ObjectiveTo investigate the effect of auricular plaster therapy plus umbilical application of Chinese herbal medicine on opioid analgesic action and adverse reactions.MethodOne hundred and twentypatients with cancer pain treated with opioids were randomly allocated to treatment and control groups. The treatment group received auricular plaster therapy plus umbilical application of Chinese herbal medicine. Opioid dosage and adverse reactions were observed in the two groups.ResultOpioid dosage, the severities of nausea, vomiting and defecation and the incidences of dizziness, somnolence, itching, urinary voiding difficulty and Respiratory inhibition decreased significantly and the quality of life improved significantly in the treatmentgroup compared with the control group.ConclusionAuricular plaster therapy plus umbilical application of Chinese herbal medicine can improve opioid analgesic effect and reduce the incidences and severities of adverse reactions.

Objective To observe the effects of Jiuxieling granules on rats with ulcerative colitis and explore its mechanism.Methods Ulcerative colitis models were established by immune method combined with local stimulation of 5% acetic acid.Wistar rats were randomly divided into control,model,positive control and low,middle,high dose Jiuxieling granules group.Rats in positive control group were treated with sulfasalazine,and those in three treatment groups were treated with low,middle and high dosage of Jiuxieling granules respectively for 14 days continuously.24 hours after last administration,the rats were killed to obtain colon mucous membrane to test GSH-Px,SOD,MDA,NO,tNOS and iNOS concentration of the homogenate of colon mucous membrane.Results Jiuxieling granules could markedly improve the levels of GSH-Px and SOD activity,whereas decrease the levels of MDA,NO,TNOS and iNOS.Conclusion Jiuxieling granules exert its marked curative effects on rats of ulcerative colonitis by resisting oxygen free radical lesion,inhibiting the generation of NO and NOS,and enhancing the activity of antioxidase.

Objective To study the antibacterial constituents of Senecio cannabifolius.Methods The antibacterial effect of all the extracts was tested in vitro with Staphylococcus aureus,Bacillus subtilis(Gram-positive bacilli),and Escherichia coli(Gram-negative bacillus).Chemical constituents were isolated by chromatography.Physicochemical characters and spectroscopic analyses were employed for their structural identification.Results Five compounds were isolated and their structures were elucidated by chemical and spectral methods as follows: methyl-5-hydroxy-2-pyridinecarboxylate(E-13),6-hydroxy-7,7a-dihydro-2(6H)-benzofuranone(E-14),2-(1,4-dihydroxycyclohexanyl)-acetic acid(E-16),3-hydroxycyclohexanecarboxylic acid(E-17),and 4-hydroxy benzaldehyde(B-14).Conclusion All of them are isolated from the plants of Senecio L.for the first time.

Three compounds were isolated from the extract of Taxus cuspidta Sibe et Zucc with the column chromatography on silica gel and preparative HPLC methods. Their structures were identified according to the physicochemical properties and spectral analysis, and they were identified as (E)-1-methoxy-2-O-(p-coumaroyl)-myo-inositol (1), 2-deacetoxy-7beta, 9a, 10beta-trideacetyltaxinine J (2) and (3aS, 4aR, 6S, 8S, 8aS, 9R, 10R, 10aS)-benz[f]azulene-6, 8, 9, 10 (3H)-terol, 3a, 4, 4a, 5, 6, 7, 8, 8a, 9, 10-decahydro-10a-(1-hydroxyl-1-methylethyl)-1, 8a-dimethyl-5-methylene (3). Among them, compound 1 was a new compound, and compounds 2, 3 were two novel natural products.

Objective To prepare ?-elemene solid lipid nanoparticles and investigate their particle size diameter and entrapment efficiency.Methods The ?-elemene solid lipid nanoparticles were prepared by film ultrasonic wave dissolving techniques.And the optimum formula was selected through orthogonal design test according to the entrapment efficiency.Results The optimizing technique was ?-elemene(20 ?L),stearic acid(90 mg),lecithin(90 mg),Tween80(2.5 %,5 mL) and Poloxamer 188(2.5 %,5 mL).Conclusion The technique of preparing ?-elemene by film-ultrasonic wave dissolving technique is feasible.

