OBJECTIVETo observe the effects of combined treatment with sansanmycin and macrolides on Pseudomonas aeruginosa and formation of biofilm.

METHODSMicro-dilution method was used to determine the minimal inhibitory concentrations (MICs) of sansanmycin, gentamycin, carbenicillin, polymyxin B, roxithromycin, piperacillin, and tazobactam. PA1 and PA27853 biofilms were observed under optical microscope after staining and under SEM after treatment with sansanmycin at different dosages and combined treatment with sansanmycin and roxithromycin. Viable bacteria in PA1 and PA27853 biofilms were counted after treatment with sansanmycin at different dosages or combined treatment with sansanmycin and roxithromycin.

RESULTSThe MIC of sansanmycin was lower than that of gentamycin and polymyxin B, but was higher than that of carbenicillin. Roxithromycin enhanced the penetration of sansanmycin to PA1 and PA27853 strains through biofilms. PA1 and PA27853 biofilms were gradually cleared with the increased dosages of sansanmycin or with the combined sansanmycin and roxithromycin.

CONCLUSIONSub-MIC levels of roxithromycin and sansanmycin substantially inhibit the generation of biofilms and proliferation of bacteria. Therefore, combined antibiotics can be used in treatment of intractable bacterial infection.

Animals ; Anti-Bacterial Agents ; administration & dosage ; pharmacology ; Bacterial Adhesion ; drug effects ; Biofilms ; growth & development ; Cercopithecus aethiops ; Drug Therapy, Combination ; Macrolides ; administration & dosage ; pharmacology ; Microbial Sensitivity Tests ; Oligopeptides ; administration & dosage ; pharmacology ; Pseudomonas aeruginosa ; drug effects ; physiology ; ultrastructure ; Uridine ; administration & dosage ; analogs & derivatives ; pharmacology ; Vero Cells

AIMTo study the chemical constituents from Corallodiscus flabellata.

METHODSFresh plant of Corallodiscus flabellata was extracted twice with boiling water, filtered to remove insoluble materials, concentrated under reduced pressure at temperature 55 degrees C to a small volume. The concentrated liquor was subjected to solvent-solvent partitioning using ether, ethyl acetate, and n-butanol (saturated with water). The fraction of ethyl acetate extract was chromatographed over macroporous adsorption resin (Diaion HP-20) eluted with a mixture of H2O and MeOH in increasing MeOH content. Their fractions from resin were repeatedly chromatographed over Sephadex LH-20, Toyopearl HW-40, gel MCI, Gel CHP-20 and silica gel column. Structures of compounds obtained were identified on the basis of their spectral data, hydrolysis and chemical correlation.

RESULTSTwo phenylethanoid glycosides (I, II) and three phenolic acids were obtained from the EtOAc fraction of water-extracts. Their structures were identified as 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl (1-->2)]-beta-D-glucopyranoside (I), 3,4-dihydroxyphenylethanol-8-O-[(5-O- Vanilloyl)-beta-D-apiofuranosyl(1-->2)]-beta-D-glucopyranoside (II), vanillic acid (III), syringic acid (IV) and ferulic acid (V).

CONCLUSIONI and II are new compounds. Compounds III, IV and V were isolated from this plant for the first time.

Disaccharides ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; Magnoliopsida ; chemistry ; Molecular Structure ; Plants, Medicinal ; chemistry ; Vanillic Acid ; chemistry ; isolation & purification

OBJECTIVETo study the chemical constituents from Corallodiscus flabellata.

METHODThe compounds were isolated with macroporous adsorption resin, silica gel column chromatography and identified on the basis of their physiochemical and spectral data.

RESULTSix compounds were obtained and identified as vanillic acid, 3,4-dihydroxyphenylethyl-8-O-beta-D-glucopyranoside, syringic acid, caffeic acid, isoacteoside, ferulic acid.

CONCLUSIONAll the compounds were isolated from this plant for the first time.

Gallic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Magnoliopsida ; chemistry ; Phenols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Vanillic Acid ; chemistry ; isolation & purification

AIMTo study the chemical constituents of the pine needles of Pinus massoniana lamb..

METHODSVarious chromatographic techniques were used to separate and purify. Their physico-chemical properties and spectral data (UV, IR, MS, 1H-1 H COSY, HMQC, DEPT, HMBC and ORD ect.) were measured for structure elucication.

