Objective To explore working of concentration of PDL used in primary culture of rat hippocampal neurons.Methods 3 pregnant wistar rats were executed by cervical dislocation, the embryos were taken out and the hippocampal tissue was dissected quickly.Then the tissue was digested by trypsin and planted into dishes with proper concentration(300000/3.5cm vessle) which were coated by different PDL solution in different concentration (0.01,0.05,0.1,0.25,0.5,0.75,1,2 mg/mL).The state of the cultured neurons was observed to determine the most suitable concentration of PDL solution in coating dishes.Neurons activity was observed by CCK-8 Kit.Results Most neurons had adhered in 4 h.Protrusion of neurons began to grow in 1 d.The connection between neurons appeared in 4 d.Neurons matured and the network-connection was set up in 7 d.The best working concentration of PDL is between 0.25 and 0.75 mg/mL, neurons grew well and activity was optimum during this concentration.Conclusion The working concentration of PDL is important for the hippocampal neuron culture and this work is worth being applied.

Adult ; Female ; Hemoperfusion ; Humans ; Male ; Neurotoxicity Syndromes ; etiology ; therapy ; Organophosphate Poisoning ; Pesticides ; poisoning ; Retrospective Studies ; Treatment Outcome

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma ; metabolism ; pathology ; Cyclophosphamide ; therapeutic use ; Dendritic Cell Sarcoma, Follicular ; drug therapy ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Doxorubicin ; therapeutic use ; Head and Neck Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Lung Neoplasms ; drug therapy ; secondary ; Male ; Melanoma ; metabolism ; pathology ; Middle Aged ; Neoplasm Recurrence, Local ; Prednisone ; therapeutic use ; Receptors, Complement 3b ; metabolism ; Receptors, Complement 3d ; metabolism ; Vincristine ; therapeutic use

Objective:To investigate the incidence rate and risk factors of acute kidney injury (AKI) after off-pump coronary artery bypass grafting (CABG),and to compare the effects of AKI on complications after operation and major adverse cardiovascular and cerebrovascular events (MACCE) after 3 years' follow-up.Methods:In the study,299 consecutive patients who underwent scheduled off-pump CABG from January 2010 to March 2012 were included.The patients were divided into AKI group with AKI and control group without AKI after operation.The data during perioperative stage were compared,and multivariable Logistic regression modeling was used to identify the risk factors of AKI.The complications were compared after surgery and the patients were followed up for 3 years to observe the difference of MACCE between the two groups.Results:AKI occurred in 37.1％ patients (111/299).The elevated serum creatinine levels (Wald =9.276,P =0.002,95 ％ CI 1.006-1.028),chronic obstructive pulmonary disease(COPD) (Wald =3.469,P =0.063,95％ CI 0.950-7.630),decreased left ventricular ejection fraction (LVEF) (Wald =4.414,P =0.036,95 ％ CI 0.965-0.999),and implantation of intraaortic balloon pump (IABP) before or after operation(Wald =6.745,P =0.009,95％ CI 1.336-7.925)were risk factors of AKI in multivariable logistic regression modeling.More complications occurred in AKI group,such as the duration of mechanic ventilation,the time of ICU and the length of stay post operation,reintubation,pulmonary infection,stroke,hemorrhage of digestive tract,the volume of blood transfusion (plasma and red blood cell) and renal replacement therapy(P ＜ 0.05).The difference of mortality rate had no statistical significance between the AKI group and the control group,but two patients died in the AKI group.The difference of MACCE between the two groups had no statistical significance after 3 years' follow-up either.Conclusion:The incidence of AKI was high (37.1％) after off-pump CABG.The elevated serum creatinine levels,COPD,decreased LVEF,and implantation IABP before or after operation were independent risk factors of AKI in multivariable Logistic regression modeling.More complications occurred in AKI group during perioperative period,but the difference of MACCE between the two groups after 3 years' follow-up had no statistical significance.

