OBJECTIVE:To establish capillary GC method for the determination of residual organic solvents in artesunate,such as methanol and ethyl acetate.METHODS:Capillary GC was adopted and the temperature of flame ionization detector was 250℃.RESULTS:The linear range were 0.03～0.605 mg?mL-1 for methanol(r=0.999 8) and 0.05～1.002 mg?mL-1 for ethyl acetate(r=0.999 7).The average recovery rates were 98.7% for methanol(RSD=3.0%) and 99.1% for ethyl acetate(RSD=2.1%).Only the ethyl acetate was detected in samples.CONCLUSION:Established method is simple and accurate for the content determination of residual organic solvents in artesunate.

To review the research of block copolymer micelles as delivery system for poorly soluble(antineoplastic) carrier over the last 10 years,including the composition,preparation methods and factors which influence the loading efficiency,physicochemical properties,and targeting characteristics of block copolymer micelles.Block copolymer micelles are self-assembled structures formed from amphiphilic block copolymers by dispersing in aqueous media.The hydrophilic blocks of the copolymer form the outer shell of the micelle,while the hydrophobic blocks form the inner core,and the proper coreshell micellar architecture was constituted.Block copolymer micelles have a whole set of unique properties,such as small sizes,narrow particle size distribution,high drug loading capacities,and available disposition characteristics in the body.Block copolymer micelles have been found as promising drug carriers due to making poorly soluble antineoplastic lysis,toxicities and side effects decrease,bioavailability increase,and targeting the drugs to specific sites in a passive way or attaching ligands in an active way,which can be specifically(recognized) by receptors onto the surface of copolymers.

Objective To investigate the relationship between chronic gastropathy with intestinal metaplasia (IM) and caudal type homeobox genes 2 (Cdx2),tumor necrosis factor-α (TNF-α),Helicobacter pylori (Hp) infection.Methods Onehundred and thirty patients underwent gastroscopy were divided into 2 groups by typical pathological type:gastritis group (30 cases) and IM group [100 cases,including mild IM group (30 cases),moderate IM group (35 cases),severe IM group (35 cases)].Hp infection was confirmed.The expressions of Cdx2 and TNF-α protein was evaluated by immunohistochemistry.Results The positive expression rates of Cdx2 and TNF-α protein in IM group were significantly higher than those in gastritis group [78.0％(78/100) vs.6.7％(2/30),60.0％(60/100) vs.16.7％(5/30)],and there were statistical differences (P ＜ 0.01).The positive expression rates of Cdx2 and TNF-α protein in positive and negative Hp infection in IM group were significantly higher than those in gastritis group [Cdx2 protein:79.4％(50/63) vs.1/10,75.7％(28/37) vs.5.0％(1/20);TNF-α:74.6％(47/63) vs.4/10,35.1％(13/37)vs.5.0％ (1/20)],and there were statistical differences (P ＜ 0.01).The positive expression rate of Cdx2 protein in moderate IM group and severe IM group were significantly higher than those in mild IM group [80.0％ (28/35) and 97.1％ (34/35) vs.53.3％ (16/30)],in severe IM group were significantly higher th.an those in moderate IM group,and there were statistical differences (P ＜0.05).There were no statistical differences in the positive expression rate of Cdx2 protein of difference Hp infection among the 4 groups (P＞0.05).There were no statistical differences in the positive expression rate of TNF-α protein between mile IM group and moderate IM group (P ＞ 0.05).The positive expression rate of TNF-α protein in severe IM group was significantly higher than that in mild IM group and moderate IM group [80.0％(28/35) vs.40.0％(12/30) and 57.1％(20/35)],and there were statistical differences (P ＜ 0.05).The positive expression rate of TNF-α protein of positive Hp infection patients in gastritis group,mild IM group,moderate IM group and severe IM group were significantly higher than those of negative Hp infection patients (4/10 vs.1/20,47/63vs.13/37,10/18 vs.2/12,15/21 vs.5/14,22/24 vs.6/11),and there were statistical differences (P ＜ 0.05).Conclusions The expressions of Cdx2 and TNF-α protein and Hp infection are closely related with IM.Therefore,testing the level of Cdx2 and TNF-α protein expressions can help to judge the degree of IM,which provides a basis for reversing IM.

