Aged ; Coronary Artery Disease ; Female ; Heart Ventricles ; Humans ; Vascular Fistula

OBJECTIVE: To establish a HPLC - UV method for determination of hydroxycamptothecin(HCPT) in rabbit tissues (liver, kidney, stomach, lung, spleen, heart, intestine) .METHODS: After homogenization, tissue samples and internal standard, eamptothecine, were precipitated with CH3OH-CH3CN( 1 : 1) and then centrifugalized.20?l of supernatant was injected and measured by HPLC - UV method.The chromatographic column was Lichrospher C18 column: (250mm ? 4.6mm, 5?m), CH3CN - 0.075mol/L NH4AC buffer(pH6.4) (30 : 70, contain 5mol/L tyiethylamine) served as mobile phase with flow rate of 1.0ml/ min .Detection wavelength was 384nm .RESULTS: The retention time of hydroxyeamptothecin was 4.5min.A good linearity was shown in the concentration range of (40- 1 600)ng/ml .The recovery was between 95.24% and 107.58% . The intra day RSD was less than 7.70% and the inter - day RSD was less than 6.69% .CONCLUSION: This method is simple,pratical and accurate.It could be applied to pharmacokinetic study of hydroxyeamptothecin.

Aim To investigate the role of phospho-lipase C epsilon 1 ( PLCE1 ) in modulating the apoptot-ic mechanism in esophageal cancer ( Eca ) cells. Methods Eca cell lines, OE33 and CP-C cells were cultured to assess the expression of PLCE1 . siRNA suppress expression of PLCE1 . The expression of PLCE1 and p53 was evaluated by quantitative real time PCR and Western blot. Methylation analyses of p53 were performed by bisulfite conversion of genomic DNA. Apoptosis was assessed by flow cytometry. Results OE33 and CP-C cells expressed high levels of PLCE1 . Knockdown of PLCE1 markedly increased the expression and hypomethylation of p53 , and in-creased the frequency of apoptosis. Conclusion PLCE1 suppresses apoptosis of Eca cells via promoting p53 promoter methylation and inhibiting expression of p53 .

Aim To investigate the activation of prote-ase-activated receptor 2 ( PAR2 ) in regulation of the expression of epidermal growth factor receptor ( EGFR) and apoptosis of lung cancer ( LC) cells. Methods LC cells A549 and its EGFR-silenced counterpart were incubated in the medium added with tryptase. Quanti-tative RT-PCR and Western blotting were used to de-tect the expression of EGFR in A 5 4 9 cells . The apop-tosis and Bax/Bcl-xL of cells were also recorded. Re-sults Treating A549 cells with tryptase in the culture for 48 hrs resulted in a marked increase in the expres-sion of EGFR in A549 cells. Marked decreases in a tryptase dose-dependent manner in apoptotic A549 cells were detected in the presence of tryptase. Expo-sure to tryptase markedly decreased the levels of Bax and increased the levels of Bcl-xL in A549 cells, which were not shown in EGFR-deficient A549 cells. Conclusion Tryptase can increase the expression of EGFR in LC cell line, A549 cells, which protects A549 cells from apoptosis, increases Bcl-xL, and sup-presses Bax in A549 cells.

The transesterification reaction conditions of lard with methyl acetate with combined use of immobilized lipases as catalysts were conducted. Initially, according to single factorial experiments, the studies on Lipozyme TL IM and Novozym 435 respectively catalyzed transesterification of lard showed that the optimal parameters of transesterification reaction were: the molar ratio of methyl acetate to oil of 14∶1, 40% enzyme added based on oil weight, temperature 50℃. Combined use of Lipozyme TL IM and Novozym 435 was proposed further to improve the catalytic performance by the response surface method (RSM). Herein, a 5-level-3-factor central composite rotated design was employed to evaluate the effects of lipase loading, the proportion of the two lipases and amount of methyl acetate. The optimum conditions were as followings: 40% lipase loading based on oil weight, 50%/50% the proportion of lipases (Novozym 435/Lipozyme TL IM), and the molar ratio of methyl acetate to oil of 14∶1. And under the optimal conditions, the highest biodiesel yield of 97.6% could be attained, which was higher than the biodiesel yield with each single one of the two lipases. The results suggested that the technics of combined use of certain immobilized lipases catalyzed transesterification reaction of lard for biodiesel production with methyl acetate as the acyl acceptor could raise the FAME yield and save the production cost.

Objective To assess early and medium outcomes of pathologic N2 disease unexpectedly detected in patients undergoing total video-assisted thoracic surgery lobectomy for non-small cell lung cancer.Methods Between Sep.2006 and Dec.2010,348 patients with Non-small cell lung cancer underwent total video-assisted thoracic surgery lobectomy,and within them,35( 10.1％ ) were found to have pathologic N2 disease after operation.We retrospectively reviewed the clinical and pathologic features of patients with unexpected N2 disease after video-assisted thoracic surgery lobectomy and their early and medium outcomes,including survival and recurrence pattern.Results No perioperative mortality was noted.26 patients received a lobectomy directly,and the other 9 patients after a wedge resection.All the patients had R0 resection.The medium operation time was 190 minutes and medium blood loss was 200ml.The medium stations and numbers of dissected N2 lymph nodes in operation were 4 and 10,respectively.And the medium stations and numbers of metastatic N2 Lymph nodes were 1 and 2,respectively.Among patients with pathologic N2 disease,18 (51.4％) had single-station involvement.The median duration of chest tube placement was 8 days.The median length of hospital stay was 11 days.15 complications occurred in 12 (34.3％) patients.All of the patients underwent adjuvant chemotherapy with platinum postoperatively.The median follow-up time was 23 months.The 1 - and 2-year overall survival (OS) was 80.9％ and 67.9％,and the medium OS was not reached.During follow-up,16 (45.7％) patients had a recurrence.The pattern of recurrence was locoregional in 5,distant in 11.The 1 - and 2-year disease-free survival (DFS) was 71.9％ and 44.2％,and the medium DFS was 20 months (95％,8.1 to 31.9 months).Divided the patients with pathologic N2 disease into two groups considering single-station involvement or not,the 1-and 2-year OS and DFS for the single-station group and for the multiple-station group were 87.7％,78.9％ ; 88.9％,49.4％and 67.6％,59.1％ ; 55.3％,39.5％.The medium DFS for both the two groups was 23 and 16 months respectively.Conclusion For non-small cell lung cancer with N0 disease confirmed by an exactly preoperative staging workups,if it is feasible in technology,a total video-assisted thoracic surgery lobectomy should be recommended.Even if N2 lymph node metastasis is unexpectedly detected postoperatively,the metastasis was mostly micro- or single-station involved,and a similar outcome with conventional thoracotomy can be achieved.

