Objective To investigate the effect of reptroperitoneal laparoscopic operation on the parameters of platelet, D-dimer and thrombomodulin(TM). Methods Forty cases were divided into two groups according to the operative way, retroperitoneal laparoscopic operation (n= 20) and open operation (n=20). Blood samples were taken preoperatively and at the end of the surgery. The following parameters were measured and compared within each group and between groups: platelet count (PLT), mean platelet volume (MPV), platelet distributionwidth(PDW), D-dimer, TM. ResultsThere were no significant differences for the PLT, PDW, MPV, TM and D-dimer between before and after operation in each group. There was no difference between 2 groups either for all these indicators.No patients from either group suffered thrombosis or abnormal bleeding as a pastoperative complication. Conclusion Compared with the conventional operation, retroperitoneal laparoscopic operation dioesd not induce more change on parameters of platelet, D-dimer and TM.

OBJECTIVEThe purpose of the present study was to investigate the in vitro effects of baicalin on induction of apoptosis in human prostate cancer cell line DU145.

METHODHuman prostate cancer cell line DU145 was treated with different concentration of baicalin in vitro. The apoptosis rate was determined by FACS analysis, cell cycle distribution was detected by flow cytometry, morphological changes and protein analysis were determined by means of electron microscope techniqueand immunohistochemical techniquerespectively.

RESULT50micromol x L(-1) and 125 micromol x L(-1) of baicalin dose-dependently induced apoptosis and inhibited the proliferation of prostate cancer cell DU145 in a dose and time-dependent manner. DNA flow cytometric analysis indicated that baicalin induced a arrest in G1 phase, showing a typical apoptosis peak. Electron microscopy detected a characteristic appearance of the apoptotic cells morphology. Immunohistochemical analysis revealed that induction of apoptosis by ways of inhibition of the bcl-2, loss of the Bax, and upregulation of Fas.

CONCLUSIONThe results indicate that baicalin may induce apoptosis and inhibit proliferation of prostate cancer cells, and has direct anti-tumor effects on human prostate cancer cells.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Flavonoids ; isolation & purification ; pharmacology ; G1 Phase ; Humans ; Male ; Plants, Medicinal ; chemistry ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Scutellaria ; chemistry ; bcl-2-Associated X Protein ; fas Receptor ; metabolism

Objective To observe the inhibition of piperlongumine in vitro on platelet aggregation and blood coagulation tests for children,to provide the experimental basis for clinical medication.Methods Venous blood samples from 30 children were randomly devided into 5 groups,and was centrifuge to separate platelet-rich plasma (PRP).After storing in 37 ℃ thermostat water bath for 5 minutes,the PRP which have been added DMSO as blank group,and added Aspirin (10 μmol/L)as control group,and added PL (20 μmol/L),PL(100 μmol/L),PL(200 μmol/L) as different concentrations of PL groups respectively,were induced by the addition of adenosine diphosphate (10 μmol/L),collagen(2.5 μg/mL) and the arachidonic acid(500 μg/mL).Then the platelet aggregation rate of the PRP from 5 groups could be measured by turbidimetry.Blood plasma isolated from venous blood was divided into 5 groups.In the PL groups,blood plasma were mixed up with PRP concentrations of which were 5,10,20 μmol/L.In the bland group,blood plasma were mixed up with DMSO (1％).In the control group,blood plasma were mixed up with heparin sodium(35 U/mL).After storing in 37 ℃ thermostat water bath for 5 minutes,fibrinogen(FIB),prothrombin time(PT),thrombin time(TT) and activated partial thromboplastin time of different groups were detected.Results Compared to the control group,the groups which were add PL with different concentrations (20,100,200 μmol/L) showed significant inhibition on platelet aggregation induced by AA and collagen(P<0.05).PL with concentrations of 100 μmol/L and 200 μmol/L showed significant inhibition on platelet aggregation induced by ADP(P<0.05).The PT,APTT,TT of blood plasma from children had been significantly prolonged by the intervention of PL 10 μmol/L and PL 20 μmol/L(P<0.05),however,no significant change of FIB was observed.Conclusion There are inhibitory effects of PL on platelet aggregation of blood plasma from children and anticoagulant activity in this study.

