Effect of short hairpin RNA lentivirus-mediated silencing of pleomorphic adenoma gene like-2 on proliferation, migration and invasion of hepatocellular carcinoma cells / 解剖学报

Objective To investigate the effect of short hairpin RNA(shRNA)-mediated pleomorphic adenoma gene like-2 (PALAG2) silencing on the malignant behavior of hepatocellular carcinoma cells and its mechanism. Methods Real-time PCR and Western blotting were used to detect the expression level of PLAGL2 in liver cancer tissues and adjacent tissues. Hepatoma cells MHCC97-L were cultured in vitro, the lentiviral vector plasmid PLAGL2-shRNA and control NC-shRNA were constructed, transfected into MHCC97-L cells, and stable transfected strains were selected with puromycin. CCK-8 and Transwell chamber assay detected the proliferation activity and the number of migration and invasion of MHCC97-L cells after silencing PLAGL2. Western blotting was used to detect the expression of p-PI3K and p-Akt proteins. The PI3K/ Akt signaling pathway activator was used to treat MHCC97-L cells to detect cell proliferation, migration and invasion. Results The expression of PLAGL2 was significantly increased in liver cancer tissue (P < 0. 05). Transfection of 9 strains of MHCC97-L cells with PLAGL2-shRNA could significantly reduce the expression level of PLAGL2, and the ability of proliferation, migration, and invasion of MHCC97-L cells was also weakened (P<0. 05), and the expression levels of p-PI3K, and p-Akt were inhibited (P<0. 05), PI3K/ Akt activator could obviously reverse the above phenomenon. Conclusion shRNA lentiviral vector pathway can effectively silence the expression of PLAGL2 gene in hepatocarcinoma cells. Silencing of PLAGL2 can significantly inhibit the malignant behavior of proliferation, migration and invasion of hepatocarcinoma cells, and its mechanism may be related to the inhibition of PI3K / Akt signaling pathway activation.

Baicalin protects against 17α-ethinylestradiol-induced cholestasis via Sirt1/HNF-1α/FXR pathway / 中国药理学与毒理学杂志

OBJECTIVE Baicalin is a major flavonoid component of Scutellaria baicalensis, and has been used in the treatment of liver diseases for many years. However, the role of baicalin in estrogen-induced cholestasis (EIC) remains to be elucidated. This present study explored the protective effect of baicalin against estrogen-induced liver injury and further elucidated the mechanisms involved both in vivo and in vitro. METHODS We conducted a series of experiments using 17α-ethinylestradiol (EE) induced cholestatic rats and cultured HepG2 cells. Serum, bile, and liver samples were collected for biochemical and histological analyses. Bile acid composition in liver was analyzed by LC-MS/MS. The mechanisms underlying the hepatoprotective of baicalin were investigated by RT-PCR, Western blotting analyses and immunohistochemistry. RESULTS Baicalin showed obvious hepatoprotective effects in EIC rats by reducing serum bio?markers and increasing the bile flow rate, as well as by alleviating liver histology and restoring the abnormal composition of hepatic bile acids (BAs). In addition, baicalin protected against EE induced liver injury by up-regulation of the expres?sion of hepatic efflux transporters and down-regulation of hepatic uptake transporters. Furthermore, baicalin increased the expression of hepatic BA synthase (CYP27A1) and metabolic enzymes (Bal, Baat and Sult2a1) in EIC rats. We showed that baicalin significantly inhibited hepatic inflammatory responses in EIC rats through reducing elevated levels of TNF-α, IL-1β, IL-6 and NF-κB. Finally, we confirmed that baicalin maintains BA homeostasis and alleviates inflamma?tion through Sirt1/HNF-1α/FXR signaling pathway. CONCLUSION Baicalin protects against estrogen-induced cholestatic liver injury, and the underlying mechanism involved is related to activation of the Sirt1/HNF-1α/FXR signaling pathway.