Adolescent ; Adult ; Bleomycin ; therapeutic use ; Child ; Child, Preschool ; Female ; Hemangioma ; drug therapy ; Humans ; Laryngeal Neoplasms ; drug therapy ; Male ; Middle Aged ; Pharyngeal Neoplasms ; drug therapy ; Young Adult

This study is to investigate the inhibitory effect of kaempferol on inflammatory response of lipopolysaccharide(LPS)-stimulated HMC-1 mast cells. The cytotoxicity of kaempferol to HMC-1 mast cells were analyzed by using MTT assay and then the administration concentrations of kaempferol were established. Histamine, IL-6, IL-8, IL-1β and TNF-α were measured using ELISA assay in activated HMC-1 mast cells after incubation with various concentrations of kaempferol (10, 20 and 40 µmol.L-1). Western blot was used to test the protein expression of p-IKKβ, IκBα, p-IκBα and nucleus NF-κB of LPS-induced HMC-1 mast cells after incubation with different concentrations of kaempferol. The optimal concentrations of kaempferol were defined as the range from 5 µmol.L-1 to 40 µmol.L-1. Kaempferol significantly decreased the release of histamine, IL-6, IL-8, IL-1β and TNF-α of activated HMC-1 mast cells (P<0.01). After incubation with kaempferol, the protein expression of p-IKKβ, p-IKBa and nucleus NF-κB (p65) markedly reduced in LPS-stimulated HMC-1 mast cells (P<0.01). Taken together, we concluded that kaempferol markedly inhibit mast cell-mediated inflammatory response. At the same time, kaempferol can inhibit the activation of IKKβ, block the phosphorylation of IκBα, prevent NF-KB entering into the nucleus, and then decrease the release of inflammatory mediators.
Cells, Cultured ; Histamine ; metabolism ; Humans ; I-kappa B Kinase ; metabolism ; I-kappa B Proteins ; metabolism ; Inflammation ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Kaempferols ; pharmacology ; Lipopolysaccharides ; Mast Cells ; drug effects ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Aim Toinvestigatewhetherepidermal growth factor receptor ( EGFR )-containing exosomes could induce tumor-specific regulatory T cells, and the effects of those T cells on tumor protein-specific CD8+Tcells.Methods TheexosomeswithEGFRwerepu-rified from NSCLC tumor, which modulated tolerogenic property of DCs. Then the induced TolDCs generated tumor-specific Tregs, with which the tumor protein-specific CD8 + T cells were suppressed. Results 80%exosomeswereEGFRpositivefromLCpatients while less than 2% exosomes were EGFR positive from control lung tissue. After exposed to the exosomes in the culture for 7 days, the IDO+ DCs proportion was much higher than that in control group ( 80. 8% ± 3. 2% vs 65. 6% ± 6. 4%, P <0. 05 ) . The induced Tregs was also higher ( 24. 1% ± 5. 2% vs 4. 2% ± 2. 3%,P<0. 01 ) , which suppressed the proliferation of CD8+ T cells(5. 4% ± 0. 2% vs 86. 7% ± 9. 3%, P<0.01).Conclusion Thepurifiedexosomesin-duce tolerogenic DCs. Coculture of the tolerogenic DCs and Th0 cells generates the tumor antigen-specific reg-ulatory T cells. The Tregs could suppress the tumor an-tigen specific CD8+ T cells.

To establish a UFLC-MS/MS method for the determination of valproate acid in human plasma. Methods:The sample was precipitated by methanol. The analysis of valproate acid and diclofenac sodium ( internal standard) was carried out on a Shim-pack XR-ODS II C18 column (2. 0 × 75 mm,2. 2 μm). Gradient elution was adopted with acetonitrile and water (containing 5 mmol·L-1 ammonium acetate) at a flow rate of 0. 3 ml·min-1. The detection was performed with multiple reactions monitoring (MRM) using nega-tive electrospray ionization (ESI) at m/z 142. 8→142. 8 for valproate acid and m/z 294. 0→249. 8 for diclofenac sodium. Results: The calibration curve of valproate acid was linear over the range of 8.4-168.0 μg·ml-1(r=0.997 5). Inter- and intra-day RSDs were less than 15%, and the analysis was proven to be stable. Totally 30 samples determined by EMIT were assayed by the method. And the results of the two methods analyzed by independent t-test showed no statistical significance. Conclusion:The method is rapid,sensitive and spe-cific,which can be applied in the determination of valproate acid in human plasma.