In situ meta-omic insights into the community compositions and ecological roles of hadal microbes in the Mariana Trench.

The low temperature and elevated hydrostatic pressure in hadal trenches at water depths below 6000 m render sample collection difficult. Here, in situ hadal water microbial samples were collected from the Mariana Trench and analysed. The hadal microbial communities at different depths were revealed to be consistent and were dominated by heterotrophic Marinimicrobia. Thirty high-quality metagenome-assembled genomes (MAGs) were retrieved to represent the major hadal microbes affiliated with 12 prokaryotic phyla. Most of the MAGs were newly reported and probably derived from novel hadal inhabitants as exemplified by a potentially new candidate archaeal phylum in the DPANN superphylum. Metabolic reconstruction indicated that a great number of the MAGs participated in nitrogen and sulfur cycling, in which the nitrification process was driven sequentially by Thaumarchaeota and Nitrospirae and sulfur oxidization by Rhodospirillales in the Alphaproteobacteria class. Moreover, several groups of hadal microbes were revealed to be potential carbon monoxide oxidizers. Metatranscriptomic result highlighted the contribution of Chloroflexi in degrading recalcitrant dissolved organic matter and Marinimicrobia in extracellular protein decomposition. The present work provides an in-depth view on the hadal microbial communities regarding their endemism and element cycles.

Genomics insights into ecotype formation of ammonia-oxidizing archaea in the deep ocean.

Various lineages of ammonia-oxidizing archaea (AOA) are present in deep waters, but the mechanisms that determine ecotype formation are obscure. We studied 18 high-quality genomes of the marine group I AOA lineages (alpha, gamma and delta) from the Mariana and Ogasawara trenches. The genomes of alpha AOA resembled each other, while those of gamma and delta lineages were more divergent and had even undergone insertion of some phage genes. The instability of the gamma and delta AOA genomes could be partially due to the loss of DNA polymerase B (polB) and methyladenine DNA glycosylase (tag) genes responsible for the repair of point mutations. The alpha AOA genomes harbour genes encoding a thrombospondin-like outer membrane structure that probably serves as a barrier to gene flow. Moreover, the gamma and alpha AOA lineages rely on vitamin B12 -independent MetE and B12 -dependent MetH, respectively, for methionine synthesis. The delta AOA genome contains genes involved in uptake of sugar and peptide perhaps for heterotrophic lifestyle. Our study provides insights into co-occurrence of cladogenesis and anagenesis in the formation of AOA ecotypes that perform differently in nitrogen and carbon cycling in dark oceans.

Laser speckle contrast imaging and Oxygen to See for assessing microcirculatory liver blood flow changes following different volumes of hepatectomy.

OBJECTIVE: Portal hyperperfusion after extended hepatectomy or small-for-size liver transplantation may induce organ dysfunction and failure. This study was designed to monitor and characterize the hepatic microcirculatory perfusion following different volumes of hepatectomy in rats by using laser speckle contrast image (LSCI) and Oxygen to See (O2C), a spectrometric device. METHODS: The microcirculatory liver blood flow of the rats that underwent 68%, 85% and 90% hepatectomy (68PH, 85PH and 90PH) was monitored with LSCI and O2C before and following the hepatectomy. The portal venous flow (PVF) and hepatic arterial flow (HAF) were measured with an ultrasonic flowmeter. Liver regeneration, liver injury, histologic evaluation and gene expression were also assessed at 12h, 24h, 3d and 7d post hepatectomy. RESULTS: All the 68PH and 85PH rats survived, and 57% of the 90PH rats survived. After hepatectomy, both PVF and HAF decreased transiently, with the PVF of the 85PH and 90PH rats significantly lower than that of the 68PH rats. In contrast, the PVF and HAF per gram of liver weight were greatly increased after liver resection and were proportional to the volume of resected liver. Correspondingly, the microcirculatory liver blood flow of the 68PH, 85PH and 90PH rats, as assessed by both LSCI and O2C, were increased after hepatectomy, and the 90PH group was significantly higher than the 68PH and 85PH groups. The hyperperfusion continued for approximately 3days and returned to baseline following the completion of liver regeneration. The liver venous oxygen saturation of the three groups decreased immediately after hepatectomy and returned to baseline from 24h after hepatectomy. The 90PH rats also showed delayed liver regeneration and the most severe liver injury, as reflected by increased serum ALT, AST and TBIL levels, hepatocellular vacuolization, and inflammatory and endothelial constriction gene expressions (TNF-α, IL-1ß, MIP-1α, ET-1 and TM-1). CONCLUSION: Hepatic microcirculation hyperperfusion resulting from major and extended liver resection could be assessed by LSCI and O2C methods. The 90PH in rats led to extraordinary sinusoidal hyperperfusion, severe endothelial injury and liver failure. Monitoring the changes of hepatic microcirculation perfusion following extended hepatectomy or small-for-size liver transplantation may help to analyze the extent of hyperperfusion.

