Seven compounds were isolated from Onychium japonicum by macroporous resin, silica gel, ODS, Sephadex LH-20 column chromatography and semi-preparative HPLC. Their structures were identified by NMR, MS and other spectroscopic methods as onychone A (1), quercetin (2), quercetin-3-O-α-L-rhamnoside (3), kaempferol-7-O-β-D-glucopyranoside (4), kaempferol-3-O-α-L-rhamnopyranoside (5), (-)-prunin (6), and norathyriol (7). Compound 1 is a novel macrocyclic flavonoid, and all the others are reported from this plant for the first time. In vitro cytotoxic activities of compounds 1-7 were evaluated by MTS testing with five cancer cell lines. Compound 7 exhibited weak cytotoxicity against tumor cell lines A549, SMMC-7721, and SW480.

Objective To investigate the clinical characteristics of incomplete Kawasaki disease(KD).Methods Clinical data includi-ng test results,therapeutic methods were analyzed retrospectively in 579 patients with Kawasaki disease.They were divided into classic KD and incomplete KD and made a compared analysis.Results There were no significant differences in gender,age,symptom and laboratory examination between classic and incomplete KD.But the rate of coronary artery lesions was higher in incomplete KD(18.4%) than that of classic KD(11%).Conclusion The rate of coronary artery lesions was higher in incomplete KD,and it should be paid more attention to earlier diagnosis and earlier treatment.

Objective To explore the causes of twisted or tailed DNA bands in agarose gels after electro-phoresis .Methods Prestained and poststained methods were employed to exam 3 kinds of DNA marker bands in agarose gel with GeneGreen and Ethidium Bromide ,respectively .Results Three kinds of DNA marker bands in agarose gel with 1:10000 and 1:5000 concentration of GeneGreen showed obvious distortions and trailing phenomena compared with the same concentration of Ethidium Bromide w hen the prestained method was used for electrophoresis ,which were improved when the poststained method was used in agarose gel with GeneGreen and Ethidium Bromide ,respectively .Conclusion Nucleic acid dye GeneGreen can affect the quali-ty of DNA bands in agarose gel electrophoresis .When the quality of DNA itself ,agarose gel quality ,electro-phoretic fluid quality and voltage are excluded ,the quality of nucleic acid dye should be taken into considera-tion if the bands twisting or tailing occurs when the DNA nucleic acid electrophoresis is carried out by pres-tained method .The quality of DNA electrophoresis bands can be improved by using poststained method or re-placing nucleic acid dye .

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 identify pathogenic mutations of ANTXR2 gene in a patient with juvenile hyaline fibromatosis.

METHODSGenomic DNA was extracted from peripheral venous blood sample from the patient. All coding exons (exons 1-17) and splicing sites of the ANTXR2 gene were amplified with PCR. Potential mutations were detected with direct sequencing of the PCR products. 100 unrelated healthy subjects were used as the controls. CLUSTALX (1.81) was employed to analyze cross-species conservation of the mutant amino acid. Impact of the mutations was analyzed with software including SIFT, PolyPhen-2 and MutationTaster.

RESULTSA compound heterozygous mutation c.1074delT/c.1153G>C, was identified, among which c.1153G>C has not been reported previously and was predicted to be probably damaging. Both mutations were not found among the 100 healthy controls.

CONCLUSIONThe patient's condition may be attributed to the compound heterozygous mutations of c.1074delT and c.1153G>C of the ANTXR2 gene. Above results has facilitated molecular diagnosis for this patient.

Child, Preschool ; Female ; Heterozygote ; Humans ; Hyalinosis, Systemic ; diagnosis ; genetics ; Mutation ; Receptors, Peptide ; genetics

Objective To investigate the prevalence of unintended pregnancy (UP) and exploring the risk factors of UP for married women of child-bearing age from Qingshan district,Wuhan.Methods A cross-sectional study was adopted in this study.Cluster sampling method was used with 3256 women recruited,in 2010.Information on history and risks related to social-demographic factors of UP were collected,using a self-administered questionnaire.Results Of the 3256 participants,over half of them (53.8％) reorted ever having had the history of UP and 9.1％ reported UP in the past year.Rate of UP in the past year for different age cohorts (18-30,31-40,41-49 years) were 31.8％,10.5％ and 1.8％ respectively.The most frequently reported reason for UP across all the age cohorts was "Did not use any contraceptive methods",with proportions on the reason that reported by women at 18-30,31-40 and 41-49 year-olds,were 69.7％,51.1％ and 42.4％ respectively.The second frequently reported reasons for UP were "Failure of traditional contraception" for younger cohort ( 18-30 years:13.0％ ) and "IUD dropped or pregnancy with IUD" for older-age cohorts (23.4％ at 31-40 year-olds and 37.0％ at the 41-49 year-oplds).The most frequently cited reason for "Did not use any contraceptive methods" was "Believe we were lucky so far,not to get pregnant" (59.6％).The risk factors of UP were being at older age,experiencing sex debut at younger age and got married at younger age.Conclusion The prevalence of lifetime UP history was high among women at child-bearing age from Qingshan district,Wuhan.Reproductive health services and interventions should be taken according to the needs from different age cohorts of women.Younger cohort of women should receive more attention.

BACKGROUNDHuman metapneumovirus (hMPV) was discovered by scientists in the Netherlands as a novel respiratory virus in 2001 and had been found in children with acute respiratory tract infections (ARTI) in China. The objective of this study was to determine the importance of hMPV infection in children in Beijing and the genotypes of the circulating virus by the surveillance during a four-consecutive-year period.

