OBJECTIVE:To determine plasma concentration of caffeine,dapsone and chlorzoxazone in rats,and to calculate pharmacokinetic parameters. METHODS:6 rats were given the mixture of caffeine,dapsone and chlorzoxazone intragastrically, 1.5,2 and 3 mg/kg,respectively. 0.2-0.3 ml blood were collected before medication and 0.5,1,2,3,4,6,8,12,24 h after medication.The plasma sample was treated with solid phase extraction. The plasma concentration of caffeine,dapsone and chlorzoxa-zone were determined by HPLC using N-(2-Hydroxyethyl) phthalimide as internal standard. The pharmacokinetic parameters were calculated using DAS 2.0 software. RESULTS:The linear ranges of caffeine,dapsone and chlorzoxazone were all 0.2-30 μg/ml (r were 0.996 4,0.996 1,0.998 8,respectively). The limit of quantitation were 0.2 μg/ml. The recoveries of low-concentration, medium-concentration and high concentration were(84.8±3.6)%-(111.4±10.2)%(RSD were 4.3%-9.8%,n=3),(107.0±13.3)%-(113.5±8.1)%(RSD were 7.1%-14.0%,n=3),(104.2±10.8)%-(111.1±12.2)%(RSD were 8.0%-11.0%,n=3). Pharmacoki-netic parameters were as follows as tmax(1.70±0.99),(1.50±1.00),(1.92±0.80)h;t1/2(0.73±0.22),(2.77±1.35),(2.78±2.34) h;cmax (2.60 ± 0.50),(5.78 ± 1.19),(9.76 ± 1.37) mg/L;AUC0-t (8.43 ± 0.79),(20.68 ± 1.91),(26.71 ± 2.45) mg·h/L(n=6). CONCLUSIONS:The method is simple,sensitive and accurate,and can be used for the plasma concentration determination and pharmacokinetic study of caffeine,dapson and chlorzoxazone.

Objective To evaluate the sensitivity of three different clinical consciousness scales to patients with disorders of consciousness.Methods A total of 84 patients in vegetative state (VS) were selected and scored by the consciousness scales including Glasgow Coma Scale (GCS),Coma Recovery Scale-Revised (CRS-R) and Chinese Vegetative State Scale (CVSS) respectively.The patients were followed every 2 weeks for 12 weeks or every 4 weeks for 12-24 weeks.The assessment finished in 6 months or till the time when patients emerged from minimally conscious state ( MCS.The diagnostic sensitivity and the sensitivities of each sub-scale in CRS-R were evaluated when the patients with MCS emerged from VS.The diagnostic consistency with other scales was observed and analyzed by Spearman correlation as well.The Spearman -relate was used to analyze the correlations among the total scores of three scales in the patients on admission (36 patients with acute and 48 patients with chronic phase).Results Of 84 VS patients,there were 47 emerged to MCS.CRS-R was the most sensitive in the diagnosis of MCS (P ＜ 0.05 ) among all scales.The visual subscale is the most sensitive among all the CRS-R subscales ( P ＜0.01 ).Among the 84 patients,the remaining 37 were diagnosed as VS by all three scales.The total scores correlation analysis showed that the score of CRS-R significantly correlate with that of GCS in acute and chronic phase ( P ＜0.01 ) and significantly correlate with that of CVSS only in the acute stage ( P ＜ 0.05 ).Conclusions It is indicated that CRS-R,especially the visual subscale of CRS-R is the more sensitive tool for detecting.MCS during recovery process in the disorders of consciousness.As to the patients with deep unconsciousness and with no significant recovery,the three instruments are in good agreement to determine the degree of disorders of consciousness.It is suggested that CRS-R can be widely used to assess the level of consciousness in various stages.

OBJECTIVE:To establish a method for the content determination of total flavonoids in Morus alba. METHODS:UV-visible spectrophometry was performed with Al(NO2)3-NaNO2-NaOH color-test at the wavelength of 510 nm with the reference of rutin. RESULTS:The linear range of rutin was 0.031 2-0.156 mg/ml(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recovery was 95.7%-101.0%(RSD=2.1%,n=6). CONCLUSIONS:The method is simple,sta-ble and reproducible,and can be used for the content determination of total flavonoids in M. alba.