Genomic characterization of symbiotic mycoplasmas from the stomach of deep-sea isopod bathynomus sp.

Deep-sea isopod scavengers such as Bathynomus sp. are able to live in nutrient-poor environments, which is likely attributable to the presence of symbiotic microbes in their stomach. In this study we recovered two draft genomes of mycoplasmas, Bg1 and Bg2, from the metagenomes of the stomach contents and stomach sac of a Bathynomus sp. sample from the South China Sea (depth of 898 m). Phylogenetic trees revealed a considerable genetic distance to other mycoplasma species for Bg1 and Bg2. Compared with terrestrial symbiotic mycoplasmas, the Bg1 and Bg2 genomes were enriched with genes encoding phosphoenolpyruvate-dependent phosphotransferase systems (PTSs) and sodium-driven symporters responsible for the uptake of sugars, amino acids and other carbohydrates. The genome of mycoplasma Bg1 contained sialic acid lyase and transporter genes, potentially enabling the bacteria to attach to the stomach sac and obtain organic carbons from various cell walls. Both of the mycoplasma genomes contained multiple copies of genes related to proteolysis and oligosaccharide degradation, which may help the host survive in low-nutrient conditions. The discovery of the different types of mycoplasma bacteria in the stomach of this deep-sea isopod affords insights into symbiotic model of deep-sea animals and genomic plasticity of mycoplasma bacteria.

Hepatoma-derived growth factor promotes growth and metastasis of hepatocellular carcinoma cells.

We aimed to elucidate the effects of hepatoma-derived growth factor (HDGF) on growth and metastasis of hepatocellular carcinoma (HCC) cells. Tissue microarrays with 236 HCC specimens and 18 extrahepatic metastases were utilized to detect the HDGF expression by immunohistochemistry. Meanwhile, HDGF expressions in HCC cell lines with different metastatic potentials were examined using immunofluorescence staining, real-time PCR and western blotting. After HDGF silencing, the growth and metastatic potentials of HCC cells were evaluated by soft agar assay, invasion assay, together with tumorigenicity assay in nude mice. The gelatin zymography was performed by detecting MMP-2 and MMP-9 levels. Additionally, western blotting was conducted to determine the levels of total and phosphorylated ERK1/2, JNK, p38 and Akt. The results showed that HDGF was overexpressed in HCC metastasis tumour, and the expression increased with the differentiation degree of tumours (Grade I 44.0%, Grade II 48.4% and Grade III 65.6%). Consistently, HDGF levels were positively associated with the metastatic capability of HCC cells (MHCC97L < MHCC97H < HCCLM3). The growth and metastasis were suppressed by HDGF-siRNA. Gelatinolytic activities were enhanced in the three metastatic HCC cell lines, but had no significant difference among them. The tumourigenicity and metastatic capability of HCCLM3 cells in nude mice were inhibited after silencing HDGF. Meanwhile, HDGF-siRNA specifically suppressed the total and phosphorylated protein levels of ERK1/2, while not JNK, p38 and Akt. In conclusion, HDGF was overexpressed in HCC patients and cells, and HDGF might be closely correlated with HCC metastasis via regulating ERK signalling pathway. Copyright © 2016 John Wiley & Sons, Ltd.

[Comparison of the Predictive Values of Eight Staging Systems for Primary Liver Cancer in Prognosis of Combined Hepatocellular-cholangiocellular Carcinoma Patients after Surgery].

