[Efficacy Evaluation of Three Endoscopic Therapies of Isolated Gastric Varices with Modified Tissue Adhesive].

Objective: To evaluate the efficacy of three endoscopic therapies of isolated gastric varices (IGV) with modified tissue adhesive. Methods: A retrospective analysis was conducted with the clinical data of 73 IGV patients who were treated between January 2008 and December 2019 at Beijing Ditan Hospital. Patient clinical data on age, sex, etiology, biochemistry findings, Child-Pugh classification, the type of spontaneous shunt, preoperative bleeding history, and the presence or absence of liver cancer were collected. The three therapies evaluated were endoscopic intravenous injection of tissue glue combined with lauromacrogol, endoscopic clip-assisted intravenous injection of tissue glue combined with lauromacrogol, and endoscopic clip and LOOP-assisted intravenous injection of tissue glue combined with lauromacrogol. Their respective clinical treatment outcomes, including ectopic embolism rate, survival rate, rebleeding rate, amount of lauromacrogol and tissue glue used, the number of endoscopic clips used, and the number of times of the procedure the patient underwent, were evaluated. Results: In the patient baseline data, Child-Pugh grade, preoperative thrombus formation, and the presence or absence of liver cancer, showed significant difference between the three therapies ( P<0.05). There was no significant difference in the rates of ectopic embolism among the three methods ( P>0.05), but no ectopic embolism occurred after endoscopic clip-assisted intravenous injection of tissue glue combined with lauromacrogol, or after endoscopic clip and LOOP-assisted intravenous injection of tissue glue combined with lauromacrogol. There was no significant difference in the survival rate, the rebleeding rate, amount of lauromacrogol and tissue glue used for the three therapies, but there was significant difference in the number of endoscopic clips used and the number of times the procedure was conducted within one year ( P<0.05). Conclusion: The two endoscopic therapies of intravenous injection of modified tissue glue, one assisted by clip and the other assisted by clip and LOOP, can help reduce the number of procedures IGV patients undergo within one year.

Unsedated transnasal upper gastrointestinal endoscopy has favorable diagnostic effectiveness, cardiopulmonary safety, and patient satisfaction compared with conventional or sedated endoscopy.

BACKGROUND: To assess the diagnostic effectiveness, cardiopulmonary safety, and patient comfort of transnasal endoscopy (TNE), compared with conventional endoscopy (CES) and sedated endoscopy (SES), and to compare procedural risks and patient satisfaction/preference. METHODS: In this prospective, randomized, and controlled protocol, eligible patients (n = 397) in an outpatient clinic were randomized to CES (n = 133), SES (n = 134), or unsedated TNE (n = 130) due to upper gastrointestinal (GI) complaints. Patients were continuously monitored for systolic/diastolic blood pressure (SBP/DBP), pulse rate (PR), and SpO(2) throughout the endoscopy. All subjects (n = 392) completing their assigned endoscopy were asked to evaluate endoscopy satisfaction, pain, and nausea/vomiting on visual analog scales. Patient preference for the assigned endoscopy was assessed against previous endoscopy experience or by willingness to repeat the assigned endoscopy. RESULTS: Endoscopic outcomes for the esophagus, stomach, and duodenum were comparable among the three groups. SBP/DBP and PR were more stable in patients undergoing TNE than in those undergoing CES or SES, while SpO(2) remained stable and above 95% among all three groups. Patients were more satisfied with TNE than with CES and experienced less pain and nausea/vomiting. Patients exhibited a high preference for SES, whereas 67.6% of patients who previously underwent SES and were randomly assigned to TNE were willing to undergo TNE again. CONCLUSIONS: TNE has comparable diagnostic effectiveness to CES and SES, but is less stressful on cardiopulmonary function, indicating that TNE is a more comfortable, preferred, and cost-effective endoscopic technique than CES and SES.

[Effect of baicalin on liver fatty acid binding protein in oxidative stress model in vitro].