RESULTSThree compounds were isolated from the n-BuOH fraction of water-extracts. Their structures were identified as massonianoside A (4), massonianoside A: (7S, 8R)-3, 4, 9'-trihydroxyl-3-methyoxyl-7, 8-dihydrobenzofunan-1'-propanolneoligan-9-O-alpha-L-rhamnopyranoside, massonianoside C (5), (7S, 8R)-9,9'-dihydroxyl-3,3'-dimethyoxyl-7,8-dihydrobenzofunan-1'- propanolneoligan-4-O-alpha-L-rhamnopyranoside and cedrusin-4-O-beta-glucoside (6), (7S, 8R)-3',9,9'-trihydroxyl-3-methoxyl-7,8-dihydrobenzofunan-1'- propanolneoligan-4-O-beta-D-glucopyranoside.

CONCLUSIONCompound 4 and 5 are new compounds.

Benzofurans ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Pinus ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry

AIMTo study the chemical constituents from Corallodiscus flabellata.

METHODSThe compounds were isolated and purified by macroporous adsorption resin, silica gel column chromatography and identified on the basis of their physiochemical and spectral data.

RESULTSThree phenylethanoid glycosides (I-III) were obtained from the n-BuOH fraction of water-extracts. Their structures were elucidated as 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl (1-->3)]-beta-D-glucopyranoside (I), 3,4-dihydroxyphenylethanol-8-O-[4-O-trans-caffeoyl-beta-D-apiofuranosyl (1-->3)-beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (II) and 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl(1-->3)-beta- D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (III).

CONCLUSIONCompounds I, II and III are new compounds.

Caffeic Acids ; chemistry ; isolation & purification ; Disaccharides ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Magnoliopsida ; chemistry ; Molecular Structure ; Plants, Medicinal ; chemistry

OBJECTIVETo summarize and analyze the clinical characteristics and diagnostic and therapeutic measures for the first human case of H5N1 avian influenza pneumonia in mainland of China.

METHODSThe clinical data of the first case of H5N1 avian influenza virus infection in China were analyzed and summarized.

RESULTSThe case is a 9-year old boy, who developed acute symptoms of a light common respiratory infection, including fever and dry cough without obvious catarrh. On the 7th day after onset, his temperature reached 40 degrees C, tachypnea occurred, distinct rales could be heard and large areas of consolidation were seen in the lungs on chest X-ray. The patient's peripheral blood leukocyte count was 2.81 x 10(9)/L and neutrophils dominated. After comprehensive therapeutic approaches, including antiviral therapy (amantadine) and use of low-dosage glucocorticoid, the patient's temperature returned to normal on the 3rd hospitalization day, chest X-ray showed absorbed inflammatory change on the 5th day after admission, and leukocyte count became normal on the 6th day. No complication occurred during the whole course. The case was diagnosed by the 4 fold raised antibody to the H5N1 influenza virus in recovery stage serum because the H5N1 nucleic acid test in early stage was negative. The case was cured and discharged after 3 weeks comprehensive treatment.

CONCLUSIONSIt is very important for clinicians to pay enough attention to epidemiological history, especially history of exposure to avian influenza virus contaminated material, which will be very helpful for early detection, early diagnosis of the disease, and also very important for effective treatment and better prognosis.

Amantadine ; therapeutic use ; Animals ; Antibodies, Viral ; blood ; immunology ; Antiviral Agents ; therapeutic use ; Birds ; Child ; China ; Glucocorticoids ; therapeutic use ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; isolation & purification ; Influenza in Birds ; transmission ; Influenza, Human ; complications ; diagnosis ; Male ; Pneumonia ; diagnosis ; drug therapy ; physiopathology ; virology ; Treatment Outcome

Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.

OBJECTIVETo investigate the relationship between the mRNA expression levels of p53-mediating DNA damage and repair genes in the peripheral blood lymphocytes of workers and their exposures to benzene in their working environment.

METHODSThe mRNA expression levels of p53 and related genes were determined by SYBR Green I chimeric fluorescence quantitative real-time RT-PCR analysis in peripheral blood lymphocytes of 72 workers, who were classified into group A (46 direct exposure to benzene) and group B (26 indirect exposure to benzene) based on their positions, and 29 controls. The differences of gene expression levels were analyzed by software REST 2005. Meanwhile, the peripheral blood leukocytes, hemoglobin and platelet of workers and controls were counted. Benzene content was measured in the samples of toluene, used as raw material, and spraying agents and benzene, toluene and xylene concentrations in the air of workplaces were monitored.

