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.

Objective:To investigate the correlation between CD133 expression and clinicopathological features in triple negative breast cancer (TNBC) patients, and the impact of CD133 on prognosis in these patients.Methods:Data of 70 patients who received surgical treatment in our center from Jan. 2008 to Dec. 2012 were collected. Immunohistochemistry was used to examine the expression of CD133. Patients were divided into two groups according to CD133 expression. Univariate analysis, Cox and Logistic regression multivariate analysis were used in order to investigate the correlation between CD133 expression and clinicopathological features. Kaplan-Meier curve and Log-rank analysis were used to evaluate DFS (disease-free survival) and OS (overall survival) .Results:CD133 was expressed in cytomembrane and cytoplasm with expression rate of 95.71% (67/70) . Of which, 64.29% (45/70) of patients were low CD133 expression and 35.71% (25/70) were high expression. High CD133 expression was significantly correlated with younger age (≤50) ( P=0.007) and larger tumor size (>2 cm) ( P=0.020) . Tumor size ( P=0.035) , axillary status ( P=0.001) , Ki67 ( P=0.005) and CD133 expression ( P=0.014) were independent predictors of recurrence and metastasis in TNBC patients. Axillary status was independent predictor of death event ( P=0.008) . Increased CD133 was associated with poor prognosis. Compared with high expression, patients with low CD133 expression had better DFS ( P=0.002) and OS ( P=0.088) , while OS did not reach significant difference. Conclusion:CD133 expression was correlated with age and tumor size in TNBC patients. High expression was associated with recurrence, metastasis and poor prognosis. Thus, CD133 may be a potential biomarker in predicting prognosis in TNBC.

Objective:To systematically sort out and cluster the existing indicators of key issues in the quality of postgraduate clinical degree education based on the bibliometric study, so as to build a multidimensional quality assessment index system that integrates scientificity, rationality and representativeness, and to provide a scientific measurement tool for assessing clinical professional postgraduate education in China.Methods:By mining the related functions of UCINET6 network analysis integration software and its one-dimensional and two-dimensional data analysis NetDraw program, the social network analysis (SNA) method was used to extract and cluster the education quality problem set of clinical professional degree postgraduates.Results:A three-dimensional evaluation index system was constructed. The first dimension concluded such 8 key issues in the quality of postgraduate education in clinical medicine as ability assessment, teaching system, teaching quality assurance system, professional cognition and career prospects, assessment and evaluation system and organization, and the pulse taking and diagnosis.Conclusion:The clinical graduate education quality evaluation index system is an effective measurement tool for education quality improvement, based on a multidimensional perspective, with key issues as priority areas for intervention, providing an effective evidence-based basis for ensuring the development of professional graduate education efforts from 2020-2025.

Objective: To examine sleep quality and related factors among middle school students,and to provide a scientific reference for making the relevart interventional measures. Methods: A cross-sectional questionnaire anonymous survey was conducted among 9 560 students randomly selected from Shanxi province using multi-stage stratified cluster sampling. Pittsburgh Sleep Quality Index (PSQI) was used to collect information on sleep quality. Results: Among all the participants, 2 255 (23.6%) middle school students were reported poor sleep quality. The prevalence of poor sleep quality was significantly higher in girls (24.5%) than in boys (22.4%)(χ2=5.93, P<0.05). Significant differences were observed in the prevalence of poor sleep quality between junior middle school (12.9%), senior middle school (36.6%) and vocational school students (24.6%) (χ2=636.07, P<0.01). The global PSQI, sleep duration and sleep disturbance scores were higher in girls than in boys (t=3.76，8.38，3.47，P<0.01). There were significant differences between students with different school stages in global PSQI, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbance, use of sleeping medication, and daytime dysfunction scores (F=727.43,83.69,1 670.07,8.24,26.19,4.20,609.80,P<0.05). Multivariable Logistic regression analysis showed that gender, school stage, academic pressure, parental marital state, relationship with teachers, physical activity, smoking and drinking were associated with poor sleep quality among middle school students in Shanxi. Conclusion Sleep problems are common among middle school students, and appropriate intervention should be taken to improve sleep quality of students from individual, school-and family level.

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.

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 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 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.