Anti-Lipid-Peroxidation of Hovenia dulcis Thunb. in mice was studied. Results showed that H. dulcis Thunb. had the following functions: 1. decrease MAD contents of serum and tissucs of liver and brain. 2. increase SOD activity in tissues of liver, kidney and brain. 3.impart tolerance toward cold and heat, and prolong swimming time of mice. The results indicate tLat H. dulcis Thunb. has function of Anti-Lipid-Peroxidation and may be valuable for the retardation of senility.

Objective To investigate the effects of recombinant trichosanthin(rTCS) on methylation status and expression level of p27 gene in HeLa cells.Methods HeLa cells was treated by different concentration(20 ?g/mL,40 ?g/mL,and 80 ?g/mL) of rTCS for 48 h and then methylation-specific polymerase chain reaction(MSP) was used to detect the promoter methylation status of the p27 gene,real-time PCR was used to detect levels of p27 and DNMT1 mRNA,and Western blotting assay was used to detect expression level of p27 protein before and after treatment with rTCS.Results Low expression level and promoter methylation status of the p27 gene were detected in HeLa cells.Treatment with 40 ?g/mL rTCS totally demethylated p27 promoter.Treatment with 20 ?g/mL,40 ?g/mL or 80 ?g/mL rTCS resulted in a 2.22-,4.00-or 6.03-folds increase in p27 mRNA level,respectively,and also a great increase in p27 protein level.A high DNMT1 expression level was observed in HeLa cells and treatment with 40 ?g/mL rTCS resulted in a 78% decrease at the DNMT1 mRNA expression.Conclusion rTCS could reverse promoter hypermethylation and re-activate the expression of p27 gene by inhibiting DNMT1 expression in HeLa cells,which indicates its potential use in cancer therapy.

Aim To study the effects of polyamine analogue CPENSpm on the human lung cancer line A549 in cell proliferation and apoptosis.Methods MTS was used to assay the cell proliferation,chemical analysis methods were used to determine the activities of enzymes in the polyamine metabolism,HPLC was performed to assay the intracellular concentration of polyamines,Sub-G1 and DNA fragmentation assays were used to determine the cell apoptosis.Results Treating A549 lung cancer cells by CPENSpm resulted in:①cell-growth inhibition and cell apoptosis;②inhibition of ODC(key enzyme in polyamine synthetic pathway)and activation of SSAT and SMO(key enzymes in polyamine catabolism);③great decrease of intracellular polyamine concentrations.MDL72527,the SMO inhibitor,can antagonize the effect of CPENSpm on inhibiting the proliferation of A549 cells.Conclusion CPENSpm inhibits proliferation and induces apoptosis of human A549 lung cancer cell line by interfering the polyamine metabolism,depleting intracellular polyamine contents that are need by quick-growth of cancer cells and inducing production of H2O2.

Objective: To establish the quality standard for Erxiaqingxin Tablets(Cordyceps, Rhizoma Pinelliae, Radix Puerariae, Caulis Bambusae in Taenia, Fructus Aurantii Immaturus, Pericarpium Citri Reticulatae, etc.). Methods:TLC was performed to identify Pericarpium Citri Reticulatae, Fructus Aurantii Immaturus and Radix Puerariae. HPLC was used to determine the content of puerarin. Results:The study on the quality control showed that the characteristic of identification by TLC was distinct and hightly specific. The quantification method had the linear range of 0.04092～2.4552?g. The average recovery was 99.36% and RSD was 2.32%. Conclusion:The method for identification and quantification was simple, realizable and reproducible. It can be used effectively for the quality control of Erxiaqingxin Tablets.

Objective To investigate whether difluromethylornithine (DFMO) can be used in the treatment of human leukemia. Methods The cell proliferation was detected by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium)] assay after treatment of human lymphocyte Jurkat cells by DFMO (0 to 10 mmol/L) for 24 to 72 h. Enzyme activity of spermine oxidase (SMO) and acetylpolyamine oxidase (PAO) was determined by chemiluminesence assay. DNA fragmentation assay was used to evaluate cell apoptosis. Fluorescent dye assay was performed to determine the changes in mitochondrial membrane potential. Western blotting was used to determine Bax content. Casepase-3 enzyme activity was measured by spectrophotometric method. Results DFMO treatment inhibited the proliferation of Jurkat cells significantly in a dosage- and time-dependent manner (P