OBJECTIVE:To establish a method for the determination of residuals of petroleum ether,ethanol,xylene and ace-tic acid in ecabet sodium crude drug. METHODS:Capillary GC was performed on the column of PGE-20M capillary column at the flow rate of 1.7 ml/min,detector was hydrogen flame ionization detector,carrier gas was nitrogen with high purity,column temper-ature was 45 ℃,maintaining 4 min,it increased to 80 ℃ with rate of 10 ℃/min,then increased to 135 ℃ with rate of 30 ℃/min,maintaining 3 min,the injection mode was direct injection,inlet temperature was 250 ℃,and the volume injection was 1.0 μl. RESULTS:The mass concentration was 0.050-1.952 g/L for petroleum ether,0.050-1.941 g/L for ethanol,0.024-0.948 g/L for xy-lene and 0.050-1.947g/L for acetic acid(r=0.999 1-0.999 7);RSDs of precision,stability and reproducibility tests were no more than 1.7%;recoveries were 99.3%-101.0%(RSD=0.7%,n=9),102.3%-103.7%(RSD=0.4%,n=9),101.2%-102.1%(RSD=0.3%,n=9) and 100.3%-102.2%(RSD=0.6%,n=9),respectively. CONCLUSIONS:The method is simple and accurate,and can be used for the control of residual of organic solvents in ecabet sodium crude drug.

Objective To develop a five fluorescence-labeled multiplex amplification system for 15 loci and study genetic polymorphism in Xinjiang Uygur population. Methods The STR loci were screened. The alleles were named according to the number of repeats by sequencing. The sensitivity, species specificity, identity and stability of the five fluorescence-labeled multiplex amplification system for the 15 loci were all tested. Then, the genetic polymorphism was analyzed in Xinjiang Uygur population and compared with other ethnic groups including Xizang Tibetan, Xiuyan Manchu, and Guangzhou Han pop-ulation. Results The 15 loci multiplex amplification system was established. The sensitivity was 0.3 ng with good species specificity, identity and stability. The distributions of genotype for 13 STR loci in Uygur population were in accordance with Hardy-Weinberg equilibrium with no genetic linkage between these loci. Most loci showed statistically significant among different populations. Conclusion The estab-lished system has application value in forensic evidence. The 13 STR loci in Uygur population have high polymorphisms to be the supplements to the existing loci.

Objective To investigate the influence of donepezil on neurogenesis in the subventricular zone (SVZ) under the cerebral ischemic conditions. Methods Sixty mice were randomly assigned to a sham +vehicle group, a middle cerebral artery occlusion(MCAO) + vehicle group and a MCAO + donepezil group, 20 in each group. A model of MCAO was established. Neural stem cells/progenitor cells were labeled by Mash1, and neuroblasts were labeled by doublecortin (DCX). The expressions of DCX in the SVZ cells of each group were detected by immunofluorescence. The expressions of Mash1 and DCX in the SVZ cells of each group were quantified by Western blot. Results 10 days after MCAO was undergone, DCX-positive cells were seen in the SVZ of each group, and the expressions of DCX and Mash1 in MCAO + vehicle group were markedly increased as compared with the sham + vehicle group (P < 0.05), and that in MCAO + donepezil group were significantly increased as compared with the other two groups (P < 0.05). Conclusion As a kind of acetylcholinesterase inhibitors, donepezil can promote neurogenesis in SVZ under the cerebral ischemic conditions.

Objective To investigate the impact of hepatitis C virus (HCV)-RNA levels on human immunodeficiency virus (HIV)-1 disease progression in Chinese HIV/HCV co-infected individuals. Methods Cross-sectional analysis was performed among 391 HIV-infected patients for assessment of HCV-IgG, HCV-RNA, HIV-RNA, CD4 cell counts and cell surface markers of immune activation, to compare the difference of viral and immune indexes between HCV-RNA high group and HCV-RNA low group, and to elucidate the association between HCV-RNA, HIV-RNA and CD4 cell counts in HIV/HCV co-infected patients. Results (1) The percentage of anti HCV-IgG positive of former plasma donor group (93%) and drug-injection group (97.5%) were significantly higher than that of sexual transmission group (20.1%). The percentage of HCV-RNA positive of drug-injection group (89.9%) was significantly higher than that of former plasma donor group (48.3%) and sexual transmission group (62.5%), P<0.01, respectively. (2) HCV-RNA levels were positively correlated to HIV-RNA levels (r=0.237,P

