Liu Ca

Ultrasound-targeted microbubble destruction mediated herpes simplex virus-thymidine kinase gene treats hepatoma in mice

ObjectiveThe purpose of the study was to explore the anti-tumor effect of ultrasound -targeted microbubble destruction mediated herpes simplex virus thymidine kinase (HSV-TK) suicide gene system on mice hepatoma.MethodsForty mice were randomly divided into four groups after the models of subcutaneous transplantation tumors were estabilished: (1) PBS; (2) HSV-TK (3) HSV-TK+ ultrasound (HSV-TK+US); (4) HSV-TK+ultrasound+microbubbles (HSV-TK+US+MB). The TK protein expression in liver cancer was detected by western-blot. Applying TUNEL staining detected tumor cell apoptosis. At last, the inhibition rates and survival time of the animals were compared among all groups.ResultsThe TK protein expression of HSV-TK+MB+US group in tumor-bearing mice tissues were significantly higher than those in other groups. The tumor inhibitory effect of ultrasound-targeted microbubble destruction mediated HSV-TK on mice transplantable tumor was significantly higher than those in other groups (p < 0.05), and can significantly improve the survival time of tumor-bearing mice.ConclusionUltrasound-targeted microbubble destruction can effectively transfect HSV-TK gene into target tissues and play a significant inhibition effect on tumors, which provides a new strategy for gene therapy in liver cancer.

Astragaloside IV Protects Against Ischemia Reperfusion in a Murine Model of Orthotopic Liver Transplantation

AIM The aim of this study was to determine whether Astragaloside IV (AST-IV) inhibited the transcriptional activity of nuclear factor-κB (NF-κB), attenuating liver transplant ischemia-reperfusion injury (IRI). METHODS Sprague Dawley (SD) rats were randomly divided into 2 groups: donors given AST-IV (1.5 mL; 100 μg/mL, intravenous [IV]) 1 hour before surgery (n = 32), versus controls treated with 1.5 mL physiological saline (n = 32). Orthotropic liver transplantation was performed according to the Kamada technique. Eight animals in each group were followed for seven days after surgery to assess survival. The remaining hosts in each group were divided into 3 subgroups (n = 8) to be examined at 3, 6, and 24 hours after portal vein reperfusion. We analyzed levels of alanine aminotransferase (ALT), tumor necrosis factor (TNF)-α, and NF-κB transcriptional activity and performed a morphological study of liver tissues, NF-κB, and glucocorticoid receptor (GR) expression in Kupffer cells (KCs). RESULTS Pretreatment with AST-IV significantly improved survival rates and liver function, attenuating liver parenchymal cell damage by down-regulating TNF-α levels and NF-κB expressions, inhibiting NF-κB transcriptional activity, up-regulating GR expression. CONCLUSION AST-IV attenuated hepatic IRI by inhibiting NF-κB transcriptional activity. The mechanism may relate to up-regulation of GR expression.

Suppression of BCRP expression and restoration of sensitivity to chemotherapy in multidrug-resistant HCC cell line HEPG2/ADM by RNA interference.

BACKGROUND/AIMS Breast cancer resistance protein (BCRP) is an ATP-binding cassette multidrug transporter that confers resistance to various anticancer drugs like Adriamycin. Overexpression of BCRP confers multidrug resistance (MDR) in hepatocellular carcinoma cells and is a frequent impediment to successful chemotherapy. METHODOLOGY We evaluated a new approach using RNA interference for the specific knockdown of BCRP in hepatocellular carcinoma cells. To overcome the BCRP-mediated atypical multidrug drug resistance, one small interfering RNA construct (RNAi) targeting one special region of BCRP gene were designed to inhibit the atypical MDR expression by transfecting them into HepG2/ADM cell lines. RESULTS We found that the overexpression of BCRP gene was suppressed efficiently by the introduction of small interfering RNA, which caused sequence specific gene silence. The level of BCRP mRNA reduced to 22.55% after transfected by pSUPER-BCRPs compared with those of the controls. Similarly, the level of BCRP decreased too. Furthermore, the sensitivity to Adriamycin of pSUPER-BCRPs-treated HEPG2/ADM cells is increased 3.55-fold compared to their control (p<0.05). The relative reverse rate of HepG2/ADM cell to Adriamycin is 72.2%. CONCLUSIONS These data indicated that pSUPER-BCRPs could modulate MDR and may present a new approach to overcome BCRP-mediated drug resistance in HEPG2/ADM cells. It may reverse multidrug resistance phenotype and therefore provide promising therapeutic modalities in the treatment of hepatocellular carcinoma.

Diagnosis and cure experience of hepatolithiasis-associated intrahepatic cholangiocarcinoma in 66 patients.

BACKGROUND The management of hepatolithiasis combined with intrahepatic cholangicarcinoma (IHHCC) remains a challenge due to poor prognosis. The aim of this study was to summarize our diagnosis and cure experience of IHHCC over the recent 10 years. METHODS From January 1996 to January 2006, 66 patients with IHHCC were reviewed retrospectively. RESULTS Of the 66 patients, 52 underwent surgical resection (radical resection in 38 and palliative in 14) and 8 patients abdominal exploration, while the other 6 cases received endoscopic retrograde biliary internal drainage and stent implantation. In this series, correct diagnosis of advanced stage was made during operation in 8 cases (8/60, 13.3%) and all of them (underwent unnecessary abdominal exploration, among them the positive rate of CA19-9 was 100%, and the positive rate of CEA was 87.6% (7/8), incidence rate of ascites was 100% and short-term significant weight loss was 100%, with median overall survival of only 4 months. CONCLUSION Radical resection is mandatory for IHHCC patient to achieve long-term survival, the CT and MR imaging features of IHHCC being concentric enhancement. Patients with IHHCC have significant higher CA199 and significant higher CEA and short-term significant weight loss and ascites should be considered with advanced stage of IHHCC and unnecessary non-therapeutic laparotomies should be avoided.