Aim To evaluate the effect of expression inhibition of spermine oxidase(SMO)on the actitumor activity of polyamine analogue CPENSpm (N~1-cyclopropylmethyl-N~(11)-ethylnorspermine).Methods siRNA technique was used to inhibit expression of SMO in human lung cancer line A549.QT-RT-PCR and enzyme activity assay was performed to determine the expression level of SMO.The cell proliferation was detected by MTT assay.The apoptosis of A549 cells were evaluated by DNA degradation and Sub-G_1/flow cytometry assay.Results The A549 cell line with silenced SMO expression was successfully obtained.Basic SMO mRNA and enzyme activity levels in the SMO-siRNA plasmid transfected cells were 0.53% and 14% lower than that in the control cells respectively. Treating A549 control cells by 10 μmol·L~(-1) CPENSpm for 24 hours resulted in a 10-folds up-regulation of SMO in mRNA level and 20-fold increase in enzyme activity,but this drug-induced SMO expression was obviously prevented in SMO-siRNA plasmid transfected cells.MTT assay demonstrated that SMO expression inhibition decreased the sensitivity of A549 cells to CPENSpm exposure(0～20 μmol·L~(-1)).DNA degradation and sub-G_1 assay proved a deceased ability of CPENSpm to induce apoptosis in SMO-siRNA plasmid transfected cells.Conclusion Up-regulation of SMO by CPENSpm is possibly one of the molecular basics for its antitumor activity.

AIM: To prepare recombinant human spermine oxidase (SMO) and polyclonal antibody against human SMO by gene recombination techniques. METHODS: Human SMO cDNA was amplified from total RNA of A549 cells through reverse transcription PCR. The cDNA was then cloned into pET-15b to construct SMO prokaryotic expression vector. After transforming, the vector was induced to express recombinant SMO by IPTG in E. coli BL21 (DE_3). Recombinant SMO was purified by Ni-NTA resin under denaturing condition and then was dialyzed to renature. The enzyme activity of recombinant SMO was analyzed by chemical fluorescent method. SMO polyclonal antibody was prepared by using recombinant human SMO protein purified by polyacrylamide gel electrophoresis as antigen to inoculate rabbit intradermally. The titer and specificity of anti-sera were determined by ELISA, Western blot and Immune Cell Chemistry. RESULTS: Purified and dialyzed recombinant human SMO has the specificicity of oxidizing the spermine. The polyclonal antibody has high titer and specificity against human SMO. CONCLUSION: This research established a method for prokaryotic expression, purification and polyclonal antibody preparation of human SMO. The method lays a foundation for the future functional research of SMO.

Objective To evaluate the safety and feasibility of injection test which is used to locate esophageal balloon catheter.Methods A prospective study was conducted. The patients undergoing invasive mechanical ventilation (MV) admitted to general intensive care unit (ICU) of Beijing Tiantan Hospital Affiliated to Capital Medical University from May 2015 and March 2017 were enrolled. The commercially available esophageal balloon catheter was modified to perform injection test. The catheter was withdrawn step by step and the injection test was repeated until the presence disturbance wave presented, which indicated that the balloon had just entered the esophagus. The position where disturbance wave appears was named 0 cm. End-expiratory occlusions were performed at the positions of+15,+10,+5, 0, -5, -10 and -15 cm, respectively, and the changes of esophageal pressure (Pes) and airway pressures (Paw) were measured in the spontaneous breathing and passive ventilation, and the ratio between the changes (ΔPes/ΔPaw) was calculated.Results A total of 20 patients were enrolled, of which 15 patients finished both the spontaneous and the passive ventilation parts, and 2 patients finished only the spontaneous part and 3 patients finished only passive part. ① Disturbance waves could be induced by injection test in all patients. The average depth of disturbance wave in spontaneous breathing was deeper than that in passive ventilation (cm: 42.4±3.8 vs. 41.8±3.3), but there was no significant difference between the two ventilation settings (P = 0.132). No adverse events occurred during the study period. ② Pes increased with the stepwise withdraw of esophageal catheter, reached the maximal value at+5 cm, and then decreased when the catheter was further withdrawn, no matter in the spontaneous or the passive ventilation. In spontaneous breathing, the ΔPes/ΔPaw was within the ideal range (0.8-1.2) at the positions of 0, -5 and -10 cm. The ΔPes/ΔPaw was closest to unity at the positions of 0 cm (0.98±0.15). The ΔPes/ΔPaw at -15 cm (0.66±0.26) was significantly lower than that at 0 cm (P < 0.05). For passive ventilation, the ΔPes/ΔPaw was within the ideal range at the positions of -5 cm and -10 cm, and the ΔPes/ΔPaw was closest to unity at the positions of-10 cm (0.94±0.12). The ΔPes/ΔPaw at 0 cm and -5 cm was significantly higher than that at -10 cm (1.43±0.31 and 1.12±0.14, respectively); while the ΔPes/ΔPaw at -15 cm (0.68±0.23) was significantly lower than that at -10 cm (allP < 0.01).Conclusions Ideal position of the esophageal balloon catheter could be determined quickly and easily by using injection test. The method is safe and clinically feasible.Clinical Trial Registration Clinical Trials, NCT02446938.