Mechanism of Calculus Bovis Sativus in inhibiting hepatocyte lipid deposition based on serum pharmacology / 中国中药杂志

The aim of this paper was to investigate the molecular mechanism of Calculus Bovis Sativus( CBS) in alleviating lipid accumulation in vitro by serum pharmacology. The CBS-containing serum of mice was obtained by serum pharmacology method to evaluate its effect on the proliferation of LO2 hepatocytes. The lipid reducing effects of CBS-containing serum through Nrf2 was evaluated by fructose-induced LO2 hepatocyte steatosis model,nuclear factor erythroid 2 related factor 2( Nrf2) agonist oltipraz combined intervention,cell oil red O staining and intracellular triglyceride( TG) content. The effects of CBS-containing serum on lipid peroxidation and hepatocytes apoptosis were evaluated by reactive oxygen species( ROS) and apoptosis assay,respectively. Real-time quantitative polymerase chain reaction( PCR) was used to detect the relative expression of lipid synthesis-related genes and apoptosis-related genes.RESULTS:: showed that CBS drug-containing serum had no significant effect on LO2 hepatocyte proliferation. As compared with the model group,CBS-containing serum could effectively reduce the formation of lipid droplets in fructose-induced LO2 hepatocytes,significantly reduce intracellular TG and ROS levels,and significantly reduce hepatocyte apoptosis rate( P < 0. 05). As compared with the model group,carbohydrate responsive element binding protein( ChREBP),sterol regulatory element binding protein-1 c( SREBP-1 c),fatty acid synthase( FAS),acetyl-CoA carboxylase 1( ACC1),stearoyl-CoA desaturase 1( SCD1),Bax and caspase-3 mRNA levels were significantly reduced in CBS drug-containing serum treatment group( P<0. 05). All of the above effects could be reversed by oltipraz.In conclusion,CBS-containing serum can significantly inhibit the fructose-induced LO2 liver fat deposition,and the mechanism may be related to reducing intracellular ROS level through the Nrf2 pathway and improving intracellular peroxidation state to reduce apoptosis.

Changes of inflammatory factors after hepatic cystic echinococcosis surgery and intervention of ulinastatin in Qinghai area / 中国血吸虫病防治杂志

Objective To observe the changes of inflammatory factors after the hepatic cystic echinococcosis surgery and explore the intervention effect of ulinastatin on postoperative inflammatory factors. Methods Sixty patients with hepatic cystic echinococcosis were selected and randomly divided into a control group and ulinastatin intervention group according to whether or not use ulinastatin. The peripheral venous blood was extracted in all the patients and the levels of IL-6, IL-8, IL-9, and IL-10 were detected by the ELISA method on the day before operation, 1 day, 3 days, 5 days and 7 days after operation, respectively. The data was statistical analyzed to detect the relationships between/among the inflammatory factors mentioned above and ulina-statin and time. Results The variation of the levels of IL-6, IL-8, IL-9, and IL-10 were changed by the intervention of ulina-statin at different time. The differences of the levels of IL-6, IL-8, IL-9, and IL-10 between the ulinastatin intervention group and the control group were not significant on the day before operation, 1 day and 3 days after operation (t = -1.15 to 1.82, all P > 0.05), but the levels of IL-6, IL-8, IL-9, and IL-10 of the ulinastatin intervention group were significantly lower than those of the control group and there were statistically significant differences 5 days and 7 days after the operation (t = 3.22 and 23.51, both P<0.05) . Conclusion Ulinastatin has a good effect in inhibiting the inflammatory factors and can protect and repair the postoperative hepatic injury as well in patients with hepatic cystic echinococcosis.

Changes of inflammatory factors after hepatic cystic echinococcosis surgery and intervention of ulinastatin in Qinghai area / 中国血吸虫病防治杂志

Objective To observe the changes of inflammatory factors after the hepatic cystic echinococcosis surgery and explore the intervention effect of ulinastatin on postoperative inflammatory factors. Methods Sixty patients with hepatic cystic echinococcosis were selected and randomly divided into a control group and ulinastatin intervention group according to whether or not use ulinastatin. The peripheral venous blood was extracted in all the patients and the levels of IL-6, IL-8, IL-9, and IL-10 were detected by the ELISA method on the day before operation, 1 day, 3 days, 5 days and 7 days after operation, respectively. The data was statistical analyzed to detect the relationships between/among the inflammatory factors mentioned above and ulina-statin and time. Results The variation of the levels of IL-6, IL-8, IL-9, and IL-10 were changed by the intervention of ulina-statin at different time. The differences of the levels of IL-6, IL-8, IL-9, and IL-10 between the ulinastatin intervention group and the control group were not significant on the day before operation, 1 day and 3 days after operation (t = -1.15 to 1.82, all P > 0.05), but the levels of IL-6, IL-8, IL-9, and IL-10 of the ulinastatin intervention group were significantly lower than those of the control group and there were statistically significant differences 5 days and 7 days after the operation (t = 3.22 and 23.51, both P<0.05) . Conclusion Ulinastatin has a good effect in inhibiting the inflammatory factors and can protect and repair the postoperative hepatic injury as well in patients with hepatic cystic echinococcosis.