Adult ; Chorea ; etiology ; Circulatory Arrest, Deep Hypothermia Induced ; Diffusion Magnetic Resonance Imaging ; Endarterectomy ; adverse effects ; methods ; Humans ; Male ; Pulmonary Embolism ; surgery

OBJECTIVESTo investigate whether the human PC-3 cell infected with recombinant Ad-PTEN and Ad-p27Kip1 can steadily produce PTEN and p27Kip1 protein and change the biologic behaviors such as cell proliferation, cell cycle and apoptosis. The synergistic effect of PTEN and p27Kip1 on the therapy for prostate cancer has also been investigated.

METHODSWe constructed recombinant adenovirus vector of human tumor suppressor gene PTEN and p27Kip1. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and p27Kip1 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-p27Kip1 were determined by RT-PCR and Western blot respectively. MTT assay was used to determine the effect of PTEN and p27Kip1 on growth and proliferation of PC-3 cell; the change of cell cycle and apoptosis was examined by flow cytometry, and to compare between the combined therapy group and single gene therapy group.

RESULTSThe viral titers of Ad-PTEN and Ad-p27Kip1 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. After infected by adenovirus, it had been verified that the mRNA and protein expression of PTEN and p27Kip1 were steady in human PC-3 cell. Ad-PTEN and Ad-p27 Kip1 inhibited the growth and proliferation of PC-3 cells. The progression of cell cycle of PC-3 cell was arrested in G(0)-G(1) phase, meanwhile the apoptosis rate of PC-3 was also affected after Ad-PTEN or/and Ad-p27 Kip1 infected. There was significant difference between combined therapy group and single gene therapy group.

CONCLUSIONThe recombinant Ad-PTEN and Ad-p27Kip1 vector were constructed successfully and the expression of specific PTEN and p27Kip1 was high, steadily in PC-3 cell line. These results suggested that combination of PTEN with p27Kip1 has an application value in treatment of prostate cancer in future.

Adenoviridae ; genetics ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Cell Line, Tumor ; Chromosomes, Human, Pair 10 ; genetics ; Cyclin-Dependent Kinase Inhibitor p27 ; Gene Deletion ; Genetic Therapy ; Genetic Vectors ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; pharmacology ; Male ; PTEN Phosphohydrolase ; genetics ; pharmacology ; Prostatic Neoplasms ; genetics ; physiopathology ; therapy ; Transfection

Objective CD4 +IL-17 +cells are a group of newly discovered effector CD4 +T cells, which may play a key role in the pathogenesis of cancer.This study aims to investigate the expres-sion of CD4 +IL-17 +cells in pancreatic cancer and its correlation with the clinicopathological characteristics and prognosis of the dis-ease as well as the clinical significance of the cells in the microenvironment of pancreatic cancer. Methods We collected tumor tis-sue and tumor-adjacent normal tissue samples from 51 pancreatic cancer patients.We determined the expressions of CD34 and vascular endothelial growth factor ( VEGF) and measured the proportion of IL-17 +cells in the cancer tissue using immunohistochemistry and the fluorescence activated cell sorter, respectively, followed by analysis of their correlation with tumor angiogenesis, clinicopathological pa-rameters, and survival time of the patients. Results The percentage of CD4 +IL-17 +cells in tumor tissue was positively correlated with microvessel density (r =0.534, P<0.05) and the expression of VEGF in the tumor tissue (r=0.356, P<0.05).IL-17 +cells were expressed more highly in the tumorous than in the tumor-adjacent normal tissue (P<0.05), and the expression level was correla-ted with the stage of tumor, node, and metastasis (TNM) and lymph node metastasis (P<0.05), but not with the patients′gender or age, tumor size, tumor location, histological grade, or local invasion (P>0.05).Fifty (98.0%) of the patients were successfully followed up for 2-67 months, which revealed a median survival time of 16.6 ±4.8 months, significantly longer in those with a higher expression of intratumoral IL-17 +cells (P<0.05).Univariate analysis showed an association of the survival rate with the tumor size, TNM stage, lymph node metastasis, and level of intratumoral IL-17 +cells, while multivariate analysis showed the TNM stage to be an independent prognostic factor for the survival of the pancreatic cancer patients. Conclusion The expression of CD4 +IL-17 +cells in the tumor tissue is positively correlated with tumor angiogenesis, while that of IL-17 +cells with the clinicopathological parameters and survival time of the patients and therefore may serve as an important immune indicator for the prognosis of pancreatic cancer.

