Analysis on Components Absorbed into Blood and Cerebrospinal Fluid of Schisandrae Chinensis Fructus Based on Sequential Metabolism / 中国实验方剂学杂志

ObjectiveTo identify the prototypical components and metabolites absorbed into blood and cerebrospinal fluid of Schisandrae Chinensis Fructus（SCF） based on sequential metabolism combined with liquid chromatography-mass spectrometry. MethodBlood and cerebrospinal fluid samples of integrated metabolism， intestinal metabolism and hepatic metabolism were collected from male SD rats after gavage and in situ intestinal perfusion administration， and ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry（UPLC Q-Exactive Orbitrap MS） was used to analyze and compare the differences in the spectra of SCF extract， blank plasma， administered plasma， blank cerebrospinal fluid and administered cerebrospinal fluid with ACQUITY UPLC BEH Shield RP18 column（2.1 mm×100 mm， 1.7 µm）， the mobile phase was acetonitrile（A）-0.1% formic acid aqueous solution（B） for gradient elution（0-7 min， 95%B； 7-12 min， 95%-35%B； 12-17 min， 35%-15%B； 17-20 min， 15%-12%B； 20-22 min， 12%-5%B； 22-23 min， 5%B； 23-25 min， 5%-95%B； 25-28 min， 95%B）. And heated electrospray ionization（HESI） was used with positive and negative ion modes， the scanning range was m/z 100-1 500. The prototypical constituents and their metabolites absorbed into blood and cerebrospinal fluid of SCF were identified according to the retention time， characteristic fragments， molecular formulae and the information of reference substances. ResultA total of 42 chemical components were identified in the extract of SCF， including lignans， flavonoids， amino acids， tannins， and others， of which lignans were the main ones. A total of 27 prototypical components and 14 metabolites were identified in plasma samples from different sites. A total of 15 prototypical components and 9 metabolites were identified in cerebrospinal fluid. The main metabolic reactions involved in the formation of metabolites were mainly demethylation， methylation， demethoxylation and hydroxylation. ConclusionThrough the systematic identification of the prototypical components and metabolites of SCF in rats， it provides data support for further better exploring the material basis of SCF in the treatment of central nervous system diseases.

Effects comparison of chemical hypoxia induced by cobalt chloride and hypoxia environment on rat pulmonary arterial fibroblasts / 重庆医学

Objective To compare the effects of cobalt chloride (CoCl2) induced chemical hypoxia and hypoxia environment on the proliferation,migration and phenotype transformation of rat primary pulmonary artery fibroblasts (PAFs).Methods Primary PAFs were isolated and cultured.Cells were stimulated by CoCl2,or hypoxia cell culture (1% O2) was used to stimulate and induce PAFs.Then the effects of CoCI2 and hypoxia environment on PAFs were compared by CCK-8 assay,scratch assay,transwell assay,phenotype marker protein expression and PI3K/Akt signaling pathway protein expression.Results Compared with the control group,100 μmol/mL CoCl2 stimulation had no significant effect on the cell proliferation activity,cell migration ability and phenotype transformation ability of PAFs (P>0.05);while 1% O2 could significantly improve the cell proliferation and migration activities of PAFs as well as the upregulation of α-SMA expression (P<0.05).Conclusion There exist differences of effects between CoCl2 induced chemical hypoxia and hypoxia environment on promoting cell proliferation, cell migration and phenotype transformation in PAFs.

Mechanism of Poly(I∶C)-induced apoptosis in lung adenocarcinoma A549 cells / 国际肿瘤学杂志

