Gecko crude peptides inhibit migration and lymphangiogenesis by down regulating the expression of VEGF-C in human hepatocellular carcinoma cells and human lymphatic endothelial cells / 中国药理学与毒理学杂志

OBJECTIVE To explore the role of gecko crude peptides (GCPs) in the proliferation, apoptosis, migration and lymphangiogenesis of human hepatocellular carcinoma cells (HepG2) and human lymphaticendothelial cells (HLECs) in vitro. METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the anti- proliferative effect of GCPs and siRNA-VEGF-C on HepG2 cells, Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis. The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay. The protein and mRNA expressions of vascular endo?thelial growth factor-C (VEGF-C) and CXC chemokine receptor-4 (CXCR4) were detected by q-PCR, immunofluorescence, Western blot. The protein expressions of the extracellular signal regulated kinase (ERKI/2), c-Jun N-terminal kinase (JNK), p38-mitogen activated protein kinases (p38 MAPK), serine/threonine kinase (Akt) and phosphatidylinositol- 3- kinase (PI3K) were detected by western blot. The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay. The protein and mRNA expressions of vascular endothelial growth factor receptor-3 (VEGFR-3) and stromal cell-derived factor-1 (SDF-1) were detected by q-PCR, Western blot. RESULTS GCPs and siRNA-VEGF-C inhibited HepG2 proliferation, invasion and migration, and the most obvious inhibitory effect was both synergistic effects. Thus, GCPs suppressed HLECs proliferation, migration and tube-like structure formationin a dose- dependent manner, and had inhibitory effect of tumor- induced lymphangiogenesis in vitro. Additionally, we found that GCPs and siRNA- VEGF- C decreased the expressions of MMP-2, MMP-9, VEGF-C, CXCR4, phospho-ERK1/2, phospho-P38, phospho-JNK and PI3K in HepG2 cells. Moreover, GCPs had a dose-dependent depressive effecton the expressions of VEGFR- 3, SDF- 1 in HLECs. CONCLUSION The low expression of VEGF- C mediated by siRNA-VEGF-C and GCPs inhibit tumor proliferation, invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C, and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.

Differentially expressed genes of HepG2 cells treated with gecko polypeptide mixture / 中国药理学与毒理学杂志

OBJECTIVE In order to investigate the possible anti-tumor molecular mechanisms of gecko polypeptide mixture (GPM). METHODS RNA-seq technology was used to identify the differen?tially expressed genes of human hepatocellular carcinoma (HCC) HepG2 cells treated with or without GPM. The HepG2 cells were treated with different concentration of GPM (0, 0.1, 0.2, 0.3, 0.4 mg·mL-1) for 6 h, 12 h and 24 h, respectively. MTT assay was used to detect the viability of HepG2 cells. DAPI fluorescence staining was performed to observe nucleus morphological changes of HepG2 cells. Western blot analysis was applied to observe the expression of apoptosis-related proteins in HepG2 cells. RESULTS The results showed that GPM could induce HepG2 cells apoptosis and influence HepG2 cells proliferation in a dose-dependent manner. We applied many analysis methods, including differen?tially expressed genes analysis, Gene Ontology (GO) enrichment analysis, KEGG pathway enrichment analysis, protein- protein interaction network analysis to screen out possible molecular mechanisms. ER-nucleus signaling pathway, cellular response to stress and apoptotic processes were identified the potential anti-cancer molecular biological process of GPM. GPM may also induce apoptosis in HepG2 cells via endoplasmic reticulum stress pathway. The mechanism is closely related to ERs, which might be beneficial for clinical therapy of HCC. CONCLUSION GPM can inhibit cells proliferation and induce apoptosis in HepG2 cells. The gene expression profile of GPM in HepG2 cells was obtained. The present study revealed the potential anti-tumor mechanism of GPM.

Study on excretion of 20 (S) -protopanaxadiolocotillol type epimers in rats / 中国中药杂志

Gindenosides are the active ingredients of Panax ginseng. 20 (S) -protopanaxadiolocotillol type epimers are the main metabolites of 20 (S) -protopanaxadiol. The previous studies showed that there are stereoselectivity difference in pharmacodynamics and pharmacokinetics between 24R-epimer and 24S-epimer. The purpose of this study was to explore the excretion of the epimers in bile, feces and urine of rat. Liquid chromatography tandem mass spectrometry method has been performed for determination of 24R-epimer and 24S-epimer in bile, feces and urine. 24R-epimer or 24S-epimer was intragastric administered to rats at a single dose of 10 mg x kg(-1). Results showed that after administration the recovery of 24R-epimer and 24S-epimer in feces was 17.69% and 17.09%, respectively, while both of the two epimers were hardly detected in urine. The 48 h cumulative biliary excretion rate of 24R-epimer was 8.01% after administration, while only 1.47% for 24S-epimer. It indicated that there are stereoselectivity in biliary excretion of the epimers with intragastric administration.

