Objective: To established a method for the quality standard of Yupingfeng Oral Liquid(Radix Astragali, Radix Saposhnikoviae and Rhizoma Atractylodis Macrocephalae). Methods:Radix Astragali, Radix Saposhnikoviae and Rhizoma Atractylodis Macrocephalae in oral liquid were identified by TLC. The content of astragaloside IV was determined by HPLC-ELSD. Results:The study showed that spots of samples on TLC can be well separated and the method had strong specificity. The average recovery was 97.75% and RSD was 1.31%. Conclusion:The method is simple, sensitive and accurate. It can be used for quality control of Yupingfeng Oral Liquid.

Objective To investigate the association between polymorphisms of thyroid-stimulating hormone receptor(TSHR)gene intron 1(rs179247, rs12101261)and Graves′ disease(GD)in the China Han population from Xuzhou city, Jiangsu Province. Methods Total 1 066 GD patients and 1 107 control subjects were recruited for genotyping by Taqman probe technique on Fluidigm EP1 platform. Meanwhile, serum concentrations of thyroid hormone and TSH receptor antibodies(TRAb)were determined. Results The rs179247_A, rs12101261_T were significantly associated with GD risk(OR=1.35, 95%CI 1.19-1.54, P=5.92×10-6; OR=1.32, 95%CI 1.16-1.50, P=2.22×10-5). Logistic regression identified that rs179247 was an independent susceptibility locus of GD. Serum TRAb concentration showed a significant difference(P=0.015)among rs179247_AA, AG, and GG genotypes. Conclusion rs179247 and rs12101261 in TSHR intron 1 are both associated with GD, and rs179247 may contribute risk to GD independently. The polymorphism is associated with TRAb, but not with serum concentration of thyroid hormones, age of onset, diffused thyroid goiter, ophthalmic signs, and relapse.

Objective To evaluate the efficiency of using nested PCR in restriction fragment length polymorphism analysis (P1-RFLP) for genotyping Mycoplasma pneumonia (M.pneumonia) in clinical specimens.Methods Based on the gene sequence of RepMp4 and RepMp2/3 in P1 gene of reference strains M129 (type 1) and FH (type 2),two sets of inner primers were designed with a HaeⅢ restriction enzyme site (GGCC).The nested PCR was set up to detect the target DNA in clinical specimens.The amplification products were mixed and digested with Hae Ⅲ enzyme.The genotypes were analyzed by comparing with various restriction maps and the results were verified by sequencing analysis.The concentration of DNA extracted from standard and clinical strains were detected by ten-fold dilution to evaluate the sensitivity of nested PCR-P1-RFLP and P1-RFLP.M.pneumonia-positive specimens isolated from Beijing in 2012 were analyzed by the nested PCR-P1-RFLP and the results were compared with those by P1-RFLP analysis.Results The nested PCR-P1-RFLP could effectively genotype M.pneumonia in clinical specimens and the results were consistent with those by sequencing analysis.The sensitivity of new assay was 103 times higher than that of the original P1-RFLP.Of the 115 M.pneumoniae positive clinical specimens,97.4％ (112/115) were type 1 and the rest were type 2.Conclusion The nested PCR-P1-RFLP shows high efficiency for genotyping of M.pneumonia in clinical specimens.It might be useful for the surveillance of M.pneumoniae infection.

Objective To inspect the source of an outbreak with Mycoplasma pneumoniae (Mp).Methods We carried out real-time PCR to analyze specimens collected from pediatric patients in Beijing during January 2010 to May 2012,diagnosed as pneumonia or a respiratory infection according to clinical symptoms.These positive samples were analyzed by the M-P typing system(M:multiple-locus variable-number tandem-repeat analysis,MLVA; P:P1-restriction fragment length polymorphism analysis,P1-RFLP).Results Sixty-nine specimens were tested positive to Mp by the real-time PCR in 446 specimens from pediatric patients.The infection rate was 11.69％,15.56％ and 20.00％ respectively in 2010,2011 and the first half of 2012.According to the M-P system,11 distinct genotypes were identified from 69 positive specimens,M43562P1 and M53562P1 were the two main genotypes that showed an increasing trend from 2010 to 2011,and M33562P1 and M63562P1 showed an increasing trend from 2011 to 2012 in China.Conclusion During this international Mp epidemic,the infection rate of Mp was also increase in Beijing in 2011,and M43562P1 and M53562P1 were the two main genotypes.Among them,M43562 were consistent with pop genotypes in Europe,and M53562 were consistent with pop genotype in Israel.The M-P system would be valuable to monitor the epidemic of Mp in different countries in the world.

