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1.  The Arp2/3 complex has essential roles in vesicle trafficking and transcytosis in the mammalian small intestine 
Molecular Biology of the Cell  2015;26(11):1995-2004.
The Arp2/3 complex has essential functions in the intestinal epithelium. Loss of ArpC3 results in vesicle-trafficking defects that prevent transcytosis of immunoglobulins and efficient absorption of lipids but does not affect levels of cortical F-actin.
The Arp2/3 complex is the only known nucleator of branched F-actin filaments. Work in cultured cells has established a wide array of functions for this complex in controlling cell migration, shape, and adhesion. However, loss of Arp2/3 complex function in tissues has yielded cell type–specific phenotypes. Here we report essential functions of the Arp2/3 complex in the intestinal epithelium. The Arp2/3 complex was dispensable for intestinal development, generation of cortical F-actin, and cell polarity. However, it played essential roles in vesicle trafficking. We found that in the absence of ArpC3, enterocytes had defects in the organization of the endolysosomal system. These defects were physiologically relevant, as transcytosis of IgG was disrupted, lipid absorption was perturbed, and neonatal mice died within days of birth. These data highlight the important roles of the Arp2/3 complex in vesicle trafficking in enterocytes and suggest that defects in cytoplasmic F-actin assembly by the Arp2/3 complex, rather than cortical pools, underlie many of the phenotypes seen in the mutant small intestine.
PMCID: PMC4472011  PMID: 25833710
2.  Biocompatibility Assessment of a Long-Term Wearable Artificial Pump-Lung in Sheep 
Artificial organs  2013;37(8):678-688.
The purpose of this study was to assess the biocompatibility of a newly developed long-term wearable artificial pump-lung (APL) in a clinically relevant ovine animal mode. The wearable APL device was implanted in five sheep through a left thoracotomy. The device was connected between the right atrium (RA) and pulmonary artery (PA) and evaluated for 30 days. Three sheep were used as the sham control. Platelet activation was assessed by measuring platelet surface P-selectin (CD62P) expression with flow cytometry and plasma soluble P-selectin with an enzyme-linked immunosorbent assay (ELISA). Thrombotic deposition on the device components and hollow fiber membranes (HFM) were analyzed with digital imaging and scanning electron microscopy (SEM). Surface P-selectin of the APL and sham groups changed significantly over the study period, but without significant differences between the two groups. Soluble P-selectin for the two groups peaked in the first 24 hours after the surgery. Soluble P-selectin of the APL group remained slightly elevated over the study period compared to the pre-surgical baseline value and was slightly higher compared to that of the sham group. Plasma free hemoglobin (PFH) remained in the normal ranges in all the animals. In spite of the surgery related alteration in laboratory tests and elevation of platelet activation status, the APL devices in all the animals functioned normally (oxygen transfer and blood pumping) during the 30 day study period. The device flow path and membrane surface were free of gross thrombus. Electron microscopy images showed only scattered thrombi on the fibers (membrane surface and weft). In summary, the APL exhibited excellent biocompatibility. Two forms of platelet activation, surgery related and device induced, in the animals implanted with the wearable APL were observed. The limited device-induced platelet activation did not cause gross thrombosis and impair the long-term device performance.
PMCID: PMC3675162  PMID: 23452221
Biocompatibility; Platelet activation; Artificial lungs; Integrated Pump-Oxygenators; Extracorporeal membrane oxygenation
3.  Cell-Cell Adhesions and Cell Contractility Are Upregulated upon Desmosome Disruption 
PLoS ONE  2014;9(7):e101824.
Desmosomes are perturbed in a number of disease states – including genetic disorders, autoimmune and bacterial diseases. Here, we report unexpected changes in other cell-cell adhesion structures upon loss of desmosome function. We found that perturbation of desmosomes by either loss of the core desmosomal protein desmoplakin or treatment with pathogenic anti-desmoglein 3 (Dsg3) antibodies resulted in changes in adherens junctions consistent with increased tension. The total amount of myosin IIA was increased in desmoplakin-null epidermis, and myosin IIA became highly localized to cell contacts in both desmoplakin-null and anti-Dsg3-treated mouse keratinocytes. Inhibition of myosin II activity reversed the changes to adherens junctions seen upon desmosome disruption. The increased cortical myosin IIA promoted epithelial sheet fragility, as myosin IIA-null cells were less susceptible to disruption by anti-Dsg3 antibodies. In addition to the changes in adherens junctions, we found a significant increase in the expression of a number of claudin genes, which encode for transmembrane components of the tight junction that provide barrier function. These data demonstrate that desmosome disruption results in extensive transcriptional and posttranslational changes that alter the activity of other cell adhesion structures.
PMCID: PMC4090201  PMID: 25006807
4.  Mesenchymal Stem Cells Induce Granulocytic Differentiation of Acute Promyelocytic Leukemic Cells via IL-6 and MEK/ERK Pathways 
Stem Cells and Development  2013;22(13):1955-1967.
