Advances in mechanisms of immunity enhancement induced by thermal ablation therapy in hepatocellular carcinoma

Image-guided tumor thermal ablation plays a key role in the management of hepatocellular carcinoma. Thermal ablation for hepatic tumors not only generates directly killing effect on liver tumors, but also improves the level of anti-tumor immunity of the host. Thermal ablation potentially affects the anti-tumor immunity in the following ways: (1) causing necrosis that is a source for intracellular antigens, and also inducing local inflammation, which further stimulates the antigen-presenting cells; (2) removing the tumor burden; (3) undergoing the process of heat shock in surviving tissue, leading to increase in the expression of heat shock proteins; (4) activating and enhancing tumor-specific T-cell responses. This review discusses these possible mechanisms.     

Inhibition of Ang-2 and RGS-5 expression by nanogold results in normalization of vascellum in hepatic tumor

AIM: To observe the vascular normalization effect of nanogold on hepatic tumor by inhibiting angiopoietin-2 (Ang-2), regulator of G-protein signaling-5 (RGS-5) during certain time window. METHODS: H22 cells, the hepatocellular cancer cell line, were subcutaneously injected into the right armpits of 48 BALB/c nude mice. When the size of transplanted tumor reached 3~4 mm, the mice were divided into 2 groups: 36 mice in experiment group and 12 mice in control group. The mice in experimental group underwent injection of nanogold into the tumor once a day, and the mice in control group were injected with normal saline. After continuous treatment with nanogold for 3 days, 7 days and 11 days, the mice were sacrificed, the liver tumors were taken out to measure the size and weight. The expression of Ang-1,Ang-2 and RGS-5 in the tumor was detected by the method of immunohistochemical staining. The normalizing shapes of tumor vessels and the pericytes were observed under electronic microscope. RESULTS: With nanogold treatment for 3 days, 7 days and 11 days, the positive rates of Ang-1 were 16.7％, 50.0% and 16.7%, respectively. The positive rates of Ang-2 were 33.3%, 16.7% and 41.7%, respectively. The expression of Ang-1 in experiment group was higher than that in control group, especially at the 7th day in experiment group. The expression of Ang-2 in experiment group was lower than that in control group (58.3%), especially at the 7th day in experiment group. With nanogold treatment for 3 days, 7 days and 11 days, the positive rates of RGS-5 were 33.3%, 16.7% and 50.0%, respectively. The immature pericyte coverage indexes (IMPI) were 19.6%±4.3%, 32.5%±7.9% and 41.2%±9.1% respectively. At the 7th day in experiment group, the positive rates of RGS-5 and IMPI were lower than those in other experiment groups and control group. After treated with nanogold for 7 days, the pericytes in the parietal wall of the blood vessels in the tumor showed the tendency to grow normally in morphology and were completely covered by endothelial cells. However, the pericytes in the parietal wall of the blood vessels in control group showed differences in size, impaired integrity and only a few of the pericytes covered by endothelial cells. CONCLUSION: During the time window of nangold treatment for 7 days, the chemical can normalize the blood vessels in liver cancer by inhibiting the expression of Ang-2 and RGS-5.     

Effect of siRNA-mediated Smad3 silence on proliferation and apoptosis in activated hepatic stellate cells

AIM: To investigate the effects of siRNA-mediated Smad3 silence on proliferation and apoptosis in activated hepatic stellate cells (HSCs).METHODS: HSCs-T6 cells were divided into 3 groups: blank group, negative control group and siRNA-Smad3 transfection group. The siRNA-Smad3 was transfected into HSCs-T6 cells. At different time points after transfection, cell proliferation was measured by CCK-8, cell apoptosis was detected by flow cytometry, and protein levels of P53 and Bcl-2 were determined by immunocytochemistry.RESULTS: HSCs proliferation was significantly inhibited at the time points of 24 h, 48 h and 72 h after transfection. Meanwhile, the apoptosis of HSCs was significantly increased in siRNA-Smad3 transfection group (P<0.01). Compared to the control cells, the protein expression of P53 was significantly increased while Bcl-2 protein was significantly decreased 48 h after transfection in siRNA-Smad3 transfection group (P<0.01).CONCLUSION: The siRNA-mediated Smad3 silence significantly inhibits HSCs proliferation and induces apoptosis by up-regulating the P53 expression and down-regulating the Bcl-2 expression in HSCs.     

