Objective To verify the validity of the sucrose density gradient ultracentrifugation method for lipid rafts from cerebral cortex. Methods Extract lipid rafts from cerebral cortex in mouse were extracted by the sucrose density gradi-ent ultracentrifugation method. The properties of lipid rafts were detected by Western blotting method, double enzyme and light scattering methods. HPLC MS/MS proteomics and bioinformatics were used to locate proteins of lipid rafts in cells. Re-sults Lipid rafts from cerebral cortex were provided with the model properties of lipid rafts such as high light scattering and cholesterol and high expression of Flotillin-1. HPLC MS/MS proteomics identified total 647 proteins. Most of these pro-teins were from plasma membrane, endoplasmic reticulum, cytoskeleton and cytosol, however, there were 21% proteins among total 647 proteins were from nucleus, mitochondria and ribosomes. Conclusion The sucrose density gradient ultra-centrifugation method is a effective method to extract lipid rafts from cerebral cortex, however, the properties of mixture should be considered.

Objective To establish the quality standard of Qingshen Granules. Methods Thin-layer chrmatography was used to identify Rheum officinale Baill., Hedyotis Diffusa Wild. and Coptis chinensis Franch.. The contents of rhein, emodin and chrysophanol were determined by HPLC. The HPLC consisted of Kromasil C18 (4.6 mm×250 mm, 5 μm), methanol-0.1% phosphonic acid (70∶30) as mobile phase, flow rate of 1.0 mL/min, detection wavelength at 245 nm, and the column temperature was 30 ℃. Results The results of TLC showed that relevant spots were clear without interference against the negative sample. The calibration curves for rhein, emodin and chrysophanol were found to be linear within the range of 0.003 3-0.42 μg (r =0.999 9), 0.008 0-0.51 μg (r=1.000 0), 0.009 9-0.32 μg (r=1.000 0), respectively. The average recoveries were 98.84%, 99.04% and 100.35% with RSD of 1.30% (n=6), 1.34% (n=6) and 1.89% (n=6), respectively. Conclusion The methods are accurate and quick in qualitative identification and quantitative assay, and can be used for the quality control of Qingshen Granules.

Objective To investigate the role of p53 in the regulation of heat shock protein(Hsp)84 and 86,and the correlation of their functional imbalances with accelerated brain aging and with suppressed tumorigenesis in SAMP8 mice(senescence accelerated mouse prone 8).Methods The mRNA and protein expressions of Hsp84 and Hsp86,and protein expressions of p53 pathway-related proteins(p21 and MDM2)in hippocampus of SAMP8 mice and their control SAMR1(senescence accelerated mouse resistant 1)mice were determined.Murine Neuro-2a cells were treated with 20 μmol/L Aβ25-35,and then mRNA expressions of p53,Hsp84 and Hsp86 in these cells were detected.Neuro-2a cells were co-transfected with p53 siRNA and pHsp84-Luc or pHsp86-Luc plasmid and treated with 20 μmol/L Aβ25-35,then promoter activity of Hsp84 and Hsp86 were detected in these cells.After co-transfection with pcDNA3.1-p53 or pcDNA3.1-p53DD and pHsp84-Luc or pHsp86-Luc plasmids,the neuro-2a cells were treated with 20 μmol/L Aβ25-35.Then promoter activity of Hsp84 and Hsp86 were detected in these cells at different concentrations of p53.Results The mRNA levels of Hsp84 and Hsp86 in the hippocampus of SAMP8 mice were significantly declined,which were 13.51% and 16.13% of SAMR1 mice,respectively(all P<0.01).Compared with the SAMR1 mice,the protein expressions of Hsp84 and Hsp86 in the hippocampus of SAMP8 mice were obviously declined(all P<0.01).Whereas,p53 pathway-related protein p21 expression was increased and MDM2 expression was decreased(all P<0.01).The mRNA expression of p53 in AD cells was significantly increased by 58%(P<0.01),whereas Hsp84 and Hsp86 mRNA levels were significantly decreased by 32% and 41%,respectively as compared with the normal cells(all P<0.05).Inhibition of p53 in AD cells could increase promoter activity of Hsp84 and Hsp86 significantly in a concentration-dependent manner(both P<0.05),whereas overexpression of p53 in the cells could lead to decreased promoter activity of them in a concentration-dependent manner(both P<0.05).Conclusions The p53 can negatively regulate the expressions of Hsp84 and Hsp86.The activity of p53/p21 pathway is increased,while Hsp84 and Hsp86 are inhibited in the brain of SAMP8 mice.Functional imbalance between p53 and Hsp84/86 might be the part of reasons causing accelerated aging and suppressed tumorigenesis in SAMP8 mice.

Objective To explore the true feelings and emotional needs of the patients with androgen insensitivity syndrome (AIS) during diagnosis and treatment. Methods In this qualitative study, the phenomenological research method was conducted among 9 patients with androgen insensitivity syndrome. The data were analyzed by using Colaizzi analysis method. Result The AIS patients during the diagnosis and treatment experienced six psychological feelings: eagerness to understand disease knowledge, gender cognition disorder inferiority, self-pity, anger and resentment, feeling at a loss about future, general symptoms, and anxiety for social acceptance. Conclusions Nurses should provide relevant information guidance and emotional support to alleviate patients'negative emotions and enhance their confidence in the treatment. The nurses should encourage family members of the patients to participate in psychological counselling, expand social support through multiple channels, correct patients'psychological biases, and restore their self-confidence.