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Industrial pure titanium (TA2) and Ti-6Al-4V (TC4) have been widely available for chemical equipment. However, the corrosion resistance changes during their post-fire performance. In this research, the electrochemical properties of industrial pure titanium and Ti-6Al-4V after heat treatment and different cooling methods were analyzed by AC impedance technique and dynamic polarization method. The results support two conclusions. Firstly, when the temperature is approximate to the phase transition point, for pure titanium and its alloys, water-cooling can obtain better corrosion resistance than air-cooling. With regard to pure titanium, its best corrosion resistance can be obtained at 800°C under water-cooling. For Ti-6Al-4V, its best corrosion resistance can be obtained when the temperature reached 910 °C under water cooling condition. Secondly, the corrosion current density under acid corrosion condition has risen compared with neutral corrosion condition. These results can provide some experimental data and theoretical basis for post-fire anticorrosion performance of titanium materials, and also support the safety, risk evaluation of titanium equipment.

Cortactin, a member of the actin-binding protein family, plays an important role in cell movement involving the cytoskeleton, as cell movement mediated by cortactin may induce the epithelial-mesenchymal transition. Cortactin participates in tumor proliferation, migration, and invasion and other related disease processes by binding to different proteins and participating in different pathways and mechanisms that induce the occurrence of these disease processes. Therefore, this article reviews the correlations between cortactin, the actin cytoskeleton, and the epithelial-mesenchymal transition and discusses its clinical importance in tumor therapy.

MAPK signaling pathways play critical roles in plant immunity. Here, we silenced multiple genes encoding MAPKs using virus-induced gene silencing mediated by Bean pod mottle virus to identify MAPK genes involved in soybean (Glycine max) immunity. Surprisingly, a strong hypersensitive response (HR) cell death was observed when soybean MAPK KINASE KINASE1 (GmMEKK1), a homolog of Arabidopsis (Arabidopsis thaliana) MEKK1, was silenced. The HR was accompanied by the overaccumulation of defense signaling molecules, salicylic acid (SA) and hydrogen peroxide. Genes involved in primary metabolism, translation/transcription, photosynthesis, and growth/development were down-regulated in GmMEKK1-silenced plants, while the expression of defense-related genes was activated. Accordingly, GmMEKK1-silenced plants were more resistant to downy mildew (Peronospora manshurica) and Soybean mosaic virus compared with control plants. Silencing GmMEKK1 reduced the activation of GmMPK6 but enhanced the activation of GmMPK3 in response to flg22 peptide. Unlike Arabidopsis MPK4, GmMPK4 was not activated by either flg22 or SA. Interestingly, transient overexpression of GmMEKK1 in Nicotiana benthamiana also induced HR. Our results indicate that GmMEKK1 plays both positive and negative roles in immunity and appears to differentially activate downstream MPKs by promoting GmMPK6 activation but suppressing GmMPK3 activation in response to flg22. The involvement of GmMPK4 kinase activity in cell death and in flg22- or SA-triggered defense responses in soybean requires further investigation.

Noni (Morinda citrifolia L.) is an edible and medicinal plant distributed in Hainan, China. The antibacterial activities of the extracts of water (WE), petroleum ether (PEE), ethyl acetate (EAE), chloroform (CE), and n-butanol (BE) were assayed by the disk diffusion method. The results showed that the extracts from Noni leaves possessed antibacterial effects against Bacillus subtilis, Escherichia coli, Proteus vulgaris, and Staphylococcus aureus. Among 5 different extracts, the BE produced the best antibacterial activity. The samples were first extracted by ethanol, and the primary compounds in the BE fraction of ethanol extract was further isolated and identified. Six phenolic compounds, including 5, 15-dimethylmorindol, ferulic acid, p-hydroxycinamic acid, methyl 4-hydroxybenzoate, methyl ferulate, and methyl 4-hydroxycinnamate, were identifiedby NMR. The results indicated that the phenolic compounds might significantly contribute to antibacterial activities of Noni leaves.

The present study investigates the overcharge cycling effect on thermal behavior, structure, and electrode material of lithium-ion batteries (LIB) with a Lix(Ni0.3Co0.3Mn0.2)O2 cathode. The thermal behavior of LIBs with different overcharged degrees was studied using vent sizing package 2 and differential scanning calorimetry. Changes in the internal composition of the batteries during overcharge were observed using a scanning electron microscope and an inductively coupled plasma optical emission spectrometer. The results indicate that the overcharge process can be divided into four stages. A battery was more likely to trigger thermal runaway during overcharge under adiabatic conditions than in an ambient environment. During overcharge, the surface temperature of a 150% state-of-charge (SOC) battery was approximately 3.0 °C higher than that of a 100% SOC battery. Because of generated gas and lithium deposition during overcharge cycling, the battery thickness increases and coulombic efficiency decreases. The apparent exothermic onset temperature ( T 0 ) of a battery with 150 % SOC was 87 °C below that of a 100 % SOC battery. Cathode material becomes highly reactive when the battery was overcharged to 150 % SOC. Here, the released heat was 1026.4 J/g. These results provide feasible support for understanding the overcharge mechanism and battery management system.