OBJECTIVE: To compare the predictive values of eight staging systems for primary liver cancer in the prognosis of combined hepatocellular-cholangiocellular carcinoma (cHCC-CC) patients after surgery. METHODS: The clinical data of 54 cHCC-CC patients who underwent hepatectomy or liver transplantation from May 2005 to Augest 2013 in Chinese PLA General Hospital were collected. We evaluated the prognostic value of the Okuda staging system, Cancer of the Liver Italian Program (CLIP) score, French staging system, Barcelona Clinic Liver Cancer (BCLC) staging system, 7th edition of tumour-node-metastasis (TNM) staging system for hepatocellular carcinoma and intrahepatic cholangiocarcinoma (ICC), Japan Integrated Staging (JIS) score, and Chinese University Prognostic Index. The distribution, Kaplan-Meier method, Log-rank test, and area under a receiver operating characteristic curve were used to compare the prognosis-predicting ability of these different staging systems in 54 cHCC-CC patients after surgery. RESULTS: The TNM staging system for ICC and JIS score had a better distribution of cases. The 12-and 24-month survivals of the entire cohort were 65.5% and 56.3%, respectively. A Log-rank test showed that there was a significant difference existing in the cumulative survival rates of different stage patients when using TNM staging system for ICC (stage 1 vs. stage 2, P=0.012; stage 2 vs. stage 3-4, P=0.002), Okuda staging system (stage 1 vs. stage 2, P=0.025), and French staging system (stage A and stage B, P=0.045). The 12-and 24-month area under curve of TNM staging system for ICC, BCLC staging system, JIS score, and CLIP score were 0.836 and 0.847, 0.744 and 0.780, 0.723 and 0.764, and 0.710 and 0.786, respectively. CONCLUSION: The 7th edition of TNM staging system for ICC has superior prognostic value to other seven staging systems in cHCC-CC patients undergoing surgical treatment.

Portal vein clamping alone confers protection against hepatic ischemia-reperfusion injury via preserving hepatocyte function in cirrhotic rats.

BACKGROUND: Chronic liver diseases always increase the risk of liver failure after hepatectomy. We aimed to explore the protective effect of portal vein clamping without hepatic artery blood control (PVC) on a cirrhotic rat liver that underwent ischemia and reperfusion. METHODS: Carbon tetrachloride-induced cirrhotic rats were randomly assigned to four groups as follows: cirrhotic control, PVC, portal triad clamping (PTC), and intermittent portal triad clamping (IC). After 45 min of portal vascular clamping, hepatic injury and liver function were investigated by assessing the 7-d survival rate, liver blood loss, serum alanine aminotransferase, liver tissue malondialdehyde, liver tissue adenosine triphosphate, indocyanine green retention rate, and morphology changes of the rat liver. RESULTS: The 7-d survival rates in the PVC and IC groups were much higher than in the PTC group. The PVC group had more liver blood loss during the hepatectomy than the PTC group, but had much less than the cirrhotic control group (P < 0.01). In addition, there were no differences between the IC group and PVC group. The PVC rats had a significantly higher adenosine triphosphate level in the liver tissue and a markedly lower indocyanine green retention rate than the PTC and IC rats (P < 0.05). At 1, 6, and 24 h after reperfusion, the alanine aminotransferase and malondialdehyde levels in the PTC group were much higher than those in the PVC and IC groups (P < 0.05). Based on the histopathologic analysis, hepatic injury in the PVC and IC groups were similar but less prominent than in the PTC group. CONCLUSIONS: Although both PVC and IC can confer protection against hepatic ischemic-reperfusion injury in cirrhotic rats, the PVC method is more efficient in preserving the energy and function of hepatocytes than the IC method, suggesting better prognosis after hepatectomy.

mTOR-Dependent Suppression of Remnant Liver Regeneration in Liver Failure After Massive Liver Resection in Rats.

