Cardiovascular System ; Forkhead Transcription Factors ; Humans

Objective: To study the effects of deguelin on proliferation and apoptosis of human breast cancer cell line MCF-7 and on phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway. Methods: After treatment with 0, 1, 5, 10, 15 and 20 μmol/L of deguelin for 6, 24, 48 and 72 hours, the proliferation inhibition rate of MCF-7 cells was measured by cell counting kit-8 assay. Apoptosis rate of MCF-7 cells was detected with Annexin V-fluorescein isothiocyanate/propidium iodide double staining by flow cytometry and the apoptotic morphology was observed under a transmission electron microscope. After treatment with 0, 1 and 5 μmol/L of deguelin for 6 hours, 5 proteins involved in the PI3K/Akt signaling pathway were examined by Western blot analysis. Results: Deguelin at doses of 5, 10, 15 and 20 μmol/L inhibited the proliferation of MCF-7 cells at 6, 24, 48 and 72 hours. There was a significant difference in each group compared with the control group (P<0.01). The inhibitory effect was more marked with increasing concentration and duration of treatment. There were statistical differences (P<0.05) among 5, 10, 15 and 20 μmol/L groups. However, 1 μmol/L of deguelin had no obvious effects on the proliferation of MCF-7 cells at 6, 24, 48 and 72 hours, showing no significant difference compared with control group (P>0.05). Deguelin at doses of 5, 10, 15 and 20 μmol/L induced apoptosis of MCF-7 cells at 6 hours. There were significant differences (P<0.01) in the early and late apoptosis rate between the treated groups and the control group. The typical apoptotic MCF-7 cells were observed under the transmission electron microscopy. However, 1 μmol/L of deguelin had no apparent effect in inducing apoptosis of MCF-7 cells at 6 hours. After treatment with 5 μmol/L of deguelin for 6 hours the expression of phosphorylated phosphatase and tensin homologue deleted on chromosome 10 (PTEN) (Ser380), phosphorylated 3-phosphoinositide-dependent protein kinase 1 (PDK1) (Ser241), phosphorylated Akt (Thr308) and phosphorylated glycogen synthase kinase-3β (GSK-3β) (Ser9) proteins were significantly reduced in MCF-7 cells, while there was no significant change in the expression of total Akt protein. However, after treatment with 1 μmol/L of deguelin for 6 hours, there was no apparent change in the expression of these 5 proteins. Conclusion: Deguelin can inhibit the phosphorylation of GSK-3β (Ser9) via inhibition of the phosphorylation of PTEN (Ser380) and PDK1 (Ser241) pathway, thus inducing apoptosis and inhibiting proliferation of MCF-7 cells.

OBJECTIVE:To establish the HS-GC method for the content determination of menthol in Qiangli Pipa syrup,and the content of menthol in 56 batches of Qiangli Pipa syrup commercially available was determined. METHODS:HS-GC was per-formed on the column of Agilent HP-INNOWAX,column temperature was 130 ℃(maintaining 7 min),FID was used as detector, inlet temperature was 200 ℃,detector temperature was 250 ℃,carrier gas was high-purity nitrogen,flow rate was 3 ml/min,split ratio was 10∶1 and the injection volume was 1 000 μl. RESULTS:The linear range of menthol was 0.007 07-0.141 40 mg/ml(r=0.999 1);RSDs of precision,reproducibility and stability tests were no more than 3.37%;average recovery was 94.3%-99.6%(RSD=1.86%,n=6). There was significant difference in the contents of menthol in 56 batches Qiangli Pipa syrup. CONCLU-SIONS:The method is simple,sensitive,accurate and reliable,and can be used for content determination of menthol in Qiangli Pi-pa syrup. The sampling results show it is necessary to update the detecting items for the content of Qiangli pipa loquat and strength-en the quality control of it.

Objective:Investigate the relation between the phosphorylation of FOXO1 and the apoptosis and the proliferation of lymphoma cells and to clarify its specific mechanism.Methods:The lymphoma cells Namalwa and Jurkat were treated with PI3K inhibitor wort mannin or etoposide or Wortmannin plus etoposide for different times-pan and at different concentration.The inhibition rates for cell growth of lymphoma cells were examined by XTT assay.Apoptosis were detected by flow cytometry.The expressions of p-Akt,p-FOXO1,FOXO1 and Bim were determined by Western blot analysis.Results:Wortmannin induced apoptosis of Jurkat cells and Namalwa cells and inhibited their survival effectively.The growth inhibition rate and the apoptosis rate of lymphoma cells induced by Wortmannin plus etoposide were higher than those induced by etoposide alone.After treated with Wortmannin,phosphorylation of FOXO1 remarkably reduced and bim markedly increased.Conclusion:The dephosphorylation of FOXO1 inhibits proliferation of Jurkat cells and Namalwa cells,promotes their apoptosis and enhanced the sensitivity of Non-Hodgkin lymphoma cells to etoposide.Bim activated by FOXO1 promotes cell apoptosis.