Objective To study the effects of Poly(I:C) on lung adenocarcinoma A549 cells viability and illuminate the mechanism of Poly (I:C)-induced apoptosis in A549 cells.Methods A549 cells were transfected with the complex of Poly(I:C) and lipofectamine 3000.The viability of A549 cells was tested by methyl thiazolyl tetrazolium (MTF) method.The apoptotic cells were tested by flow cytometry.The caspase proteins were tested by Western blotting and the expressions of interferon-β (IFN-β) and CXCL-10 were assayed by real-time PCR.After employing the pan-caspase inhibitor Z-VAD-FMK and caspase-8 inhibitor Z-IETD-FMK,the variation of Poly (I:C) proapoptosis in A549 cells was observed.RNA interfering experiments were employed to knock down melanoma differentiation related antigen 5 (MDA5) or retinoic acidinduced gene Ⅰ (RIG-Ⅰ),and the above indexes were tested.Results The viability of A549 cells was significantly reduced to 74.92％ ±--6.24％ after 200 ng/ml Poly (I:C) transfection compared with that before transfection (95.32％ ± 3.05％,t =2.883,P =0.041).The apoptotic rates induced by 100,200,400 ng/ml Poly(I:C) were 9.97％-± 0.88％,23.63％-± 1.41％,32.57％-± 2.39％,respectively.All of them were higher than that in the control group (0.74％-± 0.15％),with significant differences (t =4.489,P =0.002;t =11.616,P =0.000;t =16.932,P =0.000).Besides,the death receptor pathway proteins such as TNF-related apoptosis inducing ligand (TRAIL),cleaved-caspase-8 and cleaved-caspase-3 increased obviously.MDA5/RIG-Ⅰ pathway was also activated dramatically and the expressions of IFN-β,CXCL-10 were significantly up-regulated.The apoptotic rates reduced to 3.17％ ± 0.66％,5.35％ ± 0.64％ with pancaspase inhibitor Z-VAD-FMK and caspase-8 inhibitor Z-IETD-FMK pretreatment,compared with the control group (15.87％ ±0.93％),and the differences were statistically significant (t =8.643,P =0.001;t =6.824,P =0.002).Moreover,the expressions of TRAIL,IFN-β and CXCL-10 induced by Poly (I:C) were inhibited with MDA5 or RIG-Ⅰ depletion.Conclusion Poly(I:C) can reduce the survival rate of A549 cells and promote the apoptosis mainly by activating the death-receptor pathway mediated by MDA5/RIG-Ⅰ probably,which may involve in IFN-β,CXCL-10.

Research and application of NVP-BEZ235 in lung cancer / 国际肿瘤学杂志

NVP-BEZ235 is a newly developed dual phosphatidyl linositol kinase 3-kinase (PI3K) and mammalian target of rapamycin (mTOR) inhibitor.Studies have revealed that NVP-BEZ235 combinate with other drugs were effective in the treatment of lung cancer.NVP-BEZ235 might have a potential application value for the treatment of lung cancer of EGFR mutation subtype and KRAS mutation subtype.

Effects of NVP-BEZ235 on pulmonary perivascular mast cells in hypoxia rats / 药学实践杂志

Objective To assess the effects of PI3K/mTOR dual inhibitor NVP-BEZ235 on the distribution and degranulation of perivascularmast cells in the lung of hypobaric hypoxia rats.Methods SD male rats in the hypoxia group were raised in the specific hypoxic incubator (50.5 kPa).The intervene group received NVP-BEZ235 training (35 mg/kg) intervention once every two days.The control group was fed in the conditions comparable to the other groups at local pressure and normoxia environment.After 21 days, all rats were sacrificed with injection of 5％ pentobarbital sodium.Lung tissues were fixed and embedded in paraffin for further hematoxylin-eosin staining and toluidine blue staining.Results Compared with the control group, the number of pulmonary perivascular mast cells in the hypoxia group increased significantly.Hypoxia group had more mast cell accumulation and larger degranulation ratio around different diameters of pulmonary vessels (less than 50 μm, 50-100 μm, larger than 100 μm) than the control group (P< 0.05).The mast cell numbers decreased in NVP-BEZ235 intervention group compared with hypoxia group (P<0.05).The degranulation ratio of pulmonary vascular mast cells around 50-100 μm in the intervention group was smaller than the hypoxia group (P=0.000 3).Conclusion NVP-BEZ235 inhibits the aggregation of pulmonary perivascular mast cells and degranulation ratio of median diameter pulmonary perivascular mast cells in hypobaric hypoxia rats.

Effect of hand hygiene promotion on hand hygiene of health care workers / 中国感染控制杂志

Objective To evaluate the effect of hand hygiene (HH)promotion on awareness,correctness and compliance rates of HH among health care workers(HCWs).Methods HH promotion was started to carry out on May 8,2014,on-site questions,theoretical examination,and direct observation were adopted to survey the percent-age of HH awareness,correctness of six-step hand-washing method,and compliance to HH among HCWs before and at the phase five of promotion implementation,results before and after implementing promotion were compared. Results After implementing HH promotion,percentage of HH awareness,correctness of six-step hand-washing method,and compliance to HH among HCWs increased from 42.50%,35.45%,and 24.00% to 87.12%, 68.60%,and 43.20% respectively,differences were all significant (all P <0.05 ).Conclusion HH promotion has strengthened HCWs’HH awareness,standardized HH method,and enhanced compliance to HH.