For the particularity of rock and gas outburst in layered composite rock mass, taking the large outburst accident of the 21st down slope at +0 horizon in Zhengzhou Daping coal mine for example, we study the dynamic response characteristics of layered system and the whole process of rock and gas outburst under explosion load based on secondary development of ABAQUS user subroutine, and then innovatively put forward the new pattern “structural surface damage reinforcement” and “ribbon outburst of rock and gas”. The results are well consistent with 10.20 accident, which indicate the coupling relationship between rock damage and gas outburst under complicated geological conditions: (1) The weak structural surface damages dramatically and easy to form high pressure and high gradient of gas enrichment zone near the tunnel face under conditions of gas damaging erosion and blasting vibration; (2) The gas will break through the rock impedance and flow rapidly which result in rock burst once high pressure gas along structural face breakdown the safe rock pillar between fault and tunnel face.

Dust samples were collected for the heavy metals pollution survey from Xiqing District in Tianjin, including forty-three sites of Outer-ring West Road, Jinlai Road, Gong No.1Road and Xiuchuan Road. After collecting and pre-treatment about road surface dust, particle size, pH and heavy metal content were also measured and analyzed. Results show that the distribution of road dust particles in Xiqing District is irregular, most of them are between 106 ~ 250 and the average diameter of low-grade road dust particles is larger than that of high-grade road dust particles. The collected road dust samples imply that it is alkaline. Heavy metal concentrations witness a downward trend with increasing dust particle size. The average content of heavy metals in road dust is listed in the sequence of Gong No.1Road, Outer-ring West Road, Jinlai Road, and Xiuchuan Road, and the order of heavy metals is: Zinc (Zn); Chromium (Cr); Copper (Cu); Lead (Pb); and Cadmium (Cd) which are higher than those in Tianjin Soil background values, especially Cadmium (Cd) and Lead (Pb) which are 10 times more. Those Heavy metals including Cadmium (Cd), Lead (Pb), Copper (Cu), Chromium (Cr), and Zinc (Zn) are mainly associated with anthropogenic activities, such as transportation and industrial emissions. The sources of heavy metal Cadmium (Cd) are complex and may result from the wear and tear of automobile tires, brakes and so on. The potential ecological risk index method is implemented to evaluate ecological risk of heavy metals in road dust which shows that the area for study is at slight ecological risk.

Adsorption-desorption characteristics of E. coli by quartz sands of five different particle size (351、319、217、71、27μm) were studied using batch equilibrium method. The results showed that quartz sands had great adsorption quantity of E. coli. The adsorption ratio could reach above 95%,when the initial microbial concentration was 109cfu/mL. With the decreased of the quartz sand particle size,the adsorption quantity increased,and adsorption reached balance faster. The Pseudo-second-order rate equation is optimal to describe adsorption kinetics of E. coli by quartz sands, and the isothermal adsorption process is suitable to describe using Henry equation. Quartz sands had small desorption ratio of E. coli,the desorption ratio increased as quartz sands particle size increased, the largest desorption ratio was only 3.6%.The desorption dynamic processes and desorption isothermal processes also can be decreased using Pseudo-second-order rate equation and Henry equation respectively. Desorption of E. coli on quartz sands existed obvious hysteresis. It indicated there existed irreversible adsorption between E. coli and quartz sands, which the E. coli adsorbed was hard to desorb. The average particle size of quartz sands have some relevance with the parameters of Henry equation and Pseudo-second-order rate equation. This shows that particle size is one of the important factors that affect the adsorption and desorption.

This paper aims to the study of dedusting effects in confined workplace using finite volume method. First, the diffusion phenomenon of dust flow is described by discrete phase model, and the numerical results are compared with the experimental data. Second, the concentration of dust flow, which mixes with SiO2 and air, is calculated by three different dedusting methods, such as the exhaust fan method, the window ventilation method and the humidifying spray method. In terms of comparison results, the exhaust fan method has the best dust removal effect in confined workplace, and the humidifying spray method has the worst effect. The paper provides a technical support for dedusting design of confined workplace in the occupational hazards prevention field.

A safety evaluation index system regarding to the current safety situation of large recreation facilities in China is established. 13 secondary standard items are built by considerring human factor, equipment factor, environment factor and management factor. The existing safety evaluation of large recreation facilities are conducted by qualitative evaluation methods with highly fuzziness. The evaluation results are uncertain. After the network training, a safety evaluation model based on BP neural network is built. It can reduce the subjectivity of qualitative evaluation effectively with more scientific and objective results. Through the model based on BP neural network, the present safety situation of one large amusement facility is evaluated. The evaluation result is consistent with the actual situation. The method based on BP neural network in the paper provides a new method for safety evaluation of large recreation facilities.