All-trans retinoic acid (ATRA) induces clinical remission in most acute promyelocytic leukemia (APL) patients by inducing terminal differentiation of APL cells toward mature granulocytes. Here we report that human umbilical cord-derived mesenchymal stem cells (UC-MSCs) are capable of inducing granulocytic differentiation of the APL-derived NB4 cell line as well as primary APL cells and also cooperate with ATRA in an additive manner. Transwell coculture experiments revealed that UC-MSCs' differentiation-inducing effect was mediated through some soluble factors. Differentiation attenuation by IL-6Ra neutralization and induction by addition of exogenous IL-6 confirmed that IL-6 secreted by UC-MSCs was at least partially responsible for this differentiation induction process. Moreover, we found that UC-MSCs activated the MEK/ERK signaling pathway in promyelocytic cells and pharmacological inhibition of the MEK/ERK pathway reversed UC-MSC-induced differentiation, indicating that UC-MSCs exerted effect through activation of the MEK/ERK signaling pathway. These results demonstrate for the first time a stimulatory effect of MSCs on the differentiation of APL cells and bring a new insight into the interaction between MSCs and leukemic cells. Our data suggest that UC-MSCs/ATRA combination could be used as a novel therapeutic strategy for APL patients.
PMCID: PMC3685326  PMID: 23391335
5.  Statistical Experimental Design Guided Optimization of a One-Pot Biphasic Multienzyme Total Synthesis of Amorpha-4,11-diene 
PLoS ONE  2013;8(11):e79650.
In vitro synthesis of chemicals and pharmaceuticals using enzymes is of considerable interest as these biocatalysts facilitate a wide variety of reactions under mild conditions with excellent regio-, chemo- and stereoselectivities. A significant challenge in a multi-enzymatic reaction is the need to optimize the various steps involved simultaneously so as to obtain high-yield of a product. In this study, statistical experimental design was used to guide the optimization of a total synthesis of amorpha-4,11-diene (AD) using multienzymes in the mevalonate pathway. A combinatorial approach guided by Taguchi orthogonal array design identified the local optimum enzymatic activity ratio for Erg12:Erg8:Erg19:Idi:IspA to be 100∶100∶1∶25∶5, with a constant concentration of amorpha-4,11-diene synthase (Ads, 100 mg/L). The model also identified an unexpected inhibitory effect of farnesyl pyrophosphate synthase (IspA), where the activity was negatively correlated with AD yield. This was due to the precipitation of farnesyl pyrophosphate (FPP), the product of IspA. Response surface methodology was then used to optimize IspA and Ads activities simultaneously so as to minimize the accumulation of FPP and the result showed that Ads to be a critical factor. By increasing the concentration of Ads, a complete conversion (∼100%) of mevalonic acid (MVA) to AD was achieved. Monovalent ions and pH were effective means of enhancing the specific Ads activity and specific AD yield significantly. The results from this study represent the first in vitro reconstitution of the mevalonate pathway for the production of an isoprenoid and the approaches developed herein may be used to produce other isopentenyl pyrophosphate (IPP)/dimethylallyl pyrophosphate (DMAPP) based products.
PMCID: PMC3835790  PMID: 24278153
6.  Combinatorial Engineering of 1-Deoxy-D-Xylulose 5-Phosphate Pathway Using Cross-Lapping In Vitro Assembly (CLIVA) Method 
PLoS ONE  2013;8(11):e79557.
The ability to assemble multiple fragments of DNA into a plasmid in a single step is invaluable to studies in metabolic engineering and synthetic biology. Using phosphorothioate chemistry for high efficiency and site specific cleavage of sequences, a novel ligase independent cloning method (cross-lapping in vitro assembly, CLIVA) was systematically and rationally optimized in E. coli. A series of 16 constructs combinatorially expressing genes encoding enzymes in the 1-deoxy-D-xylulose 5-phosphate (DXP) pathway were assembled using multiple DNA modules. A plasmid (21.6 kb) containing 16 pathway genes, was successfully assembled from 7 modules with high efficiency (2.0 x 103 cfu/ µg input DNA) within 2 days. Overexpressions of these constructs revealed the unanticipated inhibitory effects of certain combinations of genes on the production of amorphadiene. Interestingly, the inhibitory effects were correlated to the increase in the accumulation of intracellular methylerythritol cyclodiphosphate (MEC), an intermediate metabolite in the DXP pathway. The overexpression of the iron sulfur cluster operon was found to modestly increase the production of amorphadiene. This study demonstrated the utility of CLIVA in the assembly of multiple fragments of DNA into a plasmid which enabled the rapid exploration of biological pathways.
PMCID: PMC3818232  PMID: 24223968
7.  Combining Genotype Improvement and Statistical Media Optimization for Isoprenoid Production in E. coli 
PLoS ONE  2013;8(10):e75164.
