In order to identify the chemical constituents of Yushu tablets comprehensively, we studied the chemical constituents of CHCl3 extract from Yushu tablets by the ultra performance liquid chromatography-electrospray ionization-ion trap-time of flight mass spectrometry (UPLC-ESI-IT-TOF/MS). It showed that there were more than 100 compounds separated, and forty-nine peaks among these were identified on the basis of high resolution mass spectrometry data and literature data reported. Determination of twelve peaks was further confirmed by standard substances. These components assigned to the different plant sources mainly included phenylpropanoids, triterpenoids, quinones and m-trihydroxybenzene compounds. By analyzing the chemical components of CHCl3 extract from compound Chinese medicine Yushu tablets comprehensively, this study provided the foundation for studying chemical components, pharmacodynamic substance and quality control of Yushu tablets.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Drugs, Chinese Herbal ; analysis ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; Tandem Mass Spectrometry

This article describes the preparation of salmon calcitonin ultra-flexible liposomes and their hypocalcemia effect after intranasal administration in rats. Both the conventional liposomes and ultra-flexible liposomes were prepared by rotary evaporation-sonication and extrusion. The morphology of ultra-flexible liposomes was observed with transmission electronic microscope. The size and size distribution and their zeta potential were determined by dynamic light scattering. The mean size of ultra-flexible liposomes with DC-Chol was no more than 120 nm, while the mean size of the conventional liposomes was 256.5 nm. The results showed the content of sodium deoxycholate have significant effect on the mean particle size of liposomes. The ultra-flexible liposomes were intranasal administrated at the dose of 5.0 microg x kg(-1); the concentration of serum calcium was determined by OCPC method. The results showed that the salmon calcitonin solution only slightly lowered serum calcium levels and the conventional liposomes could improve the effect of decreased serum calcium level (D%), and the ultra-flexible liposomes had the best effect on the decreased serum calcium level, and the hypocalcemia effect was correlated with the content of sodium deoxycholate which was present in the liposomes. Moreover the ciliotoxicity of ultra-flexible nanoliposomes on nasal mucocilia was investigated with the electron microscope scanning. The results showed that the ultra-flexible liposomes markedly reduced the ciliotoxicity of sodium deoxycholate on nasal mucous. Thereby the ultra-flexible liposomes significantly enhanced the hypocalcemia effect of serum calcium after intranasal administration in rats. The ultra-flexible liposomes could be an effective carrier for intranasal delivery of the peptide and protein drugs.
Administration, Intranasal ; Animals ; Calcitonin ; administration & dosage ; pharmacology ; Calcium ; blood ; Liposomes ; Male ; Particle Size ; Rats ; Rats, Sprague-Dawley

Animals ; Disease Models, Animal ; Liver Cirrhosis, Biliary ; Mice

OBJECTIVETo provide the foundation for reasonable utilization by analyzing the essential oils of Ligularia virgaurea.

METHODThe essential oils were extracted by using steam distillation and separated with GC capillary columns. The components were quantitatively determined with normalization method, and were identified with GC-MS.

RESULT41 components were identified, which took up 72.73% of the essential oils.

CONCLUSIONThe main components of essential oils were 4-methyl-1-(1-methylethyl)-3-cyclohexen-1-ol(14.369%), crotonic acid, 2,2-dimethyl-butanoic acid, 1-methyl-3-(1-methylethyl)-benzene, (1s-endo)-1,7,7-trimethyl-bicyclo[2,2,1]heptan-2-ol, trans-1-methyl-4-(1-methylethyl)-2-cyclohexen-1-ol, alpha-cadinol and alpha,alpha,4-trimethyl-3-cyclohexene-1-methanol.

Asteraceae ; chemistry ; Butyrates ; analysis ; Crotonates ; analysis ; Gas Chromatography-Mass Spectrometry ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Terpenes ; analysis