In order to increase the effectiveness of remediation on oil-contaminated soil, culture techniques and molecular biological methods were adopted to analyze community structure enriched and domesticated by different ways. The results showed that the bacterial concentration cultured in a medium of inorganic salts and oil was 1.30×1012cfu/ml, while the bacterial concentration cultured in a medium of LB and oil was 3.05×108cfu/ml; Within the perspective of structure, the microbial community cultured in a medium of inorganic salts and oil contained 11 kinds of bacterial, Meanwhile, the microbial community cultured in a medium of LB and oil was composed of 4 kinds of bacteria. Based on the results of this study, it can be inferred that the cultured medium has a significant influence on the composition of microbial communities. When domesticated oil-contaminated soil was cultured in a medium of inorganic salts and oil, the microbial concentrations and diversities were relatively higher than those cultured in LB and oil.

The present study explored changes of the SIRT6/NF-κB pathway in myocardial hypoxia/reoxygenation induced injury and the effects on mitochondrial damage and myocardial damage by regulating SIRT6. SIRT6 expression decreased and NF-κB expression increased in H9c2 cells during hypoxic injury. Cell death and mitochondrial defects paralleled mPTP opening, and a decrease in ΔΨm occurred in hypoxic myocytes compared with normoxic control cells in annexin V and propidium iodide staining and TUNEL results. These effects were suppressed in cells overexpressing SIRT6, but reemerged in cells expressing the SIRT6 mutant. We also found that NF-κB p65 increased in both the cytoplasm and nuclei, which could be repressed by SIRT6 overexpression. The expression level of NF-κB was significantly and negatively correlated with the SIRT6 mRNA level. Our data demonstrated that SIRT6/NF-κB changed during hypoxic injury and SIRT6 overexpression averted mitochondrial defects through inhibition of NF-κB in hypoxic H9c2 cells. Activation of SIRT6 may be a potential method for hypoxia/reoxygenation injury therapy.

To observe the expression of aquaporin 4 (AQP4) in diffuse brain injury (DBI) of rats and to explore the corresponding effect of AQP4 for brain edema.The rat model of DBI was established using Marmarou's impact-compression trauma model. Brain water content was measured by dry-wet weight method. Blood-brain barrier permeability was evaluated by Evans blue (EB) staining. Immunohistochemical method was used to observe the expression of AQP4.Brain water content increased after 3 h and peaked at 24 h after DBI. Brain EB content significantly increased and peaked at 12 h after DBI. The expression of AQP4 significantly increased after 3 h and peaked at 24 h after DBI, and the number of AQP4 positive astrocytes increased.The increment of the permeability of blood-brain barrier and the expression of AQP4 may contribute to the development of brain edema in rat DBI. The change of AQP4 expression in astrocytes may also contribute to determine DBI.

Patients with Crohn's disease (CD) or ulcerative colitis (UC) undergo various therapies, including antibiotic therapy. This meta-analysis of controlled clinical trials was conducted to evaluate whether the use of antibacterial therapy improves the clinical symptoms of inflammatory bowel disease (IBD). The Medline and Scopus databases were searched and a systematic review was performed. Randomized, controlled trials in which antibiotic therapy was compared with placebo were investigated. A total of 10 randomized, placebo-controlled clinical trials for CD were included in the meta-analysis. The pooling of the data from these trials yielded an odds ratio (OR) of 1.35 [95% confidence interval (CI), 1.16-1.58] for antibiotic therapy compared with placebo in patients with CD. Furthermore, nine randomized placebo-controlled clinical trials for UC matched our criteria and were included in the analysis. The pooling of the data from these trials yielded an OR of 2.17 (95% CI, 1.54-3.05) in favor of antibiotic therapy. These results suggest that antibiotics improve clinical outcomes in patients with IBD.

Through the research on stress metamorphism character of the II1 coal seam in Ludian gliding structure, the stress effecting factors of metamorphism and hydrocarbon generation process of tectonic coal were studied. It is shown that different tectonic stress has different coal evolution effects. Compared with tensional stress, compressive stress, which has apparent anisotropic characteristics, has greater stimulative effect against the increase of coal metamorphic degree, the abscission of functional group and side chains of basic structural unit, and the formation of gas primarily composed of methane.

Noni (Morinda citrifolia L.) is an edible and medicinal plant distributed in Hainan, China. The antibacterial activities of the extracts of water (WE), petroleum ether (PEE), ethyl acetate (EAE), chloroform (CE), and n-butanol (BE) were assayed by the disk diffusion method. The results showed that the extracts from Noni leaves possessed antibacterial effects against Bacillus subtilis, Escherichia coli, Proteus vulgaris, and Staphylococcus aureus. Among 5 different extracts, the BE produced the best antibacterial activity. The samples were first extracted by ethanol, and the primary compounds in the BE fraction of ethanol extract was further isolated and identified. Six phenolic compounds, including 5, 15-dimethylmorindol, ferulic acid, p-hydroxycinamic acid, methyl 4-hydroxybenzoate, methyl ferulate, and methyl 4-hydroxycinnamate, were identifiedby NMR. The results indicated that the phenolic compounds might significantly contribute to antibacterial activities of Noni leaves.

