Alkaloids from Macleaya cordata and their cytotoxicity assay / 中国中药杂志

<p><b>OBJECTIVE</b>To study the alkaloids of Macleaya cordata and their anti-tumor activities.</p><p><b>METHOD</b>Alcohol and liquid-liquid extraction were used methods were used to extract the alkaloids constituents, and silica gel, reverse-phase octadecylsilyl (ODS), sephadex LH-20 chromatographic methods and HPLC were applied to isolate and purify compounds. MS, NMR spectroscopic methods were used to determine their structures. Furthermore, the cytotoxicity of these chemical components for MCF-7 and SF-268 cell lines was measured by MTT method.</p><p><b>RESULT</b>Twelve alkaloids were isolated from the fruits of M. cordata, and their structures were identified as: maclekarpine E (1), 6-acetonyldihyrochelerythrine (2), cavidilinine (3), 6-acetonyldihyrosanguinnarine (4), O-methylzanthoxyline (5), 6-methoxy-dihydrosanguinarine (6), spallidamine (7), 6-hydroxyldihydrochelerythrine (8), arnotianamida (9), dihydrosanguinarine (10), protopine (11), and cryptopine (12).</p><p><b>CONCLUSION</b>Compounds 1, 3, 7-9 were isolated from M. cordata for the first time, and compound 5 is a new natural product. The results of cytotoxic assay indicated that compound 6 showed strong cytotoxicity against MCF-7 and SF-268 cell lines with IC50 values of 0.61 μmol · L(-1) and 0.54 μmol · L(-1), respectively.</p>

Chemical constituents of flavonoids and their glycosides in Melastoma dodecandrum / 中国中药杂志

The chemical constituents of 95% ethanol extract of Melastoma dodecandrum were isolated and purified by chromatography on silica gel, Sephadex LH-20, and HPLC, to obtain thirteen compounds eventually. On the basis of their physico-chemical properties and spectroscopic data, these compounds were identified as quercetin (1), quercetin-3-O-β-D-glucopyranoside (2), quercetin-3-O-(6"-O-p-coumaroyl) -β-D-glucopyranoside (3), kaempferol (4), kaempferol-3-O-β-D-glucopyranoside (5), kaempferol-3-O- [2",6"-di-O-(E)-coumaroyl]-β-D-glucopyra-noside (6), luteolin (7), luteolin-7-O-(6"-p-coumaroyl) -β-D-glucopyranoside (8), apigenin (9), apigenin-7-(6"-acetyl-glucopyranoside) (10) , naringenin (11), isovitexin (12), and epicatechin-[8,7-e] -4β-(4-hydroxyphenyl)-3,4-dyhydroxyl-2(3H)-pyranone (13). Eight compounds(3,5,6,8-11 and 13) were obtained from M. dodecandrum for the first time.

Virulence genes and pathogenicity of Shigella flexneri Xv isolated in Beijing / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the biochemical characteristics, virulence genes and pathogenicity of Shigella flexneri Xv isolated in Beijing.</p><p><b>METHODS</b>61 strains of S. flexneri Xv isolated from diarrhea patients in Beijing were systematically determined through biochemical reactions and serological tests. Application of PCR technique in detection of virulence genes on ipaH, sen, virF, ial and pulsed-field gel electrophoresis (PFGE) was used to identify the related characteristics and on rat lung slices to determine its pathogenicity.</p><p><b>RESULTS</b>All of the S. flexneri Xv could ferment glucose, mannitol, melibiose and arabinose. Using serum agglutination, we found that the antigen structure was (IV: 7, 8). IpaH, sen, virF and ial that carried rates of virulence genes appeared to be 100%, 81.97%, 75.41% and 80.30%, respectively. Among 61 strains of S. flexneri Xv, the PFGE typing of Shigella bacteria could be divided into 25 belt types while the results from rat lung slices showed inflammatory change of Xv.</p><p><b>CONCLUSION</b>S. flexneri Xv was found that it carried high rate of Shigella virulence genes, exhibiting genetic polymorphism and highly invasive.</p>

Progress in research on multilocus sequence typing technique / 军事医学科学院院刊

Multilocus sequence typing (MLST) is a molecular genotyping method based on nucleotide sequencing. The procedure of this method characterizes isolates of bacterial species using the DNA sequencing of multiple housekeeping genes(usually seven). For each housekeeping gene, the different sequences present within a bacterial species are assigned as distinct alleles.For each isolate, the alleles at each of the loci define the allelic profile or sequence type (ST). MLST has the advantages of being robust (based on genetic data) and electronically portable to generate data that allow rapid and global comparisons between different laboratories. In this paper, the principle, method, data analysis, application, advantages and flaws of MLST are introduced.

