Modified Hepatic Venous Plane: A Key Factor for Improving Preoperative MDCT Donor Volume Prediction in Living-Donor Liver Transplantation.

OBJECTIVE: The aim of this work was to present our experience using a modified hepatic venous plane in multidetector computerized tomography (MDCT) for reducing the discrepancy between preoperative liver volume estimation and intraoperative weight (IOW) measurement in living-donor liver transplantation (LDLT). METHODS: We retrospectively reviewed the medical records of 57 consecutive living donors with the use of MDCT as a modality for volumetric assessment for LDLT from May 2007 to January 2015. We divided living donors into 2 groups according to surgical methods: right hepatectomy (RH) and left hepatectomy (LH). Initial liver volumetric measurement (group I) was assessed. After discussions with radiologist, the transplantation surgeon used a modified hepatic venous plane for surgical significant middle hepatic venous variants (>5 mm) in 16 living donors and applied the initial surgical plane in the remaining for the modified donor liver volumetric measurement (group II). We then compared the correlations of these 2 groups with the use of IOW. RESULTS: The overall correlation (r) between group I and IOW was 0.947. The correlations (r) between group I and IOW were 0.872 and 0.955 for RH and LH, respectively. Compared with group I, group II showed better correlation with IOW: r = 0.949 and 0.981 for RH and LH, respectively. The overall correlation (r) between group II and IOW was 0.980, and the error ratio was 5.95 ± 5.05%. CONCLUSIONS: Our study showed that using a modified hepatic venous plane in preoperative MDCT, after good communication between transplant surgeon and radiologist, can provide more accurate liver volume estimation and achieve a better correlation with IOW in LDLT.

Long-Term Outcome of Liver Transplant Recipients After the Development of Renal Failure Requiring Dialysis: A Study Using the National Health Insurance Database in Taiwan.

BACKGROUND: The aims of this study were to identify the incidence of renal failure requiring dialysis and to investigate the long-term outcome after renal failure in liver transplantation (LT) patients. METHODS: The primary database used was the Taiwan National Health Insurance Research Database. Subjects with LT from 1997 to 2009 were included. Patients were grouped into the dialysis cohort if they once received hemodialysis owing to any pattern of renal failure during peri-transplantation periods or after LT. Otherwise, they were categorized into the nondialysis cohort. We conducted a retrospective observational study on the correlation of renal failure requiring dialysis and its effect on LT recipients. RESULTS: The analysis included data of 1,771 LT recipients with a mean follow-up time of 3.8 ± 2.9 years. The mean age was 43.2 ± 19.3 years, and 69.4% were male. Overall patient survival was 86.2% at 1 year, 82.2% at 3 years, and 80.5% at 5 years. Renal failure requiring dialysis had developed in the 323 patients (18.2%). Among them, 26 individuals (1.5%) had progressed to end-stage renal disease without renal recovery after perioperative hemodialysis. Individuals who developed renal failure requiring dialysis had a higher mortality compared with LT recipients never requiring dialysis (hazard ratio, 8.75; 95% confidence interval, 7.0-10.9). CONCLUSIONS: Renal failure requiring dialysis development after LT is common and carries high mortality in Chinese liver allograft recipients. Recognizing risk factors permits the timely institution of proper treatment, which is the key to reducing untoward outcomes.

Folate and vitamin B-6 status are not associated with homocysteine, oxidative stress and antioxidant capacities in patients with hepatocellular carcinoma.

