Hepatocellular carcinoma(HCC)is one of the most common malignancies worldwide.Due to the difficulty of diagnosis in the early stage of HCC, most HCCs are diagnosed in intermediate-advanced stage.Moreover, the high invasion, metastasis and recurrence rate of HCC result in the high mortality of HCC.MicroRNAs(miRNAs) are a class of highly conserved, endogenous, small, non-coding ,single stranded RNA with the length of 22 nucleotides.There are plentiful of miRNAs in liver.MiRNAs not only can regulate the growth and development of liver, but also are closely related to the formation of HCC.In the process of HCC formation, miRNAs could function as oncogenes or tumor suppressor genes to regulate multiple biological processes related to HCC, including cell differentiation,proliferation,tumorigenesis,angiogenesis,invasion,and metastasis.With the intensive study of molecular mechanisms of miRNAs in the process of HCC formation, increasingly studies have revealed that miRNAs could become sensitive biomarkers and effective therapeutic targets for HCC.

Objective To investigate the effects of preoperative jaundice relieving on hemihepatectomy of hilar cholangiocarcinoma.Methods The clinical data of 18 patients who received preoperative percutaneous transhepatic cholangiography and drainage (PTCD) or endoscopic nasobiliary drainage (ENBD) before hemihepatectomy at the Tongji Hospital of Huazhong University of Science and Technology from January 2007 to January 2012 were retrospectively analyzed.The condition of the 18 patients (jaundice relieving group) was compared with that of 24 patients (non-jaundice relieving group) who did not receive PTCD or ENBD before hemihepatectomy.The differences in the pre-and postoperative blood loss,blood transfusion,operation time and postoperative incidence of complications between the 2 groups were analyzed.All data were analyzed using the t test or chi-square test.Results After PTCD or ENBD,the levels of total bilirubin (TBil),direct bilirubin (DBil),alanine aminotransferase (ALT) were (27 ± 5) μmol/L,(22 ± 6) μmol/L and (52 ± 42) U/L,which were significantly lower than (287 ± 120)μmol/L,(212 ± 86)μmol/L,and (267 ± 180)U/L before PTCD or ENBD in the jaundice relieving group (t =4.33,6.61,4.19,P ＜0.05).In the jaundice relieving group,left hemihepatectomy was performed on 14 patients,and right hemihepatectomy on 4 patients,and the radical resection rate was 16/18.In the nonjaundice relieving group,left hemihepatectomy was performed on 11 patients,and right hemihepatectomy on 13 patients,and the radical resection rate was 83.3％ (20/24).There was no significant difference in the radical resection rate between the 2 groups (x2 =1.09,P ＞ 0.05).The operation time,volume of intraoperative blood loss,volume of blood transfusion were (5.0 ± 0.8) hours,(562 ± 207) ml and (430 ± 317) ml in the jaundice relieving group,and (6.3 ± 1.5)hours,(815 ± 463)ml and (750 ± 146)ml in the non-jaundice relieving group,with significant differences between the 2 groups (t =4.77,7.80,4.65,P ＜ 0.05).The incidences of postoperative complications,bleeding and postoperative hepatic failure were 3/18,1/18 and 1/18 in the jaundice relieving group,and 75.0％ (18/24),33.3％ (8/24) and 33.3％ (8/24) in the non-jaundice relieving group,with significant differences between the 2 groups (x2=5.14,7.58,7.58,P ＜ 0.05).Conclusion Preoperative jaundice relieving could shorten the operation time and reduce the volume of intraoperative blood loss and the incidence of postoperative complications.

Objective To explore the anti-hepatic-fibrosis mechanism of Baogan Ning(BN).Methods Fifty Wistar rats were randomized into 5 groups: normal group,model group,high-and low-dosage BN groups(35.4 and 17.7 g/kg respectively),and colchicine group(0.11mg/kg).Rat models of hepatic fibrosis were induced by multiplex factors.Hepatic protein was extracted,and the expression of hepatic leptic receptors of OB-Rb,JAK2 and STAT3 was detected with Western Blot method.Results The protein expression was not obvious in the normal group,and the color of protein band in the medication groups was light but dark in the model group.Compared with the normal group,the expression of OB-Rb,JAK2 and STAT3 increased in the model group.However,the expression of OB-Rb,JAK2 and STAT3 decreased in the medication groups as compared with the model group.And the expression of OB-Rb,JAK2 and STAT3 decreased in the BN groups as compared with the colchicines group.Conclusion The anti-hepatic-fibrosis mechanism of Baogan Ning is probably related with the inhibition of OB-Rb expression,thus inhibit the JAK2/STAT3 message pathway.