BACKGROUND AND AIMS: Massive hepatectomy often leads to fatal liver failure because of a small remnant liver volume. The aim of this study was to investigate the potential mechanisms leading to liver failure. METHODS: Sprague-Dawley rats had performed a sham operation, 85 % partial hepatectomy (PH) or 90 % PH, and all had free access to water with or without supplemented glucose. Liver function and survival were evaluated. Liver parenchymal injury was assessed by evaluating hepatic pathology, blood biochemistry, and apoptotic and necrotic alterations. The regeneration response was assessed by the weight gain of the remnant liver, hepatocyte proliferation markers, and regeneration-related molecules. RESULTS: The 90 % hepatectomy resulted in a significantly lower survival rate and impaired liver function; however, no significant more serious liver parenchymal injuries were detected. TNF-α, HGF, myc and IL-6 were either similarly expressed or overexpressed; however, the increase in remnant liver weight, mitotic index, and the presence of Ki-67 and PCNA were significantly lower in the 90 %-hepatectomized rats. mTOR, p70S6K and 4EBP1 were not activated in the remnant liver after a 90 % hepatectomy as obviously as those after an 85 % hepatectomy, which was concomitant with the higher expression of phospho-AMPK and a lower intrahepatic ATP level. Glucose treatment significantly improved the survival rate of 90 %-hepatectomized rats. CONCLUSIONS: Suppression of remnant liver regeneration was observed in the 90 % PH and contributed to fatal liver failure. This suppressed liver regenerative capacity was related to the inhibited activation of mTOR signaling.

The monitoring of microvascular liver blood flow changes during ischemia and reperfusion using laser speckle contrast imaging.

OBJECTIVE: The recovery of microvascular liver blood flow (LBF) after ischemia is an important determinant of the degree of hepatocellular injury. Laser speckle contrast imaging (LSCI) was recently suggested to be a suitable instrument for monitoring the LBF. This study was designed to evaluate LSCI in monitoring the LBF changes during liver ischemia and reperfusion (IR). METHODS: A rat model with 120-min ischemia and 60-min reperfusion to 90% of the liver (entire liver except the caudate lobe, which was kept as portal blood bypass) was used. The LBF of the sham operation (SO) group and the IR group was measured with LSCI at the following time points: before ischemia (Baseline), 5 min after the start of ischemia (I-5 min), 5 min before the end of ischemia (I-115 min) and 5 and 60 min after the start of reperfusion (R-5 min and R-60 min). The reproducibility among different rats or repeated measurements, the liver histopathology, the liver biological zero (BZ) and the influence of liver movement on the LSCI measurements were investigated. RESULTS: The entire exposed liver surface after laparotomy was suitable for full-view LSCI imaging. Establishing many circular or oval regions of interest (ROIs) on the LSCI flux image was a simple and convenient method for calculating and comparing the LBF of different ROIs and different liver lobes. There was good-to-moderate intra-individual and inter-individual reproducibility for the LSCI measurements of the LBF in the rats of the SO group. In the IR group, the total blood inflow occlusion resulted in a notable drop of the LBF from the baseline (P<0.05) that remained for the 120 min of ischemia. The LBF decreased further after the reperfusion (P<0.05), reflecting the IR-induced liver microcirculation dysfunction. The histopathological examination revealed severe hepatic sinus congestion and damaged hepatocytes in the IR group. The no flow BZ and liver movement contributed to the LBF values. CONCLUSIONS: LSCI technology is a simple, convenient and accurate method for the real-time monitoring of microvascular LBF changes during ischemia and reperfusion, regardless of the contribution of biological zero and liver movement. This finding suggests the possible application of LSCI for monitoring the microvascular LBF changes intraoperatively.

Preserving hepatic artery flow during portal triad blood inflow occlusion reduces the outgrowth of hepatocarcinoma in mice after ischemia-reperfusion.

AIM: To investigate the effect of different hepatic vascular occlusion maneuvers on the growth of hepatocarcinoma after liver ischemia-reperfusion (I/R) injury. METHODS: A mice hepatocarcinoma model was established by portal vein injection of H22 hepatoma cells. After 3 days, the mice underwent sham operation, occlusion of portal triad (OPT), portal vein (OPV), or intermittent clamping (INT) operation. The hepatic I/R injury, pathological changes, hepatic replacement area, proliferative cell nuclear antigen expression, and extracellular signal-regulated kinase (ERK) 1/2 activation were assessed 5 days after reperfusion. RESULTS: Alanine aminotransferase and aspartate aminotransferase levels in the OPV group were significantly lower than those in the OPT and INT groups at 24 h after reperfusion. The hepatic injury of clamped liver lobes in the OPV group, represented by histopathological alterations and myeloperoxidase activity, was much slighter than that in the OPT and INT groups. The values of hepatic replacement area in the sham operation, OPT, OPV, and INT groups were 7.661 2.55%, 35.61 1 4.23%, 9.02 1 3.01%, and 19.95 1 4.10%, respectively. Proliferative cell nuclear antigen expression and ERK1/2 activation of tumor cells were the highest in the OPT group, and the lowest in the OPV and INT groups. CONCLUSION: Preserving hepatic artery flow during portal triad blood inflow occlusion substantially inhibits the outgrowth of hepatocarcinoma via attenuating hepatic I/R injury in a murine liver tumor model. These results suggest a better prevention of hepatic tumor outgrowth after hepatectomy by using the selective portal vein clamping method in liver cancer patients.