Isoprenoids are a large and diverse class of compounds that includes many high value natural products and are thus in great demand. To meet the increasing demand for isoprenoid compounds, metabolic engineering of microbes has been used to produce isoprenoids in an economical and sustainable manner. To achieve high isoprenoid yields using this technology, the availability of metabolic precursors feeding the deoxyxylulose phosphate (DXP) pathway, responsible for isoprenoid biosynthesis, has to be optimized. In this study, phosphoenolpyruvate, a vital DXP pathway precursor, was enriched by deleting the genes encoding the carbohydrate phosphotransferase system (PTS) in E. coli. Production of lycopene (a C40 isoprenoid) was maximized by optimizing growth medium and culture conditions. In optimized conditions, the lycopene yield from PTS mutant was seven fold higher than that obtained from the wild type strain. This resulted in the highest reported specific yield of lycopene produced from the DXP pathway in E. coli to date (20,000 µg/g dry cell weight). Both the copy number of the plasmid encoding the lycopene biosynthetic genes and the expression were found to be increased in the optimized media. Deletion of PTS together with a similar optimization strategy was also successful in enhancing the production of amorpha-1,4-diene, a distinct C15 isoprenoid, suggesting that the approaches developed herein can be generally applied to optimize production of other isoprenoids.
PMCID: PMC3790805  PMID: 24124471
8.  Draft Genome Sequence of the Sponge-Associated Strain Bacillus atrophaeus C89, a Potential Producer of Marine Drugs 
Journal of Bacteriology  2012;194(16):4454.
Bacillus atrophaeus C89, isolated from the marine sponge Dysidea avara, is a potential producer of bioactive compounds, such as neobacillamide A and bacillamide C. Here, we present a 4.2-Mb assembly of its genome. The nonribosomal peptide synthetases (NRPSs) make it possible to produce the bioactive compounds.
PMCID: PMC3416270  PMID: 22843588
9.  30-Day In-vivo Performance of a Wearable Artificial Pump-Lung for Ambulatory Respiratory Support 
The Annals of thoracic surgery  2011;93(1):274-281.
The purpose of this study was to evaluate the long-term in-vivo hemodynamics, gas transfer and biocompatibility of an integrated artificial pump-lung (APL) developed for ambulatory respiratory support.
The study was conducted in an ovine model by surgically placing the APL between the right atrium and pulmonary artery. Nine sheep were implanted. Heparin was infused as an anticoagulant. The device flow, gas transfer and plasma free hemoglobin (PFH) were measured daily. Hematological data, platelet activation and blood biochemistry were assessed twice a week. After 30 days, the sheep were euthanized for necropsy. The explanted devices were examined for gross thrombosis.
Five sheep survived for 29 to 31 days and were electively terminated. Four sheep expired or were terminated early due to mechanical failure of IV lines or device. The APL devices in the five long-term animals were capable of delivering an oxygen transfer rate of 148±18 ml/min at a flow rate of 2.99±0.46 l/min with blood oxygen saturation of 96.7±1.3%. The device flow and oxygen transfer were stable over 30 days. The animals had normal end-organ functions except for surgery-related transient alteration in kidney function, liver function, and cell and tissue injury. There was no hemolysis. The device flow path and membrane surface were free of gross thrombus.
The APL exhibited the capability of providing respiratory support with excellent biocompatibility, long-term reliability and the potential for bridging to lung transplant.
PMCID: PMC3244576  PMID: 22115337
Artificial organs; Device; Extracorporeal membrane oxygenation, ECMO; Lung; Transplantation, Lung
10.  Enhancing solubility of deoxyxylulose phosphate pathway enzymes for microbial isoprenoid production 
Recombinant proteins are routinely overexpressed in metabolic engineering. It is well known that some over-expressed heterologous recombinant enzymes are insoluble with little or no enzymatic activity. This study examined the solubility of over-expressed homologous enzymes of the deoxyxylulose phosphate pathway (DXP) and the impact of inclusion body formation on metabolic engineering of microbes.
Four enzymes of this pathway (DXS, ISPG, ISPH and ISPA), but not all, were highly insoluble, regardless of the expression systems used. Insoluble dxs (the committed enzyme of DXP pathway) was found to be inactive. Expressions of fusion tags did not significantly improve the solubility of dxs. However, hypertonic media containing sorbitol, an osmolyte, successfully doubled the solubility of dxs, with the concomitant improvement in microbial production of the metabolite, DXP. Similarly, sorbitol significantly improved the production of soluble and functional ERG12, the committed enzyme in the mevalonate pathway.
This study demonstrated the unanticipated findings that some over-expressed homologous enzymes of the DXP pathway were highly insoluble, forming inclusion bodies, which affected metabolite formation. Sorbitol was found to increase both the solubility and function of some of these over-expressed enzymes, a strategy to increase the production of secondary metabolites.