The purpose of this study was to study the effects of serum folate and plasma pyridoxal 5'-phosphate (PLP) on plasma homocysteine, oxidative stress and antioxidant capacities in 44 hepatocellular carcinoma (HCC) patients and 56 healthy controls. The responses of folate, vitamin B-6, homocysteine, oxidative stress and antioxidant enzyme activities in HCC patients before and after tumor resection were also determined. Patients with HCC before tumor resection had significantly lower folate, PLP, homocysteine, glutathione peroxidase and superoxide dismutase levels, but higher malondialdehyde, total antioxidant capacity and glutathione S-transferase activity when compared with healthy controls. Oxidative stress was significantly decreased to a level similar to that of healthy controls after tumor resection in the HCC group. There were no associations of folate and PLP with plasma homocysteine, indicators of oxidative stress and antioxidant capacities. Serum folate and plasma PLP were not significant factors affecting plasma homocysteine, oxidative stress and antioxidant capacities in patients with HCC.

The G-protein coupled estrogen receptor, GPER: The inside and inside-out story.

GPER possesses structural and functional characteristics shared by members of the G-protein-coupled receptor (GPCR) superfamily, the largest class of plasma membrane receptors. This newly appreciated estrogen receptor is localized predominately within intracellular membranes in most, but not all, cell types and its surface expression is modulated by steroid hormones and during tissue injury. An intracellular staining pattern is not unique among GPCRs, which employ a diverse array of molecular mechanisms that restrict cell surface expression and effectively regulating receptor binding and activation. The finding that GPER displays an intracellular predisposition has created some confusion as the estrogen-inducible transcription factors, ERα and ERß, also reside intracellularly, and has led to complex suggestions of receptor interaction. GPER undergoes constitutive retrograde trafficking from the plasma membrane to the endoplasmic reticulum and recent studies indicate its interaction with PDZ binding proteins that sort transmembrane receptors to synaptosomes and endosomes. Genetic targeting and selective ligand approaches as well as cell models that express GPER in the absence of ERs clearly supports GPER as a bonafide "stand alone" receptor. Here, the molecular details that regulate GPER action, its cell biological activities and its implicated roles in physiological and pathological processes are reviewed.

A nationwide population-based study of the risk of tuberculosis in different solid organ transplantations in Taiwan.

Advances in immunosuppressants for solid organ transplantation (SOT) have improved prevention and treatment of acute rejection as well as reduced the risk of chronic graft damage. However, SOT recipients are prone to developing opportunistic infections because of their long-term immunosuppressed status. Tuberculosis (TB) is a serious opportunistic infection that is associated with increased morbidity and mortality in SOT recipients. However, nationwide population-based research specifically focused on the associations between kidney transplantation (KTx), liver transplantation (LTx), and heart transplantation (HTx), and subsequent TB infection is lacking. This study was conducted using Taiwan's National Health Insurance Research Database, which provided claims data for SOT recipients from 2000 to 2009. Clinical features, treatment, and outcomes were analyzed to determine the risk for TB after SOT. In total, 153 (3.2%) RTx, 19 (1.1%) LTx, and 26 (2.8%) HTx recipients became infected with TB. Compared with non-TB patients, HTx recipients with TB had significantly higher prevalence of older age (P = .037), hypertension (P < .001), and coronary artery disease (CAD) (P = .002). There were also greater percentages of male sex (P = .018), diabetes (P = .029), hyperlipidemia (P = .016), CAD (P < .001), and chronic obstructive pulmonary disease (COPD) (P < .001) in RTx recipients with TB than in those without. In conclusion, posttransplantation TB is a serious problem worldwide, and a high index of suspicion is warranted to ensure early diagnosis and prompt initiation of treatment for TB among SOT patients. In this preliminary study, KTx recipients had a higher risk of TB infection than LTx and HTx recipients, and the high-risk factors were male sex, diabetes, hyperlipidemia, CAD, and COPD. The use of optimal immunosuppressive agents to minimize acute rejection, monitoring of high-risk recipients, prompt diagnosis, and appropriate treatment are required for the management of TB infection in endemic areas such as Taiwan.

Cytomegalovirus disease after liver transplantation: a nationwide population-based study.