Objective:To discuss the mechanism of Baoganning anti-liver fibrosis.Methods:liver fibrosis rat models were established by complex factors.Automatic biochemical analyzer was used to detect TC、TG and RIA isotope analysis was used to detect serum leptin.To observe the influence of Baoganning on TC、TG and leptin level of liver fibrosis model rats. Results:Compared with the normal group,levels of serum TC,TG decreased apparently in model and every drug groups(P0.05).Conclusion:Baoganning can effectively prevent and treat liver fibrosis in rats,which may be closely related to the improvement of liver function,adjustment of lipid and reduction of serum leptin.

ObjectiveTo investigate the relationship between Dicer expression and clinicopathological characteristics and prognosis by detecting the expression of Dicer in hilar cholangiocarcinoma tissues and cells.MethodsThe expression of Dicer in tissues was detected using immunohistochemistry.Western blotting and RT-PCR were used to investigate Dicer expression in QBC939 and HIBEpic cells.The relationship between Dicer expression and clinicopathological characteristics was analyzed.A Kaplan-Maier analysis was performed to analyze the disease-free survival (DFS) and overall survival (OS) after radical surgical resection of hilar cholangiocarcinoma.ResultsWhen compared to control,Dicer was significantly down-regulated in hilar cholangiocarcinoma tissues (P＜0.05) and in QBC939 (P＜0.05).The expression of Dicer was higher in well differentiated adenocarcinoma than poorly and moderately differentiated tumours. Univariate analysis showed low expression of Dicer protein was significantly correlated with short disease-free survival and overall survival of patients with hilar cholangiocarcinoma after radical surgical resection (P＜0.01). Multivariate analysis revealed that the expression of Dicer was the most important factor for predicting prognosis after radical surgical resection of hilar cholangiocarcinoma (P＜0.05).ConclusionsDicer could be used as a prognostic marker for hilar cholangiocarcinoma.

AIM To study the effects of Biejiajian Pills (Colla Carapacis Trionycis,Asini Corii Colla,Nidus Vespae,etc.) on NF-κB,p65,p50 and IκB in NF-κB signaling pathway and target gene expression in HSC-T6 cells of rats.METHODS HSC-T6 cells were cultured with Biejiajian Pills drug serum for 24 hours,the expressions of p65,p50,VEGF and TIMP-1 mRNA were determined by qPCR;the expression of p65 was measured by immunofluorescence;the expressions of IκBα,IκBβ and α-SMA were determined by Western blot.RESULTS The Biejiajian Pills middle-,high-dose and positive control groups showed significantly lower expressions of p65,VEGF and TIMP-1 mRNA as compared with the blank control group and negative control group,the expressions of p50 mRNA among various groups showed no significant differences.But immunofluorescence showed that the expression of p65 in cytoplasm was decreased.Meanwhile,Biejiajian Pills showed significantly higher IκBα protein expression and obvious down-regulation of α-SMA expression in a dose-dependent manner,but had no significant influence on the expression of IκBβ.CONCLUSION Biejiajian Pills' therapeutic effects on hepatic fibrosis may be related to influencing NF-κB signaling pathway and inhibiting the expression of down-stream target gene.

OBJECTIVETo study the effect of Biejiajian Pills on Wnt signal pathway and the mechanisms underlying its action to suppress the invasiveness of hepatocellular carcinoma.

METHODSHepG2 cells cultured in the serum of rats fed with Biejiajian Pills for 48 h were examined for β-catenin expression using immunofluorescence, β-catenin/TCF4 complex activity with luciferase, and expressions of the downstream proteins cyclin D1 and MMP-2 using qRT-PCR.

RESULTSBiejiajian Pills-treated sera significantly reduced the expressions of cytoplasmic and nuclear β-catenin protein, cyclin D1 and MMP-2 proteins and lowered the activities of β-catenin/TCF4 complex.

CONCLUSIONBiejiajian Pills may serve as a potential anti-tumor agent, whose effect might be mediated by inhibiting the Wnt/β-catenin pathway.

