The changes of bioactive constituents were analyzed for Puhuang-Wulingzhi before and after compatibility and the antiplatelet and antithrombin activitives were evaluated in order to elucidate the scientific and reasonable of Puhuang-Wulingzhi compatibility. UPLC-QTOF-MA-Markerlynx, principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis were used for data analysis and tracking changes of chemical composition during the decocting process. In vitro platelet aggregation induced by ADP, thrombin time(TT) and prothrombin time (PT) were investigated for Puhuang-Wulingzhi before and after compatibility. The results showed that significant differences were found between the mixed decoction and codecoction of Wulingzhi and Puhuang. Five compounds changed obviously were identified as typhaneoside, naringenin, isorhamnetin-3-O-ruinoside, quercetin-3-O-neohesperidoside, kaempferol-3-O-neohesperidoside. The codecoction, comparing with the single decoction, was more significant in antiplatelet aggregation and could prolong thrombin time. In the same crude drug dose, the thrombin time (TT) elongation were greater. These data could provide references for elucidation of bioactive components for this herb pair.
Animals ; Antithrombins ; chemistry ; pharmacology ; Blood Platelets ; drug effects ; physiology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Humans ; Molecular Structure ; Platelet Aggregation ; drug effects ; Rabbits ; Thrombin Time

Objectives To study the level and development of serum specific antibody against severe acute respiratory syndrome coronavirus(SARS-CoV)of different populations in SARS pestilence district after SARS outbreak in a general hospital.Discuss SARS sub-clinical infection and protective action of the IgG antibody.Methods Seroepidemiology method,enzyme-linked immunosorbent assay (ELISA)and indirect immunfluorescence assay(IFA)were employed to investigate the changing level of serum antibody to SARS-associated coronavirus in non-SARS population in SARS pestilence district during and after SARS outbreak.The development of IgM and IgG antibody in patients with SARS in 6 weeks after the onset of SARS was studied qualitatively.The level changing of IgG antibody in con- valescent patients with SARS in 82 weeks after the onset was observed dynamically.Results The ELISA test outcome of IgG antibody was negative in 200 non-SARS people who were random samples of normal mass in SARS pestilence district and common community.The positive rate was 0.41% in 487 SARS high risk population tested by ELISA,but showed negative when retested by IFA.The A value level of IgG antibody existed significant difference in non SARS mass during and after SARS outbreak and the later's was higher them the former's(P

In order to evaluate the effect and mechanism of the mulberry leaf alkaloid, flavones, and polysaccharide intervention on diabetes, the overall metabolite profiling characteristics for the plasma of diabetic mouse was performed by using an ultra-performance liquid chromatography/electrospray-tandem mass spectrometry (UPLC-ESI-MS). The 8 potential biomarkers were found in diabetic mice plasma based on the data of MS/MS characteristics obtained from the UPLC-OrbitrapMS analysis, which mainly involved in sphingolipids, amino acid metabolic pathway. The principal component analysis showed that the normal group and model group were obviously distinguished and implied that metabolic disturbance was happened in diabetic mice plasma. The extracts of mulberry leaf flavonoids, polysaccharide, alkaloid had exhibited the effects of callback function for diabetic mice through regulating the amino acid metabolism and sphingolipid metabolism.
Alkaloids ; chemistry ; Amino Acids ; metabolism ; Animals ; Biomarkers ; blood ; Chromatography, High Pressure Liquid ; Diabetes Mellitus, Experimental ; drug therapy ; Flavones ; chemistry ; Flavonoids ; chemistry ; Metabolic Networks and Pathways ; Metabolomics ; Mice ; Morus ; chemistry ; Plant Leaves ; chemistry ; Sphingolipids ; metabolism ; Tandem Mass Spectrometry