Against the particularity of stratum-structure in “three soft” mine areas, according to rock indoor test and on-site sucking experiment, discussed the characteristics of argillization, compression fracture and sucking technique of soft coal with low permeability. It is clearly pointed out that the gas can be highly effectively sucked only by compression fracture along the occurrence of the coal seam, creating inter-seams crack belt because of the difference of bulgy deformation. After the flooding experiment in the 24080 workface of Pingdingshan No.10 mine, the average single-bore volume of gas increases from 77 m3 to 7 893 m3, while decay cycle extended from 7 days to 80∼90 days. Also, the single-bore extracting rate of gas increases to 33%.

Based on the basic principles of hydrogeology and soil mechanics, studied the genesis mechanism and control factors of settling of ground surface caused by the drainage of the aquifer in the construction of coal mines, and put forward a corresponding calculating model demonstrated by practical example. The study provides mining areas, which are covered with a very thick Quaternary soil layer and abundant ground water, with a theoretical basis aimed at forecasting the settling of ground surface.

OBJECTIVE: To investigate combined chemotherapeutic effects of rofecoxib in combination with 5-fluorouracil (5-FU), cisplatin (DDP) and etoposide (VP-16) in vitro, and to explore the potential mechanisms in modulating multidrug resistance (MDR) expression. MEHTODS: The BGC-823 gastric cancer cell line was incubated for 48 h with 0.1 µmol/L rofecoxib, 5-FU, DDP and VP-16 (1 µg/mL, 10 µg/mL and 100 µg/mL) alone, and combined with rofecoxib, respectively. Methyl-thiazolyl-tetrazolium and the terminal deoxynucleotidyl transferase–mediated 2′-deoxyuridine 5′-yriphosphate nick-end labeling assays were performed to calculate inhibitory rates and apoptotic index. Middle effects principles (CI values) were used to determine the interaction between rofecoxib and chemotherapeutic agents. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis were employed to determine expression of MDR1, multidrug resistance-associated protein 1 (MRP1), glutathione S-tranferase-π (GST-π) mRNA and protein in gastric cancer cells administered by rofecoxib, respectively. RESULTS: Both anticancer drugs such as 5-FU, DDP and VP-16 and rofecoxib inhibited the cells' proliferation and induced apoptosis in a dose-dependent manner, and a more significant inhibition was achieved when the cells were co-treated with anticancer drugs and rofecoxib. There was a synergetic role when different concentrations of chemotherapeutic agents were combined with rofecoxib (all CI < 1, P < 0.01 or 0.05). RT-PCR analyses of MDR gene families in BGC-823 gastric cancer cells revealed a strong expression in MRP1 and GST-π mRNA, but MDR1 mRNA was undetectable. After administration with different concentrations of rofecoxib (0.1, 1.0, 10 µmol/L), significant downregulation of MRP1 and GST-π mRNA was observed (MRP1: from 0.984 ± 0.093–0.513 ± 0.098; GST-π: from 1.078 ± 0.201–0.472 ± 0.084, P < 0.01 or 0.05). In addition, MRP1 and GST-π protein expression induced by rofecoxib were also reduced (P < 0.01 or 0.05). CONCLUSION: Rofecoxib, a specific cyclooxygenase-2 inhibitor, plays a chemotherapeutic sensitizer role in various anticancer agents on the BGC-823 gastric cancer cell line, which could be partly explained by its ability to reverse the intrinsic MRP1 and GST-πin vitro.

Gas outbursts from “three-soft” coal seams (soft roof, soft floor and soft coal) constitute a very serious problem in the Ludian gliding structure area in western Henan. By means of theories and methods of gas geology, structural geology, coal petrology and rock tests, we have discussed the effect of control of several physical properties of soft roof on gas preservation and proposed a new method of forecasting gas geological hazards under open structural conditions. The result shows that the areas with type III or IV soft roofs are the most dangerous areas where gas outburst most likely can take place. Therefore, countermeasures should be taken in these areas to prevent gas outbursts.

Field experimental studies on surge flow irrigation with sediment-laden water in northwestern China showed that a sealing layer was formed on the soil surface as a result of the sediments in the irrigation water going through a ‘trapping’, ‘settling’ and ‘sealing’ processes in the irrigated field. The sealing layer reduced the surface roughness and intake rate of the surface soil, and thus sped up the flow advance. Therefore, higher irrigation efficiency can be achieved with less irrigation water requirement. Analytical studies on water saving efficiencies indicated that the water saving effect of surge irrigation with sediment-laden water was achieved by an interactive impact of sediment content and the intermittent feature of surge flow, but the sediments effect was more pronounced.