Objective To establish a method for very long chain fatty acids( VLCFA )with liquid chromatography-tandem mass spectrometry( LC-MS/MS ). Methods One hundred and one healthy cases and 35 suspected ALD patients collected from April to June in 2009 were enrolled into this study. Quantitative analyzed the concentrations of VLCFA in serum was performed using liquid chromatography-tandem mass spectrometry. The precision, accuracy and recovery were analyzed, and the stability of VLCFA concentration of sample under room temperature and repeated freeze-thawing were also investigated. Serum levels of VLCFA in 101 normal cases were determined and analyzed statistically. The results for the 35 randomly chosen serum samples were compared with those from MDI in Germany. Results Serum VLCFA were separated well under these gradient condition with small interference. The linear range of C22:0 was from 2 mg/L to 64 mg/L, the recovery was 99. 92% -102. 05%, and the relative standard deviation ( RSD ) of intra-day and inter-day was less than 6% and 9% respectively. For C24:0 they were 2-64 mg/L. 95. 12%-100. 44%. ≤6%, ≤7%,respectively. For C26:0, they were 0-8 mg/L, 92.21%-103.71%, ≤7%, ≤8%, respectively. The accuracy of C22: 0,C24:0 and C26:0 were among 85% to 115%. The samples could be stable within 12 h at room temperature and repeated 10 times freeze-thawing. The values of VLCFA in 101 normal cases followed a normal distribution and the measured values were C22:0 =( 19. 43 ±4.43 ) mg/L,C24:0 =( 19. 10 ±4. 58 )mg/L, C26:0 = ( 0. 21 ± 0. 11 ) mg/L, the ratio of C24: 0/C22:0 and C26:0/C22: 0 were ( 0. 99 ± 0. 13 )and ( 0. 01 ±0. 01 ) respectively. The statistical analysis showed the concentration of C26:0 in adults ( 0. 18±0. 10 ) mg/L and children ( 0. 21 ± 0. 08 ) mg/L, C24: 0/C22:0 in adults ( 1.01 ± 0. 10 ) and children ( 0. 99 ±0. 14 ) has no significant( t values were 1. 439,0. 806, respectively, all P ＞ 0. 05 ); the ratio of C24:0/C22:0 in male (1.05 ± 0. 10 ) and female (0.97 ± 0. 10 ) has significant difference ( t =3. 394,P =0. 001 ). Compared the values determined by MDI laboratory, the results of C22: 0( 16. 93 ±4. 30 ) mg/L,C24: 0( 19. 57 ± 6. 40 ) mg/L by this method and C22:0 ( 13.85 ± 3. 17 ) mg/L, C24:0( 16. 10 ±5.84 ) mg/L by MDI have significant differences( t = 8. 401 ,P =0. 000;t =9. 914,P =0. 000 ),but C26:0( 0.68 ±0.48 ) mg/L, C24:0/C22:0( 1.20 ±0.40 ), C26: 0/C22:0 ( 0.04 ±0.04 )by this method and C26: 0( 0. 65 ± 0. 67 ) mg/L, C24:0/C22: 0( 1.19 ± 0. 43 ), C26:0/C22: 0 ( 0. 05 ± 0. 05 )by MDI have no differences( t values were 0. 372,0. 317,0. 945 ,respectively ,all P ＞0. 05 ). Conclusions The quantitative analysis method for serum very long chain fatty acid using LC-MS/MS is accurate, sensitive,specific and stable. It could provide important biochemistry information for diagnosis in clinic.

Objective To establish a LC-MS/MS method for the determination of MMA in serum,and provide a assay for the diagnosis and screening of methylmalonic academia in the clinic. Methods MMA was extracted from 205 serum samples from healthy controls and 146 serum samples from patients with liquidliquid extraction method with MTBE as the extraction solvent. The supernatant was transferred to a tube and dried with nitrogen gas. Then the residual was derived with HCI-BuOH mixed agent to give a product, which was analyzed directly by LC-MS-MS system with a gradient elution, selective reaction monitor, a Discovery C18 column (50 mm × 2. 1 mm ,5 μm) as the isolation column and a mobile phase consisting of methanol and water (0. 1% formic acid, V/V), respectively. The concentration of MMA was detected with the isotope internal standard method. The stand curve was employed with a series of calibrators. The recovery was estimated with the 3 serum samples with the concentrations of 2, 25, 80 μg/L respectively. The accuracy,precision and stability were also tested with quality control samples. Moreover, the range of concentrations in healthy people were detected to investigate the influence of hemolysis on the detection results. Thirteen samples were randomly tested according to the digital chart. The testing results were compared with the results provided by Medical Diagnositic institution (MDI) in Germany. The paired t-test was applied to statistical analysis. Results The linear range of this method was 2-100 μg/L, and the correlation coefficient (R2 ) was more than 0. 995. The retention time of MMA derivative was 10. 5 min. Succinic acid and MMA were not found to interfere with each other. The within-run RSD was less than 6. 4%, and the between-run RSD was less than 5.0%. The recovery rates were from 96. 42% to 103. 33%. The limit of quantification was 1 μg/L.The accuracies of the method were form 94. 2% to 108. 2%. The samples were stable for 6 h at room temperature and stable for 70 d even keep at - 20 ℃. The samples were stable after 10 freeze-thaw cycles. The derivatives of MMA were found to be stable at least for 5 d at 4 ℃. The medians of the hemolysis group and the normal group were 102.53 (13.84-302.33) μg/L and 39.52 (11.94-203.08) μg/L,respectively. There was significant difference between the 2 groups ( T = 8, P ＜ 0. 05 ). The medians of comparison test in our laboratory and the MDI were 32. 82(24. 50-100. 42) μg/L and 32. 20(26. 65-93. 30)μg/L There was no significant difference between the 2 groups( T=7 ,P ＞0. 05 ). The mean value (-x± s)of 158 healthy adults( 18-58 years old) and 47 healthy teenages( 1-17 years old)were ( 18.46 ± 10.49 )μg/L and (22. 38 ± 11.45) μg/L, respectively. Conclusions A LC-MS/MS method for analysis of MMA in serum is established successfully. The quantitative method is simple and accurate with good sensitivity,specificity and repeatability. The method can be applied for diagnosis, screening and monitoring of methylmalonic acidemia.

