Application of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Analysis in Identification of Fritillariae Cirrhosae Bulbus / 中国药学杂志

OBJECTIVE: To identify 70 samples of Fritillariae Cirrhosae Bulbus from 16 cities such as Beijing, Tianjin, Changchun,Jilin and so on by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis kit.METHODS: The genomic DNA of Fritillariae Cirrhosae Bulbus was extracted by kit method, and PCR reaction and RFLP identification were carried out. The method of the Chinese Pharmacopoeia of 2015 was used for comparison and re-examination.RESULTS: The purity of the genomic DNA of Fritillariae Cirrhosae Bulbus was very good, the OD260/OD280 value was between 1.90-2.1, and the concentration could reach the requirement of PCR. Seventy samples were tested, of which 37 were positive and 33 were counterfeit, and the false rate was 47.1%.CONCLUSION: The purity and concentration of the nucleic acid meet the requirements of PCR and RFLP. The test results of the samples from 16 cities show that the counterfeit rate of Fritillariae Cirrhosae Bulbus is high in the market. Relevant government departments should strengthen the quality management of the traditional Chinese medicine market.

Affecting Factors of Secondhand Smoke Exposure in Korea: Focused on Different Exposure Locations

Exposure to secondhand smoke (SHS) not only can cause serious illness, but is also an economic and social burden. Contextual and individual factors of non-smoker exposure to SHS depend on location. However, studies focusing on this subject are lacking. In this study, we described and compared the factors related to SHS exposure according to location in Korea. Regarding individual factors related to SHS exposure, a common individual variable model and location-specific variable model was used to evaluate SHS exposure at home/work/public locations based on sex. In common individual variables, such as age, and smoking status showed different relationships with SHS exposure in different locations. Among home-related variables, housing type and family with a single father and unmarried children showed the strongest positive relationships with SHS exposure in both males and females. In the workplace, service and sales workers, blue-collar workers, and manual laborers showed the strongest positive association with SHS exposure in males and females. For multilevel analysis in public places, only SHS exposure in females was positively related with cancer screening rate. Exposure to SHS in public places showed a positive relationship with drinking rate and single-parent family in males and females. The problem of SHS embodies social policies and interactions between individuals and social contextual factors. Policy makers should consider the contextual factors of specific locations and regional and individual context, along with differences between males and females, to develop effective strategies for reducing SHS exposure.

A study on gyrA mutant in Levofloxacin-resistant Brucella melitensis induced resistant strains in vitro / 中国地方病学杂志

Objective To study on the drug-resistance mechanism of Brucella resistance to Quinolone antibiotics to guide the selection and use of antimicrobial agents in clinical practice. Methods Six strains of Brucella melitensis(Bru1, Bru2, Bru3, Bru4, Bru5, Bru6) were selected to be induced resistance to levofloxacin in vitro respectively. The MICs of the 6 strains of Brucella melitensis and induced resistant strains were measured by agar dilution method. The sensitivity to Quinolone antibiotics (Levofloxacin, Ciprofloxacin, Lomefloxacin, Norfloxacin, Fleroxacin, Ofloxaein) of 6 strains of Brucella melitensis and induced resistant strains was measured by K-B method. The gyrA of the 6 strains of Brucella melitensis and induced resistant strains was amplified by PCR, then the nucleotide sequence of the genes were analyzed. Results The MICs of Bru1,Bru2,Bru3,Bru4, Bru6 were 0.50 μg/ml and Bru5 was 0.25 μg/ml. The strains Bin3, Bru4 were induced into drug-resistant strains by Levofloxacin, then were named LEVR3 and LEVR4 respectively. The MICs of LEVR3 and LEVR4 were 64,128 μg/ml with 128 and 256 times higher than that of the parental strains. The 6 strains of Brucella melitensis were sensitive to Quinolone antibiotics, LEVR3 and LEVR4 were resistant to Quinolone antibiotics. Neucleotide sequence analysis and comparison of the derived amino acid sequence revealed that Quinolone resistance-determing region of GyrA had a substitution at position Ala87 and Asp91 in laboratory resistant strains. Conclusion The results of in vitro experiments show that acquired resistance of Brucella melitensis strains to Levofloxacin could beinduced when exposed to high level of some antibacterial agents for short term. Two drug-resistant strains occur mutations in gyrA and have cross-resistance to other Quinolones.

Gene Mutation Analysis of A Child with Familial Hypercholesterolemia and His Family / 实用儿科临床杂志

Objective To screen the mutation of certain gene of a 10-years-old boy with multiple xanthomas and very high level of cholesterol who could be diagnosed as homozygous familial hypercholesterolemia (FH),to explore the relationship between the genotype and phenotype,and to discuss the molecular pathologic mechanism.Methods The basic information of life styles were asked from the boy and his familial members.The blood was drown to examine the lipid and genes.The boy was examined with electrocardiogram examination,ultrasonography and coronary CT angiography (CTA) to evaluate the degree of atherosclerosis.Peripheral blood DNA of the boy and his parents were extracted by phenol-chloroform method and investigated for mutations of promoter and all 18 exons of low density lipoprotein receptor(LDLR) gene.Screening was carried out by using Touch-down polymerase chain reaction (PCR) and single strand conformation polymorphism(PCR-SSCP),combined with DNA sequence analysis.In addition,the apolipoprotein B100 gene(apoB100) for known mutations (R3500Q) which caused familial defective apoB100 was screened by PCR-DNA sequence analysis.Results 1.The level of cholesterol of his parents were higher than the normal.2.Several clinical manifestations of atherosclerosis were detected from that boy.Increased intima-media thickness and plaques were detected in the common carotid artery.Mitral valve regurgitation was found by echocardiography.Coronary stenosis was confirmed by CTA.3.No mutations R3500Q of apoB100 was observed.4.A homozygous mutation in exon13 of the LDLR gene (D601Y) were identified in the boy and his parents harbour D601Y heterozygous mutation due to a single base pair substitution of G for T in the codon for residue 1864.Conclusions The final diagnosis of the boy with multiple xanthomas was homozygous FH.His disease was caused by D601Y homozygous mutation in exon13 of the LDLR gene inherited from his heterozygous parents.