OBJECTIVE: To investigate the effect of baicalin on liver fatty acid binding protein in oxidative stress model in vitro. METHODS: (1) Cellular oxidative stress in vitro was induced by incubating cells with 400µmol/L hydrogen peroxide (H2O2) for 20 minutes at 37 degrees C in the dark. After Chang liver cell line was treated with different dose of baicalin for 24, 48 and 72 hours. MTT assay was employed to detect cell viability, and then the hydrogen peroxide (TC50) of the different dose of baicalin was calculated. (2) Based on MTT assay, cells were treated with three different doses of baicalin (25, 50, 100 µmol/L) for 24 and 48 hours before being exposed to 400 µmol/L H2O2 for 20 minutes at 37 degrees C. Then, reactive oxygen species (ROS) assay and activity assays of superoxide dismutase (SOD) and reduced glutathione hormone (GSH) were evaluated. (3) Realtime PCR and Western blotting were applied to explore the influence of baicalin on the expression level of L-FABP. (4) One-way ANOVA was used for results statistical analysis. RESULT: (1) MTT assay showed baicalin treatment at 25, 50, 100 µmol/L for 24 and 48 hours was feasible (83.60% ± 3.47%, 72.36% ± 2.18%, 70.16% ± 2.04% for 24 hours; 84.93% ± 3.11%, 76.16% ± 2.45%, 72.72% ± 2.31% for 48 hours, P > 0.05, F = 386.24, 475.92 respectively). Meanwhile, we found by the linear regression model that the median toxic concentration of baicalin for 48 hours was 170.6 µmol/L, and the median toxic concentration of baicalin for 24 hours was 153.2 µmol/L. (2) ROS assay showed dichlorofluorescin in all baicalin-treated cells after stress was significantly reduced (37.0 ± 3.30, 22.90 ± 3.84, 29.60 ± 2.52 for 24 hours respectively, P < 0.05, F = 70.06; 35.77 ± 2.35, 21.80 ± 3.10, 23.87 ± 1.98 for 48 hours respectively, P < 0.05, F = 110.92) as compared with the H2O2-treated cells. Moreover, 50 µmol/L baicalin treatment for 48 hours was the optimal condition against ROS generation (21.80 ± 3.10, P < 0.01, F = 110.92). Furthermore, the activities of intracellular SOD and GSH was increased significantly (51.53 ± 1.91 µg/mg for SOD, P < 0.05, F = 93.81; 49.85 ± 1.45 U/mg for GSH, P < 0.05, F = 92.51). (3) Although realtime PCR analysis indicated 50 µmol/L baicalin treatment for 48 hours could have no changes of the level of L-FABP expression under the oxidative stress condition, western blotting analysis indicated 50 µmol/L baicalin treatment for 48 hours could increase up to about 80% for the level of L-FABP expression. CONCLUSION: Baicalin was suggested to be able to enhance both L-FABP expression and activity of intracellular SOD and GSH, and therefore protected hepatocytes from oxidative stress.

[The expression and significance of osteopontin in the development of nonalcoholic fatty liver fibrosis in rats].

OBJECTIVE: To investigate the changes of osteopontin (OPN) in the liver tissues during nonalcoholic fatty liver fibrosis in rats and to explore the effect of OPN in the development of nonalcoholic fatty liver fibrosis. METHODS: Fifty-six male Wistar rats were randomly divided into a control group (8 rats) and a high-fat diet group. The high-fat diet group was divided into 6 subgroups (8 rats in each subgroup) with high-fat feedings for 4, 8, 12, 16, 20 or 24 weeks. Conventional histochemical, HE, Masson-trichrome and immunohistochemical staining for alpha-smooth muscle actin (a-SMA) were performed with the liver histological preparations. The expression of OPN was detected with reverse transcription and polymerase chain reactions and Western blot. RESULTS: Levels of OPN in liver tissues in rat nonalcoholic fatty liver fibrosis induced by high-fat diet were significantly increased over those in the control group (F=7.15, P less than 0.01). OPN expressions were closely correlated with a-SMA and nonalcoholic fatty liver fibrosis, and correlation coefficients of the two groups were 0.94 and 0.82, and both P values were less than 0.01. CONCLUSION: Expression of OPN increases dramatically in the livers during the development of nonalcoholic fatty liver fibrosis, and OPN may play an important role in this event.

[The significance and effects of liver X receptor alpha in nonalcoholic fatty liver disease in rats].

OBJECTIVE: To explore liver X receptor alpha (LXR alpha) gene changes and their significance in nonalcoholic fatty liver disease (NAFLD) in rats. METHODS: A rat model of nonalcoholic fatty liver disease was produced with a fatty diet regime (feeding group, FG). Rats fed with normal diet served as controls (CG). The mRNA and protein expressions of LXR alpha in liver tissues were detected by reverse transcription and polymerase chain reaction (RT-PCR) and Western blot. RESULTS: The concentration of free fatty acid (FFA) in the sera of GF rats started to increase to 0.33 mmol/L after 4 weeks of fat diet feeding, while the FFA of the CG was just 0.24+/-0.03 mmol/L, and the difference was significant (P<0.05). The concentration of ALT and AST in sera of the FG rats started to increase to 75.8 U/L and 138.9 U/L at the 8th week, much higher than those of the CG (P<0.01), and at the 12th week they increased further (P<0.01). Meanwhile, the mRNA and protein expressions of LXR alpha at the 2nd week was significantly increased to 0.62 (P>0.01) and its peak was reached at the 12th week (P<0.01). There was a significant positive correlation between the expression of LXR alpha and the degree of NAFLD. CONCLUSION: The changes of LXR alpha gene are closely related to the development of nonalcoholic fatty liver disease.

Lutein prevents alcohol-induced liver disease in rats by modulating oxidative stress and inflammation.