Objective To explore a more effective way to enhance the clinical efficacy of cluster immunotherapy for perennial allergic rhinitis. Methods A total of 60 perennial allergic rhinitis patients were evenly randomized into treatment goup and control group. The two groups were given cluster immunotherapy, and the treatment group was given daily vesiculating moxibustion additionally . Before and after treatment, the scores of traditional Chinese medical constitution and the rhinitis quality of life ( QOL) were observed, and one year after treatment, the therapeutic effect and safety were evaluated. Results ( 1) One year after treatment, the treatment group showed better total clinical efficiency than the control group ( 96.67% vs 80.00%, P<0.05). ( 2) After treatment, the average scores of Qi deficiency constitution, yang deficiency constitution and special constitution were significantly lower than those before treatment in the treatment group ( P<0.01). The improvement of Qi deficiency constitution and yang deficiency constitution in the treatment group was superior to that in the control group ( P<0.05). ( 3) The total QOL scores and the scores of each dimension of QOL scale were improved in both groups, and the differences were significant except for the emotion dimension ( P<0.01). Insignificant differences were shown between the two groups after treatment ( P>0.05). ( 4) Two cases had grade 1 general adverse reaction and 5 cases had local adverse reaction during cluster immunotherapy. During vesiculating moxibustion, 3 cases had blistering and the blistering disappeared after symptomatic treatment. Conclusion Daily vesiculating moxibustion combined with cluster immunity therapy is effective for the treatment of allergic rhinitis, showing good effect on improving Qi deficiency constitution, yang deficiency constitution and special constitution as well as the quality of life of the patients.

Objective To set up the rolling circle amplification (RCA) system for detecting hepatitis B virus (HBV) covalently closed circular DNA (cccDNA), and to evaluate the specificity and sensitivity of this system. Methods Plasmids containing full-length of wild-type HBV genome were treated with restriction enzyme and T4 DNA ligase, and then were concentrated. The DNA fragments were recovered by the nucleic acid purification kit and severed as standard HBV cccDNA. Total DNA was extracted from hepatic tissues of seven chronic hepatitis B patients. RCA method was used to amplify genomes from tissue samples. Standard HBV cccDNA, 3.2 kb liner HBV DNA, normal hepatic tissue samples and 15 serum samples of patients with chronic HBV infection were used as controls to determine the specificity of RCA. Ten-fold serial dilutions of standard HBV cccDNA were used for determining the sensitivity. Results The standard HBV cccDNA was successfully constructed and could be detected by RCA method. HBV cccDNA could be amplified from 2 mg hepatic tissue samples at least of HBV infected patients, and could be detected as low as 1 ×102 copy/μL. cccDNA was not detected in 3.2 kb liner HBV DNA, normal hepatic tissue samples and 15 serum samples of chronic HBV infected patients. Conclusion RCA method can be used for rapid and simple detection of HBV cccDNA with high specificity and sensitivity.

Objective:To study the pharmacokinetics,thissue distribution and secretion of nerve growth factor(NGF)in mice.Methods:The conecntration of NGF in various body fluids and tissue were determined by isotope tracer combined SDSPAGE method.Results:The plasma concenmtration-time curve was in accordance with the two-compartment pharmacokinetic model.The elimation half-life(t1/2β)was 3.1.The half-life of distribution(t1/2ka)was 5min.Tpeak was 25 min.AUC was 72.4 mg·kg-1·h-1.The concentrations of NGF were high in thyroid,blood,submaxillary glands,superior cervical ganglion,adrenasl and kidneys.Conclusion:NGF has a wide distribution,high tissue concentrationa nd excrtet mainly through the urine.