Objective:To establish a method for isolating tumor infiltrating dendritic cells(TIDC)from mice models of Lewis lung cancer by positive selection using anti-CD11c magnetic beads.Methods:A total of 1.0?10~6 Lewis lung cancer cells were subcutaneously injected into a C57BL/6 mice at the lateral abdominal wall to establish the mouse model of lung cancer.TIDC were isolated positively using anti-mouse CD11c magnetic beads;they were labeled by anti-mouse CD11c,and then the purity of the isolated cells was tested by FACScan flow cytometer.The cells were also double labeled by PE-conjugated MHC-ⅡmAb and FITC-conjugated CD83 mAb or FITC-conjugated CD86 mAb to analyze the phenotype of cells by FACScan flow cytometer.Results:The positive selection using anti-CD11 c magnetic beads isolated(1.73?0.31)?10~6 TIDC from each gram of Lewis lung cancer tissue,which accounted for(2.18?0.29)%of the cells in the tumor tissues.The purity of TIDC was 96.49%.Electron microscope showed that the isolated TIDC had the typical character of DC cells.The positive rates of MHC-Ⅱ,CD83 and CD86 molecules in TIDC surface were(51.25?4.21)%, (3.48?0.34)%and(3.07?0.65)%,respectively.Conclusion:The positive selection using anti-CD11c magnetic beads is highly effective,simple,and economic,and is worth popularizing.

Objective To analyze the relationship between the expression of MHC class I chain-related gene A/B(MICA/B) on different human cancer cells and their sensitivity to NK cell cytotoxicity.Methods Hie expression MICA/B in K562,Bcap37,769P and A549 cells was measured by FACS.Isolation of NK cells were obtained by the modified RosetteSep~((R)) procedure.Cytotoxicity of NK cells at different ef0.50)%,(90.72±0.64)%,(55.59±0.55)%,and(3.84±0.10)% respectively.The purity of NK cells obtained by the modified RosetteSep~((R)) procedure was(96.52±2.42)%,whereas the cytotoxicity of NK cells against K562,Bcap39 and 769P was much higher than that of A549(P<0.01).The expression of MICA/B was significantly correlated with the cytotoxicity of NK cells at E:T ratios of 5:1,10:1 and 20:1 respectively(P<0.01).Conclusion The MICA/B expression on cancer cell lines was correlated with NK cell-mediated cytolysis and influenced the cytotoxicity of NK cells.

Objective To extract exosomes from dendritic cell (DC)and investigate antitumor effect of the special cytotoxic T lymphocytes activated by exosomes against 769-P.Methods Monocytes from peripheral blood of healthy donors were cultured to induce DC in intro and their phenotypes were analyzed by FACS.The exosomes were extracted from DC by ultrafiltration and ultracentrifugation and observed under electric microscope.Proliferation of T cells and the cytotoxicities of CTL against 769-P induced by exosome were measured by MTT.Results The exosomes could be separated from culture supernatant of DC by ultrafiltration and ultracentrifugation.The combination of the exosomes from DC and the 769-P antigen could effectively stimulate T cell proliferation and enhance their cytotoxicity, the absorption value of T cell proliferation(1.68±0.22), the cytotoxicity of CTL(38.23±2.16)%.Conclusion The exosomes extracted by ultrafiltration and ultracentrifugation can present tumor antigen to T cells and induce the cytotoxicity of CTL.

Objective To analyze the MSCT imaging of vascular and biliary ducts affected by hepatic cystic enchionococcosis disease(HCD),and to evaluate the 3D image reconstruction of MSCT.Methods Forty four cases with HCD confirmed by radical surgery were collected in this study.All cases underwent multi-phase scanning with 64-slices CT,and 3D image reconstruction of hepatic vessels was performed.Three cases suspected of bile duct complications were examined with negative CT cholangiography.All the imaging data were acquired before surgery.The spatial relationships between the cysts and the neighbor vessels and(or)bile ducts were compared with the findings during operation,then consistency was tested.Results Altogether 46 lesions were detected by MSCT before surgery.All lesions were found and diagnosed correctly,including 3 lesions rupturing into biliary duct Compared with surgery findings,the spatial relationships between lesions and the vessels were entirely consistent in first grade portal vein(14 cases)and inferior macular vein(18 cases).The Kappa values were 0.8805,0.6087,0.7751,0.7392 for first grade hepatic artery,second grade hepatic artery,second grade portal vein and hepatic vein.The relationships between bile ducts and lesions also matched in 3 bile-duct fistula cases.Conclusions MSCT with its 3D technique showed a high accuracy in the assessment of spatial relationship between vessels,biliary duct and lesions.It provides comprehensive imaging information for surgery.