Background The iso-osmolar contrast agent iodixanol may be associated with a lower incidence of cardiac events than low-osmolar contrast media (LOCM), but previous trials have yielded mixed results. Objective To compare the risk of total cardiovascular events of the iso-osmolar contrast medium, iodixanol, to LOCM. Methods Medical literature databases were searched to identify comparisons between iodixanol and LOCM with cardiovascular events as a primary endpoint. A random-effects model was used to obtain pooled odds ratio (OR) for within-hospital and 30-day events. Results A total of 2 prospective cross-sectional studies and 11 randomized controlled trials (RCTs) (covering 6859 subjects) met our criteria. There was no significant difference in the incidence of within-hospital and 30-day cardiovascular events when iodixanol was compared with LOCM, with pooled OR of 0.72 (95%CI 0.49–1.06, p = 0.09) and 1.19 (95%CI 0.70–2.02, p = 0.53), respectively. Subgroup analysis showed no relative difference when iodixanol was compared with ioxaglate (OR = 0.92, 95%CI 0.50–1.70, p = 0.80) and iohexol (OR = 0.75, 95%CI 0.48–1.17, p = 0.21). However, a reduction in the within-hospital cardiovascular events was observed when iodixanol was compared with LOCM in the RCT subgroup (OR = 0.65, 95%CI 0.44–0.96, p = 0.03). Sensitivity analyses revealed that three studies had a strong impact on the association of within-hospital cardiovascular events between iodixanol and LOCM. Meta-regression analysis failed to account for heterogeneity. No publication bias was detected. Conclusions This meta-analysis demonstrates that there is no conclusive evidence that iodixanol is superior to LOCM overall with regard to fewer cardiovascular events.

OBJECTIVE: To investigate the effects of different doses of angiotensin II (AngII) on the proliferation of hepatic stellate cells (HSC) and collagen synthesis in rats. METHODS: Pronase E and collagen I were used to isolate the HSC, and 3H-TdR,3H-Leu and 3H-pro incorporation methods were used to evaluate the effects of the different doses of AngII on HSC DNA, protein and proline synthesis. RESULTS: It was found that 10-9−10−6 mol/L AngII could increase the 3H-TdR incorporation rate of the HSC (P

To evaluate the impact of resveratrol on coronary collateral circulation in pigs suffered from experimental acute coronary occlusion.Eighteen healthy pigs were randomly divided into 3 groups: resveratrol group, nitroglycerin group and control group. Animal model of acute coronary occlusion was established through PTCA method, and the blood flow spectrum in the left circumflex artery (LCX) was detected using intracoronary Doppler ultrasound.The average peak velocity (APV) in infarction correlation artery (IRA) was significantly decreased immediately after coronary occlusion [(0.85 ± 0.25) cm/s vs. (24.83 ± 3.43) cm/s, P0.05]. The APV remained unchanged during 0, 30 and 60 minutes after the occlusion. Reversed or bidirectional blood flow was observed and the APV increased significantly [(9.22 ± 0.80) cm/s vs. (0.84 ± 0.21) cm/s, (8.93 ± 1.28) cm/s vs. (0.86 ± 0.26) cm/s respectively, P0.05] after the coronary injection of resveratrol (2 mg) or nitroglycerin (0.3 mg). There was no significant difference in peak APV between the resveratrol and nitroglycerin groups. The duration of increased APV was significantly longer in resveratrol group than that in nitroglycerin group [(58.83 ± 6.15) min vs. (21.80 ± 5.79) min, P0.05].The collateral circulation after acute coronary occlusion was obviously insufficient in pigs. Resveratrol could significantly improve the blood flow in coronary collateral circulation after acute occlusion in this model.

China's national economy will soon face "a low-carbon economy" historic opportunity for the new situation. So it is very important to synthetically use coal bed methane in "three soft" mine areas. According to research results and practical work experience , the author discusses the characteristics of argillization,compression fracture and sucking technique of soft coal with low permeability .It is clearly pointed out that the gas can be highly effectively sucked only by compression fracture along inter-seams crack belt.

In order to discuss the relation between rock burst and stress field in folding structure areas, the law the changing law of local stress area induced by mining was studied by FLAC2D simulation procedure and the targeted prevention measure was proposed from a strategic defense view based on the characteristics of rockburst in fold areas. The results show that the higher elastic energy will be accumulated easily and hazard of rockburst will be more serious when advancing downhill from anticline to syncline and mining the coalface near the wing parts firstly, then mining the coalface near the syncline. The direction of roadway should be parallel to the maximal horizontal stress or with a small angle. The most benefit direction of mining coal face should be advancing from syncline to anticline and the optimal layout should be mining the coalface firstly near the syncline.

The coal mines of Henan province are surely abundant water mining areas in China. The problem of the waste groundwater is much serious too. Through synthetical analyses of the water chemical characteristics , water chemical environment and the water chemical origin of acid groundwater, this paper proposes the principle of chemical sulfer erasing technique which can process mine acid water and the compound coagulant applying method. Some corresponding technology parameters are discussed. The weight ratio of Ca(OH) 2 and 4CaO·Al 2 O 3 ·Fe 2 O 3 is about 1:1.3~1:1.5. Quick lime quantity put in must diminish 50 mg/L when the temperature is increased by 10°C and diminish 25mg/L when pH value is increased by one unite. The processing of mine water with the method of chemical sulfer erasing technique is practicable.