PMCID: PMC3545872  PMID: 23148661
Isoprenoids; Protein solubility; Deoxyxylulose phosphate pathway; Activity analysis; Metabolic engineering
11.  Metabolite Profiling Identified Methylerythritol Cyclodiphosphate Efflux as a Limiting Step in Microbial Isoprenoid Production 
PLoS ONE  2012;7(11):e47513.
Isoprenoids are natural products that are all derived from isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). These precursors are synthesized either by the mevalonate (MVA) pathway or the 1-Deoxy-D-Xylulose 5-Phosphate (DXP) pathway. Metabolic engineering of microbes has enabled overproduction of various isoprenoid products from the DXP pathway including lycopene, artemisinic acid, taxadiene and levopimaradiene. To date, there is no method to accurately measure all the DXP metabolic intermediates simultaneously so as to enable the identification of potential flux limiting steps. In this study, a solid phase extraction coupled with ultra performance liquid chromatography mass spectrometry (SPE UPLC-MS) method was developed. This method was used to measure the DXP intermediates in genetically engineered E. coli. Unexpectedly, methylerythritol cyclodiphosphate (MEC) was found to efflux when certain enzymes of the pathway were over-expressed, demonstrating the existence of a novel competing pathway branch in the DXP metabolism. Guided by these findings, ispG was overexpressed and was found to effectively reduce the efflux of MEC inside the cells, resulting in a significant increase in downstream isoprenoid production. This study demonstrated the necessity to quantify metabolites enabling the identification of a hitherto unrecognized pathway and provided useful insights into rational design in metabolic engineering.
PMCID: PMC3487848  PMID: 23133596
12.  Effect of combined function of temperature and water activity on the growth of Vibrio harveyi 
Brazilian Journal of Microbiology  2012;43(4):1365-1375.
Vibrio harveyi is considered as a causative agent of the systemic disease, vibriosis, which occurs in many biological fields. The effects of temperatures (12.9–27.1 °C) and water activity (NaCl% 0.6%-3.4%) on V. harveyi were investigated. The behavior and growth characteristics of V. harveyi was studied and modeled. Growth curves were fitted by using Gompertz and Baranyi models, and the Baranyi model showed a better fittness. Then, the maximum growth rates (μmax) and lag phase durations (LPD, λ) obtained from both Gompertz and Baranyi model were modeled as a combination function of temperature and water activity using the response surface and Arrhenius-Davey models for secondary model. The value of r2, MSE, bias and accuracy factor suggest Baranyi model has better fitness than Gompertz model. Furthermore, validation of the developed models with independent data from ComBase also shown better interrelationship between observed and predicted growth parameter when using Baranyi model.
PMCID: PMC3769047  PMID: 24031965
Vibrio harveyi; modelling; temperature; water activity
13.  Novel reference genes for quantifying transcriptional responses of Escherichia coli to protein overexpression by quantitative PCR 
BMC Molecular Biology  2011;12:18.
Accurate interpretation of quantitative PCR (qPCR) data requires normalization using constitutively expressed reference genes. Ribosomal RNA is often used as a reference gene for transcriptional studies in E. coli. However, the choice of reliable reference genes has not been systematically validated. The objective of this study is to identify a set of reliable reference genes for transcription analysis in recombinant protein over-expression studies in E. coli.
In this study, the meta-analysis of 240 sets of single-channel Affymetrix microarray data representing over-expressions of 63 distinct recombinant proteins in various E. coli strains identified twenty candidate reference genes that were stably expressed across all conditions. The expression of these twenty genes and two commonly used reference genes, rrsA encoding ribosomal RNA 16S and ihfB, was quantified by qPCR in E. coli cells over-expressing four genes of the 1-Deoxy-D-Xylulose 5-Phosphate pathway. From these results, two independent statistical algorithms identified three novel reference genes cysG, hcaT, and idnT but not rrsA and ihfB as highly invariant in two E. coli strains, across different growth temperatures and induction conditions. Transcriptomic data normalized by the geometric average of these three genes demonstrated that genes of the lycopene synthetic pathway maintained steady expression upon enzyme overexpression. In contrast, the use of rrsA or ihfB as reference genes led to the mis-interpretation that lycopene pathway genes were regulated during enzyme over-expression.
This study identified cysG/hcaT/idnT to be reliable novel reference genes for transcription analysis in recombinant protein producing E. coli.
PMCID: PMC3110127  PMID: 21513543
14.  L-carnitine and taurine synergistically inhibit the proliferation and osteoblastic differentiation of vascular smooth muscle cells 
Acta Pharmacologica Sinica  2010;31(3):289-296.
To investigate the synergistic action of L-carnitine (LC) and taurine (TAU) on the proliferation and osteoblastic differentiation of vascular smooth muscle cells (VSMCs).
DNA and protein synthesis of VSMCs were assessed using scintillation counting. Alkaline phosphatase (ALP) activity and calcium content were determined to investigate the effects of LC and TAU on the osteoblastic differentiation and mineralization of VSMCs. TAU uptake by VSMCs was assayed. RNA interference was used to down-regulate the expression of the TAU transporter (TAUT) in rat VSMCs.
