Objective This study analyzes the expression and clinical significance of Gli1 and Gli3 proteins in hepatocellular carcinoma.Methods 36 patients with hepatocellular carcinoma were studied.The expressions of Gli1 and Gli3 in the carcinoma and adjacent non-tumor tissues were detected with immunohistochemical assay,and their correlations with clinicopathological factors were statistically analyzed.Results Expression rates of Gli1 in hepatocellular carcinoma and adjacent nontumor tissues were 75 ％ and 36.1％,respectively.Expression rates of Gli3 in hepatocellular carcinoma and adjacent non-tumor tissues were 58.3％ and 30.6％,respectively.Expression rates of Gli1 and Gli3 in hepatocellular carcinoma were significantly higher than in adjacent non-tumor tissues (P＜0.05),and a positive correlation was found between the expression of Gli1 and Gli3 (r=0.423,P＜0.05).There was no association between the expression of Gli3 and clinicopathological factors such as age,tumor size,tumor differentiation and lymphatic metastasis.The expression of Gll1 was not related witha patient's age and tumor size,hut there were significant associations with tumor differentiation and lymphatic metastasis.Conclusions Therefore,the expression rate of Gli1 was positively correlated with tumor malignancy,which makes the detection of Gli1 and Gli3 valuable for the diagnosis and prognosis of hepatocellular carcinoma.

Objective To study the expression of insulin-like growth factor-Ⅱ receptor(IGF-Ⅱ R) and its relationship with the prognosis of human epidermalgrowth factor receptor 2 (HER2) positive breast cancer.Methods The expression of IGF-Ⅱ R in 104 specimens of HER2 positive breast cancer was evaluated by immunohistochemistry.The expression was rated on 2 scales:negative and positive.The age,pathological type,tumor size,lymph node involvement,status of estrogen receptor(ER),and progesterone receptor(PR) of all the cases were reviewed.The relationship between the expression of IGF-Ⅱ R and the above pathological parameters were analyzed by statistical methods.The relationship between the expression of IGF-Ⅱ R and the disease-free survival and overall survival was studied.Results The positive expression rate of IGF-Ⅱ R was 43.3％ (45/104).The positive rate of IGF-Ⅱ R was significantly higher in the lymph node positive group than in the lymph node negative group (61.5 ％ vs 25.0％,P =0.000).No significant correlation was observed between the expression of IGF-Ⅱ R and age,pathological type,tumor size,status of ER,or PR.The disease-free survival and overall survival was significantly lower in IGF-Ⅱ R positive group than in IGF-Ⅱ R negative group.(The 5-year disease-free survival:62.6％ vs 84.7％,P =0.022; The 5-year overall survival:71.5％ vs 89.6％,P =0.024).Cox regressive analysis showed that the lymph node status was an independent risk factor of disease-free survival and overall survival,while the IGF-Ⅱ R was not.Conclusion The positive expression of IGF-Ⅱ R in HER2 positive breast cancer indicates poor prognosis.The expression of IGF-Ⅱ R may be regulated by insulin-like growth factor-Ⅱ (IGF-Ⅱ).

Background:Gastric cancer(GC)is the fourth commonly seen cancer in the world. The role of Notch signaling pathway in the pathogenesis of tumors has become a focus in related studies,however,the underlying mechanism of Notch1 in GC has not yet been fully understood. Aims:To investigate the expression and significance of Notch1 in human GC tissue. Methods:A total of 115 patients with GC admitted at Jiaxing First Hospital from October 2011 to July 2014 were enrolled. Expression of Notch1 in cancerous tissue and para-cancer noncancerous tissue was determined by immunohistochemistry. The correlation between expression of Notch1 and clinicopathological characteristics of GC was analyzed. Results:Expression rate of Notch1 in cancerous tissue was significantly higher than that in para-cancer noncancerous tissue(73. 9% vs. 3. 5% ,P < 0. 001). Notch1 expression in cancerous tissue was significantly correlated with depth of vascular involvement,lymph node metastasis and TNM stage of GC(P < 0. 05). Conclusions:Notch1 as a cancer-promoting gene is involved in the pathogenesis of GC,and has a close association with tumor invasion and metastasis.