BACKGROUND: Cytomegalovirus (CMV) disease is a significant complication after liver transplantation. The estimated incidence varies among studies, which have been conducted in single regional centers and with small cohorts. In this study, we investigated the occurrence of CMV disease among liver transplant recipients in a national cohort in Taiwan. METHODS: This retrospective study used data from the Taiwan National Health Insurance Research Database. All liver transplant recipients in the catastrophic illness database from 2000 to 2009 were enrolled. Cases of CMV disease were identified from the admission database with the use of the ICD-9-CM code 078. RESULTS: The national cohort consisted of 1,721 liver transplant recipients (1,200 men and 521 women) with a mean age of 43.9 ± 8.9 years at the time of transplantation. The mean follow-up duration was 3.6 ± 2.7 years. The mortality rate was 14.9% at 1 year and 20.5% at 5 years. During the study period, 84 patients (4.9%) were diagnosed with CMV disease. The overall prevalence of CMV disease was 14.5 per 100 person-years. The cumulative incidences of post-transplantation CMV infection at 3 months, 6 months, 1 year, 2 years, 5 years, and 10 years were 1.2%, 2.7%, 3.8%, 4.2%, 4.8%, and 4.9%, respectively. The most common CMV-related diseases were colitis, hepatitis, and pneumonia. CONCLUSIONS: The risk of CMV disease was significantly elevated in the first 6 months after liver transplantation in the Taiwanese cohort.

Second and third hepatectomies for recurrent hepatocellular carcinoma are justified.

BACKGROUND: Liver resection is the main curative treatment for hepatocellular carcinoma (HCC), but recurrence rates are high. The remnant liver is the most common site of recurrence, but the role of repeat hepatectomy in the treatment of recurrent HCC is controversial. METHODS: Patients who underwent curative hepatectomy for HCC and subsequent repeat hepatectomy for recurrent HCC between 1990 and 2007 were reviewed retrospectively. Clinicopathological characteristics, and early- and long-term outcomes of patients who had a first, second, third and fourth hepatectomy were compared. RESULTS: Some 1177 patients underwent a first hepatectomy for HCC, and 149, 35 and eight patients respectively had a second, third and fourth hepatectomies for recurrence. There were no significant differences in early postoperative outcomes after first and repeat hepatectomies. Five-year disease-free and overall survival rates after first, second and third hepatectomies were 43.6, 31.8 and 33.8 per cent (P = 0.772), and 52.4, 56.4 and 59.4 per cent (P = 0.879), respectively. Patients undergoing second and third hepatectomies for recurrence had better survival rates than those who did not have a repeat hepatectomy, but not those after fourth hepatectomy. CONCLUSION: Second and third hepatectomies seem justified for hepatic recurrence of HCC. The role of fourth hepatectomy needs further investigation.

Liver resection for hepatocellular carcinoma in patients with cirrhosis.

BACKGROUND: Although liver resection is now a safe procedure, its role for hepatocellular carcinoma (HCC) in patients with cirrhosis remains controversial. METHODS: This study compared the results of liver resection for HCC in patients with cirrhosis over two time intervals. One hundred and sixty-one patients had resection during period 1 (1991-1996) and 265 in period 2 (1997-2002). Early and long-term results after liver resection in the two periods were compared, and clinicopathological characteristics that influenced survival were identified. RESULTS: Tumour size was smaller, indocyanine green retention rate was higher, patients were older and a greater proportion of patients were asymptomatic in period 2 than period 1. Operative blood loss, need for blood transfusion, operative mortality rate, postoperative hospital stay and total hospital costs were significantly reduced in period 2. The 5-year disease-free survival rates were 28.2 and 33.9 per cent in periods 1 and 2 respectively (P = 0.042), and 5-year overall survival rates were 45.9 and 61.2 per cent (P < 0.001). Multivariate analysis identified serum alpha-fetoprotein level, need for blood transfusion and Union Internacional Contra la Cancrum tumour node metastasis stage as independent determinants of disease-free and overall survival. CONCLUSION: The results of liver resection for HCC in patients with cirrhosis improved over time. Liver resection remains a good treatment option in selected patients with HCC arising from a cirrhotic liver.