Seroepidemical study of Coxsackievirus A 16, in four provinces, China, 2005 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the situation of 1- 5-years-old children's antibody against Coxsackievirus A group 16 strain (CVA16) in Guangdong, Heilongjiang,Yunnan Province and Xinjiang Uygur Autonomous Regions, China, 2005, it can offer scientific evidences for preventing and controlling CVA16 causative hand-food and mouth disease.</p><p><b>METHODS</b>Using microneutrilization test, to study 503 serum samples randomly selected from sera collected in 2005.</p><p><b>RESULTS</b>Positive rate of anti-CVA16 antibody were 41.90%, 9.40%, 40.00% and 34.40% in Guangdong, Heilongjiang,Yunnan and Xinjiang, respectively. Antibody titer was relative low (average, 1: 6.1) and there was no statistical difference of geometry mean of antibody titer (GMT) among Guangdong, Heilongjiang, Yunnan (F = 0.97, 0.40, 1.06, respectively; P > 0.05), while there had statistical difference of GMT between Heilongjiang and other three regions( F = 10.61, P < 0.00).</p><p><b>CONCLUSIONS</b>There had probably existed local epidemic in some regions of Guangdong, Heilongjiang, Yunnan Province and Xinjiang Uygur Autonomous Regions, China, 2005 or even before, but the area and degree of transmission and epidemic had difference. Children aged from 1- 5-years-old were relatively susceptible population of CVA16 infection.</p>

Effect of mastoparan-1 on lipopolysaccharide-induced acute hepatic injury in mice / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the effect of mastoparan-1 (MP-1) on lipopolysaccharide (LPS)-induced acute hepatic injury in mice and probe into its possible mechanism.</p><p><b>METHODS</b>One hundred and four BALB/c mice were randomly divided into healthy control group (n = 8, without treatment, HC), LPS group (n = 48, with injection of LPS 5 mg/kg via tail vein), and MP-1 group (n = 48, with injection of LPS 5 mg/kg and MP-1 3 mg/kg via tail vein). Mice in LPS group and MP-1 group were sacrificed at 2nd, 6th, 12th, 24th, 48th and 72nd post injection hour (PIH), 8 mice at each time point in each group. Blood samples were collected for determination of plasma levels of LPS by kinetic turbidimetric limulus test, TNF-alpha and IL-6 by ELISA, serum levels of ALT and AST by automatic biochemistry analyzer respectively. Hepatic tissue samples were collected for determination of TLR4, TNF-alpha and IL-6 mRNA by real-time fluorescent quantitation reverse transcription polymerase chain reaction, along with the observation of pathological changes in hepatic tissue at each time point. Above-mentioned examinations were also performed in HC group.</p><p><b>RESULTS</b>Compared with those of HC group, plasma levels of LPS and TNF-alpha in LPS group significantly increased at 2nd PIH (18,320.50 +/- 2782.50 EU/mL and 988 +/- 130 ng/L, respectively), then decreased gradually to 1.80 +/- 0.80 EU/mL and 150 +/- 44 ng/L at 72nd PIH, which was close to those of HC group. The values of IL-6, ALT and AST peaked at 12th PIH, which declined to the levels close to those of HC group at 72nd PIH. Meanwhile, the expressions of TLR4, TNF-alpha and IL-6 mRNA in liver were remarkably up-regulated after injection, and the pathological changes in hepatic tissue pronounced significantly at 12th, 24th and 48th PIH. Compared with those of LPS group, the levels of LPS, cytokines, ALT and AST decreased in MP-1 group in different degrees after injection (P < 0.05 or P < 0.01), genes expression (P < 0.05 or P < 0.01) and pathological changes was respectively suppressed and alleviated in hepatic tissue.</p><p><b>CONCLUSIONS</b>MP-1 can alleviate LPS-induced acute hepatic injury in mice, which may be associated with its neutralization of LPS and attenuation of synthesis and release of inflammatory mediators.</p>