Objective: : Nicotinamide mononucleotide adenylyl transferase 2 (NMNAT2) is a crucial factor for the survival of neuron. The role of NMNAT2 in damage following traumatic brain injury (TBI) remains unknown. This study was designed to investigate the role of NMNAT2 in TBI-induced neuronal degeneration and neurological deficits in rats. Methods: : The TBI model was established in Sprague-Dawley rats by a weight-dropping method. Real-time polymerase chain reaction, western blot, immunofluorescence, Fluoro-Jade C staining, and neurological score analyses were carried out. Results: : NMNAT2 mRNA and protein levels were increased in the injured-side cortex at 6 hours and peaked 12 hours after TBI. Knocking down NMNAT2 with an injection of small interfering RNA in lateral ventricle significantly exacerbated neuronal degeneration and neurological deficits after TBI, which were accompanied by increased expression of BCL-2-associated X protein (Bax). Conclusion : NMNAT2 expression is increased and NMNAT2 exhibits neuroprotective activity in the early stages after TBI, and Bax signaling pathway may be involved in the process. Thus, NMNAT2 is likely to be an important target to prevent secondary damage following TBI.

Objective To investigate the effect of HCV RNA on virological and immunological response to highly active antiretroviral therapy (HAART),liver function and blood lipid levels in HIV/HCV co-infected patients.Methods In a cohort study,275 HIV/HCV co-infected former blood donors receiving HAART were followed up every six month in Henan province in China.HCV RNA,HIV RNA,CD+4 T cell counts,indexes of liver function and lipid levels were periodically tested.The differences of HIV viral load suppression,immunological response,liver injury and blood lipid levels between HCV RNA positive group and negative group were compared by x2 test and two independent-samples tests.Result There was no significant difference of HIV viral load suppression between HCV RNA positive group and HCV RNA negative group six-month treatment (45.6% vs.38.5% ,X2=1.150,P>0.05) and CD+4 T cell counts before (286 cells/μ1 vs.209 cells/μ1,Z=0.734,P=0.463)and after 6-month (310 cells/μ1 vs.362 cells/μl,Z=0.562,P=0.574) ,12-month(378 cells/μ1 vs.289 cells/μ1,Z=1.091,P=0.275),18-month(363 cells/μ1 vs.288 cells/μl,Z=1.435,P=0.151) ,24-month(413 cells/μ1 vs.348 cells/μ1,Z=0.939,P=0.348) HAART.The mean levels of serum ALT (55.0 U/L vs.29.5 U/L,Z=6.789,P<0.01),AST(46.0 U/L vs.33.0 U/L,Z=4.890,P<0.01)、TBIL(9.3 mmol/L vs.7.2 mmol/L,Z=3.748,P<0.01)were significantly higher in HCV RNA positive group than that in HCV RNA negative group.HCV RNA was the independent variables associated with liver injury after HAART (aOR=3.8,P<0.01).The serum triglyceride level was higher in HCV RNA positive group than that in HCV RNA negative group(1.2 mmoL/L vs.1.4 mmol/L,Z=1.936,P=0.043) .The serum HDL level was higher in HCV RNA positive group than that in HCV RNA negative group (1.5 mmol/L vs.1.3 mmol/L,Z=2.251,P=0.024).Conclusions HCV RNA does not affect HIV virological responses to HAART and CD+4 T recovery.HCV RNA is an independent risk factor associated with liver injury in HIV/HCV co-infected patients receiving HAART,but appears to provide significant protection against HAART-ieduced hyperlipidemia.

Objective To assess the pain of patients with disorders of consciousness(DOC) and e-valuate the reliability and validity of Chinese version of Nociception Coma Scale- revised ( NCS-R). Meth-ods Sixty-two patients with disorders of consciousness were assessed by NCS-R and the internal reliability, inter-rater reliability,test-retest reliability and concurrent validity was analyzed. Results Cronbach's alpha (α) of internal reliability was 0. 611, test-retest reliability for the NCS-R total scores by spearman was 0. 884. For the subscales,the reliability for the motor,verbal and facial subscale scores by Cronbach's kappa were 0. 856,0. 870 and 0. 856 respectively. Inter-rater reliability for the NCS-R total scores by spearman was 0. 662. For the subscales,the reliability for the motor,verbal and facial subscale scores by Cronbach's kappa were 0. 962,0. 867 and 0. 671 respectively. The spearman coefficient of concurrent validity was 0. 560 (all P<0. 01). Conclusion The NCS-R is a good valid scale and available for Chinese clinician and researchers to assess the pain of DOC patients.