Study on fatigue of pilots during simulated flight training based on electroencephalogram / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study EEG during simulated flight training, then to explore the situational character of mental fatigue of the pilots.</p><p><b>METHODS</b>Fifty male pilots were randomly choice to be recorded their EEG including rest,flight and recovery phase. Then the band waves of EEG were analysed among the three phase.</p><p><b>RESULTS</b>(1) During flight, the amplitude of alpha, beta, SMR, theta, gamma waves tended to increase. These changes seemed obviously in SMR, theta waves. (2) After flight, all the amplitude of the waves declined except for beta wave of the right lobe,and most of them were still higher than those before flight (P < 0.05); (3) The right brain showed a higher cerebration than the left, especially in theta wave.</p><p><b>CONCLUSION</b>Mental fatigue is started during the flight training, and the right lobe has a severe mental fatigue than the left. After flight, mental fatigue showed a slow recovery.</p>

Correlation analysis between nutritive components of Whitmania pigra and Bellamya purificata / 中国中药杂志

The dried Whitmania pigra is used for the treatment of cardiovascular and cerebrovascular diseases in traditional Chinese medicine. Bellamya purificata is widely distributed in the Chang Jiang River basin, it is natural diets of W. pigra. Current study was conducted to compare and analyze the nutritional ingredient in W. pigra, body fluid and flesh of B. purificata. Results showed that the contents of protein, crude fat and total sugar in W. pigra, body fluid and flesh of B. purificata were significantly different (P < 0.05). Protein content in W. pigra accounts up to 65.01%. The contents of inorganic elements and amino acid were abundant in W. pigra, body fluid and flesh of B. purificata. The content of essential amino acids in them were 32.6, 221.59, 40.78 mg x g(-1), respectively. The content of flavor amino acid in them were 27.51, 14.5, 32.03 mg x g(-1), while the coresponding content of antioxidant amino acid were 8.81, 5.91, 9.73 mg x g(-1), respectively. The individual amino acids of high content in them were Glu, Asp and Leu. Macro elements Ca, P, Mg and trace elements Zn, Si, Fe were abundant. It could be speculated that W. pigra may be a promising novel food, and the present results provide a foundation to develop artificial feed for W. Pigra.

Ultrasound assessment of left ventricular segmental myocardial transmural displacement characteristics in patients with hypertrophic cardiomyopathy / 中华医学超声杂志（电子版）

Objective To evaluate the segmental myocardium of left ventricular wall in patients with myocardial hypertrophic cardiomyopathy (HCM) by TDI-Q, explore whether the segmental myocardium contractile function is changed or not and determine the myocardial mechanics parameters variation. Methods Thirty-two healthy volunteers and twenty-one patients with hypertrophic cardiomyopathy were included and the standard dynamic two-dimensional tissue Doppler imaging (TDI) of mitral, papillary muscle and apical short axis view were collected in three consecutive cardiac cycles. The mechanical parameters variation and characteristics of systolic radial peak displacement (RD) and time to peak in left ventricle subendocardial, mid-myocardium and epicardial myocardium at different level and segment were analyzed.Results In healthy control group, at left ventricular basal, apical and papillary muscle level, there was no significant difference for time to peak and systolic radial peak displacement (F=0.74, 1.28 and 1.79, all P>0.05). In patients with HCM, time to peak of systolic RD at left ventricular different level was asynchronous. Time to peak of RD in septum at papillary muscle levels and apical lateral wall were longer than those of other segments. In healthy control group, except for apical inferior and lateral wall, RD of subendocardial myocardium was significantly greater than that of epicardial myocardium at different segments (t=-1.903, 4.574,-3.552,-2.614,-1.728,-1.790,-1.836,-2.794 and 2.733, all P<0.05 ). In patients with HCM, RD of subendocardial myocardium was significantly greater than that of epicardial myocardium in posterior wall, septum at basal level and in inferior wall, posterior wall and lateral wall at papillary muscle level (t=-2.305,-2.148, 3.550,-1.182 and-3.602, all P < 0.05). At the same segment, transmural RD of subendocardial and subepicardial myocardium in healthy subjects were greater than that in patients with HCM. In inferior wall, posterior wall, lateral wall and septum at basal level, in inferior wall, posterior wall and septum at papillary muscle level, and in lateral wall and septum at apical level, differences of transmural RD were statistically significant (t=-3.787,-2.983,-4.325,-6.972,-2.352, 2.823,-3.274,-1.338 and-2.857, all P<0.05). Conclusions In patients with HCM, synchrony of left ventricular motion at different level was abnormal and transmural RD of endocardial and epicardial myocardium was decreased, which suggested regional systolic dysfunction. Ultrasound assessment of left ventricular segmental transmural mechanics can further reveal left ventricular mechanical characteristics in patients with hypertrophic cardiomyopathy.