In order to establish HPLC fingerprint of Liuwei Dihuang soft capsule, and to provide certain reference for an quality control of it, the HPLC method was performed on an Agilent C18 (4.6 mm x 250 mm, 5 μm) column with acetonitrile-0.02% trifluoroacetic acid as mobile phase, gradient elution volume flow of 1.0 mL x min(-1), column temperature was 30 degrees C, detection wavelength: 0-60 min, 238 nm, 60-70 min, 210 nm. The software for chromatographic fingerprint was applied to analysis different batches of Liuwei Dihuang soft capsule samples. Sixteen mutual peaks were selected as the fingerprint peaks in 12 samples with loganin as the reference peak, and all of the detected peaks were separated effectively. Cluster analysis (HCA) and similarity analysis (SA) were done based on data of 12 samples clustering analysis of 12 batches of samples were divided into 2 categories. Including 7 for the first class, the rest was second, similarities calculated by SA were all above 0.92, indicating a good similarity between the reference and twelve batches of samples, also, the analysis results of HCA and SA basically the same. This method is simple with good precision, repeatability and stability, and provides the basis for Liuwei Dihuang soft capsule quality control.
Capsules ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Quality Control

OBJECTIVERotavirus is the single most common cause of severe dehydrating diarrhea in young children worldwide, but the pathogenesis and immunity against this disease are not completely understood. A prospective study was conducted to assess gene expression of toll-like receptors (TLR) in children with acute rotavirus diarrhea.

METHODSSeventy-five children with acute rotavirus diarrhea and 38 control children were enrolled in this study from Sep. 2004 to Jan. 2005. All the 75 patients had detailed records of clinical characteristics. Rotavirus antigen was detected by ELISA from stools. Peripheral blood mononuclear cells (PBMC) were separated by Ficoll reagent and RNA was extracted by Trizol. The levels of mRNA for five TLRs in PBMC were examined by fluorescent quantitative RT-PCR.

RESULTSPatients with acute rotavirus infection had elevated mean levels of TLR 2, 3, 4, 7, 8 mRNA expressions in PBMC within 3 days since onset of the disease, P less than 0.05. But only TLR 2, 3, 8 mRNA levels remained increased in patients within 7 or 14 days since onset (P less than 0.05). Mean levels of mRNA for TLR 4 in PBMC was higher in patients with more severe diarrhea including longer duration of diarrhea, more episodes of diarrhea per day and higher severity scores (P less than 0.05).

CONCLUSIONManifold TLR may play roles in the start-up and regulation of immune responses in children with acute rotavirus diarrhea. These findings will be helpful to further recognize immune response in Chinese children with rotavirus diarrhea and, consequently, may provide directions and insights that could prove critical to the prevention or treatment of this important disease.

Acute Disease ; Antigens, Viral ; analysis ; Child, Preschool ; Diarrhea ; genetics ; virology ; Enzyme-Linked Immunosorbent Assay ; Feces ; virology ; Gene Expression ; Host-Pathogen Interactions ; Humans ; Infant ; Leukocytes, Mononuclear ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; immunology ; physiology ; Rotavirus Infections ; genetics ; virology ; Toll-Like Receptors ; genetics

To investigate the optimum harvesting time and utilization of mulberry leaves during different growth periods based on the content of alkaloids and flavonoids, 88 samples of 11 species of mulberry leaves were collected and analyzed. UPLC-TQ/MS method was applied and the results showed that the ingredients of alkaloids and flavonoids in mulberry leaves are quite different in different growth periods and different species. There was a sharp decline of the average content of alkaloids in all samples from October, while the content of flavonoids dropped either from October but with less volatile. The content of flavonoids in M. atropurpurea was much higher than alkaloids, while M. australis was opposite completely. There was a sharp decline of alkaloids in M. cathayana and M. mongolica from Tuly to August, however, the content of alkaloids and flavonoids in M. alba is neither too high nor too low. In summary, it is more suitable to harvest tender mulberry leaves harvested from the end of September to beginning of October that provide a scientific evidence for rational harvest and comprehensive utilization of mulberry leaves.
Alkaloids ; analysis ; isolation & purification ; Chromatography, High Pressure Liquid ; Drug Stability ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Flavonoids ; analysis ; isolation & purification ; Mass Spectrometry ; Medicine, Chinese Traditional ; Morus ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; Reproducibility of Results ; Seasons

AIMTo establish a simple and effective method for RBCs labeling and survival assays, and the qualities of rabbit RBCs preserved in GMA solution at 4 degrees C were verified.