Objective To explore the diagnostic value and clinical significance of the combined detection of antinuclear antibody (ANA) and anti-nuclear antibody spectrum (ANAs) in systemic lupus erythematosus (SLE) .Methods 110 patients with SLE ,88 patients with other autoimmune diseases (AID) and 50 individuals with healthy physical examination were selected and detected se-rum ANA by using indirect immunofluorescence (IIF);the Western blot was adopted to detect the 15 items of ANAs .Results A-mong 110 cases of SLE ,the positive rates of serum ANA ,anti-ribonucleoprotein /Smith antibody (nRNP/Sm) ,anti-Smith antibody (Sm) ,anti-Sjogren′s syndrome A antibody (SSA) ,anti-Ro-52 antibody (Ro-52) ,anti-Sjogren′s syndrome B antibody (SSB) ,anti-scleroderma 70 antibody (Scl-70) ,anti-PM-Scl antibody (PM-Scl) ,anti-cytoplasmic group acyl-tRNA antibody (J0-1) ,anti-centro-mere antibodies (CENP B) ,anti-proliferative protein antibody (PCNA ) ,anti-double stranded DNA antibody (ds-DNA ) ,anti-nu-cleosome antibody (AnuA) ,anti-histone antibody (AHA) ,anti-ribosomal P protein antibody (ARPA) and anti-mitochondrial anti-body M2 subtype (AMA M2) were 98 .2% ,59 .1 % ,39 .1 % ,71 .8 % ,68 .2 % ,21 .8 % ,2 .7 % ,3 .6 % ,0 .9% ,9 .1% ,5 .5% , 44 .5% ,38 .2 % ,27 .3 % ,38 .2% and 15 .5% respectively ,the positive rate of above 16 kinds of autoantibody in the orther AID were64.8% ,14.8% ,0% ,37.5% ,42.0% ,11.4% ,9.1% ,0% ,5.7% ,9.1% ,1.1% ,2.3% ,1.1% ,1.1% ,5.7% and2.3% respec-tively ;ANA had the highest diagnostic sensitivity (98 .2% ) and low specificity (35 .2% ) for SLE ,the antibodies with higher speci-ficity in diagnosing SLE were anti-Sm antibody (100 .0% ) ,anti-ds-DNA antibody (97 .7% ) ,anti-AnuA antibody (99 .0% ) ,anti-AHA antibody (99 .0% ) ,anti-ARPA antibody (94 .3% ) and anti-PCNA antibody (98 .9% ) and the disease control group (50 .9% ) .In detecting ANA karyotype by IIF ,the maximum was nuclear particle type (43 .5% ) and the disease control group (50 .9% ) ,no statistically significant difference was found between them (P>0 .05) ,part of the ANA-negative patients have positive ANAs .Conclusion Multiple autoantibodies can be detected in the serum of the SLE patients .The combined detection of ANA and ANAs has important clinical significance for diagnosing and differentially diagnosing SLE .

Objective To investigate the influence of acute hypervolemic hemodilution (AHH) on pharmacodaynamics of cisatracurium in patients undergoing general anesthesia. Methods Sixty ASA Ⅰ or Ⅱ patients aged 18-60 yr scheduled for major abdominal surgery under general anesthesia were randomly allocated into 2 groups (n = 30 each): control group and AHH group. Each group was further divided into 3 subgroups according to the initial dose of cisatracurium (30, 40, 50 μg/kg) . The radial artery and right internal jugular vein were cannulated. BP, HR, CVP, SpO_2, P_(ET) CO_2 and body temperature were continuously monitored. The response of left adductor pollicis muscle to TOF stimulation of ulna nerve was monitored using TOF- Watch~R SX (Organon). Both groups received 10 ml/kg multiple electrolyte solution (plasma-Lyte A) during induction of anesthesia. In group AHH 15 ml/kg 6% hydroxyethyl starch (HES) 130/0.4 solution was infused via internal jugular vein over 30-40 min in addition to plasma-Lyte A. Five minutes after completion of plasma-Lyte A or HES, cisatracurium 30, 40 or SO fig/kg was injected iv in the respective subgroups. After the maximal T_1 block was achieved, the second dose was given to reach a total dose of 100 μg/kg. The onset time, duration of clinical action, total duration of action and recovery index were recorded. The doses for 50% , 90% and 95% T_1 depression (ED_(50), ED_(90), ED_(95)) were calculated by Probit method. Results The ED_(50), ED_(90), ED_(95) of cisatracurium were significantly higher in AHH group than in control group. The onset time of cisatracurium was significantly longer but clinical and total duration of action was significantly shorter in AHH group than in control group. There was no significant difference in recovery index between the two groups. Conclusion AHH can decrease the potency of cisatracurium.

Objective To construct and identify recombinant expression plasmid of small interfering RNA (siRNA)targeting hepatitis B virus X protein(HBx), and observe its effect on mitoehondrial function in healthy liver cell line steadily expressed HBx gene (HL-7702/HBx). Methods Two siRNA sequences containing short hairpin structure, which target on the total length HBx gene, were synthesized and cloned into the vector psiRNA-Hh1GFPzeo to eonstruct recombinant expression plasmids pX1 and pX2. Non-specific recombinant pScr plasmid served as control. After siRNA transfected into HL-7702/HBx cells line by liposome, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to identify the suppressive effect on HBx expression. Levels of intraeellular reactive oxygen species (ROS) and mitochondrial membrane potential (△ m) were determined by flow cytometry. The experimental results were compared by analysis of variance. Results Successful constructions of pX1 and pX2 were confirmed by restriction enzyme digestion and sequencing. The expressions of HBx mRNA and protein after 48 h of transfection into HL-7702/HBx cells in control group were 0.65± 0.12 and 0.62± 0.09, respectively, which were both higher than those (0.33±0.10 and 0.19±0.08, respectively) in group pX1 (t=4.73, P<0.05; t=7.53, P<0.05) and those (0.48±0.10 and 0.37±0.11, respectively) in group pX2 (t=2.39, P<0.05;t=4.43,P<0.05). But the inhibition of group pX1 was stronger than that of pX2 (t=2.28,P<0.05). Levels of ROS and △ m after RNA interference were 5.00±0.38 and 33.86±0.50, respectively, while those in control group were 72. 10±0. 55 and 3. 57±0.26, respectively (ROS: t=276.22, P<0.05; △ m: t=107.15, P<0.05). Conclusions siRNA targeting HBx can efficiently and specifically suppress the HBx expression in HL-7702/HBx cells, and decrease the level of ROS and increase the level of △ m, thus relieve cellular oxidative stress.