Expression, purification and enzymatic characterization of adenylate kinase of Thermus thermophilus HB27 in Escherichia coli / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone the gene encoding adenylate kinase of Thermus thermophilus HB27, an extremely thermophilic bacterium, express the protein in Escherichia coil and study the enzymatic characterization.</p><p><b>METHODS</b>The DNA fragment encoding adenylate kinase was obtained by PCR from the total DNA of Thermus thermophilus HB27 and cloned into the vector pET-28a. The recombinant plasmid was identified by PCR, restriction endonuclease digestion and sequence analysis. Enzymatic characterization of the expressed protein was carried out using spectrophotometric assays.</p><p><b>RESULTS</b>The gene coding for adenylate kinase from Thermus thermophilus HB27 was cloned and the protein was overexpressed in Escherichia coli BL21(DE3). The optimum reactive pH and temperature for the enzyme were 8.5 and 90 degrees celsius;, respectively. The Km of the recombinant adenylate kinase for ADP was 68.6 micromol/L, with an V(max)ADP of 0.294 mmol/(L.min). Under the condition of environmental temperature at 70, 80, 90, or 100 degrees celsius; for 7 h, the recombinant adenylate kinase still retained the enzymatic activity with high thermostability. AP5A, a specific adenylate kinase inhibitor, inhibited the enzymatic activity of the protein by 70% at the concentration of 2.0 mmol/L, with a Ki value of 46.39 micromol/L for ADP.</p><p><b>CONCLUSION</b>The gene coding for adenylate kinase of Thermus thermophilus HB27 has been successfully cloned and expressed in Escherichia coil, which provides the basis for potential use of the highly thermostable recombinant HB27 adenylate kinase.</p>

Correlating endothelin releasing function of vascular endothelial cells to tea polyphenols and angiotensin Ⅱ / 中国组织工程研究

BACKGROUND: Angiotensin Ⅱ (Ang-Ⅱ) can stimulate vascular endothelial cells to excrete endothelin, a kind of potent vasoconstrictor. The content of endothelin in blood or cell culture media directly reflects the function and injured status of vascular endothelial cells. Therefore, it is significant for strengthening vascular endothelial cells to resist the injured factors. Tea polyphenols is a mainly active component of tea, and it is considered as a reagent for anti-atherosclerosis, protecting injuries of vascular endothelial cells and preventing cardiovascular diseases. OBJECTIVE: To observe the effects of tea polyphenols in different concentrations on endothelin content in vascular endothelial cells induced by Ang-Ⅱ at various time points through establishing Ang-Ⅱ-induced vascular endothelial cell injury models and further to investigate the protective effect of tea polyphenols on vascular endothelial cells. DESIGN: Observational study.SETTING: Aerospace and Diving Medical Center, Navy General Hospital of Chinese PLA.MATERIALS: The experiment was carried out in Laboratory of Aerospace and Diving Medical Center, Navy General Hospital of Chinese PLA from March to September 2005. Main materials were detailed as follows: Ang-Ⅱ (Sigma Company), tea polyphenols (Department of Tea Science, Zhejiang University) and vascular endothelial cells (human large artery vascular endothelial cell system, CBI Company, USA).METHODS: Cultured vascular endothelial cells were divided into 4 groups: ① Control group: The normal culture media was added in the isopyknic vascular endothelial cells, and 100 μL supernatant was extracted before and at 0.5, 6 and 24 hours after filling moisturized liquid. ② Ang-Ⅱ group: Cell culture media containing 10-7 mol/L Ang-Ⅱ was added in the vascular endothelial cells, and other operations were as the same as those in the control group. ③ High-concentration tea polyphenols + Ang-Ⅱ group: Cell culture media containing 50 mg/L tea polyphenols was added in the vascular endothelial cells, and other operations were as the same as those in the Ang-Ⅱ group. ④ Low-concentration tea polyphenols + Ang-Ⅱ group: Cell culture media containing 25 mg/L tea polyphenols. 100 μL supernatant was extracted before and after 0.5, 6 and 24 hours treatment in each group. Thereafter, radioimmunoassay was used to measure the content of endothelin. MAIN OUTCOME MEASURES: Content of endothelin.RESULTS: ① Content of endothelin in Ang-Ⅱ group was higher than that in the control group (P ＜ 0.01). ② At 6 and 24 hours after high-concentration tea polyphenols incubation, content of endothelin in high-concentration tea polyphenols + Ang-Ⅱ group was lower than that in Ang-Ⅱ group (P ＜ 0.01). Moreover, the content of endothelin in low-concentration tea polyphenols + Ang-Ⅱ group was lower than that in both high-concentration tea polyphenols + Ang-Ⅱ group and angiotensin Ⅱ group (P ＜ 0.01). CONCLUSION: Tea polyphenols has inhibitory effects on endothelin releasing function of vascular endothelial cells induced by Ang-Ⅱ, suggesting that tea polyphenols has protective effect on vascular endothelial cells, and the effect of low-concentration tea polyphenols is superior to that of the high-concentration one.

