AIM:To investigate the effect of small interfering RNA(siRNA)-mediated progranulin(PGRN) gene silencing on the proliferation and migration abilities of human non-small-cell lung cancer A549 cells and its mecha-nism.METHODS:The mRNA and protein expression levels of PGRN in the A 549 cells and human bronchial epithelial (HBE)cells were detected by qPCR and Western blot.A549 cells were transfected with PGRN-siRNA by liposome meth-od.The expression of PGRN at mRNA and protein levels in the A 549 cells transfected with PGRN-siRNA was detected by qPCR and Western blot, respectively.The cell viability was measured by MTT assay.The cell proliferation ability was measured by living cells counting and crystal violet staining assays.The cell migration ability was measured by wound-heal-ing and Transwell assays.The protein levels of proliferating cell nuclear antigen(PCNA),cyclin D1,Bcl-2 and Bax were determined by Western blot.The protein levels of phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2) and phosphorylated protein kinase B(p-Akt)were also determined by Western blot.RESULTS:The expression of PGRN at mRNA and protein levels was higher in the A 549 cells than that in the HBE cells(P<0.05).The expression of PGRN at mRNA and protein levels in the A549 cells transfected with PGRN-siRNA was significantly decreased,and the cell pro-liferation and migration abilities were significantly decreased.The protein expression levels of PCNA,cyclin D1 and Bcl-2 were significantly reduced and the protein expression level of Bax was significantly increased(P<0.05).Meanwhile,the protein levels of p-ERK1/2 and p-Akt were down-regulated(P<0.05).CONCLUSION:PGRN gene silencing obviously inhibits the proliferation and migration abilities of human non-small-cell lung cancer A549 cells.The PI3K/Akt and MAPK/ERK signaling pathways may play an important role in these processes.

OBJECTIVETo assay ligustilide content in the herb of Szechwan Lovage Rhizome (Chuanxiong, CX), which is the dried rhizome of Ligusticum chuanxiong in order to assess the quality.

METHODLigustilide was quantitatively analyzed by high-performance liquid chromatography in 21 CX samples. An Alltima C18 column (4.6 mmx 150 mm, 5 microm) was used as the analytical column. The mobile phase consisted of water and acetonitrile (40:60). The flow rate was maintained at 1.0 mL x min(-1) with the column temperature at ambient conditions. The detection wavelength was set at 350 nm.

RESULTThe average content of Z-ligustilide in 21 CX samples was found to be 7.40 +/- 3.54 mg x g(-1)(x +/- s, n = 21). Therefore,the content of Z-ligustilide in CX should not be less than 0.66% (calculated on the dried basis).

CONCLUSIONThe overall analytical procedure is rapid and accuracy which is considered suitable for the quantitative analysis of ligustilide in CX. The amount of ligustilide in CX samples collected from different cultivation areas was obviously different. However, a relatively higher content of ligustilide was generally found in the CX collected from its main cultivated areas.

4-Butyrolactone ; analogs & derivatives ; analysis ; China ; Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Ligusticum ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Rhizome ; chemistry

OBJECTIVETo investigate the expressions of leptin and leptin receptor in hepatocellular carcinoma (HCC) and explore the clinicopathological significance.

METHODSThe expressions of leptin and leptin receptor were examined by immunohistochemistry in 81 HCC patients undergoing curative tumor resection. The correlations between the expression of two biomarkers and the clinicopathological factors were analyzed.

RESULTSThe overexpression rate of leptin and leptin receptor in HCC was 56.8% and 35.8%, respectively. No significant correlation was observed between their overexpression (r=0.236, P=0.034). Leptin receptor overexpression was significantly correlated to the tumor size and TNM stage (P<0.05), but not to age, body mass index, α-fetoprotein, hepatitis B surface antigen status, tumor grade, vascular invasion, or liver cirrhosis (P≥0.05). Leptin overexpression showed no significant correlations to the above clinicopathological factors (P≥0.05).

CONCLUSIONLeptin receptor overexpression may have an inhibitory effect on hepatocellular carcinoma. The expression status of leptin receptor decides the action of leptin and leptin receptor after their binding.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Leptin ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Receptors, Leptin ; metabolism

OBJECTIVETo explore the prognostic values of Raf-1 kinase (Raf-1), phosphorylated mitogen extracellular kinase 1 (pMEK1), and phosphorylated extracellular signal-regulated protein kinase 1/2(pERK1/2) in hepatocellular carcinoma (HCC) patients.

METHODSWe assessed the expressions of Raf-1, pMEK1, and pERK1/2 in HCC using immunohistochemical techniques. The relationships between the expressions of Raf-1, pMEK1, and pERK1/2 and the prognosis were explored.

RESULTSThe over-expression rates of Raf-1, pMEK1, and pERK1/2 in HCC were 38.3%, 46.7%, and 38.3%, respectively. The over-expressions of Raf-1, pMEK1, and pERK1/2 were positively correlated with each other (P>0.05), but had no significant correlation with sex, age, α-fetoprotein, hepatitis B surface antigen status, the TNM stage, size,differentiation and vascular invasion of tumor, and liver cirrhosis (P>0.05). Univariate survival analysis and COX proportional hazard regression model showed that Raf-1 over-expression was an independent prognostic factor of poor survival (P<0.05).