The poor chemical stability and solubility, easy volatile and other shortcomings of TCM volatile oils can be improved after they are included by cyclodextrin. Therefore, cyclodextrin inclusion technology is widely used in the preparation process of volatile oil. This article reviewed the domain of cyclodextrin inclusion of volatile oils, the types of cyclodextrin, inclusion technologies, characterization for inclusion, components changes before and after inclusion, and dissolution characteristics of cyclodextrin inclusion.

Objective To investigate the factors affecting the stability of the volatile oil extracted fromYinqiaosan Decoction.Methods The main chemical compositions and the extraction repetitiveness of the compound volatile oil were determined by GC-MS, and the stability of multiple extracted volatile oil was studied. Absorbance of the compound volatile oil was used as the evaluation index, and the factors affecting the stability of the of the compound volatile oil were investigated, such as illumination, temperatures and pH values of volatile oil solution and metal ions.Results The results of the GC-MS chromatograph indicated that the main chemical compositions of the compound volatile oil extracted fromYinqiaosan Decoction twice were the same. The results of the stability of the volatile oil showed that the preservation temperature and illumination affected the stability of the volatile oil to a certain extent. The absorbance values of the compound volatile oil changed slowly when it was stored at a relatively low temperature (4℃) and shielded from light, and it was less stable when stored in normal temperature and under illumination. Meanwhile, the absorbance of the compound volatile oil changed quickly in acid or alkaline solutions and was in instability. The metal ions, such as Cu2+ and Fe3+, have chemical reactions with the compositions of the compound volatile oil and there was a big change in the UV-Vis spectrum of the compound volatile oils.Conclusion The compound volatile oil should be stored at a relatively low temperature (4℃) and shielded from light. At the same time, it should be stored avoiding acids, alkaline and the metal ions, such as Cu2+ and Fe3+, to guarantee its stability. This study provides a reference for the preservation conditions and the preparation conditions of the compound volatile oil extracted fromYinqiaosan Decoction.

[ ABSTRACT] AIM:To evaluate the clinical application of single nucleotide polymorphism array ( SNP array) in prenatal diagnosis for screening the abnormality of women with Down’ s syndrome ( DS) .METHODS:The amniotic fluid samples ( n=312) collected by amniocentesis for the DS screening abnormality women were tested by karyotyping and SNP array analysis, respectively.The findings of karyotyping and SNP array analysis were compared.RESULTS:Two cases of trisomy 21 were identified by karyotyping and SNP array analysis, but SNP array analysis failed to identify 6 cases of chro-mosome balanced structural rearrangement.SNP detected 176 cases copy number variants ( CNVs) in 303 cases normal karyotype were detected by SNP, including 106 benign CNVs, 61 variants of unknown significance (VOUS), 9 de novo CNVs, and none of them was pathogenic.The distribution difference of CNVs in DS screening positive group and DS screening positive plus advanced maternal age group was not statistically significant ( P>0.05) .Furthermore, we reported 14 kinds of CNVs for the first time in population.CONCLUSION:SNP array can further assure chromosome microdupli-cation/microdeletion.In normal karyotype fetus of prenatal diagnosis, SNP can detect some clinical significant CNVs.

AIM:To investigate the expressions of methylthioadenosine phosphorylase(MTAP)and ornithine decarboxylase(ODC)in human ovarian cancer.METHODS:60 fresh samples of ovarian cancer were collected.The expressions of MTAP mRNA and protein were analyzed by using RT-PCR and Western blotting,respectively.ODC activity was measured by high performance liquid chromatography.RESULTS:The expression levels of MTAP mRNA and protein in ovarian cancer were lower than those of control.In 9 of the 60 samples(15%)there were absence of detectable MTAP mRNA and protein.No significant relevance was found between the expression of MTAP and clinical pathologic features.ODC activity in ovarian cancer was(3.82?1.03)U,which was higher than that of normal ovarian tissues(1.38?0.59)U.ODC activity was related with tumor grade.In MTAP-deficiency ovarian cancer tissues ODC activity was significantly increased when compared with that of MTAP-expressing ovarian cancer samples.CONCLUSION:Down-regulated MTAP expression and up-regulated ODC activity really exist in ovarian cancer.Activation of ODC resulting from MTAP deletion may be one of the pathogenetic factors of ovarian cancer.