SUMMARY 1 Resveratrol, a polyphenol in red wine, has a cardioprotective effect. Resveratrol-targeting protein (RTP) has been purified using a resveratrol affinity column (RAC) and has been identified as quinone reductase type 2 (NQO2). We hypothesize that NQO2 is the target protein of resveratrol in vascular smooth muscle cells (VSMC) and that resveratrol inhibits proliferation of VSMC through its action on NQO2. In the present study, we investigated the correlation between NQO2 regulation and cell proliferation in VSMC in response to resveratrol treatment. 2 The RTP was purified using RAC and was detected with a NQO2 polyclonal antibody. The VSMC were incubated with resveratrol (1, 10 and 50 µmol/L) for 24, 48 and 72 h. Cell proliferation was detected by cell counting and bromodeoxyuridine (BrdU) assay. A lentiviral vector incorporating NQO2 short interference (si) RNA of short hairpin design was constructed and transduced into VSMC. Real-time quantitative polymerase chain reactionwas used to measure NQO2 mRNA levels; NQO2 expression was determined by western blot analysis. 3 Using RAC, we extracted a 26 kDa protein from aortic smooth muscle, which was referred to as RTP-26. Proliferation of VSMC was inhibited by resveratrol in a concentration- and time-dependent manner. The mRNA and protein expression of NQO2 was also repressed by resveratrol in a concentration- and time-dependent manner. A similar pattern of inhibition was observed for cells treated with resveratrol (25 µmol/L) as for cells transduced with a lentiviral vector containing siRNA sequences against NQO2. 4 Collectively, these data indicate that the suppression of VSMC proliferation mediated by resveratrol correlates with NQO2 downregulation.

Cyclooxygenase-2 (COX-2) is related closely to the tumorigenesis of bladder cancer, and COX-2 inhibitor has potential antitumor effect. This study was to investigate the effects of selective COX-2 inhibitors on the proliferation and apoptosis of human bladder cancer cell line T24.The effects of selective COX-2 inhibitors SC-58125 and celecoxib, and nonselective COX inhibitor indomethacin on the proliferation of T24 cells were evaluated by MTT assay. Cell apoptosis was determined by flow cytometry (FCM), DNA ladder electrophoresis, and fluorescent microscopy with Hoechst33258 staining. The expression of apoptosis-related genes Bcl-2 and Bax were analyzed by reverse transcription-polymerase chain reaction (RT-PCR).Within concentrations of 12.5-200 micromol/L, SC-58125, celecoxib, and indomethacin could inhibit the proliferation of T24 cells to different extents. SC-58125 tended to be more effective than the other two. The 50% inhibition concentration (IC50) of SC-58125 was determined to be 25-50 micromol/L. The apoptosis of T24 cells was enhanced after exposure to SC-58125. When treated with 100 micromol/L SC-58125 for 6 and 12 h, the apoptosis rates of T24 cells were (7.95+/-1.88)% and (12.5+/-2.42)%, respectively, which were significantly higher than that of control cells (P0.05). But the expression of Bcl-2 and Bax genes did not change.Selective COX-2 inhibitor could inhibit the proliferation and induce the apoptosis of T24 cells.

To observe the role of PPARgamma during the activation process of hepatic stellate cells (HSC).By morphology and RT-PCR, we study the changes of expression of PPARgamma in culture-activated HSC or in vivo activated HSC induced by dimethylnitrosamine (DMN).In vitro, the expression level of PPARgamma in freshly isolated HSC (0.72+/-0.01) significantly reduced to 0.48+/-0.03 on the third day of culture (t = 19.8372, P0.01), and reduced 70% on the seventh culture-day and could not be detected after the second passage. In vivo, HSC freshly isolated from normal control rats expressed PPARgamma (0.76+/-0.01). During the development of rat liver fibrosis induced by DMN, the expression level significantly reduced to 0.46+/-0.02 after the third injection of DMN (t = 29.5318, P0.01), and reduced 66% on the end of first week and could not be detected on the end of second and third week.The expression of PPARgamma might play an important role on the maintenance of resting-form of HSC, and the reduction of expression of PPARgamma might be an early event during the activation process of HSC.

