1.Human papillomavirus genotyping:Establishment and application of DNA array method
Fangjin WANG ; Min WANG ; Ding WANG ; Kelin XIAO ; Yunshao HE
Chinese Journal of Clinical Laboratory Science 2006;0(03):-
Objective To establish a convenient, fast, economic and hypersensitive low-density DNA array method to detect the genotypes of human papillomavirus (HPV) and evaluate its application in clinic services.Methods HPV in cervical swab samples from 355 suspected female patients collected in gynaecology and obstetrics clinic were genotyped by hybrid capture (HC) II method and low-density DNA array simultaneously. HPV in 730 clinic samples from the area of Pearl River delta were genotyped by low-density DNA array.Results Among 355 suspected samples positive HPV-DNA were detectable in 211 (59.4%) samples by DNA array and 222 (62.5%) samples by HCII method. The concordance rate between the two assays was 94.1%. In the HPV genotypes 15 high-risk type and 5 low-risk type were detected by low-density DNA array. The most common high-risk types were HPV-16, 52, 58 and 56. The peak age of HPV infection was 26-30 years. The distribution of genotypes was different from various degree of cervical changes.Conclusion Either single type or multiple type of HPV infection can be detected by low-density DNA array. The combination of HPV detection with cytology detection will provide instruction for cervical cancer screening.
2.Clinical research of the distribution of H.pylori ureA, vacA s1 gene and cagA subtype distribution in patients with digestive diseases in Guangzhou
Juan LI ; Qiang ZHOU ; Hongyun JIA ; Xiaoyan DENG ; Fangjin WANG ; Yunshao HE ; Xiaoman WU
Chinese Journal of Laboratory Medicine 2011;34(7):638-642
Objective To detect the distribution of H.pylori ureA, vacA s1 gene and cagA subtype(ABC, ABD, ABAB, AAD, et al) in patients with digestive diseases in Guangzhou and investigate the relationship with the pathological findings of gastric mucosa.Methods A total of 227 randomly selected gastric mucosa from patients with digestive diseases were enrolled in the research, including 46 without pathological changes, 130 with chronic gastritis, 29 with peptic ulcer, 15 with atrophic gastritis and 7 with gastric cancer.Real-time PCR assay were used to detect Helicobacter pylori ureA gene and vacA s1 gene.EPIYA motifs in the 3′ region of cagA were amplified by conventional PCR followed by subtype sequencing. The conserved gene ureA was used to detect H.pylori infection.Results Among the 227 patients with digestive diseases, 50.7% (115/227) patients were H.pylori positive, in which 91.3%(105/115) carried vacA s1 and 78.3% (90/115) carried cagA. Four types of cagA-EPIYA subtype were detected, including ABC 17.8%(16/90), ABD 78.9%(71/90), AAD 2.2%(2/90) and ABAB 1.1%(1/90).In the non-pathological change group, 32.6% (15/46) were H.pylori positive, in which 28.3% (13/46) carried vacA s1 and 26.1% (12/46) carried cagA;in chronic gastritis group, it was 48.5% (63/130), 43.8% (57/130) and 36.2% (47/130), respectively;in ulcer group, it was 72.4% (21/29), 65.5% (19/29) and 55.2% (16/29), respectively;in atrophic gastritis group, it was 66.7% (10/15), 66.7% (10/15) and 66.7% (10/15), respectively;in gastric cancer group, it was 85.7% (6/7), 85.7% (6/7) and 71.4% (5/7), respectively.The distribution of H.pylori among the 4 groups had statistical significance (χ2=16.72;P<0.01). H.pylori was more prevalent in ulcer, atrophic gastritis and cancer group than in inflammation group and non-pathological change group (χ2=16.02;P<0.01).In patients infected by H.pylori, there was no significant difference in the distribution of vacA s1 gene as high virulence factors among non-pathological change, inflammation, ulcer, atrophic gastritis and cancer group (χ2=2.00;P=0.74), as well as cagA (χ2=3.44;P=0.49) and EPIYA subtypes (χ2=3.66;P=0.45).Conclusions H.pylori infection is significantly associated with the pathological change of gastric mucosa for patients with digestive diseases in Guangzhou, while the relationship with the pathogenicity of H.pylori with high virulence genotype is not significant.More samples and diseases reclassification are needed to make an advanced analysis of the effect of H.pylori with high virulence in gastrointestinal diseases development.
3.Analysis of effect of Epimedium on cells endogenous metabolites before and after the processing based on UPLC-Q-Exactive Orbitrap-MS
Zimin YUAN ; Mingyue YUAN ; Jing WANG ; Fangjin LU
International Journal of Traditional Chinese Medicine 2024;46(4):485-491
Objective:To analyze the effects of processed Epimedii Folium on endogenous metabolites of mouse melanoma cells (B16 cells) before and after processing based on cell metabolomics; To investigate the changes of processed Epimedii Folium before and after processing.Methods:Ultra performance liquid chromatography tandem four-stage orbital trap mass spectrometry (UPLC-Q-Exactive Orbitrap-MS) technology was used, and the endogenous small molecules of B16 cells treated with Epimedii Folium and processed Epimedii Folium were analyzed by metabolomics. The differential metabolites between groups were obtained, and relevant metabolic pathways were analyzed based on the MetaboAnalyst 5.0 database.Results:Significant changes were observed in 13 kinds of endogenous metabolites, including alanine, carnitine C3∶0, glutamic acid-1, lactic acid, isoleucine, choline, phosphatidylcholine (34∶2, 36∶2), free fatty acids, citric acid, carnitine C4∶0, lysophosphatidylcholine 16∶0 and malic acid after the intervention of Epimedii Folium and processed Epimedii Folium. And the impact of processed products on differential metabolites was stronger than that of raw products. The main pathways involved were Warburg effect, pyruvate metabolism, malate-aspartic acid shuttle, pyruvaldehyde degradation and so on.Conclusions:Epimedii Folium and processed Epimedii Folium would have certain effects on cellular metabolic pathways. The results may be related to the pharmacological effects and changes in cold and hot properties of Epimedii Folium before and after processing.