LC and TAU synergistically inhibited the proliferation and β-glycerophosphate (β-GP)-induced osteoblastic differentiation of VSMCs as evidenced by the decreased [3H]thymidine incorporation, ALP activity and calcium deposition. Furthermore, LC stimulated the TAU uptake and TAUT expression in VSMCs. Suppression of TAUT with short hairpin RNA (shRNA) abolished the synergistic action of LC and TAU in VSMCs.
The synergistic inhibitory action of LC and TAU on the proliferation and osteoblastic differentiation of VSMCs is attributable to the up-regulation of TAUT expression and TAU uptake by LC.
PMCID: PMC4002410  PMID: 20154715
L-carnitine; taurine; vascular smooth muscle cells; proliferation; osteoblastic differentiation
15.  A ZYG-12–dynein interaction at the nuclear envelope defines cytoskeletal architecture in the C. elegans gonad 
The Journal of Cell Biology  2009;186(2):229-241.
Changes in cellular microtubule organization often accompany developmental progression. In the Caenorhabditis elegans embryo, the centrosome, which is attached to the nucleus via ZYG-12, organizes the microtubule network. In this study, we investigate ZYG-12 function and microtubule organization before embryo formation in the gonad. Surprisingly, ZYG-12 is dispensable for centrosome attachment in the germline. However, ZYG-12–mediated recruitment of dynein to the nuclear envelope is required to maintain microtubule organization, membrane architecture, and nuclear positioning within the syncytial gonad. We examined γ-tubulin localization and microtubule regrowth after depolymerization to identify sites of nucleation in germ cells. γ-Tubulin localizes to the plasma membrane in addition to the centrosome, and regrowth initiates at both sites. Because we do not observe organized microtubules around zyg-12(ct350) mutant nuclei with attached centrosomes, we propose that gonad architecture, including membrane and nuclear positioning, is determined by microtubule nucleation at the plasma membrane combined with tension on the microtubules by dynein anchored at the nucleus by ZYG-12.
PMCID: PMC2717649  PMID: 19635841
16.  Assessment of hemodynamics in precancerous lesion of hepatocellular carcinoma: Evaluation with MR perfusion 
AIM: To investigate the hemodynamic changes in a precancerous lesion model of hepatocellular carcinoma (HCC).
METHODS: Hemodynamic changes in 18 Wistar rats were studied with non-invasive magnetic resonance (MR) perfusion. The changes induced by diethylnitrosamine (DEN) developed into liver nodular lesions due to hepatic cirrhosis during the progression of carcinogenesis. The MR perfusion data [positive enhancement integral (PEI)] were compared between the nodular lesions corresponding well with MR images and pathology and their surrounding hepatic parenchyma.
RESULTS: A total of 46 nodules were located by MR imaging and autopsy, including 22 dysplastic nodules (DN), 9 regenerative nodules (RN), 10 early HCCs and 5 overt HCCs. Among the 22 DNs, 6 were low-grade DN (LGDN) and 16 were high-grade DN (HGDN). The average PEI of RN, DN, early and overt HCC was 205.67 ± 31.17, 161.94 ± 20.74, 226.09 ± 34.83, 491.86 ± 44.61 respectively, and their liver parenchyma nearby was 204.84 ± 70.19. Comparison of the blood perfusion index between each RN and its surrounding hepatic parenchyma showed no statistically significant difference (P = 0.06). There were significant differences in DN (P = 0.02). During the late hepatic arterial phase, the perfusion curve in DN declined. DN had an iso-signal intensity at the early hepatic arterial phase and a low signal intensity at the portal venous phase. Of the 10 early HCCs, 4 demonstrated less blood perfusion and 6 displayed minimally increased blood flow compared to the surrounding parenchyma. Five HCCs showed significantly increased blood supply compared to the surrounding parenchyma (P = 0.02).
CONCLUSION: Non-invasive MR perfusion can detect changes in blood supply of precancerous lesions.
PMCID: PMC4146991  PMID: 17451197
Liver; Perfusion; Magnetic resonance imaging; Rat
17.  Detection of small hepatocellular carcinoma: Comparison of dynamic enhancement magnetic resonance imaging and multiphase multirow-detector helical CT scanning 
AIM: To compare the gadolinium-enhanced multiphase dynamic magnetic resonance imaging (MRI) and multiphase multirow-detector helical CT (MDCT) scanning for detection of small hepatocellular carcinoma (HCC).
METHODS: MDCT scanning and baseline MRI with SE T1-WI and T2-WI sequence combined with FMPSPGR sequence were performed in 37 patients with 43 small HCCs. Receiver operating characteristic (ROC) curves were plotted to analyze the results for modality.