OBJECTIVE: Oxidative stress and inflammation play an important role in pathogenesis of alcohol-induced liver injury. The present study was designed to investigate the protective role of Lutein against alcohol-induced liver injury. TREATMENT: Wistar rats weighing 150-200 g were divided into 3 groups, control, EtOH treatment, Lutein followed by EtOH treatment. Ethanol-treated rats received EtOH [5 g/kg body weight] by gavage every 12 hours for a total of 3 doses. For Lutein pre-treatment, Lutein at a dose of 40 mg/kg was dissolved in the EtOH and gavaged 30 mins before EtOH treatment. METHODS: Oxidative stress markers-(reactive oxygen species, lipid peroxidation, protein carbonyls and sulfhydryls content), liver markers (ALT, AST, ALP and LDH) were determined. Antioxidant enzyme activities and its master regulator Nrf-2 expression were analyzed. Further, inflammatory proteins NF-κB, COX-2, iNOS and inflammatory cytokines (TNF-α, MCP-1, IL-1ß, IL-6) were analyzed. RESULTS: The results showed significant decrease in oxidative stress markers and liver markers in the lutein pre-treatment. Lutein treatment down regulated inflammatory proteins and cytokines with concomitant up regulation in Nrf-2 levels and antioxidant enzymic activities. CONCLUSION: The present study showed that Lutein treatment exerted potent antioxidant and anti-inflammatory property and offered significant cytoprotection against alcohol-induced liver injury.

Involvement of ß-catenin in matrine-induced autophagy and apoptosis in WB-F344 cells.

Matrine, one of the main components extracted from Sophora flavescens, has exhibited pharmacological effects on the differentiation in rat liver oval cells. However, its function and mechanism have not yet been fully elucidated. To further investigate them, an in vitro model was established using a rat liver oval cell line called WB-F344 and treated with matrine. Initially, a significant increase in the number of monodansylcadaverine-positive cells and in the levels of microtubule-associated protein 1A/1B-light chain 3 (LC3)-II, which is a specific marker for detecting autophagy, was observed in matrine-treated cells. This indicated that autophagy was stimulated by matrine, which was further confirmed by transmission electron microscopy. Additionally, the apoptotic oval cells were easily detected under matrine treatment using an Annexin-V-fluorescein isothiocyanate/propidium iodide assay, indicating that autophagy and apoptosis were synchronously induced by matrine. A decrease in B-cell lymphoma (Bcl-2) mRNA expression, but an increase in Bcl2-associated X protein (Bax) mRNA expression were observed in matrine-treated cells, which led to an upregulation of the Bax/Bcl-2 ratio, a molecular marker for determining the extent of apoptosis. Next, the molecular mechanism of matrine-induced autophagy and apoptosis was analyzed in WB-F344 cells. ß-catenin degradation was downregulated by matrine and rapamycin, a foregone chemical agonist of autophagy, whereas it was upregulated by 3-methyladenine, a specific inhibitor of autophagy. Additionally, ß-catenin activation induced an increase in LC3-II levels and reversed the Bax/Bcl-2 mRNA ratio under matrine treatment, whereas inhibition of ß-catenin by RNA interference induced a decrease of the LC3-II amount and of the Bax/Bcl-2 mRNA ratio. Finally, matrine treatment attenuated p53; however, with little or no change in LC3-II levels, but a decrease in ß-catenin levels occurred in WB-F344 cells upon treatment with pifithrin-α, a chemical inhibitor of p53, revealing that p53, interfering with ß-catenin, may not be involved in matrine-induced autophagy in WB-F344 cells. These results demonstrate that ß-catenin is involved in matrine-induced autophagy and apoptosis in WB-F344 cells, while ß-catenin is negatively regulated by autophagy and positively by p53, indicating that ß-catenin may be involved in the crosstalk between autophagy and apoptosis in WB-F344 cells.

Autophagy inhibition enhances etoposide-induced cell death in human hepatoma G2 cells.

Induction of autophagy usually acts as a survival mechanism of cancer cells in response to chemotherapy. However, the function and molecular mechanism of autophagy in human hepatoma cells under drug treatment is still not clear. To address this issue, we established an experimental model in which HepG2 cells were treated with etoposide, a widely used anticancer agent. We demonstrate the etoposide-induced accumulation of GFP-LC3 dots by fluorescent microscopy, the up-regulation of LC3-II protein expression by Western blotting and the increased number of autophagic vacuoles by electron microscopy, confirming the activation of autophagy by etoposide in HepG2 cells. Inhibition of autophagy by either 3-methyladenine (3MA) or beclin-1 small interfering RNA enhanced etoposide-induced cell death. Furthermore, activation of p53 and AMPK was detected in etoposide-treated cells and inhibition of AMPK triggered apoptosis through suppression of autophagy. On the other hand, inactivation of p53 promoted cell survival through augmentation of autophagy. Collectively, these findings indicate that etoposide-induced autophagy promotes hepatoma cell adaptation and survival, and that autophagy inhibition improves the chemotherapeutic effect of etoposide. Moreover, AMPK activation is clearly associated with etoposide-induced autophagy. We conclude that manipulation of AMPK may be a promising approach of adjuvant chemotherapy for hepatocellular carcinoma.