Screening of Panax notoginsenoside water in oil microemulsion formulations and their evaluation in vitro and in vivo / 药学学报

Water in oil (W/O) microemulsion formulation was developed to enhance intestinal absorption of ginsenoside Rb1 (Rb1) of panax notoginseng (PNS). Effects of W/O microemulsions on pharmacokinetics after intraduodenal administration, membrane fluidity and membrane transport of Rb, were studied in rats, liposomes and parallel artificial membrane permeability assay (PAMPA), respectively. Soybean phospholipids/ethanol (SP/EtOH) was selected as surfactant/cosurfactant, together with PNS 400 mg x mL(-1) solution and various kinds of oils, to prepare 11 W/O microemulsions. Most of the microemulsions can enhance Rb1 intestinal absorption significantly. Besides surfactant/cosurfactant, oil also had an effect on the enhanced absorption and the order of enhancement was as follows: glyceryl laurate approximately = isopropyl myristate > isopropyl palmitate > 2-ethylhexanol palmitate. The effection of absorption enhancement by the long chain glyceride ( > C14) is lower than that by the medium chain glyceride (C8 - C14). Most of W/O microemulsions were found to enhance the membrane fluidity of liposomes to different extents. In PAMPA analysis, efficient permeability coefficient (Pe) of diluted-microemulsion (D-ME) is mostly higher than that of PNS solution, which indicated the components of microemulision can facilitate the membrane permeability of the drug. Meanwhile, linearity correlation between Pe and ratio of relative bioavailability (Fr) was acquired for undiluted microemulison (ME). Therefore, W/O microemulsions can enhance intestinal absorption of Rbr, and this effect may be attiributed to its enhancement on membrane fluidity to a certain degree. PAMPA analysis could be brought into not only the investigation of membrane transport of crude drug, but also conditioned preformulation research (e.g. absorption enhancer etc.).

Comparison between the characteristics of absorption and pharmacokinetic behavior of ginsenoside Rg1 and ginsenoside Rb, of Panax notoginseng saponins / 药学学报

To compare the characteristics of absorption and pharmacokinetic behavior of ginsenoside Rg1 (Rg1) with ginsenoside Rb1 (Rb1) of panax notoginseng saponins (PNS), bile excretion of both Rg1 and Rb1 were studied after i.v. and i.g. of PNS solution. Plasma protein binding ratios were studied using equilibrium dialysis method, and referred to pharmacokinetic parameters. It shows that (61.48 +/- 18.30)% dose of Rg1 and (3.94 +/- 1.49)% dose of Rb1 were separately excreted into bile 10 hours after i.v. administration (PNS 50 mg x mL(-1)), and (0.91 +/- 0.51)% dose of Rg1 and (0.055 +/- 0.02)% dose of Rb1 were excreted into bile 12 hours after i.g. administration (PNS 1 500 mg x mL(-1)). Plasma protein binding degrees of Rg1 and Rb1 were 6.56% - 12.74% and 80.1% - 89.69%, respectively. Stomach, intestinal and hepatic throughput efficiency (F(S), F1 and F(H)) for Rg1 were 49.85%, 13.05%, 50.56%, respectively, and 25.82%, 4.18%, 65.77% for Rb1. Therefore, poor intestinal absorption is a primary reason for the low bioavailability of both Rg1 and Rb1. Rg1 possesses relatively high bile excretion and low plasma protein binding rate, in contrast, Rb1 possesses low bile excretion and high plasma protein binding rate. Membrane permeability and elimination rate of Rb1 were lower than that of Rg1, meanwhile, longer MRT and bigger AUC could be found for Rb1.

