Effects of spautin-1 on autophagy and apoptosis of acute pancreatitis cell model induced by cerulein / 中华胰腺病杂志

Objective To investigate the potential of Spautin-1 to treat acute pancreatitis by inhibiting impaired autophagy and promoting apoptosis using an acute pancreatitis cell model induced by cerulein in vitro.Methods Pancreatic acinar cells AR42J were treated with sterile saline,100 nmol/L cerulein,and 100 nmol/L cerulein combined with 10 μmol/L spautin-1 for 24 h,respectively,and then western blot was used to detect the expression of LC3,p62 and Beclin1 in the cells to reflect autophagy;the trypsin activity in cells was detected by BGAM.Flow cytometry was used to detect the apoptosis;LDH cytotoxicity assay kit was used to detect cell necrosis.Results The expression of LC3 Ⅰ (0 h-24 h:4.32±0.46,4.68±0.41,5.22± 0.38,5.88 ±0.63),LC3 Ⅱ (0 h-24 h:0.36 ±0.02,0.64 ±0.05,0.93 ±0.08,2.43 ±0.23) and p62 (0 h-24 h:0.24 ± 0.01,0.22 ± 0.02,0.84 ± 0.09,1.25 ± 0.13) was increased in AR42J cell model treated with cerulein (P ＜ 0.05),indicated that cells undergo autophagy,but autophagy flux is blocked.The expression of LC3 Ⅰ (0.65 ± 0.06 vs 0.24 ± 0.01),LC3 Ⅱ (1.26 ± 0.15 vs 0.71 ± 0.08) and p62 (1.06 ± 0.09 vs 0.56 ± 0.06) were decreased after incubated with spautin-1,with the decreased trypsin activity (1.65 ±0.18 vs 1.13 ±0.14),increased apoptosis (6.58 ±4.01 vs 23.64 ±2.12) (P ＜0.05) and reduced cellnecrosis (27.58± 3.46vs7.64 ± 2.12) (P＜0.05).Conclusions Inthe AR42 Jcellmodel ofacute pancreatitis induced by cerulein in vitro,spautin-1 can inhibit impaired autophagy induced by cerulein,decrease trypsin activity,increase apoptosis and reduce cell necrosis to protect pancreatic cells from damage,which has a certain value for remissing and even curing acute pancreatitis.

Effects of capsaicin on oxidative stress in lipopolysaccharide-induced acute lung injury in mice / 中华急诊医学杂志

Objective To investigate the effects of transient receptor potential vanilloid 1 activation by capsaicin on the oxidative stress in lipopolysaccharide-induced lung injury in mice in order to elucidate the potential mechanisms.Methods A total of 108 specific pathogen free (SPF) ICR male mice were randomly divided into six groups:normal control group (n =18),capsaicin control group (CAP control group,n =18),capsazepine control group (CAPZ control group,n =18),acute lung injury group (n =18),capsaicin treatment group (CAP treatment group,n =18) and capsazepine treatment group (CAPZ treatment group,n =18).After modeling,superoxide dismutase (SOD),catalase (CAT) and malondiachehyche (MDA) levels in lung were measured with the method of chromatometry,and the expression of heme oxygenase 1 (HO-1) in lung tissue was assessed with enzyme linked immunosorbent assay (ELISA),while the level of NF-E2-related factor-2 (Nrf2) was determined by western blotting and the expression of Nrf2 mRNA was measured by RTPCR.Pathological changes of lung tissue were observed under light microscope.Results The activities of SOD and CAT in lung tissue at 3,8,16 h were dramatically lower in acute lung injury group than those in normal control group (P ＜ 0.05),while the level of MDA was higher.Compared with acute lung injury group,the lung levels of SOD and CAT at 8 h and 16 h were higher in CAP treatment group (P ＜0.05),while the lung level of MDA was lower (P ＜ 0.05).The levels of SOD and CAT in CAPZ treatment group were decreased at 8 h and 16 h,while the levels of MDA in this group were increased at 3,8,16 h (P ＜0.05).The pulmonary levels of HO-1,Nrf2 and expression of Nrf2 mRNA were significantly higher in acute lung injury group than those in normal control group (P ＜ 0.05).Compared with acute lung injury group,the levels of HO-1,NRF2 and expression of NRF2 mRNA were increased markedly in CAP treatment group (P ＜ 0.05)and were obviously decreased in CAPZ treatment group (P ＜0.05).At 8 h,16 h after modeling,the degree of lung damage was ameliorated in CAP treatment group compared with acute lung injury group under light microscope,while the lung damage was aggravated in CAPZ treatment group.Conclusions The activation of TRPV1 could apparently up-regulate the levels of CAT,SOD,Nrf2,HO-1,and reduce the MDA level in lung tissue of mice with acute lung injury,ultimately protecting the endotoxemia mice from oxidative stress.