Objective To study the effects of peroxisome proliferators-activated receptor (PPAR) ? on the growth of human hepatocellular carcinoma cells and explore the roles of phosphatase and tensin homologue deleted on chromosome ten (PTEN) and phospho-Akt in this process. Methods SMMC-7721 cells were treated with 15-d-PGJ2 or pioglitazone, which were two kinds of PPAR? ligands, at different concentrations. The viability of SMMC-7721 cells was evaluated by MTT assay. The cell cycle was analyzed by flow cytometry. PTEN mRNA level was determined by RT-PCR. The protein expressions of PTEN and pAkt were measured by Western blot analysis. Results It was demonstrated through MTT assay that both 15-d-PGJ2 and pioglitazone had an inhibitory effect on the growth of SMMC-7721 cells in a time- and dose- dependent manner. According to flow cytometry detection, more cells were arrested in G0/G1 phase. Increased expression of PTEN mRNA was detected in 15-d-PGJ2 or pioglitazone-treated cells through RT-PCR. Increased expression of PTEN protein and decreased expression of pAkt were confirmed by Western blot analysis. Conclusion The ligands of PPAR? could inhibit SMMC-7721 cells proliferation in a time- and dose- dependent manner. The upregulation of PTEN may be involved in the underlying mechanism.

Objective: To investigate the effect of Sirolimus on vein graft neointima hyperplasia via oral administration compared with local delivery, and find out an effective and safe way to provide support for clinical application. Methods: A rabbit external jugular vein-to-common carotid artery model was established. Twenty-four healthy rabbits were divided into 4 groups at random: blank-control group, F-127 control group, group 3 that received locally applied slow-releasing Sirolimus with F-127, group 4 that received oral Sirolimus (the commercial name Rapamune). The ratio of intima to medium thickness and re-stenosis rate (ratio of lumina to lumina plus intima area) were measured, PCNA positive cells by immunohistochemical staining were detected to indicate the degree of cell proliferation, and apoptosis cells detected by TUNEL. Results: Compared with blank-control group, neointima hyperplasia was inhibited significantly in group 3 and group 4 intima thickness were (90.11?10.99)?m versus (29.38?10.45) ?m, (18.29?9.03)?m, respectively. Re-stenosis rate was reduced (lumina area/ total area ratio were 0. 58?0.11 versus 0.80?0.16, 0.77?0.16, respectively). Proliferation of VSMC was inhibited (cell proliferation indexes were 31.03%?6.80% versus 20.32%?9.19%, 16.22%?5.85%, respectively) and cell apoptosis level raised (cell apoptosis indexes were 16.27%?6.49% versus 33.39%? 7.05%, 33.42%?7.11%, respectively). There was no significant difference between group 3 and group 4. Conclusion: Both locally applied slow-releasing Sirolimus and oral Rapamune could inhibit vein graft neointima hyperplasia; Administration via local delivery was preferred for little side-effect on the whole body. This conclusion provides support for clinical application.

Objective: To study the effects of mda-7/IL-24 on the growth, proliferation, apoptosis of different hepatic carcinoma cell lines and the related mechanisms. Methods: A recombinant adenovirus Ad-mda-7 was constructed and was used to transfect human hepatic carcinoma cell lines (HepG2, Hep3B and PLC/ PRF/5) and normal liver cell line L02. MTT assay and FACS were employed to assess the growth and apoptosis of cells; the expression of related protein expression was examined by Western blotting. The cells were treated with calpastatin I (ALLN,25 μmol/L) for 30 min to block the endoplasmic reticulum stress (ER-stress) and the above indices were examined again. Results: Treatment with Ad-mda-7 resulted in selective inhibition of cell proliferation and induced apoptosis, especially in HepG2 cells; Ad-mda-7 showed no influence on normal cells. Pretreatment with ALLN partially inhibited the above effects of Ad-mda-7. Western blotting revealed that Ad-mda-7 induced up-regulation of BiP/GRP7B and Bax protein, activation of caspase-12, caspase-3 and phosphorylation of p38 MAPK in HepG2 cells. Blocking ER-stress with ALLN down-regulated Bax, caspase-12 expression and inhibited activation of caspase-3 and caspase-12, but showed no effect on the expression of BiP/GRP78 or phosphorylation of p38 MAPK. Conclusion: mda-7/IL-24 can cause growth inhibition and promote apoptosis of hepatic carcinoma cells through the ER-stress pathway.