Supportive effects of human AGM region and fetal liver stromal cells on differentiation of murine embryonic stem cells into hematopoietic stem cells and in vivo hematopoietic reconstitution

AIM: To investigate the differentiation of murine embryonic stem cells (ESCs) into hematopoietic stem cells (HSCs) by the supportive effects of human aorta-gonad-mesonephros (AGM) region and fetal liver (FL) stromal cells.METHODS: E14 ESCs were induced into embryoid body (EB) first. Then the cells from EB were further co-cultured with human AGM region and FL stromal cells in non-contact system. On day 6, the cells derived from EB were collected for Sca-1⁺c-Kit⁺ cell analysis by flow cytometry, colony forming unit (CFU) assay and teratoma formation checking. BALB/c female mice conditioned with lethal dose of γ-ray irradiation were transplanted with EB cells from different culture systems. The survival rates, engraftment of donor cells, reconstitution of hematopoietic were monitored.RESULTS: Sca-1⁺c-Kit⁺ cells in EB cells co-cultured with human AGM region and FL stromal cells had the value of (21.96±2.54)%, and the total CFU was as (520±52)/10⁵ cells, which were statistically greater than those in EB cells only cultured with human AGM region stromal cells (P＜0.05). No teratoma was found in NOD-SCID mice after subcutaneous injection of EB cells co-cultured with human AGM region and FL stromal cells. In BALB/c female mice transplanted of EB cells co-cultured with human AGM region and FL stromal cells, the survival rate was 77.8%, and the peripheral blood cell count was obviously improved on day 14. PCR results showed the recipients all had sry gene copies from donor in bone marrow. The recipient mice transplanted with EB cells only cultured with human AGM region stromal cells all died within 15 days.CONCLUSION: Stromal cells from human AGM region and FL enhance the directed differentiation of ESCs into HSCs which can reconstruct hematopoiesis in vivo.     

Role of glucocorticoid receptor in hepatic injury in early stage after severe multiple injury

AIM: To investigate the functions of GR in the course of hepatic secondary injury after severe multiple injury. METHODS: Rat model was produced by adopting severe thoracic impact injury accompanied with mono-side femur fracture, and glucocorticoid receptor was blocked before severe multiple injury. Hepatic macropathology and alterations under light microscope were examined. Maximal binding volume of glucocorticoid receptor (GR) in hepatic tissue was assayed by radio-ligand binding assay and protein content was assayed by Western blot. RESULTS: Maximal binding volume and protein content of GR were gradually decreased in hepatic tissue after severe multiple injury, obviously lower than that in normal control at 4 h after trauma (P＜0. 01), and the lowest value was at 12 h after trauma. Maximal binding volume was 12. 9% of normal control (P＜0. 01), and protein content was 21. 9% of normal control (P＜0. 01). In addition, no significant pathological alteration in liver was found after trauma. However, the pathological analysis showed that the administration of GR blocker before severe multiple injury could cause severe hepatic congestion and infiltration of inflammatory cells in hepatic sinusoid in a dose-dependent manner. CONCLUSION: GR insufficiency could cause secondary hepatic injury in early stage after severe multiple injury, implying an important role of GR in injury and anti-injury mechanism of hepatic tissue cells.     

Effect of intestinal endotoxemia on hepatic energy metabolism in acute liver failure

AIM:To study the effect of intestinal endotoxemia(IETM) on hepatic energy metabolism in acute liver failure. METHODS:Intoxication by thioacetamide (TAA) was used to establish rat model of acute liver injury.Ketone body(acetoacetate and β-hydroxybutyrate) in arterial blood and ATP content of hepatocellular mitochondria were determined by using enzymatic fluorimetric micromethod.Colectomy was adopted in observing the changes in plasma endotoxin content and serum alanine aminotransferase (ALT) activity. RESULTS:In the TAA group,plasma endotoxin content and serum ALT activity were all significantly higher than those in the control group(P＜0.01),arterial ketone body ratio of acetoacetate to β-hydroxybutyrate (AKBR) decreased below 0.4,total ketone body in arterial blood was significantly lower than that in the control group(P＜0.01).In the TAA+colectomy group,there was no endotoxemia to be found,ATP content of hepatocellular mitochondria was significantly higher than that in the TAA group(P＜0.01), though serum ALT activity was higher than that in the control group(P＜0.05),but significantly lower than that in the TAA group(P＜0.01). CONCLUSION:IETM played a key role in the occurrence of acute liver failure,hepatic dysfunction might be caused by IETM through damaging hepatic energy metabolism.     

DETECTION OF A LIVER TUMOR IN A BEAGLE DOG USING SINGLE PHOTON EMISSION COMPUTED TOMOGRAPHY

This case report describes the detection of a liver tumor in an 11–year-old female Beagle dog using single photon-emission computed tomography (SPECT). A mass was palpated in the cranial abdomen during removal of recurrent mammary tumors. A conventional nuclear medicine liver scan, following injection of ^99mTc-sulfur colloid, was equivocal. SPECT was then performed in a second procedure, using a simple system that rotated the dog before a gamma camera. Transverse plane images clearly showed a spheric space-occupying lesion, found at gross necropsy and confirmed histologically as a hepatoma. Discrete liver tumors may be detected more readily and their size determined more accurately using SPECT than with conventional nuclear medicine approaches.     

Association of Helicobacter with cholangiohepatitis in cats

Infection with Helicobacter spp. is increasingly linked with hepatobiliary inflammation and neoplasia in people and in a variety of animals. We sought to determine if Helicobacter species infection is associated with cholangiohepatitis in cats. Deoxyribonucleic acid was extracted from tissue blocks from cats with cholangiohepatitis (32), noninflammatory liver disease (13), and cats with normal liver histology (4). Deoxyribonucleic acid was polymerase chain reaction-amplified with 2 sets of Helicobacter genus-specific primers, gel purified, and sequenced. Polymerase chain reaction-positive hepatic tissue was further examined with Steiner's stain, immunocytochemistry for Helicobacter species, and eubacterial fluorescent in situ hybridization. Gastric tissues of cats with known Helicobacter infection status served as controls for deoxyribonucleic acid extraction and sequence comparison. Helicobacter species were detected in 2/32 cats with cholangiohepatitis, and 1/17 controls. Sequences had 100% identity with Helicobacter species liver, Helicobacter pylori, and Helicobacter fenelliae/cinaedii in a cat with suppurative cholangitis, Helicobacter species liver, Helicobacter pylori, and Helicobacter nemistrineae in a cat with mild lymphocytic portal hepatitis, and Helicobacter bilis in a cat with portosystemic vascular anomaly. In contrast, sequences from gastric biopsies showed highest homology (99-100%) to "Helicobacter heilmannii," Helicobacter bizzozeronii, Helicobacter felis, and Helicobacter salomonis. Fluorescent in situ hybridization revealed a semicurved bacterium, with Helicobacter-like morphology, in an intrahepatic bile duct of the cat with suppurative cholangitis. This study has identified Helicobacter deoxyribonucleic acid in 2/32 cats with cholangiohepatitis and 1/13 cats with noninflammatory liver disease. Deoxyribonucleic acid sequences of hepatic Helicobacter species were distinct from those found in the stomach and are broadly consistent with those identified in cat intestine and bile, and hepatobiliary disease in people and rodents.     

Fall panicum (Panicum dichotomiflorum) hepatotoxicosis in horses and sheep

BACKGROUND: Fourteen horses at a boarding stable in Virginia were diagnosed with hepatic disease and locally grown hay was implicated as the cause. HYPOTHESIS: Panicum dichotomiflorum, the predominant grass species in the hay, is hepatotoxic to horses. ANIMALS: Naturally occurring cases were adult horses of various breeds. Two healthy adult horses and 2 healthy adult sheep were used in feeding trials. METHODS: Blood and liver specimens collected from affected animals during the outbreak were analyzed. Some of the affected animals were treated supportively; the main intervention was hay withdrawal. Feeding trials were not blinded and no treatments were provided. Blood and liver specimens were collected and analyzed throughout the trials. RESULTS: Five affected animals were euthanized, whereas the others recovered. One research horse was euthanized for postmortem examination, and the other research animals recovered after hay withdrawal. All affected animals had evidence of hepatic disease with abnormally high aspartate aminotransferase (AST), sorbitol dehydrogenase (SDH), gamma glutamyl transferase (GGT), and alkaline phosphatase (ALP) activity. Evaluation of liver biopsy specimens disclosed mild lymphocytic and histiocytic inflammation, mild vacuolar change (hydropic degeneration), prominently clumped chromatin, and necrosis of individual hepatocytes. CONCLUSIONS AND CLINICAL IMPORTANCE: Severe hepatotoxicosis developed rapidly after Panicum hay exposure. Patchy hepatocyte necrosis was observed, implicating apoptosis as the mechanism of hepatotoxicosis. Absence of fibrosis in the research animals indicates that immediate withdrawal of Panicum hay should allow all but severely affected animals to recover from acute exposure.     

不同剂量党参·淫羊藿提取物对青脚麻鸡肝组织的影响

[目的]研究不同剂量党参、淫羊藿对青脚麻鸡肝组织结构的影响。[方法]将420只14日龄青脚麻鸡随机均分7组,分别为饮用党参和淫羊藿提取物大(2.0 g/L)、中(1.0 g/L)、小(0.5 g/L)浓度组和对照组(饮用自来水)。于28,35,42,49,56日龄时,早晨空腹随机抽样12只解剖,迅速取肝脏标本,称量并计算肝系数,并于Bouns液中固定24 h后,观察肝脏组织结构的变化。[结果]2.0 g/L党参组在28、35、42日龄时肝血窦增大,血窦内枯否氏细胞增多;2.0 g/L淫羊藿组在42、49、56日龄时肝细胞索分界明显,血窦内枯否氏细胞增多,肝血窦增大。[结论]2.0 g/L的党参、淫羊藿提取物对青脚麻鸡肝组织结构有明显的促进作用。