AIM: To detect the micrometastasis of gastric carcinoma in peripheral blood circulation using immunomagnetic beads sorting technique and RT-PCR technique, and to discuss its significance and the difference between the two methods. METHODS: Density gradient centrifugation was used to isolate mononuclear cells from peripheral blood, immunomagnetic beads sorting technique and RT-PCR technique were used to detect the disseminated carcinoma cells. HE, immunocytochemical and immunofluorescence staining were also used to identify the characteristics of the cells separated with immunomagnetic beads sorting technique. RESULTS: Cells expressing cytokeratin were separated and enriched from the peripheral blood specimens of patients suffering from gastric carcinoma or chronic gastritis. After HE staining, two kinds of cells with little cytoplasm were found. Majority of these cells had small and round nuclei, even chromatins and the thickness of nuclear membrane was normal. Immunohistochemical staining indicated that there were CD34 and CD45 expression on the cell membrane of this kind of cells and these cells also showed expressed human telomerase reverse transcriptase by immunofluorescence staining, but the expression of carcinoembryonic antigen was absent. So, these cells might hematopoiesis precursors. Another kind of cells had larger and abnormal nuclei with thicker nuclear membranes. Massed chromatins and poly-nucleoli were found in the nuclei. These cells expressed human telomerase reverse transcriptase and carcinoembryonic antigen, but CD34 and CD45 were not found on the cell membrane. So, these cells were considered as gastric carcinoma cells escaping from the original focuses and existing in the peripheral blood circulation. Carcinoma cells were found in 25 of 60(41.7%) specimens of peripheral blood from patients with gastric carcinoma, while there were no such cells separated from the blood specimens of chronic gastritis patients. The difference of positive rates of disseminated carcinoma cells between two groups was markedly significant (P < 0.005). The expressions of CK20 mRNA in peripheral blood specimens were examinated with RT-PCR. CK20 mRNA was detected from 32 of 60(53.3%) peripheral blood specimens in the group of gastric carcinoma patients, while none of the specimens from patients suffering from chronic gastritis had CK20 mRNA. Significant difference was also found between two groups (P < 0.005). Statistic analyses also showed that there was a significant difference between the positive rates of two methods in detecting the disseminated carcinoma cells from the peripheral blood circulation of gastric carcinoma patients (P < 0.05). CONCLUSION: The results demonstrated that there were disseminated carcinoma cells in the peripheral blood circulation of some patients with gastric carcinoma. Disseminated carcinoma cells can be detected from the peripheral blood samples with immunomagnetic beads sorting technique and RT-PCR technique. The positive rate of RT-PCR technique is higher than that of immunomagnetic beads sorting technique in detecting micrometastasis.

To investigate the therapeutic effects and mechanism of octreotide on experimental hepatic fibrosis in rats.Hepatofibrotic rats models were established with carbon tetrachloride. All the experimental rats were divided into four groups: normal control group, pre-and post-treatment model group, and octreotide-treated group in which the rats were injected subcutaneously with octreotide at the dose of 50ng/100g, twice daily, for thirty days. Serum levels of hyaluronic acid (HA), laminin (LN) and pro-collagen type III peptide (PCIII) were detected by radioimmunoassay. Hepatic fibrosis scoring grade was assessed through Van-Gieson staining and observed under light microscope. Protein expression levels of alpha-smooth muscle actin (alpha-SMA) and transforming growth factor beta1 (TGFbeta1) were determined with immunohistochemical staining method. Messenger RNA (mRNA) levels of collagen type I and PCIII were detected by reverse transcription polymerase chain reaction.Serum levels of HA (ng/L), LN (microg/L) and PCIII (ng/L) in pre- and post-treatment model groups were higher than those in normal control group (121.8+/-9.5 and 110.3+/-13.4 vs. 33.1+/-3.7, 85.7+/-12.1 and 78.2+/-7.9 vs. 37.1+/-6.3, 35.9+/-3.5 and 33.7+/-2.6 vs. 15.6+/-2.8, respectively, tor = 9.41, P0.05), and there was no significant difference between the two model groups. Concentrations of HA (55.8ng/L+/-7.2ng/L), LN (43.1microg/L+/-3.4microg/L) and PCIII (27.8ng/L+/-3.4ng/L) decreased significantly in octreotide-treated group, compared with those in model groups (tor=2.76, P0.05). With histological analysis, fibrotic scoring grade in octreotide-treated group was obviously ameliorated, compared with that in model groups (chi2or = 3.97, P0.05). Imaging analysis revealed that alpha-SMA and TGFbeta1 immunohistological staining areas were markedly shrinked in octreotide-treated group (tor = 2.47, P0.05). In two model groups, PCIII and type I mRNA levels significantly up-regulated as compared with those in normal group (tor = 9.27, P0.001), and they were inhibited by octreotide markedly (tor = 2.47, P0.05).Octreotide can inhibit hepatic stellate cells transforming into myofibroblasts, down-regulate TGFbeta1, collagen type I and PCIII transcriptions, so that it has therapeutic effects on experimental hepatic fibrosis.