RESULTS: The areas below ROC curve (Az) were calculated. There was no statistical difference in dynamic enhancement MDCT and MRI. The detection rate of small HCC was 97.5%-97.6% on multiphase MDCT scanning and 90.7%-94.7% on MRI, respectively. The sensitivity of detection for small HCC on MDCT scanning was higher than that on dynamic enhancement MRI. The sensitivity of detection for minute HCC (tumor diameter ≤ 1 cm) was 90.0%-95.0% on MDCT scanning and 70.0%-85.0% on MRI, respectively.
CONCLUSION: MDCT scanning should be performed for early detection and effective treatment of small HCC in patients with chronic hepatitis and cirrhosis during follow-up.
PMCID: PMC4147003  PMID: 17451209
Hepatocellular carcinoma; X-ray computed tomography; Magnetic resonance imaging
18.  CT perfusion at early stage of hepatic diffuse disease 
AIM: To determine the validity of the non-invasive method of CT perfusion (CTP) in rat model of hepatic diffuse disease.
METHODS: Twenty-eight Wistar rats were divided into two groups. Liver diffuse lesions were induced by diethyln-itrosamine in 14 rats of test group. Rats in control group were bred with pure water. From the 1st to 12th wk after the test group was intervened, both groups were studied every week with CTP. CTP parameters of liver parenchyma in different periods and pathologic changes in two groups were compared and analyzed.
RESULTS: The process of hepatic diffuse lesions in test groups was classified into three stages or periods according to the pathologic alterations, namely hepatitis, hepatic fibrosis, and cirrhosis. During this period, hepatic artery flow (HAF) of control group declined slightly, mean transit time (MTT), blood flow (BF) and volume (BV) increased, but there were no significant differences between different periods. In test group, HAF tended to increase gradually, MTT prolonged obviously, BV and BF decreased at the same time. The results of statistical analysis revealed that the difference in the HAF ratio of test group to control group was significant. The ratio of BV and BF in test group to control group in stage of hepatitis and hepatic cirrhosis, hepatic fibrosis and early stage of hepatic cirrhosis was significantly different, but there was no significant difference between hepatitis and hepatic fibrosis. The main pathological changes in stage of hepatitis were swelling of hepatic cells, while sinusoid capillarization and deposition of collagen aggravated gradually in the extravascular Disse’s spaces in stage of fibrosis and early stage of cirrhosis.
CONCLUSION: The technique could reflect some early changes of hepatic blood perfusion in rat with liver diffuse disease and is valuable for their early diagnosis.
PMCID: PMC4316005  PMID: 15948256
Experimental animal; Hepatitis; Hepatic fibrosis; Hepatic cirrhosis; Computed tomography; Perfusion
19.  Postprocessing techniques of CT colonography in detection of colorectal carcinoma 
AIM: To evaluate the value of postprocessing techniques of CT colonography, including multiplanar reformation (MPR), virtual colonoscopy (VC), shaded surface display (SSD) and Raysum, in detection of colorectal carcinomas.
METHODS: Sixty-four patients with colorectal carcinoma underwent volume scanning with spiral CT. MPR, VC, SSD and Raysum images were obtained by using four kinds of postprocessing techniques in workstation. The results were comparatively analyzed according to circumferential extent, lesion length and pathology pattern of colorectal carcinomas. All diagnoses were proved pathologically and surgically.
RESULTS: The accuracy of circumferential extent of colorectal carcinoma determined by MPR, VC, SSD and Raysum was 100.0%, 82.8%, 79.7% and 79.7%, respectively. There was a significant statistical difference between MPR and VC. The consistent rate of lesion length was 89.1%, 76.6%, 95.3% and 100.0%, respectively. There was a statistical difference between VC and SSD. The accuracy of discriminating pathology pattern was 81.3%, 92.2%, 71.9% and 71.9%, respectively. There was a statistical difference between VC and SSD. MPR could determine accurately the circumference of colorectal carcinoma, Raysum could determine the length of lesion more precisely than SSD, VC was helpful in discriminating pathology patterns.
CONCLUSION: MPR, VC, SSD and Raysum have advantage and disadvantage in detection of colorectal carcinoma, use of these methods in combination can disclose the lesion more accurately.
PMCID: PMC4572757  PMID: 15162528
20.  Role of multiphase scans by multirow-detector helical CT in detecting small hepatocellular carcinoma 
World Journal of Gastroenterology  2003;9(10):2198-2201.
AIM: To evaluate the role of multiphasic scanning by multirow-detector helical CT (MDCT) in detecing small hypervascular hepatocellular carcinoma (SHCC).
METHODS: Multiphasic scanning was carried out in 75 patients with SHCC with Marconi MX8000 CT scanner. The early arterial phase (EAP), late arterial phase (LAP) and the portal venous phase (PVP) scans were started at 21 s, 34 s and 85 s respectively. The mean difference of CT values between tumor and liver parenchyma for each scanning phase was measured, and the sensitivity of detection of SHCC in each of these phases and in the combined phase was calculated and statistically analyzed.
