective ECG-gated group (93.22%, 99.21%, 91.64%, 99.05%) and retrospective ECG-gated group (94.55%, 98.80%, 95.86%, 98.54%) were not significantly different. CONCLUSION: though the effective radiation dose is significantly lower, the success rate, image quality and diagnosis of prospective ECG-gated 320-detector CTCA is comparable with that of retrospective ECG-gated 320-detector CTCA on patients with stable heart rates less than 65 bpm.

[Evaluation of esophageal varices and predicting the risk of esophageal varices bleeding with multi-detector CT in patients with portal hypertension].

OBJECTIVE: The objective of this study was to evaluate the performance of 320-row multi-detector CT (MDCT) in the detection and grading of esophageal varices and to evaluate the ability of MDCT in predicting the risk of hemorrhage in comparison with upper endoscopy in patients with portal hypertension. METHODS: A total of 69 patients, with clinically confirmed portal hypertension, underwent epigastric triphasic enhancement scans using 320-row MDCT 1 weeks or less before upper endoscopies were performed. Two blinded abdominal imagers retrospectively interpreted all CT images to detect the presence of esophageal varices and divided EV into large EV (≥ 5 mm) and small EV (< 5 mm). The correlation between CT measurements and endoscopic grading was assessed by kappa values. With endoscopy as standard, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value of the two radiologists in detection of EV were calculated. Correlations between CT measurements of varix size and red color sign were assessed by correlation analysis. RESULTS: Of the total of 69 patients, 56 patients had esophageal varices, 41 had large EV, and 15 had small EV according to the endoscopic findings. MDCT showed an excellent interobserver reliability with regard to the diagnosis of the EV (k = 0.94). Agreement between CT and endoscopy with regard to the grading of EV were excellent (k = 0.77). The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of radiologist 1 in the detection of EV was 95%, 69%, 87%, 93% and 75% respectively; the sensitivity, specificity, accuracy, positive predictive value and negative predictive value of radiologist 2 in the detection of EV was 93%, 77%, 87%, 95% and 71%, respectively. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of the two radiologists in the detection of large EV was 95%, 100%, 97%, 100% and 93%, respectively. CT variceal grading showed a strong correlation with endoscopic grading for both observers (P < 0.01). Correlations between CT measurements of varix size and red color sign were significant in both radiologists with a correlation coefficient r of 0.731 for radiologist 1 (P < 0.01) and 0.718 for radiologist 2 (P < 0.01). CONCLUSION: 320-row MDCT is useful for the detection and grading of EV, and it is useful in evaluation of EV for predicting a risk of hemorrhage.

[Enhanced green fluorescence protein reporter gene labeling of mesenchymal stem cells mediated by lentivirus].

OBJECTIVE: To explore the effect of enhanced green fluorescence protein (EGFP) labeling mediated by lentivirus on the biophysical properties of mesenchymal stem cells (MSC), and whether the EGFP gene expression is permanent and stable. METHODS: MSC were infected with EGFP lentivirus at different virus multiplicity of infection (MOI). EGFP positive rate was measured with fluorescent-activated cell scanning (FACS) analysis, and EGFP expression in MSC was investigated under a fluorescence microscope. Cell viability, proliferation, apoptosis and cell cycle were detected with trypan blue stain, MTT colorimetric assay, Hoechst stain and FACS analysis respectively. To evaluate the stability of EGFP expression, EGFP lentivirus infected MSC were harvested after cultured continuously in vitro for 2, 4, 8 or 16 weeks, and EGFP positive rate and fluorescence strength were detected with FACS analysis. RESULTS: After infected with EGFP lentivirus (MOI = 20) for 96 h, EGFP positive rate of MSC was 97.39% +/- 0.68%. Cell viability, proliferation, apoptosis and cell cycle of MSC infected with EGFP lentivirus were unaffected, as compared with control MSC (P > 0.05). When cultured in vitro continuously for 2, 4, 8 or 16 weeks, EGFP positive rates of EGFP-MSC were 97.50% +/- 0.54%, 97.32% +/- 0.51%, 97.39% +/- 0.11%, and 97.48% +/- 0.13% respectively, while EGFP fluorescence strength were 440 +/- 13, 445 +/- 12, 458 +/- 13 and 456 +/- 16 respectively. Both EGFP positive rate and fluorescence strength kept in a stable level. CONCLUSION: EGFP lentivirus can efficiently label MSC and has no significant effect on the biophysical properties of MSC. EGFP gene expression in MSC is permanent and stable. EGFP-MSC can be used for further cell tracing research.