Objective To explore the correlation between the levels of liver fibrosis and liver fibrosis biochemical parameters of advanced schistosomiasis patients. Methods A total of 48 advanced schistosomiasis patients were investigated and they were examined by the liver biopsy and B ultrasound imaging. At the same time,the liver fibrosis biochemical parameters,including glu-tamine transpeptidase(GGT),alkaline phosphatase(AKP),procollagenⅢ(PC-Ⅲ),collagen typeⅣ(Ⅳ-C),hyaluronic acid (HA)and laminin(LN),were detected. The liver fibrosis levels were classified by the liver biopsy and B ultrasound imaging,re-spectively,and the correlation between the levels of liver fibrosis and liver fibrosis biochemical parameters were analyzed statisti-cally. Results There was no correlation between the liver fibrosis levels classified by the liver biopsy and all the liver fibrosis bio-chemical parameters;there was a weak correlation between the liver fibrosis levels classified by the B ultrasound imaging and GGT,AKP,LN and PC-Ⅲ,respectively;there was a significant correlation between the liver fibrosis levels classified by the B ul-trasound imaging and HA andⅣ-C,respectively. Conclusions B ultrasound examination is a better,noninvasive fibrosis in-spection method. Liver fibrosis biochemical parameters combined with the B ultrasound examination may better reflect the overall condition of liver fibrosis.

Objective To investigate the incidence of occult pancreatobiliary reflux and to evaluate its relation to gallbladder epithelial dysplasia and cancer. Methods From July 2006 to Feb 2008,956 cases underwent selective biliary procedure or preoperative endoscopic retrograde cholangiopanereatography (ERCP), bile was collected and amylase was measured. All removed gallbladders were pathologically examined for dysplasia and cancer. Results Occult pancreatobihary reflux was found in 75 of 754 patients in this study, with an incidence of 9. 9%. The biliary amylase values in the patients with occult pancreatobiliary reflux and in controls were 7701±20 378 IU/L and 16±51 IU/L, respectively ( P <0. 01 ).Gallbladder dysplasia and cancer were found in 31.0% and 3.4% of the patients with occult pancreatobiliaryreflux, respectively, and both were higher than those in the patients without pancreatobiliary reflux ( P <0. 05). In the patients with occult pancreatobiliary reflux, the biliary amylase level with gallbladder dysplasia or cancer was 2388 ± 2745 IU/L and was higher than those without gallbladder dysplasia or cancer (P < 0. 01 ). Conclusions With an incidence of 9.9% in patients of normal pancreatobiliary junction,the occult pancreatobihary reflux may contribute to the pathogenesis of gallbladder epithelial dysplasia and cancer.

Objective To study the clinicopathologic characteristics of triple-negative breast cancer (TNBC) and its value in the prediction of prognosis. Method In this study,500 cases of female breast cancers were examined immunohistochcmically for the TNBC. The clinicopathologic characteristics of the 243 TNBC cases were inspected. Results TNBC accounted for 17.6% (88/500) of the 500 breast cancers. The histological types of the TNBC included mainly infihrative ductal carcinoma, metaplastic carcinoma and medullar carcinoma. Among those, histological grade Ⅲ accounted for 72.7% (64/88) of all the TNBC and was more common than that in hormone receptor positive breast cancers (HR+ group ) and Her-2 overexpression breast cancers (Her-2 group)(P=0.000). The positive rates of CK5/6 and EGFR in the TNBC were 30.7% (27/88) and 34.1% (30/88), respectively. The positive rates of ERCC1 and KIT in the TNBC were 28.4% (25/88) and 34.1% (30/88), respectively, Both of which were higher than those in the HR + group and Her-2 group, respectively (P=0.032 and P=0.026). 3-year survival rate of the TNBC was 71.5% and it was lower than that of HR group (P=0.021) and not significantly different from that of Her-2 group (P=0.474). Conclusions TNBC is the breast cancer with high aggressive pathologic futures and poor prognosis. EGFR and ERCC1 expression were positive in a portion of TNBC cases.

OBJECTIVETo investigate the role of R-Spondinl in the activation of hepatic stellate cells (HSCs).