Animals ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; Cyclin D1 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Rats ; Transcription Factor 4 ; Transcription Factors ; metabolism ; Wnt Proteins ; Wnt Signaling Pathway ; beta Catenin ; metabolism

OBJECTIVETo investigate the effect of Biejiajian Pills on the expressions of the signal molecules and target genes of Wnt signal pathway in HepG2 cells and explore the mechanisms by which Biejiajian pills suppress the invasiveness of hepatocellular carcinoma.

METHODSHepG2 cells were cultured for 48 h in the presence of serum collected from rats fed with Biejiajian Pills. The expressions of β-catenin, GSK-3β and P-GSK-3β in the cultured cells were assessed by Western blotting and the expressions of CD44v6 and VEGF were detected using immunohistochemistry.

RESULTSHepG2 cells cultured with the serum of rats fed with Biejiajian Pills showed lowered expressions of β-catenin protein both in the cytoplasm and the nuclei with also inhibition of phosphorylation of GSK-3β and reduced expression of CD44v6 and VEGF.

CONCLUSIONBiejiajian Pills can significantly reduce the expression of β-catenin by decreasing the phosphorylation of GSK-3β and blocking the Wnt/β-catenin signaling pathway to cause down-regulation of the target genes CD44v6 and VEGF, which may be one of the molecular mechanisms by which Biejiajian Pills suppress the proliferation and invasiveness of hepatocellular carcinoma.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Hep G2 Cells ; drug effects ; Humans ; Hyaluronan Receptors ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Phosphorylation ; Rats ; Vascular Endothelial Growth Factor A ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

ObjectiveTo investigate the protective effect of Danggui Shaoyaosan （DSS）-contained serum on β-amyloid （Aβ）_1-40-injured rat adrenal pheochromocytoma PC12 cells and its mechanism in regulating ubiquitin-proteasome pathway （UPP）. MethodAβ_1-40 was used to intervene PC12 cells to prepare the cell models of Alzheimer's disease （AD）， and the experiment was divided into the blank， model， and DSS-contained serum high， medium， and low-dose groups （10%， 5%， and 2.5%）. Cell viability and apoptosis were detected using cell counting kit-8 （CCK-8） method and flow cytometry， respectively. The content of Aβ and p-Tau protein was determined by enzyme-linked immunosorbent assay （ELISA）. The ubiquitin （Ub）， ubiquitin ligase E3 （E3）， 26S proteasome， ubiquitin carboxyl terminal hydrolase1 （UCHL1）， and UCHL3 protein expressions of UPP were displayed using immunofluorescence cytochemistry （ICC）， and the mRNA and protein expression levels of Ub， E3-parkin， 26S， UCHL1， and UCHL3 were determined by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultThe data of the CCK8 experiment verified that 5 μmol·L^-1 and 48 hours were the optimal conditions for modeling Aβ_1-40-injured PC12 cells. As compared with the blank group， the cell viability rate in the model group decreased （P<0.05） with an increased apoptosis rate （P<0.05）， the content of Aβ and p-Tau contents was elevated （P<0.05）， the mRNA and protein expression levels of Ub increased， and the mRNA and protein expression levels of 26S， E3， and UCHL1+3 decreased （P<0.05）. As compared with the model group， the cell viability rate in the DSS-contained medium-dose group increased （P<0.05）， whereas the apoptosis rate in each DSS-contained group decreased （P<0.05）. The content of Aβ in each DDS-contained group decreased （P<0.05）， and the content of p-Tau in the DDS-contained high and medium-dose groups decreased （P<0.05）. The mRNA expression level of Ub decreased， and that of 26S increased in each DDS-contained group （P<0.05）. The mRNA expression level of UCHL1 in the DDS-contained medium-dose group increased （P<0.05）， and the mRNA expression levels of E3 and UCHL 3 in the DDS-contained high and medium-dose groups increased （P<0.05）. The protein expression level of Ub in each DDS-contained group decreased， and the protein expression levels of 26S， E3， and UCHL1+3 in the DDS high and medium-dose groups increased. The DSS-contained serum medium-dose group exerted the optimal effect. ConclusionDSS-contained serum can increase cell viability rate， reduce cell apoptosis rate， eliminate Aβ and p-Tau protein deposits， and exert protective effects on Aβ_1-40-injured PC12 cells. Its mechanism may involve UPP via decreasing the expression of Ub and increasing that of 26S， E3， UCHL1， and UCHL3.