[The role of early hepatic artery ischemia on biliary complications after liver transplantation and hepatic arterial interventional therapy].

OBJECTIVE: To explore the influence of early hepatic artery ischemia on the occurrence and prognosis of biliary complications after orthotopic liver transplantation (OLT), and the value of early hepatic arterial interventional therapy. METHODS: In the 720 recipients who received OLT in our hospital from October 2003 to June 2007, 32 cases were detected hepatic artery stenosis (HAS, 30 cases) or hepatic artery thrombosis (HAT, 2 cases) by color Doppler Ultrasound from 4 to 65 days (mean, 25 +/- 15) after OLT. All of them were confirmed by DSA and/or CT angiography. Of the 32 patients, 20 were treated by hepatic arterial interventional therapy. The end-point of follow-up was the time of patient's death and retransplantation. RESULTS: In this study, 20 cases developed biliary complications, including the common bile duct stenosis in 2 cases, intra- and extra hepatic bile duct stenosis in 13 cases and multiple intrahepatic bile duct stenosis in 5 cases. Among them, 2 patients complicated with bile leakage, 4 with biloma and 3 with liver abscess. Of the 20 patients, 8 with HAS received successful hepatic arterial interventional therapy which was performed two weeks after HAS detected; 10 with HAS didn't receive hepatic arterial interventional therapy; 1 with HAT received successful thrombolysis; 1 with HAS received failed hepatic artery stent implantation. During a median follow-up of 262 days (range, 22 -517 days), 10 patients died, 6 underwent retransplantation, and the other 4 survived; cumulated survival rates at 6, 12 and 24 months were 60.0%, 34.9% and 0, respectively. 12 cases didn't develop biliary complications. Nine of them received successful hepatic arterial interventional therapy within 2 weeks HAS detected, 2 with acute rejection received flushing anti-rejection therapy, 1 with HAT received retransplantation because of unsuccessful thrombolysis. During a median follow-up of 952 days (range, 14 - 1398 days), 3 patients died, 1 underwent retransplantation, and the other 8 survived; cumulated survival rates at 6, 12 and 24 months were 75%, 66.7% and 66.7%, respectively. CONCLUSION: Early hepatic artery ischemia after OLT is an important agent for biliary complications. Early and successful hepatic arterial interventional therapy helped to reduce the incidence of biliary complications and improve the patients' prognosis.

[Using 1H-MRS to evaluate the effects of warm ischemia reperfusion injury on the regeneration of hepatic cells in orthotopic transplanted livers in rats].

OBJECTIVE: To investigate the prognostic value of proton magnetic resonance spectroscopy (1H-MRS) in the study of warm ischemia reperfusion injury to the regeneration of hepatic cells of the livers following their orthotopic transplantation in rats. METHODS: A rat orthotopic liver transplantation (OLT) model with warm ischemia, the experimental group, was established and the same was done with a control group but without warm ischemia of the livers. They were studied at 6 time points (6 hours, 1, 3, 7, 14, and 30 days after OLT). All rats took axial T1 weighted and T2 weighted imaging scans and 1H MR spectroscopies. RESULTS: The positive rate of proliferating cell nuclear antigen (PCNA) and the mean peak choline/water ratio in the experimental group were significantly higher than those in the control group and the peak choline /water ratio had a positive correlation with the positive rate of PCNA. Serum ALT and AST increased significantly after OLT, especially during the 6 hour to 3day period. The levels of ALT and AST were markedly higher in the experimental group compared to the control group. CONCLUSION: Warm ischemia reperfusion injury of OLT has a significant effect on the regeneration of hepatic cells, and the choline peak of 1H-MRS can be used to evaluate the regeneration of hepatic cells non-invasively.