[Transient hepatic venous occlusion induced liver hemodynamic change and reperfusion injury in rats].

OBJECTIVE: To observe the hemodynamic change and reperfusion injury cause by transient hepatic venous occlusion and transient hepatic inflow occlusion in rats. METHODS: The rat liver was divided into 3 different areas: the ischemia reperfusion (IR) area: the inflow of the right superior lobe was clamped for half an hour; the non-isolated lobe congestive reperfusion (NIL-CR) area: the outflow of the right median lobe was clamped for half an hour; and the isolated lobe congestive reperfusion (IL-CR) area: the outflow of the left lobe was clamped for half an hour. The flux value and the oxygen saturation of microcirculation were monitored before at clamping for 30 minutes, and on 1 day, 3 days ,and 7 days after reperfusion. The hepatic damage and Suzuki's score were evaluated. RESULTS: After clamping for 30 minutes, the flux value in the IR area was significantly higher than in NIL-CR area (P<0.01) and IL-CR area (P<0.01), the oxygen saturation in the IR area was significantly higher than in NIL-CR area (P<0.01) and IL-CR area (P<0.05). Compared with IR area, both NIL-CR area and IL-CR area were found having more severe liver damage in terms of Suzuki's score in early postoperative period (at clamping for 30 minutes and on 1 day, P<0.01). However, there was no significant difference between NIL-CR area and IL-CR area in flux value, oxygen saturation, and Suzuki's score (P>0.05). CONCLUSIONS: Hepatic venous occlusion can more effectively decrease the blood perfrusion and oxygen saturation; thus, compared to the IR, CR can result in more severe liver damage. The presence of normal liver tissue around the congestion area can not influence liver damage in transient hepatic venous occlusion.

Preserving hepatic artery flow during portal triad blood inflow occlusion improves remnant liver regeneration in rats after partial hepatectomy.

BACKGROUND: In the present study, we compared preservation of the hepatic artery flow during liver blood inflow occlusion with total portal triad blood flow clamping (the Pringle maneuver) to examine their effects on liver regeneration in rats after partial hepatectomy. MATERIALS AND METHODS: Male Wistar rats were randomized to a control group (without hepatic inflow occlusion), an occlusion of the portal triad (OPT) group (OPT for 30 min under portal blood bypass), and an occlusion of the portal vein (OPV) group (OPV only for 30 min under portal blood bypass). All the rats underwent partial hepatectomy at the end of hepatic blood control. Liver regeneration was assessed on days 3 and 7 after hepatectomy. Liver damage, extracellular signal-regulated kinase-1/2 activation, and cytokine expression of the remnant liver in the first 24 h after hepatectomy were also assessed. RESULTS: Significantly greater liver regeneration, at a level similar to that of the control group, as indicated by the percentage of the initial liver weight, proliferating cell nuclear antigen and Ki-67 labeling indexes, and technetium-99m galactosyl human serum albumin liver uptake, was observed in the OPV group on day 3 after hepatectomy (P < 0.05 versus the OPT group). Liver damage, as represented by alanine aminotransferase and aspartate aminotransferase measurement and histopathologic examination, was substantially alleviated in the OPV group compared with the OPT group. In contrast to the control and OPV groups, the OPT group had markedly increased extracellular signal-regulated kinase-1/2 activation, heat shock protein 70, and interleukin-6 expression in response to ischemia and partial hepatectomy. CONCLUSIONS: Our results have indicated that compared with the Pringle maneuver, clamping the portal vein while preserving the hepatic artery flow during partial hepatectomy is better for remnant liver regeneration at an early posthepatectomy stage.