The aim of this study was to obtain the capsid protein VP2 of human bocavirus (HBoV) identified in Beijing recently and construct virus-like particles (VLPs) in insect cells for further study of this virus. The full-length VP2 gene of HBoV from BJ3722 was inserted into the baculovirus expression transfer vector (pFastBac 1) to obtain the recombinant Bacmid, and generation of recombinant baculoviruses was followed by transfection of the recombinant Bacmid into insect cells. Then the recombinant VP2 protein was recognized by SDS-PAGE using Coomassie-blue staining and Western blot using hyper-immune serum against VP2 of HBoV from rabbit. The recombinant baculoviruses were harvested and amplified to gain large amounts of viruses with high titers to infect insect cells at a multiplicity of infection (MOI) of 0. 5. After 7-10 days or 4-5 days of the infection, the supernatants of culture or the cell lysates treated with lysing solution were harvested, and ultracentrifuged twice through 40% sucrose cushion to obtain purified VLPs, which were followed by Western blot and IFA for VLPs' composition and specificity analysis, by electron microscopy for VLPs' morphologic structure. The recombinant VP2 protein with molecular weight of approximately 61 kD expressed in recombinant baculoviruses was recognized by SDS-PAGE using Coomassie-blue staining and Western blot. The presence of VP2 on VLPs was demonstrated by Western blot and IFA from samples collected during the purification of VLPs from the supernatants of culture or the cell lysates, and the expression of VP2 in insect cells led to the formation of VLPs which formed the typical icosahedral appearance of parvoviruses with a diameter of approximately 20 nm. In conclusion, the recombinant baculoviruses were constructed, the HBoV VP2 protein was expressed in insect cells with high specific antigenicity and VLPs was formed successfully.
Animals ; Blotting, Western ; Capsid Proteins ; genetics ; metabolism ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Fluorescent Antibody Technique, Indirect ; Human bocavirus ; genetics ; metabolism ; Microscopy, Electron, Transmission ; Polymerase Chain Reaction ; Spodoptera ; Virion ; genetics ; metabolism ; ultrastructure

OBJECTIVETo find out the importance of human bocavirus (HBoV) as an infectious agent for population in Beijing, China, seroprevalence study was conducted by using expressed recombinant major capsid VP2 protein as an antigen.

METHODSSerum specimens collected from infants and children who visited the Children's Hospital Affiliated to the Capital Institute of Pediatrics for health check up and adults visited the Xuanwu Hospital, Beijing for diseases other than respiratory infections from April 1996 to March 1997 were used for investigation. The major capsid protein VP2 from HBoV was expressed in E. coli strain BL21 (DE3) with the transformed PET30b vector inserted with full-length VP2 gene of HBoV and the specific antigenicity of this expressed protein was validated by previous study. Western blot was used to detect specific IgG antibody against HBoV in collected serum specimens diluted to 1:200. Mock expressed protein was E. coli cells strain BL21 (DE3) with the transformed PET30b vector without insert. Anti-His monoclonal antibody and rabbit anti-HBoV VP2 polypeptides hyper-immune serum were used as positive control for antibody detection.

RESULTSOut of 677 serum specimens tested, 400 (59.1%) were positive by Western blot. About 45.3% (34/75) of the newborns under 1 month of age had anti-HBoV antibodies, and antibody positive rates were lower in the age groups of 1 and 2 months (41.4% and 31.3%, respectively) and were higher in the following ages from 6 months to 7 years (from 45.6% to 69.7%). The antibody positive rates were at a relatively constant level (about 70%) in the age groups from 7 years to 40 years and became lower (61.8% - 62.8%) in groups of age over 50 years.

CONCLUSIONThe high seroprevalence against recombinant HBoV VP2 protein and early age antibody acquisition indicate that HBoV has been circulating in Beijing, China as early as in 1996 and most of children had been exposed to HBoV by the age of 7 years. Infants under the age of 6 months were susceptible to infection with this virus.

Adolescent ; Adult ; Antibodies, Viral ; blood ; Blotting, Western ; Bocavirus ; immunology ; Capsid Proteins ; immunology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Immunoglobulin G ; blood ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Parvoviridae Infections ; epidemiology ; immunology ; Prevalence ; Seroepidemiologic Studies ; Young Adult

OBJECTIVETo study the chemical constituents from Lamium maculatum var. kansuense.

METHODThe chemical constituents were isolated and repeatedly purified on silica gel column and the structures were elucidated by the NMR spectra and physico-chemical properties.

RESULTSix compounds were obtained and identified as polypodine B (I), 5-OH-8-epiloganin (II), shlanzhiside methyl ester (III), liriodendrin (IV), quercitroside (V), uridine (VI).

CONCLUSIONCompound IV was found from genus Lamium for the first time and the rest of the compounds were found from Lamium maculatum var kansuense for the first time.