Objective To explore the clinical features, diagnosis and interventional management of acute deep venous thrombosis of lower extremity (LEDVT)combined with type Ⅱ heparin-induced thrombocytopenia (HITⅡ) and to improve the knowledge of this disease. Methods A retrospective review and analysis of the clinical data of the patients with acute LEDVT combined with HIT Ⅱ enrolled from January 2010 to June 2014. All of them underwent anticoagulation with low molecular weight heparin (LMWH) and the comprehensive interventional therapy at the beginning of treatment.When HIT Ⅱ was
identified, all forms of heparin and LMWH were avoided . Alternative anticoagulation was commenced with argatrobam. Adjustments in interventional therapy were taken while the short-term low-dose glucocorticoid treatment were used.The clinical manifestations, changes of PLT, 4Ts score (Warkentin 4T scoring system, 4Ts) , HIT antibody assay (ELISA) and response to therapy of the patients were analyzed and the treatment effect was observed . The efficacy of interventional therapy was evaluated according to the improvement clinical symptoms and venography. Results The incidence of acute LEDVT combined with HIT Ⅱ was 1.9%(8/416). There were 4 males and 4 females with a median age of 24 years in this study. The median time between their initiation exposure to heparin and onset of thrombocytopenia was 5 days (range,3 to 8 days). The median platelet counts prior to HIT Ⅱ was 218 × 109/L( range,122 × 109/L to 254 × 109/L ). Platelet counts decreased to the lowest level range from 20 × 109/L to 51 × 109/L(median 32 × 109/L). After alternative anticoagulation, the interval period which PLT recovered to the basic level was range from 3 to 7 days (median 3.5 days) . According to the score of 4Ts , there were 2 cases score 6 and 6 cases score 8. HIT antibody assay (ELISA) was detected in 6 patients which the results were positive. During heparin anticoagulation treatment, the LEDVT condition of all patients continued to deteriorate. Vein thrombosis extended in 7 patients. Among them, 5 patients occurred new thrombosis in the inferior vena cava and(or) at the vessel of catheter insertion. Another 2 patients complicated with pulmonary embolism. After underwent anticoagulation with argatrobam , with the increased of PLT the treatment efficacy of thrombolysis therapy was ameliorated. At the endpoint of interventional therapy, the curative effect evaluation was excellent in 3 cases, good in 3 cases and medium in 2 cases respectively. All patients were followed up for 12 to 20 months (median 15.5 months) with no evidence of recurrence .Conclusions The study showed that acute LEDVT combined with HITⅡdisplayed the following features:(1)an absolute drop in platelet count below the normal range (PLT ≤100 × 109/L) or as a relative decrease of 30% to 50% from baseline counts. (2) refractory venous thrombosis,during the interventional treatment of acute LEDVT, platelets counts should be monitored regularly in patients who receiving heparin anticoagulation. For patients with strongly suspected HIT Ⅱ, withdrawal of all forms of heparin and early introduction of alternative anticoagulant therapy can improve the effect of interventional therapy.

OBJECTIVETo investigate the effects of adenovirus-mediated PTEN and P27 on the invasion of PC-3 in vitro and angiogenesis, along with their synergy in the treatment of prostate cancer.