Preserving hepatic artery flow during portal triad blood inflow occlusion reduces liver ischemia-reperfusion injury in rats.

BACKGROUND: Temporary portal triad clamping (Pringle maneuver) during liver resection can reduce intraoperative blood loss, but also correlates with liver ischemia and reperfusion (I/R) injury. The hepatic artery supplies 20%-30% blood but more than 50% O(2) to the liver. In this study, we explored if preservation of hepatic artery flow when performing portal triad blood inflow occlusion could reduce liver I/R injury while not increasing the blood loss. MATERIALS AND METHODS: Three groups of rats were created: group SO (sham operation), group OPT (occlusion of portal triad under portal blood bypass), and group OPV (occlusion of portal vein under portal blood bypass). Blood flow was occluded for 90, 100, 110, and 120 min before reperfusion. Liver I/R injury was assessed by measuring the survival of rats within 7 d after operation, liver blood loss, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), liver tissue malondialdehyde (MDA), and Na(+)-K(+)-ATPase, and liver histology. RESULTS: The 7-d survival of rats in group OPV was higher than in group OPT. The safe tolerance limit was 90 min for group OPT and 110 min for group OPV. Liver blood loss in group OPT and OPV were significantly less than in group SO. However, no significant difference was observed in the amount of blood loss between group OPT and group OPV. The group OPV had significantly lower ALT, AST, and MDA values on the first hour and first day post-reperfusion than in group OPT. The Na(+)-K(+)-ATPase activity in OPV group was significantly higher than in group OPT 1 h post-reperfusion. Hepatocyte injury was significantly less in group OPV than in group OPT on histopathology. CONCLUSIONS: These data indicate that continuously clamping the portal vein while preserving the hepatic artery did not increase blood loss significantly in a rat liver I/R model, however this maneuver induced less liver I/R injury. It is therefore suggested that preserving hepatic artery inflow during portal triad blood inflow occlusion might become an alternative maneuver in liver surgery due to its ability to extend the safe tolerant time limit in normothermic hepatic ischemia.

[Effects of different hepatic inflow occlusion methods on liver regeneration following partial hepatectomy in rats].

OBJECTIVE: To explore the effects of different hepatic inflow occlusion methods on liver regeneration in rats after partial hepatectomy (PH). METHODS: Male Wistar-Furth rats were randomly assigned to three groups: control group, underwent 68% hepatectomy alone; occlusion of portal triad (OPT) group, subjected to occlusion of portal triad under portal blood bypass; and occlusion of portal vein (OPV) group, subjected to occlusion of portal vein under portal blood bypass. Blood flow was occluded for 20, 30, and 40 minutes before 68% hepatectomy. According to the 7-day survival of each group, a same occlusion time T was set. Each group was divided into two subgroups (n = 8), in which animals were killed 3 and 7 days later. Liver regeneration was calculated as a percent of initial liver weight. Immunohistochemistry for proliferating cell nuclear antigen (PCNA) and Ki-67 was performed to quantify proliferating cells. In addition, functional liver volume represented by 99Tc(m)-GSA radioactivity was assessed. RESULTS: The safe tolerance limit time was 30 minutes for OPT group and 40 minutes for OPV group. At 3 days after PH, no significant difference was observed in the regeneration rate of each group (P > 0.05). However, liver radioactive activity, PCNA labeling index, and Ki-67 index of OPV group was significantly higher than those of OPT group (P < 0.05); the latter were similar to those of control group (P > 0.05). At 7 days after PH, no significant difference was observed in all indexes among three groups (P > 0.05). CONCLUSION: Compared with Pringle maneuver, preserving the hepatic artery flow during portal triad blood inflow occlusion can promote remnant liver regeneration early after PH.

Paraventricular nucleus corticotrophin releasing hormone contributes to sympathoexcitation via interaction with neurotransmitters in heart failure.