Liver resection for hepatocellular carcinoma in patients with end-stage renal failure.

BACKGROUND AND OBJECTIVES: Surgical resection remains the main option for curing hepatocellular carcinoma (HCC). However, liver resection in patients with end-stage renal disease (ESRD) is risky. The aim of this study is to clarify the role of liver resection for treating HCC in patients with ESRD. METHODS: A retrospective review was carried out on 468 patients who underwent liver resection for HCC between 1989 and 1999. The clinicopathological characteristics and operative results of 12 patients who had ESRD (ESRD group) were compared with those of the other 456 patients who did not have ESRD (non-ESRD group). In the ESRD group, heparin-free hemodialysis using the periodic saline-rinse method was performed during the perioperative period. RESULTS: The ESRD group had lower hemoglobin and a higher serum creatinine levels. Other patient background and tumor pathological characteristics were comparable between the two groups as well. The operative morbidity and mortality between the two groups were also similar. The 5-year disease-free survival rates for ESRD and non-ESRD groups were 35.0 and 34.2% (P = 0.31), respectively, while the 5-year actuarial survival rates were 67.8 and 53.3% (P = 0.54), respectively. CONCLUSION: With improving techniques and knowledge of dialysis, liver resection for HCC is justified in selected patients with ESRD.

The effect of arsenic trioxide on the expression of Hsc and HNF4 in nuclear matrix proteins in HepG2 cells.

Anticancer agents interfere with the proliferation and survival of tumor cells by a variety of mechanisms. An important factor in the development of a cytotoxic effect by certain anticancer agents is the localization of drug-induced lesions within the cell nucleus. Drug-target interactions at the level of nuclear matrix (NM) may be critical events in the induction of cell death by some of these agents. Arsenic trioxide (As2O3) was identified as a very potent anti-leukemic agent by inducing apoptosis. The present study shows that As2O3 significantly inhibits the growth of hepatoblastoma cell line, HepG2, changes the composition of nuclear matrix proteins and reduces the expression of Hsc 70 and HNF4 in HepG2, which in turn initiate a cascade of events that compromise multiple nuclear functions and, ultimately, cell survival.

Presence of neuronal nitric oxide synthase in autonomic and sensory ganglion neurons innervating the lacrimal glands of the cat: an immunofluorescent and retrograde tracer double-labeling study.

It is generally considered that parasympathetic postganglionic nerve fibers innervating the lacrimal gland (LG) arise from the pterygopalatine ganglion (PPG), while sympathetic and sensory innervations arise from the superior cervical ganglion (SCG) and trigeminal ganglion (TG), respectively. Recently, we reported for the first time that the parasympathetic innervation of the cat LG was also provided by the otic ganglion (OG) and ciliary ganglion (CG), and that the sensory innervation was also provided by the superior vagal ganglion (SVG) and superior glossopharyngeal ganglion (SGG). To determine if nitric oxide (NO) is a neurotransmitter of the autonomic and sensory neurons innervating the LG, we injected the cholera toxin B subunit (CTB) as a retrograde tracer into the cat LG, and used double-labeling fluorescent immunohistochemistry for CTB and nitric oxide synthase (NOS). We found that NOS-/CTB-immunofluorescent double-labeled perikarya were localized in the PPG, OG, TG, SVG and SGG, but not in the CG and SCG. The highest numbers of NOS-/CTB-immunofluorescent double-labeled neurons were found in the PPG and TG. In addition, we examined the presence of nitrergic nerve fibers in the LG using NADPH-d histochemistry and found that a large amount of NADPH-d-stained nerve fibers were distributed around the glandular acini and in the walls of glandular ducts and blood vessels. This study provides the first direct evidence showing that NO may act as a neurotransmitter or modulator involved in the parasympathetic and sensory regulation of lacrimal secretion and blood circulation, but may not be implicated in the sympathetic control of LG activities, and that nitrergic nerve fibers in the LG arise mainly from parasympathetic postganglionic neurons in the PPG and sensory neurons in the TG. The present results suggest that NO plays an important role in the regulation of LG activities.