RESULTSThere were no significant differences in the mRNA expression levels of p53, Ku80, Ape1 and Mdm-2 between group A or group B and control group (P > 0.05). The expression up-regulation of p21 mRNA was found, but without significant difference (P > 0.05). However, the mRNA expression levels of Rad51, Bcl-2, Bax, Xpa and Xpc in group A and Rad51 in group B were downregulated significantly (P < 0.05 or P < 0.01). Moreover, both the counts of white blood cell, hemoglobin and platelet in group A were (4.93 +/- 1.27) x 10(9)/L, (123.97 +/- 11.80) g/L and (124.02 +/- 41.22) x 10(9)/L respectively and platelet in group B (135.80 +/- 39.44) x 10(9)/L were significantly lower than in control group (P < 0.05 or P < 0.01).

CONCLUSIONThe mRNA expression levels of some p53-mediating DNA damage and repair genes are downregulated in the workers chronically exposed to low benzene concentration. The working environment impacts on health of group A workers are greater than the ones of group B.

Adult ; Benzene ; adverse effects ; DNA Damage ; DNA Repair ; Female ; Humans ; Lymphocytes ; drug effects ; metabolism ; Male ; Occupational Exposure ; adverse effects ; RNA, Messenger ; genetics ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult

Because advantage of tissue origin and proliferation potential, the umbilical cord-derived mesenchymal stem cells (UC-MSC) and placental chorionic villous-derived mesenchymal stem cells (CV-MSC) have clinical application potential, as compared with bone marrow MSC. But whether the differences of biological characteristics exist between UC-MSC and CV-MSC, which deserve to be further explored. This study was purposed to compare the biological characteristics of UC-MSC and CV-MSC. The placental and umbilical cord were cleaned by using the sterile physiological salt, the UC-MSC and CV-MSC were separated by enzyme digestion. Short tandem repeat (STR) analysis was used to detect whether the MSC obtained from fetal tissue. MTT method was used to detect proliferation of MSC. Flow cytometry was applied to analyze cell phenotype. The different differential medium was used to detect their multi-directional differentiation capacity. After the MSC and PHA-stimulated peripheral blood mononuclear cells were co-cultured, the γ-interferon (IFN-γ) levels of the co-culture supernatant were detected using the ELISA. The results showed that these MSC were derived from fetal tissue by STR analysis. They were adherent cells with typical fibroblast morphology. Cells expressed the MSC surface markers CD90, CD73 and CD105 and CD44, not expressed CD45 and of CD11b and CD34.These cells could differentiate into osteoblasts and adipoblasts under culture with different conditioned medium, but in the adipogenic differentiation of CV-MSC, the larger lipid droplets appeared. It is concluded that these cells are obtained MSC. These MSC can inhibit peripheral blood mononuclear cells stimulated by PHA to secrete IFN-γ, and the the CV-MSC have a stronger suppression capacity, which makes the CV-MSC to have a greater advantage in the treatment of autoimmune diseases.
Cell Differentiation ; Cells, Cultured ; Female ; Flow Cytometry ; Humans ; Mesenchymal Stromal Cells ; cytology ; Placenta ; cytology ; Pregnancy ; Umbilical Cord ; cytology

This study was aimed to investigate the effect of IL-1β on hematopoietic support of human umbilical cord mesenchymal stem cells (hUC-MSC). 2×10(6) hUC-MSC were seeded in 75 cm(2) flasks, after adherence to wall for 2 h, 10 ng/ml IL-1β was added in hUC-MSC supernatant and cultured for 36 h, then the culture supernatants and cells were harvested. The effect of conditioned medium with/without IL-1β on CD34(+) cell hematopoietic support was observed, mRNA expression changes of hUC-MSC cultured in medium with/without IL-1β were monitored by real time PCR, the differences in hematopoiesis-related factors were detected by ELISA. The results showed that the conditioned culture medium of hUC-MSC with IL-1β enhanced the ability to form colony of CD34(+) cells, especially CFU-G and CFU-GM in vitro; IL-1β promoted the mRNA expression of GM-CSF, G-CSF, IL-6 on MSC; IL-1β also promoted the secretion of GM-CSF, G-CSF, and IL-6 protein from hUC-MSC. It is concluded that IL-1β enhances hematopoietic support capacity especially, capability of MSC to myeloid differentiation.
Cell Differentiation ; Cells, Cultured ; Culture Media ; Granulocyte Colony-Stimulating Factor ; secretion ; Granulocyte-Macrophage Colony-Stimulating Factor ; secretion ; Hematopoietic System ; drug effects ; Humans ; Interleukin-1beta ; pharmacology ; Interleukin-6 ; secretion ; Mesenchymal Stromal Cells ; cytology ; drug effects ; secretion ; Umbilical Cord ; cytology