METHODSThe bloods were taken through the ear arteries of the rabbits. The RBCs were labeled by fluorescein isothiocyanate (FITC), and were reinjected to the same rabbit through ear veins. The percentage of FITC labeled RBCs was assayed by FACS at a series of times after injection. The SAS software was employed to analyze the data and establish the regression equations. The 24-hour recovery and the half-life span of the labeled RBCs were calculated according to the equations.

RESULTSThe 24-hour recovery and the half-life span of the labeled RBCs in the control group were 93.76% +/- 5.40% and 22.50% +/- 4.37 days respectively, which was in agreement with the previous papers. The 24-hour recovery and the half-life span of the labeled RBCs in the GMA group were 89.13% +/- 7.10% and 11.41% +/- 1.63 days respectively, which was coincident with the infusion conditions.

CONCLUSIONCompared with other methods of RBCs labeling in vivo, FITC labeling was thought to be easier and cheaper to use, which could facilitate the analysis of the biological character of the labeled cells, and could be used to trace the fate of labeled cells.

Animals ; Blood Preservation ; methods ; Erythrocyte Aging ; physiology ; Erythrocyte Count ; Erythrocytes ; physiology ; Fluorescein-5-isothiocyanate ; Rabbits ; Software

The present study is to determine the flavonoid glycosides, terpene lactones, biflavones, gingko acid and procyanidins of ginkgo pollen. UPLC-TQ-MS technology was used for the determination of 24 kinds of resource chemical composition in ginkgo pollen qualitatively and quantitatively. The results shows that the contents of rutin, quercetion 3-O-[4-O-(α-L-rhamnosyl )-β-D-glucoside] and kaempferolis were 120.9, 114.0, 222.1 μg x g(-1). In this paper, the contents of 24 kinds of chemical components of ginkgo pollen were determinated by UPLC-TQ-MS for the first time. This method is simple and quick, which will be benefit for recycling utilization of ginkgo pollen.
Chromatography, High Pressure Liquid ; methods ; Flavonoids ; analysis ; Ginkgo biloba ; chemistry ; Mass Spectrometry ; Pollen ; chemistry ; Proanthocyanidins ; analysis ; Rutin ; analysis ; Terpenes ; analysis

OBJECTIVETo investigate the influence of three pontic types on alveolar ridge mucosal microecosystem.

METHODSSixty patients ready to accept three unit metal ceramic bridges were selected. The bacterial type and the cultivable flora were counted and the proportions of bacteria detected on the top of alveolar ridge mucosal contact area before tooth preparation and three months after bridge insertion.

RESULTSType and CFU of bacteria on the alveolar ridge mucosa under modified base-type pontics and modified ridge lap pontics increased significantly (P < 0.05); while there was no significant change under the ovata pontics (P > 0.05). Before tooth preparation and 3 months after fixed prosthesis insertion, the percentages of oral Streptococci and Neisseriae changed significantly (P < 0.05).

CONCLUSIONSThe ovata pontic had less influence on mucosal microecosystem than the other two pontics and is the appropriate pontic design for clinical dentist.

Adult ; Alveolar Process ; microbiology ; Denture Design ; Denture, Partial, Fixed ; microbiology ; Humans ; Middle Aged ; Mouth Mucosa ; microbiology

To study the dynamic changes of ultrastructure of erythrocytes in prolonged preservation of blood with preservative fluid containing superoxide dismutase (SOD), the whole blood samples were preserved at 4 degrees C in SOD-containing solution, the morphologic changes of erythrocyte were dynamically ob served by transmission microscopy after preservation for 42, 75 and 85 days, an d the blood samples preserved in GMA solution served as control. Three variance was applied to analyze the data with SAS software. The results showed that the metamorphotic rates of erythrocyte preserved in SOD-containing solution for 42, 75 and 85 days were lower than those of erythrocytes preserved in GMA solution. Most of metamorphotic rates of erythrocyte preserved in SOD-containing solution for 42, 75 and 85 days were correspond to those of erythrocytes preserved in GMA solution for 42 days, or even lower. It is concluded that SOD-containing preservative fluid might help to maintain the normal morphology of erythrocytes in prolonged preservation at 4 degrees C.
Blood Preservation ; Erythrocyte Deformability ; Erythrocytes ; ultrastructure ; Humans ; Microscopy, Electron ; Superoxide Dismutase ; pharmacology ; Time Factors