Subcellular localization and resistance to Gibberella fujikuroi of AtELHYPRP2 in transgenic tobacco / 生物工程学报

The subcellular localization and the resistance to fungal pathogen Gibberella fujikuroi of the protein encoded by Arabidopsis AtELHYPRP2 (EARLI1-LIKE HYBRID PROLINE-RICH PROTEIN 2, AT4G12500) were investigated using transgenic tobacco plants. The coding sequence of AtELHYPRP2 was amplified from genomic DNA of Col-0 ecotype. After restriction digestion, the PCR fragment was ligated into pCAMBIA1302 to produce a fusion expression vector, pCAMBIA1302-AtELHYPRP2-GFP. Then the recombinant plasmid was introduced into Agrobacterium tumefaciens strain LBA4404 and transgenic tobacco plants were regenerated and selected via leaf disc transformation method. RT-PCR and Western blotting analyses showed that AtELHYPRP2 expressed effectively in transgenic tobacco plants. Observation under laser confocal microscopy revealed that the green fluorescence of AtELHYPRP2-GFP fusion protein could overlap with the red fluorescence came from propidium iodide staining, indicating AtELHYPRP2 is localized to cell surface. Antimicrobial experiments exhibited that the constitutive expression of AtELHYPRP2 could enhance the resistance of tobacco to fungal pathogen G. fujikuroi and the infection sites could accumulate H2O2 obviously. The basal expression levels of PR1 and the systemic expression levels of PR1 and PR5 in transgenic tobacco plants were higher than that of the wild-type plants, suggesting AtELHYPRP2 may play a role in systemic acquired resistance.

Distribution and drug resistance of pathogens isolated from lower respiratory tract in patients with ventilator-associated pneumonia / 中华临床感染病杂志

Objective To investigate the distribution and antimicrobial resistance of pathogens isolated from lower respiratory tracts of early onset and late-onset ventilator-associated pneumonia (VAP)patients in intensive care unit. Methods A retrospective analysis was performed on data collected from1324 patients admitted to the ICU in the Third People' s Hospital in Wenzhou from July 2008 to December 2011 who received invasive mechanical ventilation.Patients were divided into early-onset pneumonia group (EOP,≤ 4 d mechanical ventilation ) and late-onset pneumonia group (LOP, ＞ 4 d mechanical ventilation).x2 or t test was performed to compare the pathogen distribution and antimicrobial resistance between two groups.Results In 1324 patients,552 (41.7％) suffered from VAP,including 74 ( 5.6％ )patients with EOP,382 (28.9％) patients with LOP,and 96 (7.3％) patients with both EOP and LOP.The top 6 pathogens in EOP group were Acinetobacter baumannii ( 72,22.6％ ),Pseudomonas aeruginosa (45,14.1％ ),Klebsiella pneumoniae (32,10.0％ ),Canidia albicans ( 31,9.7％ ),Burkholderia cepacia ( 31,9.7％ ) and Staphyloccocus aureus ( 28,8.8％ ) ; and in LOP group were Acinetobacter baumannii (235,21.2％ ),Canidia albicans (201,18.1％ ),Pseudomonas aeruginosa ( 111,10.0％ ),Candida glabrata ( 86,7.8％ ),Klebsiella pneumoniae ( 81,7.3％ ) and Staphyloccocus aureus ( 46,4.2％ ).Staphylococcus aureus infection in EOP group was more common than that in the LOP group (x2 =10.780,P =0.002),but the separation rate of Candida albicans in EOP group was significantly lower than that in the LOP group (x2 =12.907,P =0.000).Gram-negative bacteria isolated from EOP or LOP group were both highly resistant to most commonly used antibacterial agents,especially for Acinetobacter baumannii and Pseudomonas aeruginosa strains. Most Staphyloccocus aureus strains were methicillin-resistant strains (EOP:67.9％,19/28; LOP:63.0％,29/46).Canidia albicans and Candida glabrata were sensitive to most antifungal agents.Conclusion For both EOP and LOP groups,the majority of pathogens isolated from lower respiratory tract are Gram-negative bacteria,and drug resistance is serious.

