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 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 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.

With the development of recombinant DNA technology, genetically altered mouse models for human cancers are critical to the investigation and characterization of oncogene and tumor suppressor gene expression and function and the associated cancer phenotype. Recently, the inhibition of tumor angiogenesis becomes a new “hot spot” in cancer researches. And the genes involved in the regulation of tumor angiogenesis play an increasingly important role in the field of tumor researches. This review is about the progress of the research in transgenic tumor models and focuses on genes regulating tumor angiogenesis associated with transgenic mice model.

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

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.

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.