CONCLUSIONRaf-1 over-expression is an independent marker for the patients of HCC, which may provide new clue in the future targeted therapy.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; enzymology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; Humans ; Liver Neoplasms ; diagnosis ; enzymology ; MAP Kinase Kinase 1 ; metabolism ; Male ; Middle Aged ; Phosphorylation ; Prognosis ; Proto-Oncogene Proteins c-raf ; metabolism

OBJECTIVETo explore the role of estrogen in the regulation of the expression of claudin-6 and biological behavior in MCF-7 cells.

METHODSRT-PCR and immunocytochemistry were conducted to analyze the expression and localization of claudin-6 in MCF-7 cells treated with 17β-estradiol. CCK-8 kit assay and Scratch Test were conducted to analyze the capability of proliferation and migration of 17β-estradiol treated MCF-7 cells.

RESULTSRT-PCR analysis and immunocytochemistry showed that 17β-estradiol induced a concentration-and time-dependent effect on claudin-6. At 5 nmol/L and at 24 h, 17β-estradiol treatment led to an increased level of claudin-6, which was located in the membrane of MCF-7 cells. CCK-8 analysis showed a significant decrease in the capability of proliferation of MCF-7 cells compared with the control group (P < 0.05). Cells Scratch Test showed decreased migration capability of MCF-7 cells compared with the control group (P > 0.05).

CONCLUSIONS17β-E2 might regulate the expression of claudin-6 and inhibit the proliferation and migration of MCF-7 cells. The inhibitory effects of 17β-E2 on growth and migration of MCF-7 cells may be mediated by claudin-6 expression regulation.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Claudins ; Dose-Response Relationship, Drug ; Estradiol ; administration & dosage ; pharmacology ; Female ; Humans ; Membrane Proteins ; metabolism

Objective To understand the relationship of tendency on drug abuse and related influencing factors for middle school students.Methods A total of 603 students wcre assessed with self-made copying style scale,consciousness to resistance of smoking,drinking and resistance ability through a questionnaire.A relationship model was set up by structure equation modeling.Results Copying style and resistance ability had significantly direct influence on tendency of drug abuse(with standardized coefficients were 0.202 and 0.092 respectively).Consciousness to resistance of smoking and drinking had significantly indirect influence on tendency of drug abuse.The goodneSS of fit for the structural equation model was satisfactory,with GFI,AGFI,RMSEA as 0.981,0.972 and 0.014 respectively.Conclusion It Was meaningful to grasp some copying skills and resistance ability to lower the tendency of drug abuse in middle school students.

Objective: To investigate the clinicopathologic features of micronodular thymoma with lymphoid stroma(MNT). Methods: Five cases of MNT diagnosed from January 2007 to December 2016 in Henan Provincial People′s Hospital were collected.Hematoxylin-Eosin staining and immunohistochemistry were used to evaluate the histological and immunophenotypic characteristics in 5 MNT cases. Epstein-Barr virus (EBV) status was detected by in situ hybridization for EBV-encoded small RNA (EBER). Polymerase chain reaction was used to detect the rearrangement of immunoglobulin genes. Results: Five cases were MNT, including 3 male and 2 female patients, mean aged 59 years (from 43 to 63 years). All patients had ananterior mediastinal mass, with no myasthenia gravis and autoimmune diseases, and underwent surgical resection.Half to ten years follow-up showed no recurrence.Grossly, the tumors were solid in 4 cases, and cystic and solid in 1 case; the border was clear. Histologically, the tumors presented as a distribution of micronodules separated by abundant lymphoid stroma with prominent germinal centers. The nodules were composed of neoplastic spindle, oval cells containing bland, oval nuclei.Immunohistochemical study showed strong positivity of the tumor cells for CKpan, CK19, CK5/6 and p63. Stains for EMA, CD117, calretinin, TTF1 were negative in the tumor cells.Scattered CD3, CD1a, and TdT positive immature T lymphocytes were noted in and around tumor nodules. Many lymphocytes in the stroma, including germinal centers, were positive for CD20.The bcl-2 was also detected in lymphocytes in the stroma, mantle and marginal zone of lymphoid follicles, and in part of tumor cells. Tumor cells and lymphocytes were negative for EBER. Immunoglobulin genes rearrangement analysis showed that B lymphocytes were polyclonal. Conclusions MNT is a rare thymoma, which occurs in the elderly and has no obvious symptom. After complete resection, the prognosis is very good. The diagnosis should be based on a combination of clinicopathologic features, and other types of thymoma should be excluded.

OBJECTIVETo study the effect of polydatin on the expression level of miR-214 and liver function in atherosclerotic mice.