RESULTS: The mean difference of CT values between tumor and liver parenchyma was significant in 71 lesions ≥ 1 cm in three phases (P < 0.05). In 91 tumor foci, the detectability of SHCC was 45.1%, 83.5% and 92.3% in EAP, LAP and double arterial phases (DAP), respectively. The early arterial phase plus the portal venous phase and the double arterial phase plus the portal venous phase were 94.5%, 97.8%, respectively. Whereas the detectability in LAP plus PVP and in DAP plus PVP had no statistical difference.
CONCLUSION: The utility of faster speed and thinner slice MDCT and multiphase scanning protocol can improve the detectability of hypervascular small hepatocellular carcinoma. Among which LAP is superior to EAP in depicting the lesions.
PMCID: PMC4656462  PMID: 14562377
21.  Budd-Chiari syndrome: Diagnosis with three-dimensional contrast-enhanced magnetic resonance angiography 
World Journal of Gastroenterology  2003;9(10):2317-2321.
AIM: To evaluate the role of three-dimensional contrast-enhanced magnetic resonance angiography (3D CE MRA) in the diagnosis of Budd-Chiari syndrome (BCS).
METHODS: Twenty-three patients with BCS underwent 3D CE MRA examination, in which 13 cases were secondary to either hepatocellular carcinoma (11 cases), right adrenal carcinoma (1 case) or thrombophlebitis (1 case) and 10 suffered from primary BCS. The patency of the inferior vena cava (IVC), hepatic and portal veins as well as the presence of intra- and extrahepatic collaterals, liver parenchymal abnormalities and porto-systemic varices were evaluated. Inferior vena cavography was performed in 10 cases. The diagnosis of IVC obstruction by 3D CE MRA was compared with that demonstrated by inferior vena cavography.
RESULTS: The major features of BCS could be clearly displayed on 3D CE MRA. Positive hepatic venous signs included tumor thrombosis (9 cases), tumor compression (2 cases), nonvisualization (4 cases) and focal stenosis (2 cases). Positive IVC findings were noted as severe stenosis or occlusion (10 cases), tumor invasion (2 cases), thrombosis (3 cases), thrombophlebitis (1 case) and septum formation (3 cases). Intrahepatic collaterals were shown in 9 patients, 2 of them with “spider web” sign. The displayed extrahepatic collaterals included dilated azygos and hemi-azygos veins (13 cases) and left renal-inferior phrenic-pericardiophrenic veins (2 cases). The occlusion of the left intrahepatic portal veins was found in 2 cases. Porto-systemic varices were detected in 10 patients. Liver parenchymal abnormalities displayed by 3D CE MRA were enlargement of the caudate lobe (7 cases), heterogenous enhancement (18 cases) and complicated tumors (13 cases). Compared with the inferior vena cavography performed in 10 cases, the accuracy of 3D CE MRA was 100% in the diagnosis of IVC obstruction.
CONCLUSION: 3D CE MRA can display the major features of BCS and provide an accurate diagnosis.
PMCID: PMC4656485  PMID: 14562400
22.  Three-dimensional contrast-enhanced MR angiography in diagnosis of portal vein involvement by hepatic tumors 
AIM: To assess the accuracy of three-dimensional contrast-enhanced magnetic resonance angiography (3D CE MRA) in evaluation of the portal vein involvement in patients with hepatic tumors.
METHODS: 3D CE MRA was performed in 62 patients with hepatic tumors to assess the patency of the main, right and left portal veins before hepatic surgery. A total of 186 veins were examined for encasement, occlusion and tumor thrombosis. The results of 3D CE MRA diagnosis were then correlated with the surgical-pathological and intra-operative sonographic findings.
RESULTS: 3D CE MRA correctly detected 48 of 49 involved and 135 of 137 noninvolved portal veins with the sensitivity of 98%, specificity of 99%, positive predictive value of 96% and negative predictive value of 99%.
CONCLUSION: 3D CE MRA is accurate in evaluation of the portal vein involvement in patients with hepatic tumors.
PMCID: PMC4611385  PMID: 12717869
23.  Effect of cisapride on intestinal bacterial and endotoxin translocation in cirrhosis 
AIM: To investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats.
METHODS: All animals were assessed with variables including bacterial and endotoxin translocation, intestinal bacterial overgrowth, intestinal transit and permeability. Bacterial translocation (BT) was assessed by bacterial culture of MLN, liver and spleen, IBO by a jejunal bacterial count of the specific organism, intestinal permeability by determination of the 24-hour urinary 99mTc-DTPA excretion and intestinal transit by measurement of the distribution of 51Cr in the intestine.