METHODSClinical specimens collected from children with ARTI from January 2006 to December 2009 were tested for hMPV by RT-PCR using primers targeting the matrix (M) gene, followed by genotyping of hMPV directly from positive samples by diplex PCR with primers for glycoprotein (G) genes. Sequence analysis was used for genotyping of those un-typable samples. Common respiratory viruses in these clinical specimens were tested by virus isolation and antigen detection, in addition to hMPV detection.

RESULTSOf 4730 tested specimens, 191 (4.0%) were positive for hMPV and 62.8% of 191 were identified as genotype A. The positive rate of hMPV from hospitalized patients was higher than that from outpatients each year. Most of hMPV positive children were under five years old. The peak of hMPV activity mostly occurred in late spring and overlapped with or followed that of respiratory syncytial virus (RSV) and followed by parainfluenza virus 3. Of hMPV infected cases, 68.6% were lower respiratory tract infection, among which 79.4% were hospitalized, and upper respiratory tract infection was diagnosed for 31.4% of hMPV infected children. The 9.4% of hMPV positive samples were found to co-exist with other respiratory viruses.

CONCLUSIONShMPV was an important pathogen for ARTI in pediatric patients, especially those under five years old. Both genotypes A and B circulated simultaneously in Beijing.

Adolescent ; China ; Female ; Genotype ; Humans ; Male ; Metapneumovirus ; genetics ; pathogenicity ; Respiratory Tract Infections ; virology ; Reverse Transcriptase Polymerase Chain Reaction

Objective To explore the effect of building continuous renal replacement therapy (CRRT) professional nursing team in emergency intensive care unit (EICU).Methods To establish CRRT professional nursing team in EICU,define the duties,responsibilities and job contents,and provide theory and manipulative skill training about CRRT care,make processes and guides,accumulate cases,strengthen CRRT nursing quality control.The scores of theory knowledge and manipulative skill in CRRT and the incidence rate of CRRT nursing associated complication were respectively compared before and after the establishment of the CRRT professional nursing team.Results The scores of theory knowledge and manipulative skill in CRRT were respectively (95.53 ± 1.78),(97.00 ± 1.20) after the establishment of CRRT professional nursing team,and higher than (85.95 ±-3.82),(89.68 ± 2.85) before the establishment,and the differences were statistically significant (t =14.55,14.57,respectively; P ＜ 0.01).The incidence rate of CRRT nursing associated complication was decreased from 5.26％ to 1.44％,and the difference was statistically significant (x2 =4.59,P ＜ 0.05).Conclusions The establishment of CRRT professional nursing team in EICU can improve nurses' theory and clinical skill,standardize nurses' professional nursing behaviors,mobilize nurses' enthusiasm for the job and initiative in learning,and improve nursing quality.

OBJECTIVETo identify variations in hemagglutinin genes from influenza viruses (H3N2) isolated from infants and young children with acute respiratory infection (ARI) between March, 2004 and April 2005.

METHODSRNAs from influenza A virus strains (subtype H3) isolated from specimens collected from ARI children were extracted followed by amplification for HA1 fragments from hemagglutinin (HA) genes by RT-PCR. The sequences of the fragments were defined by direct sequencing for the PCR products or the target inserts after the PCR fragments were cloned into the TA-cloning vector pBS-T and analyzed by bioinformatic software.

RESULTSFragments of 987 bps of HA1 (encoding 329 amino acids) from a total of 32 strains of influenza A virus (subtype H3) isolated from the 2004 season and 1 from the 2003 season were amplified and the sequences were compared with vaccine reference strains recommended by WHO which were used in recent years. There were several consistent amino acid variations which involved in both antigenic epitopes A and B and receptor binding site (RBS) for isolated strains in the 2004 influenza season compared with the vaccine strains used during the recent years and the virus strains isolated in March 2004, indicated the antigenic drift of the viruses isolated in 2004 influenza season may lead to variant viruses.

CONCLUSIONThe variations of the HA genes from influenza virus (subtype H3) strains in the 2004-2005 influenza season were confirmed by sequence analysis for the HA1 regions of the hemagglutinin genes, which indicate that the antigenic drift would have been caused by the diversification of the genes and the efficacy of the recently used vaccines should be kept under close watch.

Antigenic Variation ; Child ; China ; epidemiology ; Evolution, Molecular ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H3N2 Subtype ; genetics ; immunology ; isolation & purification ; Influenza, Human ; epidemiology ; immunology ; virology ; RNA, Viral ; genetics ; Sequence Analysis, RNA

OBJECTIVETo establish a loop-mediated isothermal amplification (LAMP) method for rapid diagnosis of Vibrio cholerae.

METHODSBased on the ompW nucleic sequence of Vibrio cholerae, a pair of primers was designed for LAMP. The reaction conditions were optimized, and the specificity, sensitivity, and practicability of LAMP were tested using 47 bacterial strains and simulated contaminated sites.

RESULTSThe results of viable bacterium count showed that LAMP was capable of detecting Vibrio cholerae at a level as low as 1.6x10(2) cfu/ml. The minimal detectable concentration was 1.6+10(3) cfu/ml for simulated contaminated samples such as feces and seawater, and 1.6+10(4) cfu/ml for contaminated milk. All the 21 strains of Vibrio cholerae yielded positive results in LAMP, and the 26 strains of other bacteria all showed negative results, with a detection specificity of 100%.

CONCLUSIONThe established LAMP method has high specificity and sensitivity for detecting Vibrio cholerae and is applicable in field monitoring and epidemiological study of Vibrio cholerae.

Bacterial Proteins ; genetics ; Cholera ; diagnosis ; microbiology ; Clinical Laboratory Techniques ; methods ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Sensitivity and Specificity ; Vibrio cholerae ; genetics ; isolation & purification