BACKGROUND: Natural bilogical bone-derived materials processed with physical and chemical methods possess natural network pore system. They have good cellular compatibility, and help osteoblasts attach and grow on them, and can be used as the scaffolds of osteoblasts.OBJECTIVE: To observe the osteogenetic effect of partially deproteinised decalcified bone (PDDB) as scaffolds of osteoblasts in vivo.DESIGN: A randomized and controlled trial.SETTING: Central Laboratory of the First Hospital of Kunming Medical College.MATERIALS: Partially deproteinised decalcified bone; human embryonic periosteum-derived osteoblasts; twenty 4-week-old BALB/c nude mice,with the body mass of 25 to 28 g, of either gender.METHODS: This experiment was conducted at the Central Laboratory of the First Affiliated Hospital of Kuming Medical College from January to June 2003. PDDB composited with human embryonic periostea-induced osteoblasts were implanted into the nude mice after cultured for 1 week in vivo, 4 scaffolds in each nude mouse. Composite of scalfolds and cells implanted on the left side of the spine was set as experimental side and simple implanted material on the right side of the spine was set as blank control side. Then alkaline phosphatase activity and routine histological examination were performed 4 and 8 weeks after operation.MAIN OUTCOME MEASURES: General observation, alkaline phosphatase activity and routine histological examination were performed after the materials were taken out.RESULTS: Twenty nude mice entered the stage of result analysis. ① General observation of the implanted materials: No necrosis, ecpyesis, or fluidifying was found around the implanted materials, but ingrowth and enwrapption of soft tissues were found. ② Measurement of alkaline phosphatase activity: alkaline phosphatase activity after PDDB composited osteoblasts in vivo was stronger at week 8 than at week 4 [(22.854±6.018) nkat/g vs(11.286±4.268) nkat/g], and was much stronger than that of the simple implanted materials [(1.217±0.083) nkat/g vs (2.717±0.583) nkat/g]. ③ Results of routine histological examination: Cartilage formed at week 4 and part of cartilage formed new bone and marrow cavity at week 8 at the experimental side, cartilage and new bone formed much more as time went by, but there was no any cartilage or bone formation at the control side.CONCLUSION: Cartilage and bone form after PDDB composited with osteoblasts are implanted, and more cartilage and new bone form as time passes. PDDB can be used as the scaffolds of osteoblasts.

Objective To investigate the effects of VCAM-1 on migration and invasion of glioma cell lines . Methods The techniques of lentivirus pSGU6/GFP/Neo-based VCAM-1 shRNA and EF1 a-GFP/puro-based VCAM-1 expression vector, the scratch wound healing migration and transwell invasion assays , and the Western blot and cell staining were applied to observe the effects of VCAM-1 expression levels on migration and invasion of glioma cell line cells.There are four groups in T98G cells including control, vector, scramble and shRNA-VCAM-1 groups and three groups in U251 cells covering control, vector and VCAM-1 overexpressed groups ( n=6 per group) .Results The stabled glioma cell lines of T98 G cells with down-regulated VCAM-1 and U251 cells with VCAM-1 overexpression were established by using lentivirus-based VCAM-1 shRNA and expression vector.The ability of scratch wound healing (migration activity) decreased significantly (P<0.01) in T98G cells with lower VCAM-1 expression levels, while the migration activity was obviously improved in U251 cells with overexpressed VCAM-1 ( P <0.05 ) .Similarly, the invasion ability was significantly inhibited ( P <0.05) in T98G cells with silenced VCAM-1, as well as VCAM-1 overexpression could enhance the invasion ability of U251 cells ( P<0.01 ) .Conclusions VCAM-1 improves the migration activity and invasion ability of human glioma cell line cells.