Subcellular localization and resistance to Botrytis cinerea of a new type lipid transfer protein AtDhyPRP1 from Arabidopsis thaliana / 生物工程学报

Genetic transformation was adopted to analyze the subcellular localization and the resistance to fungal pathogens of Arabidopsis lipid transfer protein AtDHyPRP1. The coding sequence of AtDHyPRP1 amplified by PCR from Ws ecotype was used to construct the plant binary expression vector pRI101-AN-AtDHyPRP1 and the fusion expression vector pCAMBIA1302-AtDHyPRP1-GFP. Transgenic tobacco and Arabidopsis plants were produced by leaf disc and floral dip protocols, respectively. AtDHyPRP1 could improve the resistance of tobacco to Botrytis cinerea remarkably and the infection sites on transgenic tobacco leaves accumulated large amounts of H2O2. Observation under laser scanning confocal microscope showed that AtDHyPRP1 was localized to cell surface. It suggested that AtDHyPRP1 might play special function after secretion to outside of the cell and was involved in plant defense system against pathogens.

Effect of goal of blood glucose control on protein metabolism in the patients with critically cerebral disease / 肠外与肠内营养

Objective: To investigate the effect of three different goals of blood glucose control by insulin therapy on protein metabolism in the patients with critically cerebral disease.Methods: We performed a prospective,randomized,controlled study.A total of 122 patients with critically cerebral disease were randomly divided into three groups according to the target glucose-control levels:Group A: 4.4-6.1 mmol/L(n=41),Group B: 6.1-8.3 mmol/L(n=42),and Group C: 10-11.1 mmol/L(n=39).The state of protein metabolism and ratio of hypoglycemia of three groups were compared.Results: The indicators of nitrogen balance,prealbumin and transferrin in group A and group B were significantly better than that in group C(P<0.05),but the ratio of hypoglycemia in group A was significantly higher than those in group B and group C(P<0.05).Conclusion: Maintaining the blood glucose level ≤8.3 mmol/L might improve the nutrition status in the patients with critically cerebral disease by decreasing catabolism and promoting synthesis of protein.

Study on Cryopreservation of Seeds of Aquilaria sinensis(Lour.)Gilg / 广州中医药大学学报

【Objective】To explore the long-term storage method for the seeds of Aquilaria sinensis(Lour.)Gilg..【Methods】The seeds of Aquilaria sinensis(Lour.)Gilg.with different water contents were preserved with liquid nitrogen for 55 days.And then the seeds were taken out to detect their germination capacity.The effect of cryopreservation methods on the germination capacity was also observed.【Results】The seeds with water content being 16.30% lost their germination capacity completely,the seeds with water content being 13.02% and 9.34% had a lower germination capacity,and the seeds with water content being 7.35% had a higher germination capacity.Quickly-frozen seeds had a higher germination capacity than the slowly-frozen seeds.【Conclusion】Cryopreservation with liquid nitrogen is suitable for the long-term preservation of the seeds of Aquilaria sinensis (Lour.)Gilg.with water content being 7.35%.

Effect of Different Storage Methods on Germination Capacity of Aquilaria sinensis(Lour.) Gilg / 广州中医药大学学报

【Objective】To devise the optimal method and the optimal water content for the storage of seeds of Aquilaria sinensis(Lour.) Gilg..【Methods】The seeds of Aquilaria sinensis(Lour.) Gilg.were stored in plastic bag,drying apparatus,wet sand and dry sand separately.And then the germination capacity of the seeds with different water content was detected at 4℃ and room temperature(30℃) in different time.【Results】The germination capacity of Aquilaria sinensis(Lour.) Gilg.decreased with the prolongation of storage time and with the decrease of water content.The seeds with water content being 9.34% and 7.35% had a higher germination capacity when stored at 4℃ for 35 days than that stored at room temperature.The fresh seeds stored in wet sand sprouted and the seeds stored in drying apparatus and dry sand had lower germination capacity and went mouldy.【Conclusion】Low temperature and moderate water content are benificial to the storage of the seeds of Aquilaria sinensis(Lour.) Gilg..

Chemical constituents of Lignum Aquilariae Resinatum(Ⅱ) / 中草药

Objective To study the chemical constituents of Lignum Aquilariae Resinatum.MethodsThe compounds were isolated and purified by silica chromatography and their structures were identified on the basis of physicochemical constant and spectral analysis.Results The compounds were determined as 6-hydroxy-2-[2-(3'-methoxy-4'-hydroxy phenylethyl)] chromone(Ⅰ)and a triterpene,hederagenin(Ⅱ).Conclusion Compound I is a new compound and compound Ⅱ is found in this plant for the first time.