Ecdysterone ; analogs & derivatives ; chemistry ; isolation & purification ; Furans ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Plants, Medicinal ; chemistry ; Uridine ; chemistry ; isolation & purification

OBJECTIVETo obtain isolated human metapneumovirus (HMPV) strains from clinical specimens collected from infants and children in Beijing and to promote the investigation on this important respiratory pathogen.

METHODClinical specimens including throat swabs from outpatients and nasopharyngeal aspirates from hospitalized children were collected from infants and children visited the affiliated children's hospital for acute respiratory infections during May 2008 to April 2009. HMPV positive specimens identified by RT-PCR and/or direct immunofluorescent assay with monoclonal antibody against HMPV were inoculated to LLC-MK(2) cells and incubated at 37°C and 33°C, respectively. The replication of the virus in the cells was detected by direct immunofluorescent assay followed by RT-PCR. The genotypes of the isolated virus strains were identified by RT-PCR.

RESULTOut of 1092 clinical specimens, 81 were HMPV positive by RT-PCR, the positive rate was 7.4% (81/1092). Among these positive specimens, 33 were inoculated to LLC-MK(2) cells and the replication of HMPV was revealed by antigen detection and RT-PCR from 5 out of these 33 inoculates. These isolated viruses could be passed in LLC-MK(2) cells and were not cross-reacted with other common respiratory viruses, such as ADV, RSV and Parainfluenza viruses 1/2/3 by monoclonal antibodies against these viruses in direct immunofluorescent assay. The HMPV was more likely to be isolated from fresh specimens within 24 hours after the collection of specimens which were not frozen. Four of the 5 isolated strains were identified as genotype A and 1 as genotype B. Unlike other respiratory viruses, these isolated HMPV did not show specific CPE in cell culture and the replication of the virus was identified by antigen detection and RT-PCR.

CONCLUSIONHMPV of both genotypes were isolated from infants and children with acute respiratory infections in Beijing which will accelerate the investigation of this important virus.

Acute Disease ; Child ; China ; Genes, Viral ; genetics ; Genotype ; Humans ; Infant ; Metapneumovirus ; isolation & purification ; Respiratory Tract Infections ; virology

OBJECTIVETo study the chemical constituents from Lamium maculatum L. var Kansuense.

METHODThe chemical constituents were isolated and repeatedly purified by silica gel column chromatography and the structures were elucidated by the NMR spectra and physico-chemical properties.

RESULTTen compounds were obtained and they were identified as D-mannitol, beta-sitosterol, stigmasterol, rutin, 3'-methylquercetin-3-O-rutinoside, n-butyl-beta-D-fructopyranoside, daucosterol, acteoside, 20-hydroxyecdysone, allantoin.

CONCLUSIONAll the compounds were obtained from L. maculatum L. var Kansuense for the first time.

Allantoin ; chemistry ; isolation & purification ; Ecdysterone ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Mannitol ; chemistry ; isolation & purification ; Phenols ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rutin ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification ; Stigmasterol ; chemistry ; isolation & purification

BACKGROUNDHuman bocavirus (HBoV) is a newly identified human parvovirus that was originally detected in the respiratory secretions of children with respiratory infections. This study aimed to learn about the importance of HBoV infections by revealing the prevalence of serum antibodies against HBoV in Beijing population.

METHODSTwo batches of serum specimens collected in different periods were tested by Western blotting for specific IgG against HBoV using recombinant VP2 as antigen.

RESULTSOut of 677 serum specimens collected during April 1996 to March 1997, 400 (59.1%) were positive and antibody positive rate for another batch of 141 serum specimens collected in August, 2005 from adults aged from 20 years to over 60 years was 78.7% (111/141). Comparison of the sero-prevalence profiles for serum specimens collected during 1996 - 1997 to those collected in 2005 indicated that the antibody positive rate for specimens collected in 2005 was higher than that of the corresponding age groups collected during 1996 - 1997.

CONCLUSIONSThe data suggest that HBoV has been circulating in Beijing population for at least over 10 years, and most of children had been exposed to HBoV by age of 7 years. Higher HBoV antibody positive rate shown in the serum specimens collected in 2005 suggested that infections by HBoV have been increased in Beijing population in recent years.

Adult ; Antibodies, Viral ; blood ; Blotting, Western ; Bocavirus ; pathogenicity ; China ; epidemiology ; Humans ; Immunoglobulin G ; immunology ; Middle Aged ; Parvoviridae Infections ; blood ; epidemiology ; immunology ; Seroepidemiologic Studies ; Viral Proteins ; immunology ; Young Adult