The relationship between systemic inflammatory response syndrome and severity of acute pancreatitis combined with plateau erythrocythemia / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the relationship between systemic inflammatory response syndrome(SIRS) and severity of acute pancreatitis combined with plateau erythrocythemia in the high altitude.</p><p><b>METHODS</b>A retrospective analysis on the clinical data which involved acute pancreatitis combined with plateau erythrocythemia (n = 40) and without plateau erythrocythemia (n = 40) admitted from September 2006 to September 2009 was conducted. According to the unified standards, these cases were divided into plateau erythrocythemia group and no plateau erythrocythemia group. The patients in plateau erythrocythemia group were further divided into severe group and mild group according to scores of APACHEII. The data was analyzed according to the patient with (or without) SIRS, SIRS's standard indicators, diagnostic parameter and relation of severity and duration of SIRS in acute pancreatitis combined with plateau erythrocythemia.</p><p><b>RESULTS</b>There was significantly discrepancy between plateau erythrocythemia group and no plateau erythrocythemia group not only in the incidence of patients who developed SIRS, but also in two items of patients fulfilling or not fulfilling diagnostic criteria of SIRS (P < 0.05). There was significant statistical difference in three items of diagnostic parameter of SIRS between plateau erythrocythemia group and no plateau erythrocythemia group (P < 0.05). Significant difference in two and three diagnostic parameter was found on severity of SIRS in acute pancreatitis combined with plateau erythrocythemia (P < 0.05). The more severity acute pancreatitis combined with plateau erythrocythemia was, the longer duration of SIRS was.</p><p><b>CONCLUSION</b>SIRS is highly correlated with the severity of SIRS in acute pancreatitis combined with plateau erythrocythemia in the high altitude.</p>

The evaluation of total cystectomy for treatment of hepatic cystic echinococcosis / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the application of total-cystectomy in the hepatic cystic echinococcosis (HCE).</p><p><b>METHODS</b>An retrospective analysis was made in 223 cases of HCE. The observation targets included the days post operative, blood loss during operation, complications of residual cavity and local recurrence. The patients were divided into two groups: cyst-resection group and cyst ectomy group. The cyst-resection group was divided into two sub-groups, per cystectomy group and liver resection group.</p><p><b>RESULTS</b>The incidences of residual cavity and local recurrence in the cyst-resection group were lower than that in the cystectomy group (P < 0.01). The incidences of blood loss and average days in ward in the per cystectomy group were lower than that in partial liver resection group (P < 0.01).</p><p><b>CONCLUSIONS</b>HCE can decrease the complication of liver residual cavity and local recurrence effectively, total cystectomy should be selected as first procedures for treatment of hepatic cystic echinococcosis.</p>

Development of monoclonal antibodies against SARS-CoV and identification of antigenic epitopes / 生物工程学报

Based on the genomic sequence of SARS-CoV strain BJ101, antigenic immunodominant genes coding for the structure proteins of SARS-CoV were predicted by bio-informatics methods, and two chimeric genes A and B with multi-immunodominants lined up by Gly-Pro-Gly linker were synthesized. The chimeric genes were cloned into plasmid pGEX-6p-1 and expressed in E. coli with IPGT inducing. BALB/c mice were immunized with the purified recombinant fusion protein. The specificity of monoclonal antibodies were tested with a commercial ELISA kit for detecting antibody against SARS-CoV. The results showed that two peptides with molecular weights of 34kD and 35kD expressed by the two chimeric genes could be recognized by SARS patient convalescent serum in Western blot. Six positive hybridoma cell lines stably secreting monoclonal antibodies were selected. The subtype of monoclonal antibody D3C5 is IgG2a, and subtypes of all other five monoclonal antibodies are IgG1. Light chains of all monoclonal antibodies are kappa. With a commercial SARS-CoV antibodies detection ELISA kit, five out of six monoclonal antibodies were positively recognized. In western blot analysis with inactived virus cultures, D3D1 specifically recognized a band of about 180 kD. To further analyse the epitopes corresponding to the monoclonal antibodies, six oligoes (S1-S6) from S gene were synthesized and expressed. The results showed that the monoclonal antibodies D3D1 and D3C5 specifically recognized expression product of S2 and S5 oligoes, respectively. The S2 and S5 oligoes are corresponding to 447-458aa and 789-799aa of SARS-CoV S protein respectively.