Chemically sulfated natural galactomannans with specific antiviral and anticoagulant activities.

Naturally occurring galactomannans were sulfated to give sulfated galactomannans with degrees of substitution of 0.7-1.4 per sugar unit and molecular weights of M¯n=0.6×10(4)-2.4×10(4). Sulfated galactomannans were found to have specific biological activities in vitro such as anticoagulant, anti-HIV and anti-Dengue virus activities. The biological activities were compared with those of standard dextran and curdlan sulfates, which are polysaccharides with potent antiviral activity and low cytotoxicity. It was found that sulfated galactomannans had moderate to high anticoagulant activity, 13.4-36.6unit/mg, compared to that of dextran and curdlan sulfates, 22.7 and 10.0unit/mg, and high anti-HIV and anti-Dengue virus activities, 0.04-0.8µg/mL and 0.2-1.1µg/mL, compared to those curdlan sulfates, 0.1µg/mL, respectively. The cytotoxicity on MT-4 and LCC-MK2 cells was low. Surface plasmon resonance (SPR) of sulfated galactomannans revealed strong interaction with poly-l-lysine as a model compound of virus proteins, and suggested that the specific biological activities might originate in the electrostatic interaction of negatively charged sulfate groups of sulfated galactomannans and positively charged amino groups of surface proteins of viruses. These results suggest that sulfated galactomannans effectively prevented the infection of cells by viruses and the degree of substitution and molecular weights played important roles in the biological activities.

Isolation and antiviral activity of water-soluble Cynomorium songaricum Rupr. polysaccharides.

The plant, Cynomorium songaricum Rupr., is used as a traditional medicine in China and Mongolia. In the present study, two new water-soluble polysaccharides isolated from C. songaricum Rupr. were purified by successive Sephadex G-75 and G-50 column chromatographies and then characterized by high resolution NMR and IR spectroscopies. The molecular weights of two polysaccharides were determined by an aqueous GPC to be [Formula: see text] = 3.7 × 10(4) and 1.0 × 10(4), respectively. In addition, it was found that the polysaccharide with the larger molecular weight was an acidic polysaccharide. It was found that the iodine-starch reaction of both isolated polysaccharides was negative and the methylation analysis gave 2, 4, 6-tri-O-methyl alditol acetate as a main product. NMR and IR measurements and sugar analysis revealed that both polysaccharides had a (1 â†’ 3)-α-d-glucopyranosidic main chain with a small number of branches. After sulfation, the sulfated C. songaricum Rupr. polysaccharides were found to have a potent inhibitory effect on HIV infection of MT-4 cells at a 50% effective concentration of 0.3-0.4 µg/ml, a concentration that has almost the same high activity as standard dextran and curdlan sulfates, EC50 = 0.35 and 0.14 µg/ml, respectively. The 50% cytotoxic concentration was low, CC50>1000 µg/ml. In addition, the interaction between the sulfated polysaccharides and poly-l-lysine as a model protein compound was investigated by a surface plasmon resonance to reveal the anti-HIV mechanism.

Determination of Content and Related Substances of Methylthioninium Chloride Injection by HPLC / 中国药师

Objective:To establish an HPLC method for the determination of methylthioninium chloride injection and its related substances. Methods:Octadecylsilane bonded silica was used as the filler. A Waters Symmetry C18 column (150 mm × 4. 6 mm, 5μm) was used with the mobile phase of 0. 3% phosphoric acid solution-acetonitrile (70∶ 30). The detection wavelength was 246 nm and the column temperature was 30℃. The flow rate was 1. 0 ml·min-1 and the injection volume was 10μl. Results:The separation degree of methylthioninium chloride peak and impurity peaks was good. Methylthioninium chloride within the range of 2. 0-60. 0 μg· ml-1showed a good linear relationship (r=0. 9998). Impurity A within the range of 0. 2-3. 2 μg·ml-1 showed a good linear relation-ship (r=0. 999 6). The average recovery of methylthioninium chloride was 99. 75% and RSD was 0. 86% (n=9). Conclusion:The method is simple, accurate and specific with high precision. It can be used to determine the content of methylthioninium chloride injec-tion and its related substances.

Improvement of the Determination Method of Content and Related Substances of Molsidomine Tablet / 中国药房

OBJECTIVE:To improve the method for the determination of content and related substances of Molsidomine tablet. METHODS:HPLC was adopted to determine the content and related substances,HPLC-MS was adopted for the qualitative degra-dation impurities in destruction test. The column was Waters Symmetry-C18 with mobile phase of 0.05% formic acid- methanol (gradient elution) at a flow rate 1.0 ml/min and 0.2 ml/min (respectively for content and related substances determination,mass spectrometry),the detection wavelength was 210 nm,column temperature was 30℃,the injection volume was 20μl and 5μl(re-spectively for content and related substances determination,mass spectrometry). RESULTS:Each component was well separated un-der chromatographic conditions;degradation impurities were confirmed;the linear range of molsidomine was 4. 0-28.0 μg/ml(r=0.9997);RSDs of precision,stability and reproducibility tests were lower than 1%;recovery was 97.74%-100.70%(RSD=0.49%,n=9). CONCLUSIONS:The method is simple,accurate with good specific and high precision,and can improve the quali-ty control of Molsidomine tablet.

MORPHOLOGICAL STUDY OF THE PANCREAS IN HUMAN FETUS / 解剖学报

Pancreas of 53 aborted fetuses ranging from 5 to 41 weeks were fixed in Bouin's fluid and embedded in paraffin, and the sections were stained with Alcian bluephloxine and Grimelius silver technique. The A, B and D cell of islet of Langerhans were demonstrated with immunohistochemical PAP method also. 14 pancreas among them were fixed in 2% glutaraldehyde and embedded in Epon 812, and observed with TEM. Typical A, B and D islet cells were observed with LM at 12 weeks of fetal age. Undifferentiated, differentiated and degenerated cells were distinguished in the pancreatic islets at 14 to 41 weeks of fetal age. The secretory granules of the exoerine cells were observed after 14 weeks of fetal age. The functional differentiation of endocrine cells precede that of exocrine cells. The morphometrical measurement testified that the endocrine portion of fetal pancreas was obviously larger than that of adult. This study provides some morphological data for the pancreas transplantation in clinical medicine.