1.Detection and Genotyping of Treponema pallidum by a Nested PCR
Heping ZHENG ; Zhiying OU ; Yushan HU ; Jinmei HUANG ; Meiling LI ; Xingzhong WU ; Weiying ZENG ; Huiqing PAN
Chinese Journal of Dermatology 2003;0(09):-
Objective To develop a nested PCR for the detection of early syphilis and genotyping of Treponema pallidum (TP), and to investigate the distribution of genotypes of TP in Guangzhou. Methods Specimens were consecutively collected from genital ulcers of patients with suspected chancre during 2002-2004, and were detected by dark-field microscopy and nested PCR. The acidic repeat protein (arp) gene and the T. pallidum repeat (tpr) gene family were amplified with the positive specimens above. The number of repeats presented in the arp gene and the restriction fragment length polymorphism by Mse I in the tpr gene were analyzed by electrophoresis. The strains were genotyped according to Pillay's criteria. Results Out of 62 patients with suspected chancre, 33 cases (53.2%) were positive by dark-field microscopy and 54 cases (87.1%) by nested PCR. Of 47 TP-positive specimens genotyped by arp gene, 36 (76.6%) were type 14, while of 49 cases genotyped by tpr gene 39 (79.6%) were type d. By combining genotypes of arp and tpr genes, 7 genotypes were found, including 14d (31, 66.0%), 13d (5, 10.6%), 14b (4, 8.5%), 12b (3, 6.4%), 12d (2, 4.3%), 15d(l, 2.2%) and 14i (1, 2.2%). Conclusions Nested PCR shows a high sensitivity in early detection of TP. Genotype 14d seems the predominant type of TP in Guangzhou.
2.Study on relationship between sperm morphology with seminal plasma zinc and sperm acrosin activity in male infertile patients
Yanru BING ; Heguo MO ; Jianyun HUANG ; Lulin LI ; Shuilian OU ; Yongyi LI ; Xiaoqin LU ; Jinmei CAI
International Journal of Laboratory Medicine 2014;(13):1708-1709
Objective To investigate the relationship of sperm morphology with seminal plasma zinc and sperm acrosin activity in male infertile patients.Methods 455 patients with male infertility were divided into 5 groups and 8 groups according to age and sperm morphology respectively.The sperm morphology,seminal plasma zinc and sperm acrosin activity were detected by the Diff-Quik stain,modified 5-Br-PAPS and modified Kennedy methods respectively.Results The percentages of morphologically normal sperm and seminal plasma zinc concentration in the infertile men groups were not different in various age groups(P >0.05 ).The sperm acrosin activity in the age group of 36-50 years was remarkably decreased (P <0.05),while which in other age groups had no statistical difference(P >0.05).Seminal plasma zinc concentration in the infertile men groups were not different in the 8 groups (P >0.05 ).Sperm acrosin activity was decreased with the percentage decrease of morphologically normal sperm (r =0.93,P <0.01).Conclusion The percentage of morphologically normal sperm is not affectd by the age and the seminal plasma zinc,which is positively correlated with the sperm acrosin activity.
3.Content Determination of 3 Saccharides in 3 Kinds of Medicinal Dendrobii Caulis by HPLC-CAD
Xiaoqian ZHENG ; Chuanshan JIN ; Yazhong ZHANG ; Junling LIU ; Nianjun YU ; Jinmei OU
China Pharmacy 2020;31(10):1185-1189
OBJECTIVE:To establish the content determination me thod of 3 mono/disaccharides in 3 kinds of medicinal Dendrobii Caulis. METHODS :HPLC-CAD method was established. The determination was performed on Shodex Asahipak NH2P-50 4E column with mobile phase consisted of acetonitrile-water (75 ∶ 25,V/V)at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃,and sample size was 10 µL. CAD detection condition included that data acquisition frequency was 5 Hz,filter constant was 5 s,atomization temperature was 35 ℃ ,gas source was nitrogen with pressure of 4.012 × 105 Pa. RESULTS:The linear range of fructose ,D-anhydrous glucose and sucrose were 0.156 2-1.873 8 mg/mL(r=0.999 5),0.012 7- 0.152 4 mg/mL(r=0.999 7),0.277 6-3.331 2 mg/mL(r=0.999 8),respectively. The limits of quantification were 0.002 61,0.004 24 and 0.005 12 mg/mL,and the limits of detection were 0.000 78,0.001 27 and 0.001 54 mg/mL,respectively. RSDs of precision , stability,reproducibility and durability tests were all lower than 3%. The recoveries were 95.98%-98.15%(RSD=0.83%,n=6), 95.64%-98.62%(RSD=1.10%,n=6)and 97.53%-98.94%(RSD=0.53%,n=6). The contents of them were 0.28%-1.12%, 0.02%-0.13%,0.76%-2.67%,respectively. The total content was 1.38%~3.10%. The order of saccharide content in 3 kinds of Dendrobii Caulis was sucrose >fructose>D-anhydrous glucose ;the order of sucrose content and total content were Dendrobium huoshanense>D. moniliforme >D. officinale ;the order of D-anhydrous glucose content was D. huoshanense >D. officinale >D. moniliforme; the order of fructose content was D. moniliforme >D. officinale >D. huoshanense . CONCLUSIONS :Established method is sensitive ,reproducible and simple in operation ,and can be used for content determination of 3 saccharides in 3 kinds of medicinal Dendrobii Caulis. There are differences in the contents of saccharide s among 3 kinds of Dendrobii Caulis.