The effect of red wine and wine polyphenol resveratrol on endothelial function was investigated in experimental hypercholesterolemic rabbits. Endothelial function as measured by flow-mediated dilation (FMD) in the femoral artery was 19.28+/-2.81% in control animals fed a regular diet. In contrast, rabbits fed a high-cholesterol (1.5%) diet showed a reduced endothelial function, as revealed by a 25% reduction in the measured FMD. Intragastric feeding of resveratrol (3 mg/kg/day), red wine (4 ml/kg/day), dealcoholized red wine (4 ml/kg/day), for 12 weeks in hypercholesterolemic rabbits significantly mitigated the reduction in endothelial function, and resulted in FMD values of 14.52+/-0.60, 18.95+/-2.30, 17.58+/-1.43, and 18.80+/-3.94%, respectively. Measurement of plasma endothelin 1 (ET-1) and nitric oxide (NO) levels showed that feeding a high-cholesterol diet significantly increased plasma ET-1 levels (from 51.4+/-17.6 to 96.9+/-24.3 pg/ml), and decreased plasma NO concentration (from 104.6+/-18.5 to 67.7+/-16.1 pg/ml). With administration of resveratrol, red wine, or dealcoholized red wine, plasma ET-1 levels statistically decreased, in parallel with a significant elevation in NO levels. These results provide in vivo evidence suggesting that resveratrol and red wine improve endothelial function, which may be one of the mechanisms by which this red wine polyphenol exerts its alcohol-independent cardioprotective effects.

AIM:To investigate effect of losartan, an AT1 receptor antagonist, on hepatic fibrosis induced by CCl(4); and to determine whether or not AT1 receptors are expressed on hepatic stellate cells. METHODS AND RESULTS:Fifty male Sprague-Dawley rats, weighing (180 plus minus20)g, were randomized into five groups (control group, model group, and three losartan treated groups), in which all rats were given the subcutaneous injection of 40% CCl(4)(every 3 days for 6 weeks) except for rats of control group. Rats of losartan-treated groups were treated with losartan (20 mg/kg, 10 mg/kg, 5 mg/kg, daily gavage). After 6 weeks liver tissue and serum samples of all rats were examined. Serum hyaluronic acid (HA), procollagen type III (PC III) were detected by radioimmunoassays. van Giesion collagen staining was used to evaluate the extracellular matrix of rats with liver fibrosis. The expression of AT1 receptors, transforming growth factor-beta (TGF-beta), and alpha-smooth muscle actinalpha-SMA) in liver tissue were determined by immunohistochemical techniques. Compared with model group, serum ALT and AST of losartan-treated groups were significantly reduced (italict = 4.20,P0.01 and italict = 4.57,P0.01). Serum HA and PC III also had significant differences (italict = 3.53,P0.01 and t=2.20, P0.05). The degree of fibrosis was improved by losartan and correlated with the expressions of AT1 receptors, TGF-beta, and alpha-SMA in liver tissue.CONCLUSION:AT1 receptor antagonist, losartan, could limit the progression of the hepatic fibrosis induced by CCl(4). The mechanism may be related to the decrease in the expression of AT1 receptors and TGF-beta, ameliorating the injury of hepatocytes; activation of local renin-angiotensin system might relate to hepatic fibrosis; and during progression of fibrosis, activated hepatic stellate cells might express AT1 receptors.

AIM: To examine the expression of activin A, a member of the transforming growth factor (TGF-β) superfamily, recently has been reported to beoverexpressed in liver cirrhosis, in the course of carbon tetrachloride-induced rat hepatic fibrosis. METHODS: Hepatic fibrosis was induced in rats by subcutaneous injections of 40% carbon tetrachloride oily solution for a period of 1 to 7 weeks. At the end of 1, 2, 3, 4, 5, 6 and 7 weeks after carbon tetrachloride injections, the rats were killed in group (6-10 rats each time) for study. The activin A messenger RNA expression and its protein localization were assessed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. RESULTS: The normal rat liver expressed activin A mRNA and protein, and its expression was transiently decreased and became undetectable after carbon tetrachloride injections for 2 or 3 weeks and then increased gradually. After injection of carbon tetrachloride for 6 and 7 weeks, activin A mRNA and protein expressions were significantly enchanced in rat liver. Compared with that of the normal rat liver. Activin A mRNA expression levels in rats receiving carbon tetrachloride injections for 6 and 7 weeks were 1.6 and 2.2 times that of those in normal rat liver respectively (0.456 ± 0.094 vs 0.286 ± 0.0670, P < 0.01; 0.620 ± 0.134 vs 0.286 ± 0.0670, P < 0.01). Immunohistochemistry showed that activin A expressed in hepatocytes of normal liver, and its expression was decreased in rats receiving carbon tetrachloride for 2 or 3 weeks. Compared with normal liver, activin A expression distribution mode changed in fibrotic liver, being increased significantly in hepatocytes around fibrotic areas. CONCLUSION: Activin A expression was increased in late stage of hepatic fibrosis, and this may be involved in hepatic fibrosis formation in this period.

Tetrandrine (Tet) is a dibenzylisoquinoline alkaloid isolated from Stephania tetrandra S. Moore, a Chinese herbal medicine. In the past decade, lots of studies demonstrated that Tet has multiple bioactivities. It is promising to use Tet as an antifibrogenetic in liver or lung fibrosis with or without portal or pulmonary hypertension, as well as an immunomodulating and anticarcinoma drug.