Recent studies indicate that systemic administration of tumor necrosis factor (TNF)-α induces increases in corticotrophin releasing hormone (CRH) and CRH type 1 receptors in the hypothalamic paraventricular nucleus (PVN). In this study, we explored the hypothesis that CRH in the PVN contributes to sympathoexcitation via interaction with neurotransmitters in heart failure (HF). Sprague-Dawley rats with HF or sham-operated controls (SHAM) were treated for 4 weeks with a continuous bilateral PVN infusion of the selective CRH-R1 antagonist NBI-27914 or vehicle. Rats with HF had higher levels of glutamate, norepinephrine (NE) and tyrosine hydroxylase (TH), and lower levels of gamma-aminobutyric acid (GABA) and the 67-kDa isoform of glutamate decarboxylase (GAD67) in the PVN when compared to SHAM rats. Plasma levels of cytokines, NE, ACTH and renal sympathetic nerve activity (RSNA) were increased in HF rats. Bilateral PVN infusions of NBI-27914 attenuated the decreases in PVN GABA and GAD67, and the increases in RSNA, ACTH and PVN glutamate, NE and TH observed in HF rats. These findings suggest that CRH in the PVN modulates neurotransmitters and contributes to sympathoexcitation in rats with ischemia-induced HF.

Effects of partial portal vein arterialization on the hilar bile duct in a rat model.

BACKGROUND: Liver revascularization is frequently required during the enlarged radical operation for hilar cholangiocarcinoma involving the hepatic artery. Researchers have carried out a number of experiments applying partial portal vein arterialization (PVA) in clinical practice. In this study we aimed to establish a theoretical basis for clinical application of partial PVA and to investigate the effects of partial PVA on rat hilar bile duct and hepatic functions. METHODS: Thirty rats were randomly and equally assigned into 3 groups: control (group A), hepatic artery ligation+bile duct recanalization (group B), and partial PVA+bile duct recanalization (group C). Proliferation and apoptosis of rat hilar bile duct epithelial cells, arteriolar counts of the peribiliary plexus (PBP) of the bile duct wall, changes in serum biochemistry, and pathologic changes in the bile duct were assessed 1 month after operation. RESULTS: The proliferation of hilar bile duct epithelial cells in group B was greater than in groups A and C (P<0.01). No apoptotic hilar bile duct epithelial cells were detected in any of the groups. The PBP arteriolar counts of the hilar bile duct wall were similar in groups A and C (P>0.05), but the count was lower in group B than in group A (P<0.01). No statistically significant differences in alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and albumin were found in the 3 groups. The gamma-glutamyltransferase value was higher in group B than in groups A and C (P<0.01). The hepatic tissues of groups A and C showed no significant abnormality. Chronic inflammatory changes in the hilar bile duct walls were observed only in group B. CONCLUSION: Partial PVA can restore the arterial blood supply of the hilar bile duct and significantly extenuate the injury to hilar bile duct epithelial cells resulting from hepatic artery ligation.

[Experimental study of a battlefield shock resuscitation fluid with new formula].

OBJECTIVE: To propose a resuscitation fluid with a new formula for resuscitation of shock in battlefield on the basis of pathogenesis of hemorrhagic shock and clinical experiences, and to evaluate its safety and effectiveness in a rat hemorrhagic shock model. METHODS: After hemorrhagic shock was reproduced in rats, a mixture of lactate Ringer solution and hydroxyethyl starch solution at 2:1 proportion (referred to as LH) was used for resuscitation in animals of control group; LH supplemented with dexamethasone and furosemide (referred to as LHDF) was used as resuscitation fluid for experimental group. After 4 hours of infusion, blood and major organs were obtained for serum biochemical tests, lung water content measurement and histopathological observation. RESULTS: The mean arterial pressure of rats of both control and experimental groups recovered rapidly after resuscitation. There was no significant difference in the parameters of serum biochemistry between control group and experimental group. The wet/dry weight ratio of lung tissue in experimental group was significantly lower than control group (4.56 ± 0.14 vs. 4.88 ± 0.29, P <0.05). The blood alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in experimental group were also significantly lower than in control group [ALT: (73.02 ± 41.89) U/L vs. (193.85 ± 104.49) U/L; AST: (199.06 ± 108.7) U/L vs. (395.25 ± 137.08) U/L, both P <0.05). Diuretic effect was obviously observed in experimental group than control group [time of start urination: (76 ± 20) minutes vs. (153 ± 14) minutes; urine volume: (9.6 ± 5.2) ml vs. (1.5 ± 2.2) ml, P <0.01 and P <0.05] , and the amount of ascitic fluid in experimental group was significantly lower than in control group [(1.3 ± 0.6) ml vs. (5.0 ± 3.0) ml , P <0.05). Histopathological observation of the liver, lung and intestine also showed less pathological changes in experimental group than in control group. CONCLUSION: The designed battlefield anti shock fluid in this study has been shown to be effective in fluid resuscitation for hemorrhage shock in rats, with reduced tissue edema and less injury to the liver, lung and intestine.