METHODSForty male ApoE(-/-) mice were randomly allocated into 4 groups (n=10), namely the model group, low- and high-dose polydatin groups, and simvastin group, with 10 male C57BL/6J mice serving as the normal control group. Mouse models of atherosclerosis were established by feeding the ApoE(-/-) mice with a high-fat diet. After 12 weeks of treatment, blood levels of glucose, lipids, AST, and ALT and the contents of T-SOD and MDA in the liver tissue were detected. The pathologies of the liver were examined with HE staining, and miR-214 expression in the liver was detected using quantitative real-time PCR.

RESULTSCompared with the normal control mice, the mice in the model group showed significantly increased blood glucose, serum TC, TG, LDL-C, ALT, and AST levels, and MDA contents in the liver (P<0.01), with significantly decreased serum HDL-C level and SOD and miR-214 levels in liver (P<0.01). Polydatin treatment significantly ameliorated such changes in blood glucose, serum ALT, AST, TC, TG, LDL-C, and HDL-C levels, and MDA, SOD, and miR-214 contents in liver tissue (P<0.05).

CONCLUSIONs Polydatin can reduce blood glucose and lipid levels and protect the liver function in atherosclerotic mice possibly by up-regulating the expression of miR-214 and T-SOD and down-regulating MDA in the liver.

Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; drug therapy ; Blood Glucose ; analysis ; Diet, High-Fat ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Glucosides ; pharmacology ; Lipids ; blood ; Liver ; drug effects ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; MicroRNAs ; metabolism ; Stilbenes ; pharmacology ; Superoxide Dismutase ; metabolism

Objective To characterize the clinical and cardiac MRI features of dilated cardiomypathy ( DCM) and left ventricular noncompaction (LVNC). Methods Compared the clinical and MRI features between 25 patients with LVNC and 21 patients with DCM. The MRI derived diastolic left ventricular wall thickness and the number and degree of noncompaction (NC) were evaluated using the 17-segment model. Results Chest distress, shortness of breath and abnormal ECG were presented in all DCM patients, abnormal ECG was evidenced in 22 LVNC patients and 21 out of 25 LVNC patients presented similar clinical symptoms as DCM patients while the rest 4 LVNC patients were asymptomatic. Left atrial and ventricular dimensions were significantly smaller in LVNC patients compared to DCM patients. The degree of left ventricular (LV) spherical remodeling was significantly greater in patients with DCM (sphericity index, SI = 0. 81 ± 0. 06) than in patients with LVNC ( SI = 0. 74 ± 0. 11, P < 0. 05). The LV ejection fraction ( LVEF) was significantly higher in patients with LVNC (32. 7% ± 14. 2% ) than that in patients with DCM (15. 0% ±5. 1% ). The number of NC segments in LVNC patients (9 ± 1) was significantly higher than the number of hypertrabeculation segment in DCM patients (5 ±2). The left ventricular apex (the 17th segment) was unexceptionally involved in all LVNC patients, while hypertrabeculation was absent in the 17th segment of DCM patients. The NC was more common in the apical and mid segments (16th, 12th and 11th segments) than in basal and mid septal segments (2nd, 3rd, 8th and 9th segments) in both LVNC and DCM patients. The thickness of compacted myocardium of the segments associated with noncompaction appeared thin in two groups. The wall thickness of noncompaction myocardium segments was thicker in LVNC patients than in DCM patients. The end-diastolic NC/C ratio was, on average, higher in patients with LVNC (3. 3 ± 0. 6) than in patients with DCM (1.9 ± 0. 3 ) . Conclusions The clinical manifestation is similar while there are significant differences in the morphology and function of left atria and left ventricle between the LVNC and DCM patients. The different distribution and degree of NC were helpful to differentiate LVNC from DCM.

BACKGROUNDClaudin-6 is a protein component of tight junctions and its expression could downregulate the malignant phenotype of breast carcinoma. Here we investigated the mechanisms of claudin-6 induced human MCF-7 breast cancer cells apoptosis, invasion, and migration.

METHODSTerminal deoxyribonucleotide transferase-mediated nick-end labeling assay and Annexin-V/PI double stain assay were carried out to evaluate apoptosis. Inhibitors of each pathway were used to inactivate the signaling pathways. The expression of claudin-6 and phosphate p38, Erk 1/2 and Akt protein levels was confirmed by Western blotting analysis. Invasive and migratory traits of claudin-6 expressing cells were determined by Boyden chamber invasion assay and monolayer wound-healing assay.

RESULTSCells with high-level expression of claudin-6 had a higher rate of apoptosis than control cells. Western blotting assay showed that by contrast to control groups, p38 pathways were more activated in claudin-6 expressing cells. However, after inhibitor SB203580 treatment, the activation status could be significantly counteracted. Furthermore, by applying inhibitors to the apoptotic rate, invasive and migratory traits were also recovered in cells with claudin-6 expression.

CONCLUSIONClaudin-6 may function through p38 mitogen-activated protein kinase pathway, of which inhibition may reverse claudin-6-induced cell apoptosis, invasion, and migration.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Movement ; genetics ; physiology ; Claudins ; genetics ; metabolism ; Humans ; In Situ Nick-End Labeling ; MAP Kinase Signaling System ; genetics ; physiology ; MCF-7 Cells ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism