Autophagy, a cellular process of "self-eating" by which intracellular components are degraded within the lysosome, is an evolutionarily conserved response to various stresses. Autophagy is associated with numerous patho-physiological conditions, and dysregulation of autophagy contributes to the pathogenesis of a variety of human diseases including cancer. Depending on context, activation of autophagy may promote either cell survival or death, two major events that determine pathological process of many illnesses. Importantly, the activity of autophagy is often associated with apoptosis, another critical cellular process determining cellular fate. A better understanding of biology of autophagy and its implication in human health and disorder, as well as the relationship between autophagy and apoptosis, has the potential of facilitating the development of autophagy-based therapeutic interventions for human diseases such as cancer.
Apoptosis ; Autophagy ; Cell Death ; Cell Survival ; Humans ; Neoplasms ; pathology

Objective To observe the structure feature of cultured mesothelial cells, determine its anticoagulant and fibrinolytic activity, and provide a basis for selection of artificial vascular prostheses coating cells. Methods The greater omenta, aortae and subcutanenous connective tissue of SD rats have been taken for mesothelial cell, endothelial cell and fibroblast culture. The cultured mesothelial cells have been observed by light microscopy and electron microscopy. The 6 keto PGF 1? (the metabolite of prostacyclin)concentrations in medium were measured by rodioimmunoassay, Tissue plasminogen activator(t PA) activity was detected by chromogenic assay. Results The cultured mesothelial cells have many structure feature similar to endothelial cells. The average concentration of the 6 keto PGF 1? in mesothelial cell culture was higher than that in endothelial cells and fibroblasts culture, the t PA activity in mesothelial cells culture was also higher than that in fibroblasts, but there wasn't a significant difference between mesothelial cells and endothelial cells.Conclusion Mesothelial cells have similar structure and functions comparing with endothelial cells. It may be an ideal coating lining for artificial vascular impants. [

Objective To explore the effects and mechanisms of urokinase-type plasminogen activator (uPA) on high glucose-induced rat mesangial cells proliferation and phenotype transformation. Methods Rat mesangial cells were cultured and incubated in media containing either 5 mmol/L D-glucose or 30 mmol/L D-glucose with or without addition of wortmannin, or uPA (105 U/L) for different time periods. At the end of the incubation period, mesangial cells proliferation was assessed by MTT assay and flow cytometric analysis. Cyclin-dependent kinase 2 (CDK2) and p27kip1 expression and activation of Akt were evaluated by Western blotting and Akt kinase assay respectively. Furthermore, the expression and distribution of α-SMA were detected with laser confocal microscopy. Results MTT assay and flow cytometric analysis demonstrated that high glucose induced mesangial cells proliferation (P＜0.05) and an incresed proportion of cells in G2/M+S stage after 24 h incubation (P＜0.01), which were attenuated by uPA or wortmannin (P＜0.01). High glucose induced the enhance of Akt activity after 3 h (P＜0.05), and the effect was inhibited by wortmannin or uPA (P＜0.01). High glucose did not alter CDK2 expression (P＞0.05),but significantly inhibited p27kip1 expression (P＜0.05), which was attenuated by wortmannin or uPA (P＜0.01). High glucose induced the up-regulation of α-SMA expression and perinucleus location in mesangial cells after 24 h (P＜0.01), which were alleviated by wortmannin or uPA (P＜0.01). Conclusion uPA up-regulates p27kip1 expression and counteracts high glucose-induced mesangial cells proliferation and phenotype transformation via blocking PI3K-Akt signaling pathway.

Objective To analyze levofloxacin resistance in Helicobacter pylori (Hp) and the sequence difference of gyrA gene in levofloxacin resistance and sensitive Hp strains. To explore the function of gyrA gene mutation in the development of levofloxacin resistance Hp strain.Methods From July 2007 to December 2008 in Department of Gastroenterology, Jinhua People hospital of Zhejiang Province, the gastric mucosa from gastroscopy biopsy of chronic gastritis and peptic ulcer patients were cultured in Hp selective medium under microaerobic condition at 37 degrees for three to five days. The Hp strains were isolated and identified by oxidase test, catalase test,urease test and UreA gene detecting. The levofloxacin susceptibility was determined by E-test and then resistance and sensitive strains were screened. The genomic DNA of Hp strains was isolated. The gyrA gene was amplified by PCR and the sequences were analyzed. Results 38 clinical isolated Hp strains were passed the levofloxacin susceptibility E-test, among those the minimum inhibitory concentration (MIC) of 12 strains was over 1.0 μg/ml, the percentage of resistant strain was 31.58%, while the sensitive stains was 68.42%. The gyrA gene sequence result indicated 10 resistant strain with 261, 271 and 272 10 site mutation, 2 strains with C261A mutation, one strain with C261G mutation, two strains with G271A mutation, 2 strains with A272G mutation, 2 strains with C261A,G271T andA272G mutation, 1 strain with C261G and A272G mutation. However, no mutation sites were found in 26 sensitive strains. Conclusion The rate of levofloxacir-resistance in isolated clinical Hp strain was high. The drug-resistance was associated with 261,271 and 272. site mutation of gyrA gene.

Objective To study the effects of PIAS3 knocking down on the proliferation,cell cycle and apoptosis of human prostate cancer cell line DU145 in vitro.Methods PIAS3 specific short hairpin RNA(shRNA) expressing plasmid was constructed and named pSilencer4.1/PIAS3.DU145 cells were transfected with pSilencer4.1/PIAS3.The proliferation of DU145 cells was analyzed by MTT assay.Cell cycle and apoptosis of DU145 cells were analyzed by flowcytometry.Results PIAS3 shRNA expressing plasmid was succefully constructed and then confirmed by sequencing.Expression of PIAS3 in DU145 was significantly reduced after pSilencer4.1/PIAS3 transfection.MTT assay showned accelerated proliferation after PIAS3 knocking down,and showned dose-effect curve.Flowcytometry showed cells in S phase increased,cells in G0/G1 decreased and percentage of apoptotic cells decreased after PIAS3 knocking down.Conclusion Knocking down of PIAS3 expression accelerates DU145 cell proliferation and inhibit cell apoptosis in vitro.

Aim To study the mechanism of anti-tumor effect of PHⅡ-7 to K562 and K562/A02 cells.Methods The effects of individual and combined doxorubicin on K562 and K562/A02 cells were observed by MTT assay.The coefficient of drug interaction was used to analyse the synergistic effect of PHⅡ-7,obtaining the RNA from the cells stimulated by PHⅡ-7 with different doses to analyse the MDR1 gene expression level.Finally,the cumulation of doxorubicin was observed in K562 and K562/A02 cells after being coped with PHⅡ-7.Results PH Ⅱ-7 had anti-tumor effect with IC50 of (1.37?0.37) ?mol?L-1;(1.48?0.34) ?mol?L-1 for K562 and K562/A02,respectively.It could potentiate the anti-tumor effect of dororubicin with CDI of 0.22 and 0.09 for K562 and K562/A02,respectively.PHⅡ-7 could synergistically inhibit the proliferation of K562 and K562/A02 cells.The decrease of MDR1 expression level depended on the increase of dose of PHⅡ-7 acting on cells.PHⅡ-7 could also develop the cumulation of doxorubicin in cells.Conclusion PHⅡ-7 is not only a Cytotoxinic drug but also can synergistically inhibit the proliferation of K562 and K562/A02 cells with the decrease of MDR1 expression level,especially in K562/A02 cells.

OBJECTIVETo observe the effect of Yiguan Decoction (YGD) on differentiation of bone marrow mesenchymal stem cells (BMSCs) into hepatocyte-like cells in vitro.

METHODSRat BMSCs were isolated using whole bone marrow adherent method. The properties of BMSCs were identified by analyzing the expression of surface cytokines by flow cytometry. The third passage cells were differentiated into fat cells to identify their features. BMSCs were incubated with hepatocyte growth factor (HGF) plus fibroblast growth factor 4 (FGF4) or YGD containing serum YGD for 21 days. The mRNA expression of alpha-fetoprotein (alphaAFP), albumin (Alb), and hepatocyte nuclear factor 4alpha (HNF4alpha) were detected by real time PCR. Expression of AFP and cytokeratin 18 (CK18) protein was detected by cell immunofluorescence. Glycogen synthesis was observed using periodic acid-Schiff stain (PAS). CK18, Wnt 3alpha, and alphacatenin protein expressions were detected by Western blot.

RESULTSHigh expression of CD90, CD29, and CD44, and low expression of CD34 and CD11b were observed in BMSCs isolated by whole bone mar- row adherent method, and numerous lipid droplets were observed in BMSCs using oil red O staining. Both YGD containing serum and growth factor stimulated the expression levels of Alb, AFP, HNF4alpha mRNA and CK18 protein. The down-regulated expression of Wnt 3alpha and beta-catenin could be detected at 21 days after induction. The synthesized glycogen granule could be seen. Down-regulated Wnt 3alpha and beta-catenin expression could also be observed.

CONCLUSIONYGD could induce the differentiation of rat BMSCs into hepatocyte-like cells, which was related to down-regulating Wnt/beta-catenin signal pathway.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley ; Wnt Signaling Pathway

Objective To investigate the correlation between pelvic organ prolapse quantitation (POP-Q) indication points and the incidence of occult stress urinary incontinence (OSUI) and its impact on prognosis. Methods Retrospective study medical records of 93 patients with pelvic organ prolapse (POP) staged atⅢ-Ⅳ, of which underwent pelvic reconstruction operations with Prolift system from Jan. 2007 to Sept. 2012. None of these patients had clinical manifestations of stress urinary incontinence (SUI) before surgery, and in which 44 patients were included in study group (POP complicated with OSUI) because they were identified with OSUI, another 49 patients as control group (simple POP). Follow-up and collecting datas including POP-Q, stress test, urodynamic recordings, incidence of de novo SUI, statistic analyzing by logistic regression and receiver operating characteristic curve (ROC). Results (1) The study group had a much higher incidence of 30%(13/44) on de novo SUI than that of control group (4%, 2/49;P<0.01). (2) Vaginal delivery (OR=5.327, 95%CI:1.120-25.347), constipation (OR=5.789, 95%CI:1.492-22.459), preoperative OSUI (OR=13.695, 95%CI:2.980-62.944), anterior vaginal wall prolapse (OR=6.115, 95%CI:1.231-30.379) were identified as dependent risk factors for de novo SUI by logistic regression analysis. (3) For POP patients that complicated with OSUI, we chose a cutoff value of +1.5 cm for Aa point as the threshold to predicting incidence of de novo SUI according to ROC curve, area under the curve (AUC) was 0.889 (P<0.05), the sensitivity reached 88.9%and specificity was 73.9%. According to ROC curve of Ba point, a cutoff value of+2.5 cm was chosen as the threshold to predicting incidence of de novo SUI post-operation, it had a sensitivity of 66.7% and specificity of 82.6%, AUC was 0.766 (P<0.05). Conclusions Pre-operative OSUI is a dependent risk factor of de novo SUI for advanced POP patients. Aa and Ba points are correlated with preoperative OSUI, and it is worthy to be considered as a risk predictor on forecasting the incidence of de novo SUI post pelvic reconstruction surgery.

To observe the effect of calycosin on the proliferation and activation of primary hepatic stellate cells (HSCs) in rats, and prove calycosin shows the effects through peroxisome proliferator-activated receptor γ(PPARγ) and farnesoid X receptor (FXR). The results indicated that calycosin could inhibit HSC proliferation and expressions of activation marker smooth muscle actin-α and type I collagen. With the increase in HSC activation time, FXR expression reduced, but with no notable impact from calycosin. Calycosin could up-regulate PPARγ expression and its nuclear transition in a concentration-dependent manner. Its prohibitory effect on HSC activation could be blocked by PPARγ antagonist. In conclusion, calycosin could inhibit HSC activation and proliferation, which may be related with the up-regulation of PPARγ signal pathway.
Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Hepatic Stellate Cells ; cytology ; drug effects ; metabolism ; Isoflavones ; pharmacology ; Male ; PPAR gamma ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; Up-Regulation ; drug effects