[Transportal variceal sclerotherapy with n-butyl-2-cyanoacrylate for gastric fundal varices].

OBJECTIVE: To evaluate the technique, safety and clinical efficacy of transportal variceal sclerotherapy with n-butyl-2-cyanoacrylate (NBCA) for gastric fundal varices. METHODS: Twenty-one patients with gastric fundal varices confirmed by endoscopy were enrolled in this study. The causes of the gastric varices were cirrhosis caused by hepatitis virus B or C (n = 16) and hepatocellular carcinoma with portal venous obstruction (n = 5). Percutaneous transhepatic or transplenic portography were performed on all 21 patients. The gastric varices were treated with NBCA-lipiodol mixture injected via a microcatheter introduced into the varices. For 8 patients who had large gastrorenal shunts (GRS), a balloon-occluded catheter was introduced into the GRS via the right femoral and left renal veins before injecting the NBCA-lipiodol. During the NBCA-lipiodol injection, the balloon was inflated to block the flow of GRS. Follow-up evaluations included findings of the laboratory liver function tests, upper intestinal endoscopies, and the occurrences of rebleeding. RESULTS: In 20 patients (95.2%), the gastric varices were successfully obliterated with 2-8 ml of NBCA-lipiodol. In one patient with a large GRS, sclerotherapy was not successfully performed because a balloon-occluded catheter was not available during the procedure. In five patients, small amounts of NBCA-lipiodol entered into the distal pulmonary artery branches. Two of them suffered from transient irritable coughs; no patient developed severe pulmonary embolism. Embolization of portal venous branches occurred in two patients, which were not treated specifically. In comparison with the findings before the treatments, the serum alanine aminotransferase levels decreased at both 3 and 6 months after treatments (P less than 0.05); serum albumin levels increased at 6 months (P less than 0.05); the prothrombin times decreased at 6 months (P less than 0.05); but no significant changes were seen in the serum bilirubin levels. Fifteen patients were followed-up endoscopically for 3 months after the treatment. Gastric varices were completely resolved in 10 patients (66.7%) and were markedly smaller in 4 patients (26.6%). Worsening of the esophageal varices occurred in 3 patients (20%). All the patients were followed-up from 1 to 30 months [(16.7+/-8.8) months]. Rebleeding was observed in 4 patients, and the cumulative rebleeding rate at 1 year was 9.52%. CONCLUSION: Transportal variceal sclerotherapy with NBCA is a safe and effective method for treating gastric varices. Microcatheter technique and occlusion of the large gastrorenal shunt with a balloon-occluded catheter are necessary to ensure obliteration of gastric varices and prevent pulmonary embolism.

Tardus parvus waveforms in Doppler ultrasonography for hepatic artery stenosis after liver transplantation: can a new cut-off value guide the next step?