METHODSRecombinant adenovirus vectors of the human tumor suppressor genes PTEN and P27 were constructed. The replication-incompetent recombinant adenovirus was packaged and propagated in HEK293 cells. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and P27 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-P27 were determined by RT-PCR and Western blot respectively. The invasion of PC-3 cells in vitro was examined by Boyden chamber assay. MTT assay was used to testify the effect of supernatant from PC-3 infected with Ad-PTEN and Ad-P27 on the proliferation of endothelial cells ECV-304 and the CAM test was used to testify the effect of PTEN and P27 on angiogenesis. The difference between the combined therapy group and the single gene therapy group was also examined.

RESULTSThe viral titers of Ad-PTEN and Ad-P27 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. Adenovirus infection verified that the mRNA and protein expression of PTEN and P27 were steady in human PC-3 cells. The invasion in vitro of PC-3 cells was significantly inhibited by infection with Ad-PTEN or/and Ad-P27. CAM and MTT assays of ECV-304 confirmed that the supernatant from PC-3 cells infected with Ad-PTEN or/and Ad-P27 could inhibit the angiogenesis effectively. There was a significant difference between the combined therapy group and the single gene therapy group.

CONCLUSIONThe combined gene therapy of Ad-PTEN and Ad-P27 plays a synergistic role in inhibiting the invasiveness of PC-3 cells and angiogenesis.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Humans ; Male ; Neoplasm Invasiveness ; PTEN Phosphohydrolase ; genetics ; Prostatic Neoplasms ; blood supply ; pathology ; Transfection

OBJECTIVETo study the impact of the chloride channel dysfunction of the cystic fibrosis transmembrane conductance regulator (CFTR) on the cytoskeleton of Sertoli cells in the mouse.

METHODSTM4 Sertoli cells were cultured and treated with CFTR(inh)-172 at the concentrations of 1, 5, 10 and 20 μmol/L for 48 hours. Then the cytotoxicity of CFT(inh)-172 was assessed by CCK-8 assay, the expressions of F-actin and Ac-tub in the TM4 Sertoli cells detected by immunofluorescence assay, and those of N-cadherin, vimentin and vinculin determined by qPCR.

RESULTSCFTR(inh)-172 produced cytotoxicity to the TM4 Sertoli cells at the concentration of 20 μmol/L. The expressions of F-actin and Ac-tub were decreased gradually in the TM4 Sertoli cells with the prolonging of treatment time and increasing concentration of CFTR(inh)-172 (P < 0.05). The results of qPCR showed that different concentrations of CFTR(inh)-172 worked no significant influence on the mRNA expressions of N-cadherin, vimentin and vinculin in the Sertoli cells.

CONCLUSIONThe CFTR chloride channel plays an important role in maintaining the normal cytoskeleton of Sertoli cells. The reduced function and expression of the CFTR chloride channel may affect the function of Sertoli cells and consequently spermatogenesis of the testis.

Actins ; metabolism ; Animals ; Benzoates ; pharmacology ; Chloride Channels ; physiology ; Cystic Fibrosis Transmembrane Conductance Regulator ; antagonists & inhibitors ; Cytoskeleton ; drug effects ; Male ; Mice ; Sertoli Cells ; drug effects ; metabolism ; Spermatogenesis ; Thiazolidines ; pharmacology ; Time Factors

AIMTo determine peimine and peiminine in Fritillaria drug simultaneously by RP-HPLC-ELSD.

METHODSHPLC was carried out with a Waters Alliance, Model 2690, equipped with XTerra RP18 column (150 mm x 3.9 mm ID, 5 microm) and evaporated light scattering detector. The mobile phase (acetonitrile-10 mmol.L(-1) NH4HCO3 adjusted to pH 10.10 by ammonia solution) was eluted in gradient mode.

RESULTSThe recoveries of peimine and peiminine were 98.96% (n = 4), with RSD 1.01% and 98.40% (n = 4), with RSD 2.63%, respectively.

CONCLUSIONThe method is simple, sensitive and reliable. It can be used for quantitative determination of Fritillaria drug.

Cevanes ; analysis ; Chromatography, High Pressure Liquid ; methods ; Fritillaria ; chemistry ; classification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results