High-intensity focused ultrasound combined with herpes simplex virus thymidine kinase gene-loaded ultrasound-targeted microbubbles improved the survival of rabbits with VX2 liver tumor

To explore the anti‐tumor effect of high‐intensity focused ultrasound (HIFU) combined with herpes simplex virus thymidine kinase (HSV‐TK) gene‐loaded ultrasound‐targeted microbubbles on VX2 rabbit liver tumors.

NBD peptides protect against ischemia reperfusion after orthotopic liver transplantation in rats.

BACKGROUND NBD (NEMO binding domain) peptides could selectively inhibit the inflammation induced NF-κB activity, while sparing the protective functions of basal NF-κB activity. The aim of this study was to determine whether NBD peptides inhibited the transcriptional activity of nuclear factor-κB (NF-κB) during liver transplant ischemia-reperfusion injury (IRI), without affecting its basal function. MATERIALS AND METHODS Sprague Dawley (SD) rats were performed orthotropic liver transplantation according to the Kamada technique. Donors were given NBD peptides (8 mg/kg, intraperitoneal) 2 h before surgery (n = 24) and the controls were treated with the same volume of physiologic saline (n = 24). An additional 16 animals in normal condition (did not undergo any surgery) were also divided into two groups and given the same treatment as above to assess the effect of NBD peptides on basal function. We analyzed levels of alanine aminotransferase (ALT), tumor necrosis factor (TNF-α), IKK (IκB kinase) complex phosphorylation, IκBα degradation, NF-κB transcriptional activity, apoptosis, and performed a morphologic study of liver tissues at 3, 6, and 24 h after portal vein reperfusion and in normal condition (n = 8). RESULTS Pretreatment with NBD peptides significantly improved liver function, attenuating liver parenchymal cell damage, apoptosis by down-regulating TNF-α level, inhibiting IKK complex phosphorylation, IκBα degradation, and NF-κB transcriptional activity, but had no effect in normal condition. CONCLUSION NBD peptides attenuated hepatic IRI by preventing NF-κB activation, without affecting basal NF-κB activity.

Bone marrow stromal cells attenuate LPS-induced mouse acute liver injury via the prostaglandin E 2-dependent repression of the NLRP3 inflammasome in Kupffer cells

The nucleotide-binding and oligomerization domain-like receptor 3 (NLRP3) inflammasome participates in the pathogenesis of acute liver injury during sepsis. Bone marrow mesenchymal stem cells (BMSCs) attenuate sepsis through prostaglandin E 2 (PGE2) by increasing the interleukin-10 (IL-10) production of macrophages; moreover, NLRP3 inflammasome assembly is effectively regulated by IL-10 during infection. Whether BMSCs have an effect on the activation of the NLRP3 inflammasome and its underlying mechanism is unclear. Administering of BMSCs to mice or KCs after LPS stimulating have improved liver function and reduced activation of NLRP3 inflammasome in KCs. The beneficial effect of BMSCs was enhanced by over-expression of PGE2 and eliminated by silence of PGE2. Additionally, The IL-10 levels in the serum and supernatant were increased by given BMSCs and further increase by PGE2 over-expressed BMSCs, but decreased markedly by PGE2 silenced BMSCs. Furthermore, extracellular signal-regulated kinase 1 (ERK1) inhibitor reduced IL-10 production in KCs and blocked the inhibitory effect of PGE2 on the activation of the NLRP3 inflammasome. Our data reveal a novel mechanism of BMSC-mediated suppression of the activation of KCs through the secretion of PGE2 by BMSCs, which promotes KCs to secrete IL-10, leading to the inhibition of the NLRP3 inflammasome in KCs.

LXRα represses LPS-induced inflammatory responses by competing with IRF3 for GRIP1 in Kupffer cells

Liver X receptors (LXRs) in the nucleus play important roles in lipid metabolism and inflammation. The mechanism of LXR regulation of the LPS-induced Toll-like receptor 4 (TLR4) inflammatory signaling pathway remains to be elucidated. C57/BL6 mice were randomly divided into four groups: control, T0901317 (a LXRs agonist), LPS and T0901317+LPS. Additionally, Kupffer cells isolated from male C57/BL6 mice were divided into the same four groups. A decreased amount of inflammatory cells infiltrated the portal areas and the hepatic sinusoids in the livers of mice in the T0901317+LPS group than in those of mice in the LPS group. In the T0901317+LPS group, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and tumor necrosis factor alpha (TNF-α) were lower, while the serum level of interleukin-10 (IL-10) was higher. In vitro, Kupffer cells pretreated with T0901317 for 24h presented reduced TNF-α, interferon-beta (IFN-β) and interleukin-1 beta (IL-1β) levels, while the IL-10 level increased; however, the mRNA and protein expression levels of interferon regulatory factor 3 (IRF3) and glucocorticoid receptor-interacting protein 1 (GRIP1) were not significantly reduced. The co-IP data illustrated that LXRα bound to GRIP1 specifically in the T0901317+LPS group, while less IRF3 was bound to GRIP1 in the T0901317+LPS group than in the LPS group. Furthermore, the DNA-binding activity of NF-κB was decreased by pretreating Kupffer cells with T0901317 for 24h. These results suggest that activated LXRα competes with IRF3 for GRIP1 binding, thus repressing IRF3 and NF-κB transcriptional activity and inhibiting the inflammatory response initiated by LPS in Kupffer cells.