Objective To evaluate the efficacy and safety of diacerhein in the treatment of patients with knee osteoarthritis.Methods The efficacy and safety of diacerhein was randomly investigated in 42 pa- tients with knee osteoarthritis using parallel group methodology and a double-dummy technique to ensure dou- ble blind status with respect to diacerhein and control drugs diclofenac.Results Significant changes were ob- served in 20 meters walk pain,knee joint tenderness scale,WOMAC index scale,5F-36 health survey,knee joint swelling scale,compared with baseline(P＞0.05)in both diacerhein and diclofenac group respectively.No difference was found between diacerhein group and diclofenae group.The patient global assessment and physi- cian's global assessment were similar in diacerhein group and diclofenac group(P＞0.05).The side effect was similar in two groups.All of these side effects in gastrointestinal tract appeared to be transient.Conclusion Diacerhein can effectively relieve pain and swelling of knee osteoarthritis,and provides us a new effective and safe approach for treating knee osteoarthritis.

Objective To observe the expression changes of peripheral endothelial progenitor cells (EPCs) and Ang-1/Tie2 in patients with pulmonary arterial hypertension (PAH).Methods From Jun 2011 to Dec 2012,45 patients with PAH charged in Affiliated Hospital of Hangzhou Normal University were divided into 3 groups according to mean pulmonary arterial blood pressure (15 per group):mild(Group L),moderate(Group M),and severe(Group S),with another 15 normal people as control group(Group C).The EPCs were isolated from peripheral blood of every patient,number counting using fluorescence activated cell sorter (FACS),function test using cell culture in vitro.Expression of Ang-1 and Tie2 in the peripheral EPCs were measured by RT-PCR or Western-blot.Non normal data was analyzed by non parametric statistical test.Results The statistical discrepancy existed among Group L,M,S and the control in the number of EPCs [32.0 (27.0,37.0),26.0 (19.0,31.0),24.0 (22.0,26.0) vs 40.0 (37.0,51.0),P ＜ 0.05].The ability of migration [32.1 (26.5,37.5),26.8 (22.4,35.4),21.0 (17.8,34.0) vs 39.0 (33.3,42.4),P＜0.05] and adhesion of the EPCs [57.1(50.9,61.8),51.8(45.2,58.7),46.0 (37.2,55.1) vs 64.1 (56.2,75.0),P ＜ 0.05] among study groups and control group was different in statistic,the same with the proliferation activity of EPCs in different groups [0.6 (0.5,0.7),0.5 (0.4,0.6),0.4(0.3,0.5) vs 0.7(0.6,0.8),P ＜0.05].The mRNA expression of Ang-1 and Tie2 in Group M & S were significantly reduced compared with control [4.33 (2.49,4.62) and 2.89 (2.39,3.44) vs 5.31(3.78,6.22),P＜0.05],Tie2 mRNA[1.32(1.23,1.34)and 1.23(1.08,1.42)vs 1.49(1.25,1.66),P ＜ 0.05],and the protein expression of the phosphorylated Tie 2 in Group M &S were decreased [0.16 (0.15,0.24) and 0.12 (0.08,0.18) vs 0.22 (0.19,0.28),P ＜ 0.05].No significant difference of Ang-1 and Tie2 expression was observed between Group L and control [5.42 (4.72,5.95),1.54 (1.43,1.66) and0.23(0.19,0.33),P=0.674,0.867 and 0.674].Conelusion With the severity of PAH,the number and function of circulating EPCs decreased,as consistent with Ang-1 and Tie2 expression changes,suggesting that function decrease of EPCs in patients with PAH may be associated with the decrease of Ang-1/Tie2 expression.

Adult ; Female ; Hemangioma, Cavernous ; surgery ; Humans ; Nose ; surgery ; Orbit ; surgery ; Orbital Neoplasms ; surgery