METHODSTwenty-four healthy male Kunming mice were randomly divided into the following two groups:fibrosis model group (n=16) and control group (n=8). Hepatic fibrosis was induced by subcutaneous injections of CC14 (20% in olive oil) at a dose of 5 ml/kg twice per week. After 10 weeks, the animals were sacrificed by CO(2) over-exposure and liver tissues were harvested.The protein and mRNA levels of R-Spondin1, alphat-SMA,and collagen I were examined by Western blot assay and real-time PCR respectively. Additionally,HSCs were isolated from the mice liver tissues to examine the time-series expression changes of R-Spondinl, alpha-SMA, and nuclear beta-catenin.TCF activity was analyzed by luciferase reporter assay.Moreover,HSCs were cocultured with recombinant R-Spondin1 and DKK1 to evaluate dose-response.

RESULTSR-Spondinl expression was significantly higher in the fibrosis model group than in the control group (protein level:3.16 ± 0.18 vs. 0.99 ± 0.16, t =13.31, P < 0.01; mRNA level:4.36 ± 0.26 vs. 0.98 ± 0.12, t =21.46, P < 0.01).The culture-activated mouse HSCs showed up-regulated TCF activity (5.33 ± 0.34 vs. non-activated: 1.03 ± 0.09, t =20.93, P < 0.01), nuclear beta-catenin expression (4.47 ± 0.21 vs. 0.97 ± 0.14, t =25.25, P < 0.01), and R-Spondin1 expression (protein level: 4.54 ± 0.18 vs. 1.04 ± 0.12, t =31.17, P < 0.01; mRNA level:5.13 ± 0.15 vs. 1.01 ± 0.16, t=38.06, P < 0.01). Exogenous stimulation of freshly isolated mouse HSCs with recombinant R-Spondin1 induced a dose-dependent increase in both TCF activity and the expression of nuclear beta-catenin and alphat-SMA. DKK1 down-regulated activities of factors in the WNT signaling pathway and repressed activation of HSCs. Conclusion R-Spondin1 may promote HSC activation by enhancing the canonical WNT signaling pathway.

Animals ; Down-Regulation ; Extracellular Matrix Proteins ; Hepatic Stellate Cells ; Liver Cirrhosis ; Male ; Mice ; RNA, Messenger ; Wnt Signaling Pathway ; beta Catenin

Breast Neoplasms ; metabolism ; pathology ; Female ; Humans ; Ki-67 Antigen ; genetics ; metabolism

OBJECTIVE To explore mecha-nisms of imperatorin on regulating P-glycoprotein(P-gp)in blood-brain barrier(BBB)based on net-work pharmacology combined with in vitro experi-ment.METHODS Drug targets were predicted using the Pharmapper and Swiss targets data-bases;disease targets were obtained through the Genecards database;intersections between drugs and disease targets were screened by Cytoscape software;the obtained core targets were used to construct protein-protein interaction(PPI)network,gene ontology(GO)functions,and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.The effects of imperatorin(20,50,100 μ mol·L-1)on P-gp activity were monitored in hCMEC/D3in vitro BBB model,and the effects of imperatorin on the expression of target proteins were verified using Western blot method.RESULTS 55 drug targets and 3102 disease targets were obtained from the network pharmacology screening,and 37 core targets were obtained after the combination.Enrichment analysis showed that core targets were closely related to chemical synaptic trans-mission regulation,neurotransmitter receptor activity,proteinkinaseregulationactivity,G protein-coupled receptor signaling pathway,neural active ligand receptor interaction pathway,PI3K-Akt sig-naling pathway,VEGF signaling pathway,etc..In vitro experimental validation suggested that all tested concentration groups of imperatorin signifi-cantly reduced the activity and expression of P-gp,which were achieved by significantly downregu-lating the phosphorylation levels of PI3K and Akt,and repressing the expression of VEGFR2 pro-tein.CONCLUSION Network pharmacology was used to predict the core targets and signaling pathways of imperatorin on regulating P-gp in BBB and relevant validation was conducted through in vitro experiments,providing a refer-ence basis for further exploration of the mecha-nisms of imperatorin on regulating P-gp in BBB.