RESULTS: Bacterial translocation (BT) and IBO was found in 48% and 80% cirrhotic rats respectively and none in control rats. Urinary excretion of 99mTc-DTPA in cirrhotic rats with BT (22.2 ± 7.8) was greater than these without BT (10.5 ± 2.9). Intestinal transit (geometric center ratio) was significantly delayed in cirrhotic rats (0.31 ± 0.06) and further more delayed in cirrhotic rats with BT (0.24 ± 0.06) than these without BT (0.38 ± 0.11). Cirrhotic rats with IBO had significantly higher rates of intestinal bacterial and endotoxin translocation, slower intestinal transit time and higher intestinal permeability than those without IBO. It was also found that BT was closely associated with IBO and the injury of intestinal barrier. Compared with the placebo group, cisapride-treated rats had lower rates of bacterial/endotoxin translocation and IBO, which was closely associated with increased intestinal transit and improved intestinal permeability by cisapride.
CONCLUSION: These results indicate that endotoxin and bacterial translocation in cirrhotic rats may be attributed to IBO and increased intestinal permeability. Cisapride that accelerates intestinal transit and improve intestinal permeability might be helpful in preventing intestinal bacterial and endotoxin translocation.
PMCID: PMC4621577  PMID: 12632513
24.  Direct intrahepatic portacaval shunt: An experimental study 
AIM: To determine the feasibility of creating direct intrahepatic portacaval shunt (DIPS) in swine with puncture under sonographic guidance.
METHODS: DIPS was created in 10 domestic swine under sonographic guidance. Liver function, blood ammonia level and portosystemic gradient (PSG) were compared before and after the procedure. Patency of shunt was followed by portography every 7 d after DIPS.
RESULTS: DIPS was successfully established in all 10 swine without any complications. One day after procedure the alanine aminotransferase (ALT), aspartate aminotransferase (AST), and blood ammonia level (BAL) of swine rose from 5.40 ± 0.69, 16.00 ± 0.79 and 35.66 ± 4.10 to 34.20 ± 3.46, 59.70 ± 2.22 and 66.94 ± 3.44 respectively (P < 0.05). The PSG decreased from 0.59 ± 0.20 kPa to 0.24 ± 0.11 kPa after DIPS (P < 0.05). The shunt of 10 swine was kept patent from 7-28 d (median patency time was 14 d).
CONCLUSION: This initial experience demonstrated that creating intrahepatic portacaval shunt from retrohepatic segment of IVC to portal vein with puncture under sonographic guidance in swine is safe and feasible. Further studies are necessary to perform DIPS in cirrhosis patients.
PMCID: PMC4611339  PMID: 12532459
25.  Role and limitation of FMPSPGR dynamic contrast scanning in the follow-up of patients with hepatocellular carcinoma treated by TACE 
AIM: To evaluate the role and limitation of fast multiplanar spoiled gradient-recalled (FMPSPGR) MR dynamic contrast scanning in the follow-up of patients with HCC treated by transarterial chemoembolization (TACE).
METHODS: Twenty-two patients with 24 HCC lesions confirmed by biopsy or surgical resection underwent MR imaging in 4-9wks after TACE with a superconducting 1.5 T MR scanner, including SE T1WI, T2WI and FMPSPGR dynamic contrast scanning. The signal intensities of all lesions on SE T1WI, T2WI and the enhancement patterns on FMPSPGR dynamic contrast scanning were observed, and the comparison was made between MRI findings and pathological results in all the cases.
ESULTS: Of the 24 lesions, the signal intensities were various on SE T1WI and T2WI. On T1WI, 13 lesions appeared as hyperintense, 4 lesions were isointense and the other 7 lesions were hypointensese. Histologically, hyperintense lesions showed on T1WI were viable tumor or hemorrhage; isointensities were coagulative necrosis or inflammatory infiltration; hypointensities were tumor, liquified necrosis, coagulative necrosis or inflammatory infiltration. On T2WI, 15 lesions appeared as hyperintense, 3 lesions were isointense and the other 6 lesions were hypointensese. Hyperintense lesions showed on T2WI were residuals of viable tumor, hemorrhage, liquefied necrosis or inflammatory infiltration; isointense lesions were residuals of viable tumor or inflammatory infiltration; hypointense lesions were coagulative necrosis. On FMPSPGR dynamic contrast scanning, 18 of the 24 lesions enhanced on early-phase dynamic scanning corresponding to residuals of viable tumor and the other 6 lesions had no enhancement at this phase because complete necrosis were seen in the histologic examination. On delayed-phase dynamic scanning, 6 lesions had permanent enhancement appeared as inhomogeneous hyperintensity and both residuals of viable tumor and inflammatory infiltration were found by histologic examination. 18 lesions were hypointense at this phase and 8 of them coexisted with peripheral ring-like enhancement of the lesions resulting from viable tumors or inflammatory infiltration.
CONCLUSION: FMPSPGR MR dynamic contrast scanning can reflect the pathologic changes of HCC treated by TACE. Especially, early-phase dynamic scanning can evaluate accurately residuals of viable tumor and necrosis in HCC lesions. FMPSPGR dynamic contrast scanning is useful in the follow-up of patients with HCC treated by TACE combined with SE T1WI and T2WI, but it is difficult to differentiate peripheral viable tumors from inflammatory infiltration.
PMCID: PMC4656316  PMID: 12174374

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