The protective effect of high density lipoprotein on the lung function of rats with severe burns / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the protective effect of high density lipoprotein on the lung function of rats with severe burns.</p><p><b>METHODS</b>One hundred and thirty-five Wistar rats were employed in the study and were randomly divided into control (n = 15, without injury), burn (n = 60, with 30% TBSA full-thickness burn on the back) and experimental [(n = 60, with the injection of HDL (80 mg/kg) via the caudal vein immediately after burns)] groups. The rats in the latter two groups were resuscitated with intraperitoneal isotonic saline (50 ml/kg) 30 minutes after burns. The serum content of ICAM-1 and TNF-alpha as well as the blood content of PCO2 and PO2 of the rats in burn and experimental groups were determined at 12, 24, 48 and 72 post-burn hours (PBH) and in control group. The pathological changes in the lung tissue of the rats in all groups were observed under light microscope and electronic microscope at 48 PBH.</p><p><b>RESULTS</b>PCO2 and the contents of ICAM-1 and TNF-alpha in burn group were significantly higher, but the PO2 was lower than those in control group at each time-point (P < 0.05 or P < 0.01). There were no obvious differences in the above indices between the experimental and control groups (P > 0.05), but the ICAM-1 and TNF-alpha levels in experimental group were markedly decreased than those in burn group at each time-point (P < 0.05 or P < 0.01). The ICAM-1 and TNF-alpha contents in burn group at 48 PBH were (3.42 +/- 0.25) microg/L and (4. 04 +/- 0.28) ng/L, respectively, which were markedly higher than those in experimental group [(2.24 +/- 0.14) microg/L, (3.35 +/- 0.22) ng/L, P < 0.05 or P < 0.01]. Dilation of capillaries, congestion and inflammatory infiltration in the pulmonary capillaries, and loosening of conjunction between pulmonary capillary vascular endothelial cells and endothelial swelling were observed in burn group at 48 PBH. Compared with the burn group, the injury was markedly alleviated in the experiment group, and the pulmonary capillary endothelial cells showed tighter junction.</p><p><b>CONCLUSION</b>HDL exhibits a protective effect on the lung function of rats with severe burns via reducing the expression of ICAM-1 and TNF-alpha.</p>

The protective effect of high density lipoprotein on the cardiac function of rats with severe burns / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of high density lipoprotein on the cardiac function of rats with severe burns.</p><p><b>METHODS</b>One hundred and thirty-five Wistar rats were employed in the study and were randomly divided into control (n = 15, without treatment), burn (n = 60, with 30% TBSA full-thickness burn on the back) and experimental (n = 60, with the injection of HDL (80 mg/kg) via the caudal vein immediately after burns) groups. The rats in the groups with burn injury were resuscitated with intraperitoneal isotonic saline (50 ml/kg) 30 minutes after burn (PBM). The serum contents of CK, ICAM-1 and TNF-alpha of the rats of all the three groups were determined with corresponding methods. The histological changes in the cardiac muscle tissue of the rats in all groups were observed under light microscope and electronic microscope.</p><p><b>RESULTS</b>The serum contents of CK, ICAM-1 and TNF-alpha in the control group were obviously lower than those in burn group (P < 0.01), while those in experimental group were also markedly lower than those in control group (P < 0.05 or 0.01). The average reduction rate was 36.5%, 32.0% and 12.6%, respectively. The size and the structure of the cardiac muscular fiber in the control group were even and normal. Compared with the burn group, degeneration, inflammatory infiltration and mitochondrial swelling were found to be less marked in the experimental group at 48 PBH, and no focal lysis and necrosis were found, which were observed in the burn group.</p><p><b>CONCLUSION</b>High density lipoprotein can be beneficial to the protection of cardiac tissue in protecting from secondary injury in rats with severe burns.</p>

Cloned goats produced from the somatic cells of an adult transgenic goat / 生物工程学报

This study was carried out to examine the effect of different donor cell type and micro-manipulation on the development of reconstituted embryos. Cultured mural cumulus cells or fibroblast cells from an adult transgenic goat expressing human erythropoietin(rhEPO) were used as the donor cells in nuclear transfer experiments. The reconstituted eggs were generated by transferring fibroblast cells or cumulus cells into the perivitelline space of enucleated M II oocytes and then followed by electrofusion and activation. After 6 days' incubation in vivo, the reconstructed embryos developed into morulae or blastocysts were transferred into 6 foster recipients. Two of the foster-mothers were pregnant and gave birth to two offspring, which were derived from the fibroblast cell and cumulus cell, respectively. Fingerprint analysis showed that the PCR-RFLP patterns of the two offspring were identical to that of donor goats. PCR results indicated that these cloned goats carried hEPO gene as same as their donor cells.