[Experimental study of epithelial-mesenchymal transition induced by HBx protein in liver cancer cell].

OBJECTIVE: To explore whether or not HBx protein can induce epithelial-mesenchymal transition (EMT) in liver cancer cell SMMC-7721. METHODS: A recombinant adenovirus vector containing HBx gene was constructed and introduced into SMMC-7721 cell. The morphological changes were observed. The expression changes of epithelial and mesenchymal markers were investigated by RT-PCR, Western blot and immunohistochemistry. Invasive and metastatic capacity of transfected SMMC-7721 cell was evaluated through transwell migration assay. RESULTS: The recombinant adenovirus vector containing HBx gene was constructed and transfected successfully into SMMC-7721 cell. At Day 7 post-transfection, the transfected SMMC-7721 cell underwent the morphological change from a classic epithelial morphology to a spindle-like shape. In addition, the HBx protein significantly increased the expressions of N-cadherin, vimentin and fibronectin, but decreased the expressions of E-cadherin and alpha-catenin in SMMC-7721 cell. The cellular invasion assay indicated that transfection of HBx drastically enhanced the invasive potential of SMMC-7721 cell. CONCLUSION: HBx protein may induce EMT and enhance the invasive potential of SMMC-7721 cell.

Liver resection for benign hepatic lesions: a retrospective analysis of 827 consecutive cases.

AIM: To analyze the operative and perioperative factors associated with hepatectomy of benign hepatic lesions. METHODS: A total of 827 consecutive cases of benign hepatic lesion undergoing hepatectomy from January 1986 to December 2005 in the Chinese PLA General Hospital were investigated retrospectively according to their medical documentation. RESULTS: The effect of operative and perioperative factors on the outcome of patients were analyzed. Of the 827 cases undergoing hepatectomy for more than 3 liver segments accounted for 22.1%, 316 (38.21%) required transfusion of blood products during operation. The average operating time was 220.59 +/- 109.13 min, the average hospital stay after operation was 13.55 +/- 9.38 d. Child-Pugh A accounted for 98.13%. The postoperative complication rate was 13.54% and the in-hospital mortality rate was 0.24%. Multivariate analysis showed that operating time (P = 0.004, OR = 1.003) and albumin value (P = 0.040, OR = 0.938) were the independent predictors of morbidity and indicated that operating time, blood transfusion, complication rate, and LOS had a trend to decrease. CONCLUSION: Hepatectomy for benign hepatic lesions can be performed safely with a low morbidity and mortality, provided that it is carried out with optimized perioperative management and an innovative surgical technique.

[Liver resection in hepatolithiasis: 20-year's evolution].

OBJECTIVE: To analyze operative and perioperative factors associated with hepatectomy in hepatolithiasis. METHODS: 245 consecutive hepatolithiasis patients undergoing hepatectomy from January 1986 to December 2005 at Chinese People's Liberation Army General Hospital were investigated retrospectively according to medical documentation. RESULTS: Hepatolithiasis accounted for 29.6% (245/827) in all benign liver diseases treated with hepatectomy during this time period. There were 88 cases in male and 157 cases in female, the average age was (46.9 +/- 11.3) years. Cases of right liver resection and hepatic segments resection were much more than that in 1963 - 1985. Blood transfusion during operation was given in 45.3% of cases. Complication incidence was 16.3%, with infection 3.3% and bile leakage 2.4%. Length of stay after operation was (15.7 +/- 9.2) days. Perioperative mortality rate was 0.4% (1/245). CONCLUSIONS: Individualized hepatectomy is the important surgical treatment of hepatolithiasis. Hepatectomy can be performed safely with low mortality and low complication incidence, provided that it is carried out with optimized perioperative management and innovative surgical technique.