Involvement of pressure-related mechanism in activation of inducible nitric oxide synthase.

The goal of this study was to clarify the relationship between blood pressure and inducible nitric oxide synthase (iNOS) activity. Different levels of blood pressure were obtained by long-term (six days) intravenous infusion of different concentrations (0.3%-8%) of NaCl solution to normal SD rats. iNOS activity assay and measurement of urinary nitrate/nitrite (UNOx), an index of NO production of the whole body, were carried out by isotope-labeled L-arginine conversion rate measurement and Greiss Reaction respectively. Groups of normotensive and hypertensive rats including normal Wistar rats, normal Sprague-Dawley (SD) rats, high NaCl-induced hypertensive rats (NaHR) and spontaneously hypertensive rats (SHR) were used to detect the changes in iNOS protein under normotension and hypertension by Western blotting. iNOS activity of aorta and kidney tissues and UNOx increased more significantly in hypertensive animals than in the normotensive control ones. Accordingly, iNOS protein in the aortas of NaHR and SHR increased by 149% and 261% respectively. It is suggested that in addition to cytokine and bacterial products etc, blood pressure is also an effective regulatory factor involved in iNOS activation and expression.

Expression of laminin in metastatic melanoma cell lines with different metastatic potential.

Cancer metastasis is a complex multi-step process in which tumor cells leave the primary site and develop a secondary tumor in distant organs. Laminin plays an important role in this process. The expression of laminin in four melanoma cell lines with different metastatic potentials was investigated by immunohistochemistry, immunogold electron microscopy and Western blotting. Our results showed that the expression of endogenous laminin and the percentage of the positive cells are higher with increased metastatic potentials. It is, thus, suggested that endogenous laminin may contribute to the different metastatic properties in the melanoma cell line.

Alteration of nuclear matrix protein composition of neuroblastoma cells after arsenic trioxide treatment.

The aims of the present study were to assess the effects of arsenic trioxide on the nuclear matrix protein profiles of mouse neuroblastoma cells. Arsenic trioxide induces apoptosis of acute promyelocytic leukemia cells. Our results demonstrated that 2 microM As2O3 could significantly inhibit the growth of Neuro-2a cells. As early as 24 hours after As2O3 treatment, we began to observe the alteration of nuclear matrix proteins and apoptosis in tumor cells by TUNEL assay but not by DNA ladder. An increase expression of Hsc in nuclear matrix proteins of 2 microM As2O3 treated cells was also noted. Our results also showed that before a mass range of apoptosis occurred, the composition of nuclear matrix proteins had altered. Hence the alteration of nuclear matrix proteins, such as increased expression of Hsc, may be a sensitive indicator for the detection of early apoptosis.

Development of a multichannel microfluidic analysis system employing affinity capillary electrophoresis for immunoassay.

A six-channel microfluidic immunoassay device with a scanned fluorescence detection system is described. Six independent mixing, reaction, and separation manifolds are integrated within one microfluidic wafer, along with two optical alignment channels. The manifolds are operated simultaneously and data are acquired using a singlepoint fluorescence detector with a galvano-scanner to step between separation channels. A detection limit of 30 pM was obtained for fluorescein with the scanning detector, using a 7.1-Hz sampling rate for each of the reaction manifolds and alignment channels (57-Hz overall sampling rate). Simultaneous direct immunoassays for ovalbumin and for anti-estradiol were performed within the microfluidic device. Mixing, reaction, and separation could be performed within 60 s in all cases and within 30 s under optimized conditions. Simultaneous calibration and analysis could be performed with calibrant in several manifolds and sample in the other manifolds, allowing a complete immunoassay to be run within 30 s. Careful chip conditioning with methanol, water, and 0.1 M NaOH resulted in peak height RSD values of 3-8% (N = 5 or 6), allowing for cross-channel calibration. The limit of detection (LOD) for an anti-estradial assay obtained in any single channel was 4.3 nM. The LOD for the cross-channel calibration was 6.4 nM. Factors influencing chip and detection system design and performance are discussed in detail.