Objective To investigate the association between the six single nucleotide polymorphisms ( SNP),named as rs179247,rsl2101261,rs2284722,rs4903964,rs2300525,rsl7111394 in the intron 1 of thyroid stimulating hormone receptor gene (TSHR) and Graves' disease (GD).MethodsThe genotypes of the six SNPs were genotyped by Taqman probe technique on Fluidigm EP1 platform in 618 GD patients and 646 control subjects.Meanwhile,TSH receptor antibodies (TRAb) of the patients were determined.ResultsAmong the six SNPs,five S NPs were strongly associated with GD,with the most signals at rs179247_G,rs12101261_C,rs4903964 _G (P=2.85×10-10,OR=1.73,95％CI1.46-2.05;P=1.74×10-10,OR=1.73,95％CI 1.46-2.05;P=2.24×10-10,OR=1.69,95％ CI 1.44-1.99 ).The results of logistic regression analysis indicated that rs12101261 and rs4903964 were main susceptibility loci of GD in the intron 1 of TSHR.rs179247_G,rs1210126 1_C,and rs4903964_G were associated with subset of the GD patients with positive TRAb (P=4.24× 10-13,p=5.48× 10-13,P =3.89×10-12 ).Conclusionrs179247,rs12101261,and rs4903964 in TSHR intron 1 were significantly associated with GD in the Chinese Han population from Bengbu city.rs12101261 and rs4903964 were the major susceptibility SNPs associated with GD.TSHR gene may play a main role of susceptibility gene in the subset of GD patients with persistent positive TRAb.

BACKGROUNDMycoplasma pneumonia (M. pneumoniae) is one of the key pathogens of community-acquired pneumonia. A global pandemic of M. pneumoniae has occurred since 2010. The aim of this study was to survey the prevalence of M. pneumoniae in children in Beijing from 2007-2012.

METHODSA total of 3 073 clinical specimens were obtained from pediatric patients with respiratory tract infections from January 2007 to December 2012, and examined by nested polymerase chain reaction. PCR products were visualized by 2% agarose gel electrophoresis, positive products sequenced, and compared with reference sequences in GenBank. Macrolide resistance-associated mutations were also detected for some positive samples.

RESULTSOf the 3 073 specimens, 588 (19.13%) were positive for M. pneumoniae, 12.4% of which were accompanied by viral infections. Positive rates for M. pneumoniae were highest in 2007 and 2012, showing a significant difference when compared with other years. Infections tended to occur in autumn and winter and positive rates were significantly higher for children aged 3-16. The rate of macrolide resistance-associated mutations was 90.7%, and the predominant mutation was an A→G transition (89.92%) at position 2063 in domain V of the 23S rRNA gene.

CONCLUSIONSM. pneumoniae outbreaks occurred in 2007 and 2012 in pediatric patients in Beijing, which is consistent with the global prevalence of M. pneumoniae. M. pneumoniae can cause multi-system infections in children, and may be accompanied with viral infections. We determined that school-age children are more susceptible to this disease, particularly in autumn and winter. Gene mutations associated with macrolide resistance were very common in M. pneumoniae-positive specimens during this period in Beijing.

Adolescent ; Anti-Bacterial Agents ; therapeutic use ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Macrolides ; therapeutic use ; Male ; Mycoplasma pneumoniae ; pathogenicity ; Pneumonia, Mycoplasma ; drug therapy ; epidemiology ; Prevalence