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 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.

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 Mikulicz's disease (MD) was considered to be a subtype of Sj(o)gren's syndrome (SS) due to the clinical and histological similarities between them.Evidence had shown that there were differences between MD and typical SS.The purpose of this study was to investigate the correlation between MD and SS,by means of analyzing the expression of IgG4 in salivary glands and the clinical characteristics of patients who were previously considered as SS.Methods The paraffin sections of salivary glands from SS patients were stained with monoclonal antibodies to IgG4 and CD38.Patients were divided into two groups based on the pathological results.Analysis of the symptoms,the signs and the laboratory results were carried out in these patients.The difference in laboratory parameters and histopathological gradings in the two groups was analyzed.Normal and abnormal distributed data comparison was conducted using random independent samples t-test and rank sum test respectively.Two-sample rates were compared with Chi-square test.Results Based on immunohistochemistry of IgG4 distribution,the 58 patients with SS were divided into two groups:IgG4 related (9 cases) and non-IgG4 related (49 cases).Histopat-hologically,IgG4 related cases showed IgG4+ plasma cells/IgG+ plasma cells infiltration and there were more IgG4 related monoclonal antibody expressed when compared to IgG4 unrelated cases.In addition,there were also significant differences in clinical features between the two groups.IgG4 related disease was associated with male gender,higher level of plateletconnt,lymphocytes [(2.4±0.8)×109/L vs (1.4±0.7) ×109/L] count and CRP [(52±60) mg/L vs (15±17) mg/L] levels and lower titer of IgM [(1.2±0.7) g/L vs (1.8±0.8) g/L],antinuclear antibody (56％ vs 87％) and anti-SSB antibodies (13％ vs 54％) (P＜0.05),when compared with IgG4-unrelated cases.There was no significant difference in other indicators (P＞0.05).Conclusion The present study has demonstrated that some of the MD patients misdiagnosed as SS.Some of the laboratory tests such as the level of platelet and lymphocyte count,serum level of CRP,IgM,antinuclear antibody,anti-SSB antibodies,the serum levels of IgG4 and the histopathological presentations in the salivary gland are different between these two disorders.Because of good response to steroid in MD,so laboratory tests and pathological examinations for IgG4 can help to avoid misdiagnosis.

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.

Aim To investigate the effects of(S,S)ZX-5 and(R,R)ZX-5 on eNOS mRNA and protein levels.Methods Cell culture was used for the experiments in vitro.The effects of eNOS mRNA were investigated by RT-PCR.The effects of eNOS protein levels were investigated by Western blot.Results(S,S)ZX-5 in concentration of 30 mg?L-1 up-regulated eNOS expressions at protein and mRNA levels significantly in cultured ECV304(P

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 choose a best cultivar by comparing the content of total flavonoid and naringin between different cultivars of Citrus grandis 'tomentosa'.Methods The content of total flavonoids was determined by spetral photometric analysis.The content of naringin was determined by HPLC.Results The content of total flavonoid was 19.24 % and 18.06 %,and naringin content was 11.72 % and 12.38 % in the cultivars from Dachaling and Fengwei respectively,which were much higher than the other cultivars.Conclusion The contents of total flavonoid and naringin were different in different cultivars of Citrus grandis 'tomentosa'.The cultivars of Citrus grandis 'tomentosa' from Dachaling and Fengwei have the best quality.

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.