Effect of activation of peroxisome proliferator-activated receptor γ on cell cycle distribution of human gastric carcinoma / 肿瘤

Objective: To explore the effect of activation of peroxisome proliferator-activated receptor gamma (PPARγ) on cell cycle arrest of gastric carcinoma cell line MGC803. Methods: The inhibitory effect of pioglitazone (PGZ) on proliferation of MGC803 cells was analyzed by MTT assay. Cell cycle were detected by flow cytometry (FCM). The protein expression of PPARγ, cyclinD1 and cell cycle protein-dependent kinase CDK4 in MGC803 cells was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Treatment with 0.1-10 μmol/L, PGZ for 96 h significantly inhibited cell proliferation. The proportion of MGC803 cells at G1 phase was significantly increased when treated with 10 μmol/L PGZ for 48, 72 and 96 h, and showed an apparent G1 phase arrest. The expression of PPARγ was at a low level in MGC803 cells and significantly up-regulated when treated with 10 μmol/L, PGZ for 48 h (P < 0.01). The expression of CDK4 in MGC803 cells was significantly down-regulated when treated with 10 μmol/L, PGZ for 96 h (P < 0.01) and the expression of cyclinD1 was slightly down-regulated. Conclusion: Activation of PPARγ significantly induces G1 phase arrest, which is associated with down-regulation of the expression of CDK4 and cyclinD1.

Different protective effect between tea polyphenol and esmolol on vascular endothelial cells secreting endothelin induced by hypoxia combined Ang-2 / 重庆医科大学学报

Objective: To compare the protective effect of Tea polyphenol with that of esmolol on vascular endothelial cells secreting endothelin induced by hypoxia combined Ang-2. Methods: Vascular endothelial cells were divided into 4 groups that included control group,hypoxia+Ang-2 group,hypoxia+Ang-2+tea polyphenol group,hypoxia+Ang-2+esmolol group,and each group comprised eight samples. The changes of endothelin expression level were measured with radioimmunoassay at the time of 0.5 h,6 h,and 24 h. Results: ①The level of endothelin expression in hypoxia+Ang-2 group was higher significantly than that in the control group(P0.05),but there was significantly different at the time of 6 h and 24 h after hypoxia combined Ang-2(P

Clinical research of pneumonia in acute cervical spinal injury / 中国康复理论与实践

@#ObjectiveTo investigate the prevention and treatment of pneumonia in patients with acute cervical spinal cord injury (CSCI).MethodsData of 278 patients with acute traumatic CSCI admitted from 1988 to 2004 were analyzed retrospectively.Results Pneumonia was the major complication following acute CSCI and discovered by radiography during the first 3—33 days after injury. The all cases were nosocomial pneumonia and G- bacilli were main pathogens, particularly pseudomonas aeruginosa. The incidence of pneumonia of patients with score ≤6 according to the criteria of American Spinal Injury Association (ASIA) was significantly higher than those with ASIA score >6 (P<0.001).ConclusionThe high incidence of pneumonia in the CSCI is associated with the level and completeness of the injury. The G- bacilli causing nosocomial infection are main pathogens.

Synthesis and in vitro antibacterial activities of new 3,5-disubstituted oxazolidinone compounds / 药学学报

<p><b>AIM</b>To design and synthesize new oxazolidinone antibacterial agents.</p><p><b>METHODS</b>The synthetic method reported in literature has been modified and new 3,5-disubstituted oxazolidinone compounds were synthesized on the basis of SAR reported in the literature and their antibacterial activities in vitro were determined.</p><p><b>RESULTS</b>Eighteen new objective compounds were synthesized, and their structures were determined by IR, 1HNMR and FAB-MS. Within the eighteen new objective compounds, sixteen compounds showed antibacterial activity in vitro and compound 9, 10 and 10b showed better antibacterial activities in vitro than ciprofloxacin (CIP), sultamicillin (Sul) and vancomycin (VCO). Compounds 9a and 11c have no antibacterial activity in vitro at all.</p><p><b>CONCLUSION</b>Compounds 9, 10 and 10b are worthy to be intensively studied.</p>

The collecting and processing system of the sound signal of larynx / 中国医疗器械杂志

This paper presents a new kind of collecting and processing system of the sound signal of larynx where double sound cards and software filter are used. The installation of the double sound cards and processing proposal of software are mainly discussed and a new kind of ADC method of dual-channel sound signal is put forward in this paper. This system has the feature of reliable performance, simple installation and easy maintenance.