Human papillomavirus 16 E6 is associated with the nuclear matrix of esophageal carcinoma cells.

AIM: To explore the etiologic role of HPV infection in esophageal carcinoma, and the association of HPV-16 E6 with the nuclear matrix of carcinoma cells. METHODS: Two esophageal carcinoma cell lines,EC/CUHK1 and EC/CUHK2, were tested for HPV-16 E6 subgenetic fragment by polymerase chain reaction amplification of virus DNA associated nuclear matrix. RT-PCR and immunocytochemistry were also used to visualize the expression of E6 subgene in the cells. RESULTS: The HPV-16 E6 subgenetic fragment was found to be present in nuclear matrix-associated DNA, E6 oncoprotein localized in the nucleus where it is tightly associated with nuclear matrix after sequential extraction in EC/CUHK2 cells. It was not detected, however, in EC/CUHK1 cells. CONCLUSION: The interaction between HPV-16 E6 and nuclear matrix may contribute to the virus induced carcinogenesis in esophageal carcinoma.

The measurement of AgNORs in human glioma cells.

Sixty gliomas obtained by neurosurgical resections were examined. Paraffin blocks were retrieved from pathological files of the Second Affiliated Hospital in Guangzhou Medical College. The methods of argyrophilic technique for AgNORs staining, and Image Analysis System for measurement of AgNORs were used. Six parameters, which included hcount, count, narea, agnrea, agpern and agperc were used to correlated well with histopathological grades (compared grade 2 & 3, grade 3 & 4, and grade 2 & 4, respectively). We concluded that AgNORs is useful in evaluating proliferative activity and assessing the malignancy of human gliomas. It may also be used as a target for anti-neoplastic drugs in the treatment of gliomas.

Three novel neural pathways to the lacrimal glands of the cat: an investigation with cholera toxin B subunit as a retrograde tracer.

The distribution of ganglion neurons innervating the lacrimal gland (LG) was investigated following injection of cholera toxin B subunit into the LG of the cat. We report the first evidence that the otic ganglion (OG), and superior vagal and glossopharyngeal ganglia are also the sources of innervation of the LG. LG-innervating neurons in the pterygopalatine ganglion and the OG could be divided into two subpopulations: small and large neurons. They may mediate the vasodilatation and secretion, respectively.

Morphological distinction between vasodilator and secretomotor neurons in the pterygopalatine ganglion of the cat.

We investigated whether vasodilator and secretomotor ganglion neurons are morphologically distinguishable from each other in the parasympathetic ganglion of the cat. When Cholera toxin B subunit, a retrograde tracer, was injected into the palatine gland, both large and small ganglion neurons were retrogradely labeled in the pterygopalatine ganglion. On the other hand, when the tracer was injected into gland-free areas (the upper gingiva or epidural space), all neurons labeled in the ganglion were small in size. Thus, it was assumed that small and large neurons labeled in the ganglion represented, respectively, vasomotor and secretomotor and neurons [corrected].

Evidence for the collateral innervation of the esophagus and the heart from neurons in the compact formation of the nucleus ambiguus of the rat.

We investigated whether the heart receives collateral projections from the neurons which innervate the esophagus with a retrograde double-labeling method using two fluorescent tracers. Following injections of True Blue (TB) into the esophagus and Diamidino Yellow (DY) into the heart, about 21.9% of the labeled esophageal motoneurons in the compact formation of the nucleus ambiguus (AmC) were retrogradely double labeled. No single-labeled cardiac motoneurons were found in the AmC. The present results provide anatomical substrates for the esophagocardiac reflex.