PURPOSE: Considering the high false-positive diagnosis of the tardus parvus waveform (TPW) in Doppler ultrasonography (DUS) for hepatic artery stenosis (HAS) after liver transplantation (LT), this study aimed to determine clinical features and new cut-off values to help guide treatment. MATERIALS AND METHODS: This retrospective study was approved by an Institutional Review Board. A total of 171 LT recipients were included and underwent DUS and either computed tomography angiography or digital subtraction angiography with an interval < 4 weeks at least 1 month post-LT. The DUS of 69 patients exhibited TPW [defined as resistive index (RI) < 0.5 and systolic acceleration time (SAT) > 0.08 s]. A multilevel likelihood ratio (LR) analysis was used to explore new cut-off values for DUS. In addition, abnormal liver function was considered additional evidence (defined as any liver enzyme > 3-fold of the upper limit of normal level or 2-fold increased). The results were stratified into three categories, category 1 (subjects with traditional TPW), category 2 (subjects with traditional TPW and abnormal liver function), and category 3 (subjects with traditional TPW and abnormal liver function, or with new cut-off values), and the diagnostic performance of each category was analyzed. RESULTS: The LR analysis revealed new cut-off values of RI < 0.4 (LR = 10.58) or SAT > 0.12 s (LR = 16.46). The false-positive rates for categories 2 and 3 were significantly lower (7.6% vs. 18.1%, P = 0.038; 1.9% vs. 18.1%, P < 0.001, respectively) than those for category 1, while the sensitivity for category 2 was significantly lower (41.8% vs. 74.6%, P < 0.001; 41.8% vs. 61.2%, P = 0.038, respectively) than that for categories 1 and 3. CONCLUSION: Using either (1) RI < 0.4 or SAT > 0.12 s, or (2) traditional TPW (RI < 0.5 and SAT > 0.08 s) in the presence of abnormal liver functions as the DUS criteria for HAS will significantly decrease the false-positive rate compared to traditional TPW without a significant increase in the false-negative rate.

Early prediction of survival in hepatocellular carcinoma patients treated with transarterial chemoembolization plus sorafenib.

AIM: To identify clinical biomarkers that could early predict improved survival in patients with advanced-stage hepatocellular carcinoma (HCC) treated with transarterial chemoembolization combined with sorafenib (TACE-S). METHODS: We retrospectively evaluated the medical records of consecutive patients with advanced-stage HCC who underwent TACE-S from January 2012 to December 2015. At the first follow-up 4-6 wk after TACE-S (median, 38 d; range, 33-45 d), patients exhibiting the modified Response Evaluation Criteria in Solid Tumors (mRECIST)-evaluated complete response, partial response, and stable disease were categorized as early disease control. At this time point, multiple variables were analyzed to identify the related factors affecting survival. RESULTS: Ninety-five patients were included in this study, and 60 of these patients achieved early disease control, with an overall disease control rate (DCR) of 63.2%. Patients who got sorafenib at the first TACE (no previous TACE) and patients without portal vein tumor thrombus (PVTT) had a higher DCR than those who underwent previous TACE before TACE-S (72.4% vs 48.6%, P = 0.019) and those with PVTT (75.5% vs 50.0%, P = 0.010). Early disease control after TACE-S, no previous TACE, and no PVTT were the independent prognostic factors for survival in the uni- and multivariate analyses. CONCLUSION: The first follow-up 4-6 wk after TACE-S can be used as the earliest time point to assess the response to TACE-S, and patients with mRECIST-evaluated early disease control, no previous TACE, and no PVTT had better survival.

Role of interventional therapy in hepatic artery stenosis and non-anastomosis bile duct stricture after orthotopic liver transplantation.

AIM: To analyze the clinical manifestations and the effectiveness of therapy in patients with orthotopic liver transplantation (OLT)-associated hepatic artery stenosis (HAS) and non-anastomosis bile duct stricture. METHODS: Nine cases were diagnosed as HAS and non-anastomosis bile duct stricture. Percutaneous transluminal angioplasty (PTA) was performed in four HAS cases, and expectant treatment in other five HAS cases; percutaneous transhepatic bile drainage, balloon dilation, stent placement were performed in all nine cases. RESULTS: Diffuse intra- and extra-bile duct stricture was observed in nine cases, which was associated with bile mud siltation and biliary infection. Obstruction of the bile duct was improved obviously or removed. Life span/follow-up period was 13-30 mo after PTA of four HAS cases, 6-23 mo without PTA of other five cases. CONCLUSION: Progressive, non-anastomosis, and diffuse bile duct stricture are the characteristic manifestations of HAS and non-anastomosis bile duct stricture after OLT. These are often associated with bile mud siltation, biliary infection, and ultimate liver failure. Interventional therapy is significantly beneficial.

Clinical significance of multislice spiral CT scans in hepatic veins occlusion in Budd-Chiari syndrome.