BACKGROUND: Inflammation plays an important role in vessel proliferation after balloon injury. Reducing inflammatory reaction may lighten the ocurrence and development of the restenosis after angioplasty. Studies have demonstrated that PPAR_Y excitomotor has inhibitory effects on inflammation development. OBJECTIVE: To observe the changes in inflammatory factors after carotid artery balloon injury in rats and the intervention of PPARy excitomotor rosiglitazone. DESIGN, TIME AND SETTING: The randomized, controlled animal experiment was performed at the Central Laboratory of Shenzhen People's Hospital from January to June 2009. MATERIALS: Male SPF SD rats weighing about 350 g were selected to generate models of carotid balloon injury. METHODS: SD rats were equally and randomly divided into 3 groups: the control group, the balloon injury group and the rosiglitazone group. The left common carotid arteries were injured by balloon in the balloon injury group and the rosiglitazone group. The control group received sham operation. The rosiglitazone group was administered rosiglitazone daily by gavage,which began 4 days before operation and continued until harvesting.Accordingly,the control group and the balloon injury group were administered normal saline daily by gavage. MAIN OUTCOME MEASURES: All rats were executed under anesthesia at 14 days after operation, respectively to harvest left common carotid artery samples. The vessels were stained by hematoxylin-eosin, and Neointimal area (NIA) and media area (MA) as well as NIA/MA were calculated. Real time RT-PCR and Western Blot method were used to assay the expression of interleukin (IL)-6, IL-10, IL-17A mRNA and the distribution of nuclear factor (NF)-kB protein. expression levels of IL-6 and IL-17A mRNA in the rosiglitazone group were significantly lower than the balloon injury group, but higher than the Control group( P < 0.05), The expression levels of IL-10 mRNA in the rosiglitazone group were higher than the the rosiglitazone group was down-regulated, and lower than the balloon injury group, but higher than control group (P < 0.05). CONCLUSION: Rosiglitazone can regulate the expression of II-6 IL-10 IL-17A mRNA and the balance of inflammatory factors via NF-kB,inhibit the inflammatory reaction of injured vessels and may contribute to lighten the restenosis of injured vessels.

Objective To investigate the surgical strategy and outcome for occipitocervical extramedullary tumors.Methods 15 patients with occipitocervical extramedullary tumors from January 2007 to July 2010 were reviewed retrospectively.There were 7 males and 8 females with an mean age of 44.6 years (range,21-72 years).All cases were intradural tumors,including 7 cases in dorsolateral,5 cases in lateral side,and 3 cases in ventralis of spinal cord.Functional and neurological statuses were assessed using the Frankel grade and Japanese Orthopaedic Association (JOA) scale.According to Frankel grade system,there were 4 patients with Grade C,8 with Grade D,and 3 with Grade E.The mean preoperative JOA score was 10.5±3.6.Based on the tumor site,there were 5 cases in medulla oblongata-C1 and 10 cases in C1,2.Frankel grade and JOA score were used to evaluate neurological status and general health.The improvement of symptoms and characteristics of surgical strategies were analyzed.Results All cases were followed up for 2-4 years (mean 2.8 years).Complete resection of tumor and good fusion of bone graft were found in radiography postoperatively.The pathological diagnoses included 10 cases of schwannoma,3 cases of meningioma and 2 cases of neurofibroma.There was statistical difference between the preoperative and the final follow-up functional and neurological statuses including Frankel grade and JOA score.At the latest follow-up,2 cases improved from Frankel grade C to D,2from C to E,6 from D to E,but none in 2 cases with grade D.The mean JOA score at final follow-up was 15.6±1.6,and the average improvement rate was 80.7％± 17.4％.The mean preoperative C0-2 angle was 27.0°±4.1 °,and C2-7 angle was 16.8°± 12.7°.C0-2 angle was 27.6°±8.8°,and C2-7 angle was 10.2°±6.8° at the latest follow-up.However,these differences did not reach statistical significance.Conclusion Surgical treatment can effectively maintain or improve neurological function and improve quality of life.Extramedullary tumors can be resected completely by posterior approach,and spinal stabilization can be obtained satisfactorily through selecting appropriate surgical strategies.For medulla oblongata-C1 level,occipitocervical fusion is usually chosen after extirpation of tumor.For C 1,2 level,C1,2 fusion after tumor resection is useful in preventing atlantoaxial instability.