BACKGROUND: Budd-Chiari syndrome with hepatic vein occlusion (HVBCS) can induce severe portal hypertension and liver damage. We retrospectively analyzed hepatic CT features of HVBCS and evaluated the usefulness of triphasic enhancement of CT examinations and CT angiography (CTA) in its diagnosis. METHODS: Twenty-five cases with HVBCS, confirmed by digital subtraction angiography (DSA), received a triphasic enhancement CT scan within one week before DSA. The CTA images of the relevant blood vessels were reconstructed with maximum intensity projection, volume rendering and oblique reformat techniques. RESULTS: Compared with DSA, the detection rate of transverse CT and CTA images for abnormal hepatic vein were 81.7% (58/71) and 95.8% (68/71) (chi(2) = 7.044, P = 0.008), for membranous obstruction were 47.4% (9/19) and 84.2% (16/19) respectively (chi(2) = 5.729, P = 0.017), for segmental obstruction were 88.0% (22/25) and 100% (25/25) respectively (chi(2) = 1.418, P = 0.234). The detection rates for hepatic vein stenosis were 100% with each method. Diffuse hepatomegaly was found in all 6 cases in acute phase and 3 of 19 cases in chronic phase who had severe obstruction of three hepatic veins without patent intrahepatic collaterals. The other 16 cases in chronic phase had hepatatrophia to different extents related to the obstructed hepatic vein. All in acute phase and 15 in chronic phase presented typical patchy enhancement initially in caudate lobe and perihilar areas and enlarged with time delay. In all cases, parenchyma areas with atrophy, necrosis and congestion demonstrated lower and later enhancement. In all the parts, which had normal enhancement at least one patent outflow hepatic vein, accessory hepatic vein or collateral vessel was detected. CONCLUSION: Dynamic enhancement CT examination by multislice spiral CT not only could improve the diagnosis of HVBCS by CTA technique, but also could noninvasively provide anatomical information and reveal damage to the hepatic parenchyma.

[The role of multi-detector row CT in the diagnosis and hemodynamic studies of gastric varices in portal hypertension].

OBJECTIVE: To evaluate the value of multi-detector row CT (MDCT) in the diagnosis and hemodynamic studies of gastric varices (GV) in portal hypertension by comparison with endoscopy and DSA direct portography. METHODS: Thirty-six consecutive cirrhotic patients with GV confirmed by endoscopy underwent tri-phase contrast-enhanced CT scans and CT portography (CTP) within 2 weeks after endoscopy examination. Three independent experienced radiologists, who were blinded to the patients' clinical data, analyzed the CT images, including the size and location of GV as well as afferent and efferent veins of GV, separately. Interobserver agreement among the 3 radiologists with regard to the diagnosis of submucosal and perigastric GV was determined by Kappa (k) values. The findings of endoscopy were used as standards. RESULTS: Sub mucosal GV was diagnosed in 34 of the 36 patients (94.4%) and perigastric GV in all 36 patients (100%) by the observation of the 3 radiologists. MDCT showed an excellent interobserver reliability with regard to the diagnosis of submucosal GV (kappa = 0.85) and perigastric GV (kappa = 1.0). Agreement between MDCT and endoscopy with regard to the opacification of variceal size and location were 86.1% and 88.9% respectively. The sensitivity, specificity, accuracy, and positive predictive value of CTP in the opacification of afferent and efferent veins of GV were all more than 80%. The frequencies of participation of posterior gastric vein and short gastric vein in blood supply to gastric fundal varices in the isolated gastric varices and gastroesophageal varices type 2 (GEV2) were 94.1% and 70.6% respectively, both significantly higher than those in the gastroesophageal varices type 1 (GEV1, 52.6% and 31.6%, respectively, both P < 0.05). The main blood drainage route of GEV1 was via azygous system into the super vena cava (100%), whereas in the gastric fundal varices the main blood drainage route was via the gastrorenal shunts into the inferior vena cava (82.4%). CONCLUSION: MDCT can be used as an important tool for detecting submucosal and perigastric GV, and can clearly reveal the size, location, and hemodynamics of GV.